Human Biochemistry

Extraction of DNA from Whole Blood

2010-04-03T22:07:00.001-07:00

1. Introduction
There are many differing protocols and a large number of commercially available kits used for the extraction of DNA from whole blood. This procedure is one we use
routinely in both research and clinical service provision and is cheap and robust. It can also be applied to cell pellets from dispersed tissues or cell cultures (omitting the red blood lysis step.

2. Materials
This method uses standard chemicals that can be obtained from any major supplier;

we use Sigma.
1. Waterbath set at 65°C.
2. Centrifuge tubes (15 mL; Falcon).
3. Microfuge (1.5 mL) tubes.
4. Tube roller/rotator.
5. Glass Pasteur pipets, heated to seal the end and curled to form a “loop” or “hook” for spooling DNA.
6. EDTA (0.5 M), pH 8.0: Add 146.1 g of anhydrous EDTA to 800 mL of distilled water.
Adjust pH to 8.0 with NaOH pellets (this will require about 20 g). Make up to 1 L with distilled water. Autoclave at 15 p.s.i. for 15 min.
7. 1 M Tris-HCl, pH 7.6: Dissolve 121.1 g of Tris base in 800 mL of distilled water. Adjust pH with concentrated HCl (this requires about 60 mL). CAUTION: the addition of acid
produces heat. Allow mixture to cool to room temperature before finally correcting pH. Make up to 1 L with distilled water. Autoclave at 15 p.s.i. for 15 min.
8. Reagent A: Red blood cell lysis: 0.01M Tris-HCl pH 7.4, 320 mM sucrose, 5 mM MgCl2, 1% Triton X 100.
9. Add 10 mL of 1 M Tris, 109.54 g of sucrose, 0.47 g of MgCl2, and 10 mL of Triton X-100
to 800 mL of distilled water. Adjust pH to 8.0, and make up to 1 L with distilled water. Autoclave at 10 p.s.i. for 10 min .
10. Reagent B: Cell lysis: 0.4 M Tris-HCl, 150 mM NaCl, 0.06 M EDTA, 1% sodium dodecyl sulphate, pH 8.0. Take 400 mL of 1 M Tris (pH 7.6), 120 mL of 0.5 M EDTA (pH 8.0),
8.76 g of NaCl, and adjust pH to 8.0. Make up to 1 L with distilled water. Autoclave 15 min at 15. p.s.i. After autoclaving, add 10 g of sodium dodecyl sulphate.

11. 5 M sodium perchlorate: Dissolve 70 g of sodium perchlorate in 80 mL of distilled water. Make up to 100 mL.
12. TE Buffer, pH 7.6: Take 10 mL of 1 M Tris-HCl, pH 7.6, 2 mL of 0.5 M EDTA, and make up to 1 L with distilled water. Adjust pH to 7.6 and autoclave 15 min at 15. p.s.i.
13. Chloroform prechilled to 4°C.
14. Ethanol (100%) prechilled to 4°C.

3. Method

3.1. Blood Collection

1. Collect blood in either a heparin- or EDTA-containing Vacutainer by venipuncture Store at room temperature and extract within the same working day.

3.2. DNA Extraction
To extract DNA from cell cultures or disaggregated tissues, omit steps 1 through 3.
1. Place 3 mL of whole blood in a 15-mL falcon tube.
2. Add 12 mL of reagent A.
3. Mix on a rolling or rotating blood mixer for 4 min at room temperature.
4. Centrifuge at 3000g for 5 min at room temperature.
5. Discard supernatant without disturbing cell pellet. Remove remaining moisture by inverting
the tube and blotting onto tissue paper.
6. Add 1 mL of reagent B and vortex briefly to resuspend the cell pellet.
7. Add 250 μL of 5 M sodium perchlorate and mix by inverting tube several times.
8. Place tube in waterbath for 15 to 20 min at 65°C.
9. Allow to cool to room temperature.
10. Add 2 mL of ice-cold chloroform.
11. Mix on a rolling or rotating mixer for 30 to 60 min .

12. Centrifuge at 2400g for 2 min.
13. Transfer upper phase into a clean falcon tube using a sterile pipet.
14. Add 2 to 3 mL of ice-cold ethanol and invert gently to allow DNA to precipitate.

15. Using a freshly prepared flamed Pasteur pipet spool the DNA onto the hooked end .
16. Transfer to a 1.5-mL Eppendorf tube and allow to air dry .
17. Resuspend in 200 μL of TE buffer .

4. Notes
1. Autoclaving sugars at high temperature can cause caramelization (browning), which degrades the sugars.
2. As will all body fluids, blood represents a potential biohazard. Care should be taken in all steps requiring handling of blood. If the subject is from a known high risk category (e.g.,intravenous drug abusers) additional precautions may be required.
3. Rotation for less than 30 or over 60 min can reduce the DNA yield.
4. DNA should appear as a mucus-like strand in the solution phase.
5. Rotating the hooked end by rolling between thumb and forefinger usually works well. If the DNA adheres to the hook, break it off into the Eppendorf and resuspend the DNA before transferring to a fresh tube.
6. Ethanol will interfere with both measurements of DNA concentration and PCR reactions. However, overdrying the pellet will prolong the resuspension time.

7. The small amount of EDTA in TE will not affect PCR. We routinely use 1 μL per PCR reaction without adverse affects.
8. DNA can be quantified and diluted to a working concentration at this point or simply use 1 μL per PCR reaction; routinely, we expect 200 to 500 ng/μL DNA to be the yield
of this procedure.

How to prevent colon cancer

2010-04-03T10:25:00.001-07:00

Colorectal cancer, also called colon cancer or large bowel cancer, includes cancerous growths in the colon, rectum and appendix. With 655,000 deaths worldwide per year, it is the fifth most common form of cancer in the United States and the third leading cause of cancer-related death in the Western world. Colorectal cancers arise from adenomatous polyps in the colon. These mushroom-shaped growths are usually benign, but some develop into cancer over time. Localized colon cancer is usually diagnosed through colonoscopy.

Signs and symptoms

Feeling of incomplete defecation
Change in bowel habit
Reduction in size and shape of stool
Black or brown stool (bleeding)
Constipation, bowel pain, and vomiting may also be present.

Risk factors

Age. The risk of developing colorectal cancer increases with age. Most cases occur in the 60s and 70s, while cases before age 50 are uncommon unless a family history of early colon cancer is present.
Polyps of the colon, particularly adenomatous polyps, are a risk factor for colon cancer. The removal of colon polyps at the time of colonoscopy reduces the subsequent risk of colon cancer.
History of cancer. Individuals who have previously been diagnosed and treated for colon cancer are at risk for developing colon cancer in the future. Women who have had cancer of the ovary, uterus, or breast are at higher risk of developing colorectal cancer.
Heredity:
- Family history of colon cancer, especially in a close relative before the age of 55 or multiple relatives.
- Familial adenomatous polyposis (FAP) carries a near 100% risk of developing colorectal cancer by the age of 40 if untreated
- Hereditary nonpolyposis colorectal cancer (HNPCC) or Lynch syndrome
Smoking. Smokers are more likely to die of colorectal cancer than non-smokers. An American Cancer Society study found that "Women who smoked were more than 40% more likely to die from colorectal cancer than women who never had smoked. Male smokers had more than a 30% increase in risk of dying from the disease compared to men who never had smoked."
Diet. Studies show that a diet high in red meat and low in fresh fruit, vegetables, poultry and fish increases the risk of colorectal cancer. In June 2005, a study by the European Prospective Investigation into Cancer and Nutrition suggested that diets high in red and processed meat, as well as those low in fiber, are associated with an increased risk of colorectal cancer. Individuals who frequently eat fish showed a decreased risk. However, other studies have cast doubt on the claim that diets high in fiber decrease the risk of colorectal cancer; rather, low-fiber diet was associated with other risk factors, leading to confounding. The nature of the relationship between dietary fiber and risk of colorectal cancer remains controversial.
Physical inactivity. People who are physically active are at lower risk of developing colorectal cancer.
Virus. Exposure to some viruses (such as particular strains of human papilloma virus) may be associated with colorectal cancer.
Primary sclerosing cholangitis offers a risk independent to ulcerative colitis
Low levels of selenium.
Inflammatory bowel disease. About one percent of colorectal cancer patients have a history of chronic ulcerative colitis. The risk of developing colorectal cancer varies inversely with the age of onset of the colitis and directly with the extent of colonic involvement and the duration of active disease. Patients with colorectal Crohn's disease have a more than average risk of colorectal cancer, but less than that of patients with ulcerative colitis.
Environmental factors. Industrialized countries are at a relatively increased risk compared to less developed countries that traditionally had high-fiber/low-fat diets. Studies of migrant populations have revealed a role for environmental factors, particularly dietary, in the etiology of colorectal cancers.
Exogenous hormones. The differences in the time trends in colorectal cancer in males and females could be explained by cohort effects in exposure to some gender-specific risk factor; one possibility that has been suggested is exposure to estrogens.There is, however, little evidence of an influence of endogenous hormones on the risk of colorectal cancer. In contrast, there is evidence that exogenous estrogens such as hormone replacement therapy (HRT), tamoxifen, or oral contraceptives might be associated with colorectal tumors.
Alcohol. Drinking, especially heavily, may be a risk factor.

Diagnosis

Digital rectal exam (DRE): The doctor inserts a lubricated, gloved finger into the rectum to feel for abnormal areas. It only detects tumors large enough to be felt in the distal part of the rectum but is useful as an initial screening test.
Fecal occult blood test (FOBT): a test for blood in the stool. Two types of tests can be used for detecting occult blood in stools i.e. guaiac based (chemical test) and immunochemical. The sensitivity of immunochemical testing is superior to that of chemical testing without an unacceptable reduction in specifity.
Endoscopy:
- Sigmoidoscopy: A lighted probe (sigmoidoscope) is inserted into the rectum and lower colon to check for polyps and other abnormalities.
- Colonoscopy: A lighted probe called a colonoscope is inserted into the rectum and the entire colon to look for polyps and other abnormalities that may be caused by cancer. A colonoscopy has the advantage that if polyps are found during the procedure they can be immediately removed. Tissue can also be taken for biopsy.

Double contrast barium enema (DCBE): First, an overnight preparation is taken to cleanse the colon. An enema containing barium sulfate is administered, then air is insufflated into the colon, distending it. The result is a thin layer of barium over the inner lining of the colon which is visible on X-ray films. A cancer or a precancerous polyp can be detected this way. This technique can miss the (less common) flat polyp.
Virtual colonoscopy replaces X-ray films in the double contrast barium enema (above) with a special computed tomography scan and requires special workstation software in order for the radiologist to interpret. This technique is approaching colonoscopy in sensitivity for polyps. However, any polyps found must still be removed by standard colonoscopy.
Standard computed axial tomography is an x-ray method that can be used to determine the degree of spread of cancer, but is not sensitive enough to use for screening. Some cancers are found in CAT scans performed for other reasons.
Blood tests: Measurement of the patient's blood for elevated levels of certain proteins can give an indication of tumor load. In particular, high levels of carcinoembryonic antigen (CEA) in the blood can indicatemetastasis of adenocarcinoma. These tests are frequently false positive or false negative, and are not recommended for screening, it can be useful to assess disease recurrence.
Genetic counseling and genetic testing for families who may have a hereditary form of colon cancer, such as hereditary nonpolyposis colorectal cancer (HNPCC) or familial adenomatous polyposis (FAP).
Positron emission tomography (PET) is a 3-dimensional scanning technology where a radioactive sugar is injected into the patient, the sugar collects in tissues with high metabolic activity, and an image is formed by measuring the emission of radiation from the sugar. Because cancer cells often have very high metabolic rate, this can be used to differentiate benign and malignant tumors. PET is not used for screening and does not (yet) have a place in routine workup of colorectal cancer cases.
Whole-Body PET imaging is the most accurate diagnostic test for detection of recurrent colorectal cancer, and is a cost-effective way to differentiate resectable from non-resectable disease. A PET scan is indicated whenever a major management decision depends upon accurate evaluation of tumour presence and extent.
Stool DNA testing is an emerging technology in screening for colorectal cancer. Pre-malignant adenomas and cancers shed DNA markers from their cells which are not degraded during the digestive process and remain stable in the stool. Capture, followed by PCR amplifies the DNA to detectable levels for assay. Clinical studies have shown a cancer detection sensitivity of 71%–91%.

Prevention

Colon cancer can be prevented by preliminary monitoring like bowel status after 50 years. You should check your bowel status by monitoring no. of stool per day and size and structure of stool. number of stool should not more than and not less than one times in 24 hr. Even you should monitor about size and shape, it should be neither too viscous nor too loose.
Go for rectal digital examination. This will tell about formation of small nodules which is easier to remove and can controlled on primary status.
You should also go for Occult blood examination in urine which tell us about any hidden blood loss or secretion of blood in gastrointestinal track.
Take a good vitamin and high fiber diet which will improve your GIT healthiness.
Regular physical exercise for 30 minute a day will be enough.

Ask for any query

Pain relieving meditation

2010-03-25T00:26:00.001-07:00

A new research conducted by researchers of university of north Carolina, charlotte, in which they found that doing three minute meditation three times a day decrease pain sense and increases stress handling level.
That is well known that meditation is directly related to mental power and it powers up brain to handle any type of stress. Pain is also a kind of stress. Pain’s signal is received and interpreted by brain and if we just control brain or increases its resistance to cope with brain it will handle any kind of pain.

When a group of students who have never gone for any training or done any kind of mediation given a formal 1 hr training and then subjected to pain like electric shock, needle prick, that is seen that they handled this pain very well and even they graded very painful stimulus as low grade pain.
It means you don’t need to go for a fulltime training or spend any money if you are chronic pain patient because this three step training will give you relief from this. This training is called ACE.

Awareness : This is three minute procedure so this first minute goes for awareness. you need to think about what is happening inside your thought, mind and emotions.
Collection : Now second minute of your exercise need to spend on your breath. You need to collect the information and attention on your breath. This is the most important part, just concentrate on your breath. You can do it by focusing on nostril, chest or abdomen.
Spend the third minute on Expending your awareness on your physical body. Just feel perceptions like pain,tingling, warmth,pulsation and cold.

By this way you just control your pain feeling.
Source: Zaidan,F. journal of pain, march 2010, vol II, pp 199-209.

How to find blood group of your unborn child

2010-03-21T02:34:00.001-07:00

Every one wants to know that what blood group their child will have. But even you don't wants to spend any money to find this. I will tell you how to do this. First understand the theory. Blood group is genetically inherited . Blood group antigen is located on RBC, Platelet, epithelial cells and saliva secretion. These blood group antigens are made of protein and sugar moieties. These antigens are produced from DNA and DNA is located on chromosomes. There are three gene which determined blood group, I^A , I^Band I⁰. These are the gene which determines blood group. This will calculate only ABO part not the Rh part. That means you can find your child will be A, B, AB or O. Not that he/she will positive or negative. Now theory is enough lets find blood group.

First of all you need to find your gene. I will show you how. This is very simple, every human have to gene for blood group.See the box bellow and find your gene.

Blood group	Gene 1	Gene 2
A	I^A	I⁰
B	I^B	I⁰
AB	I^A	I^B
O	I⁰	I⁰

By this formula calculate your and your partners blood group gene.

Then draw a square box with four blocks in this. Like this

And add wife’s gene at top of square and husband gene on the side of square. I will show to example here

For blood group A(wife) and A(husband)

	I^A	I⁰
I^A	A	A
I⁰	A	0

By this you can find this, there is 75% chances that your child will be of A group. And 25 chances to be O group.

For blood group O(wife) and B(husband)

	I⁰	I⁰
I^B	B	B
I⁰	O	0

That means there is 50-50% probility that your child either be B or O group.

Each of square denotes 25% and Bothe of gene dominates over I⁰ .

If you have any problem please leave your comment i will answer this.

Benefits of fasting

2010-03-16T22:02:00.001-07:00

Fasting is abstaining form taking water or food far a period of time. It may involve abstaining for food,water,meat,Drinks, sex or combination of some or combination of all. Fasting is suggested by various religion and also by some orthodox but despite being religious and orthodoxy various part of world’s population is still like to do this because it has a lot of medical and emotional benefits. I am going to share you some of stuff that is about benefits of fasting and if i missed someone please add as comment. In fasting there is different kind of fasting suggested by religion, medical and dieticians. But i am going to tell you a general benefits.

Medically it refers as metabolic state who have not eaten overnight or a status that reached after complete digestion of food. But the benefits start to come out after the available blood glucose metabolized and breakdown of stored component of blood starts.
As you knows Glucose is primary source of energy for whole of body is metabolizes first and when blood glucose decrease a certain level our hormone system start to make glucose or glucose like energy source from other things like protein and lipids (fats). This metabolism utilizes glycogen from liver, protein from muscle and lipids from our muscles.
Scientist found fasting increases life span by decreasing potential reduced of cancer, cardiovascular disease, diabetes, insulin resistance, immune resistance and slowing aging.
Study conducted by DR. Mark P. Mattson in National institute of aging in US, when alternate fasting is prescribe on mouse the showed increase in age by 20-30 % and it is also conducted on small population. That means that if you start fasting it will leads to increase in life span and delay aging related problem.
According to US National Academy of Science other health benefits like stress resistance, increase in insulin sensitivity and reduced morbidity seen among fasting subject. Due to No large group and long study have been conduced so more benefits can not be estimated but in Indian mythology that have seen those saints who keep fasting only take fruits they lived for thousand of years.
As i said above long term study is not conducted but short term study shows increase in weight loss. This may be due to increase in lipid metabolism., absorption from intestine and formation of endogenous cholesterol depend on the stored fats. As scientists says that lipid is also a important component in body because it participate hormone production, cell membrane formation and some component transportation. So if your dietary supply is short the lipid requirement is fulfilled by endogenous fat by breakdown of liver, muscular and tissue fat.
Even Greek orthodox says that fasting period contributes in improvement of blood lipid profile by decreasing total and LDL cholesterol and also decreasing LDL/HDL ratio.
In Buddhism and Jainism people do not eat after noon meal. And once Buddha said that “we do not takes food in night and we all are perfect in health and never get ill. So it also proves that it increases immunity and age.
Study also subjects fasting every alternate days and eating double on other day increases insulin secretion, increases neural resistance to injury, increases blood sugar tolerance.
Study also shows it increase resistance to attenuated disease and inflammatory oxidative damage. This means you will fill little seek after fasts.
Fasting also teaches you self discipline that how to control your body, brain and thinking. When you are hungry there is fight goes place inside which your body signals your brain and brain get massage that you are fast and it start challenging your decision and after all your decision wins. It also increases self discipline, positive attitude and generates positive thoughts.
According to Lugman “when your stomach is full, your intellect begins to sleep.” It is right you also know that. Because food causes increase in blood pressure, insulin secretion and brain begins to get about you need to store food so get some rest. So if you are on fast you will get your mind fast, have more concentration and control your activity. But it do not apply for those people like student who need to more brain work, they need glucose. So never fast to increase concentration when you are student.
According to Ayurveda (ancient Indian medical science) says, increase in toxic product in your intestine, increases consequence of disease. So from fasting you just clears your stomach and have healthy life.
IT also explains that, our body is made of 80% of water and 20% of solid. So due to moon effect there is increases in emotional instability causes to feel stressed, irritation and violence so fasting acts like antidote and relaxes these problems.

Sources
Wikipedia
IF YOU THINK THAT MORE SHOULD BE ADDED PLEASE ADD AS COMMENT.

How to Cope With Having Lupus

2010-03-15T09:05:00.001-07:00

Lupus is an autoimmune disease which may affect the whole body. It is called the great pretender because lupus may seem similar to many other illnesses. It may take many years before lupus is diagnosed. Lupus affects all ages but the disease typically peaks in the 15 to 40 year-old age group. It primarily affects women. Lupus is an autoimmune disease which means the body begins to attack itself. The kidneys, heart, lungs and blood systems may all be affected. Hallmarks of the disease include skin rashes and the malar (butterfly rash) found across the nose.

Learn about lupus. The following tips and links will help to learn how to live with this chronic medical condition.
Make sure you have the right doctors and keep up with them. Lupus patients generally have a variety of doctors, including a Rheumatologist (for joints). It is essential that you keep your doctors well informed. If you don't get along well with one doctor, then find one that you do get along with.
Learn as much as you can about Lupus. Ask your doctor lots of questions and read up about it online. A good website to start is the Lupus Foundation of America, as listed below.
Take care of yourself. Get enough sleep. Drink enough. Eat enough. Remember to take some time to relax.
Pay attention to your body. Learn to recognize signs of flare ups. If you start to feel ill, try to figure out what is causing it (sun? stress? working too hard?) so that you can avoid it. Also, if you recognize that you are having a flare, you can tell your doctor so that it gets under control before it gets out of hand.
TALK to your doctor. If you have questions or concerns, call your doctor. Don't worry about "bothering them over something minor", you have a serious illness and it's their job to help you with your health. It is better to ask too often than not enough.
Coping with lupus may require a lifestyle change. The majority of lupus patients are photosensitive (sensitive to UV or sunlight). Wear sunscreen and avoid excessive sun exposure.
Keep in mind that stress can cause flare ups. Try to take things one step at a time and not overdo it.
Be aware that having lupus can be difficult for family and friends as well. They may have a difficult time understanding the disease and coping with the fact that you have it. Try to keep them well informed.
Lupus is difficult to see. It is common that people have a hard time understanding that you are ill. Try to educate them about lupus. Send them a link to a lupus website or give them a book about lupus to read.
Most importantly, it can be difficult for YOU to deal with. If you are having difficulty coping, you may want to consider seeing a psychiatrist, joining a support group (internet based or in real life), and talking to your doctor about it.
Set boundaries with family and friends about who they talk about it with. If you don't want them telling everyone about it, tell them so. It is normal for a person to want to keep his/her medical information private.

From WIKIHOW

Aspirin: A Blockbuster Therapy for Breast Cancer?!

2010-03-14T03:32:00.001-07:00

"Am I going to die?" These are the heart-sinking questions that invariably run through the minds of women who have been told that they have breast cancer. They courageously take on surgery, long and arduous chemotherapy, and radiation treatments, hoping to fend off the fate of the 40,000 women whose breast cancer will take a deadly turn this year, reappearing and relentlessly spreading throughout their body. The report just released from the Nurses' Health Study, in a rather understated way, offers 2 million American women who have had breast cancer some vital and actionable information. Taking a single aspirin tablet—a baby aspirin or one adult pill—every other day can be lifesaving. (In fact, were these aspirin tablets a hot new biotech drug, we would be popping champagne right now.) The long-term, low-dose aspirin program was initiated a year or more after the cancer diagnosis as an add-on to treatment, not as a substitute for it, to control the fate of tumor cells silently left behind. For some women, this post-treatment phase is one in which the cancer has gone underground and not entirely disappeared. It can lead to unexpected recurrence with the cancer spreading sometimes 10 to 20 years after diagnosis. Scientists seem to have stumbled upon an easy way to cut that risk. The study followed 4,164 breast cancer survivors over a period from 1976 to 2006, assessing in detail their use of aspirin. The women studied initially presented with tumors ranging from small early invasive cancers confined to the breast to more advanced ones that had spread into surrounding lymph nodes. Over more than three decades of follow-up, 400 women had a cancer recurrence with distant tumor spread, which had killed 341 of them by the time the study ended.

The power to spread is the power to kill, and what aspirin seems to be doing is interfering with that process. It was by coincidence, not by design, that almost half of the medically smart women in the NHS were diligently taking aspirin. The surprise finding: Those who made aspirin a regular habit, consuming low doses two to five times a week (mostly to help their hearts), were 71 percent less likely to have a deadly recurrence of their breast cancer compared to those who were taking little or no aspirin.
A similar trend was found with regular use of other nonsteroidal anti-inflammatory drugs, but the numbers of users over the study time were insufficient for statistical certainty. The findings are nonetheless provocative, reinforcing the notion that the beneficial effects of aspirin on cancer survival may rest with its anti-inflammatory effects. We do know that deadly cancers hijack the inflammatory system to spread and invade distant organs, but we understand woefully little about the process for any given cancer and how to treat it. Figuring out the underlying secrets of this aspirin effect will open up neglected fields of cancer study: why and how cancers ultimately claim lives.
But what does this mean for breast cancer survivors now? A lot. Aspirin has proved itself as a safe, effective, and inexpensive preventive medicine that cuts the risk for colon and prostate cancer and for years has served as a way to prevent heart disease and stroke. Here the Nurses' Health Study shows, for the first time, that breast cancer survivors can substantially lower their risk of a recurrent, deadly tangle with this cancer in the future.
The big caveat is that the study was "observational" and not a blue-chip randomized trial, making many hesitant to crow about its results. I'd say crow nonetheless. The heart disease and stroke benefits are indication enough for any woman to start a low-dose aspirin program—with her doctor's oversight.

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How to Grow Taller

2010-03-13T21:09:00.001-08:00

What you do when you're young will likely affect how big and tall you grow to be. While none of these steps can ensure you grow tall, having a healthy, low stress lifestyle will increase the odds you reach your maximum potential height

Steps

Nutrition

Include all of the following substances in your meals. They're essential for helping your body grow to its maximum natural potential.
- Vitamins- vitamins are organic substances necessary for life and essential for growth. To receive the proper vitamin intake, a well-balanced diet is recommended. However, supplements may be taken where needed but not as a substitute for food. Supplements should be taken after each square meal, or once after the largest meal for proper absorption.
  - Vitamin A (retinol, carotene) - it promotes growth and strong bones, aids in keeping skin, hair, teeth and gums healthy, and helps build resistance to infections. Carrots, liver, egg yolk, milk, green and yellow vegetables, margarine, and yellow fruits are the best sources of vitamin A. The recommended supplement is 10,000 to 25,000 IU. However, if your diet includes ample amounts of spinach, liver, sweet potatoes, or cantaloupe, it is not likely you'll need a supplement.
  - Vitamin B1 (Thiamine) - it promotes growth, aids in prevention of beriberi and nervous disorders, aids digestion and helps heart and nervous system function properly. The bet sources for it are pork, rice, whole wheat, dried yeast, peanuts, soybeans, peas and green vegetables. Supplements of 100 to 300 mg are most common. More effective when used in B complex formulas.
  - Vitamin B2 (Riboflavin) - it promotes growth, aids in reproduction, promotes healthy skin, hair and nails, and helps maintain good eyesight. You can find it in milk, liver, eggs, fish, kidney, yeast, leafy green vegetables. The recommended supplement is 100 to 300 mg per day.
  - Vitamin B6 (Pyridadne, Pyridixinal) - it reduces night muscle spasms, leg cramps, hand numbness and certain forms of neuritis, properly assimilates protein and fat, and works as a natural diuretic
    . It can be found in liver, brewers yeast, cabbage, black strap molasses, cantaloupe, beef and kidney. For best results, take supplements in equal amounts with vitamins B1 and B2. If taking a B complex formula, be sure it contains enough B6 to be effective.
  - Vitamin B12 (Cobalarnin) - it promotes growth, increases your energy level, forms and generates red blood cells, and helps balance and concentration. You can find it in cheese, liver, kidney, pork and beef. The recommended supplement dosage varies from 5 to 100 mcg., depending on deficiency.
    - Note: Common deficiencies are noted in vegetarians and high protein consumers. Vitamin B complex formula should include all the B complex vitamins so far discussed plus other important vitamins vital to the growth process. Be sure to read labels and check with your pharmacist to make sure your formula includes the proper amounts of each vitamin best suited to your needs.
  - Vitamin C (Ascorbic Acid, Cevitamin Acid) - it aids in healthy bones and teeth, prevents scurvy, helps treat and prevent common colds, enables protein cells to hold together, and aids blood vessel circulation. Plays a primary role in the growth and repair of body tissue cells. It can be found in citrus fruits, tomatoes, berries, cauliflower, raw cabbage and potatoes. The recommended dosage of supplement varies from 1,000 to 10,000 mg. per day.
  - Vitamin D (Calciferal, Viosterol, Ergosterol) - it is essential for strong bones and teeth, prevents rickets, which deteriorates bones and could cause bowed legs, knock knees and poor posture, aids treatment of conjunctivitis, and aids vitamin A. The best sources for it are milk products, fish liver oil, fish meats and sunlight. A good dosage of supplement is 400 to 1000 IU per day.
  - Vitamin E (Tocopherol) - it supplies oxygen to the body for more endurance, and helps prevent and destroy blood clots. It can be found in wheat germ, eggs, leafy greens, soy beans, spinach, whole wheat, and broccoli. A supplement of 200 to 1,000 IU per day is recommended. Due to chlorinated drinking water, in a large percentage of the population, vitamin E may be in great demand.
  - Vitamin F (Unsaturated Fatty Acids Linoleic and Arachidonic) - it aids in growth, helps prevent heart disease and cholesterol deposits in arteries. Vegetable oils, walnuts, pecans, and almonds, soybeans, linseed and sunflower oils are the best sources of vitamin F. A supplement of 100 to 150 mg is recommended. For best absorption take with vitamin E. If you consume excessive amounts of carbohydrates, you will probably need more vitamin F.
  - Vitamin K (Menadione) - it aids in proper blood clotting and helps prevent internal bleeding and hemorrhaging. It can be found in egg yolk, yogurt, fish liver oils, soybean, green vegetables and kelp. A supplement of approximately 300 mcg. is adequate. Due to an abundance of natural vitamin K, supplementation is not usually necessary. See a doctor before taking a supplement.
    - Note: Proper amounts of vitamins will differ depending on individual characteristics and diet. Certain dosages may not be suitable for all individuals and could be detrimental to your health. Please seek advice of a physician when supplementing with vitamins. All vitamins in this chapter will not be needed however,they are noted because of their ability to work well in a totally balanced diet.
- Minerals- minerals are inorganic substance found naturally in the earth. They make up a large part of our bones and teeth and help regulating other body functions. Here are some of the minerals to which you should pay special attention.
  - Calcium - it aids for strengthening soft bones and teeth, and helps maintain regular heartbeat. Milk and dairy products, sardines, soybeans, dried beans, and green vegetables contain large amounts of calcium. A supplement of 600 to 1500 mg per day is recommended.
  - Chromium - it aids in growth process, and helps prevent and lower high blood pressure. The best sources for it are meat, brewer's yeast, clams, chicken, shellfish. A supplement of 90 mcg. per day is average for adults.
  - Chlorine - it helps keep your body limber, and aids digestion by cleansing system. It can be found in table salt, kelp, and olives. A recommended supplement dosage has not been determined. However, an average daily salt intake should be more than sufficient.
  - Fluoride - it strengthens bones and reduces tooth decay. It is found in fluoridated drinking water, seafoods and gelatin. A necessary amount of supplement has yet to be determined, but most people get sufficient amounts from fluoridated drinking water. Available by prescription in areas without fluoridated drinking water. Additional fluoride should not be taken unless advised by a physician.
  - Iodide - it promotes growth, provides energy and burns excess fat. The best sources for iodide are kelp, salt, all seafood, onions, and vegetables grown in iodine-rich soil. A good amount of supplement is 80 to 150 mcg. per day for adults. Iodine aids in the proper functioning of the thyroid gland which fosters and regulates growth. Check your salt to see if it is iodized along with your vitamins or multi?mineral preparations. Too much iodine can cause a harmful effect. For additional supplements of iodine not discussed, a physician's advice is recommended.
  - Iron - it aids growth, prevents fatigue, helps form hemoglobin in blood, and aids for good skin tone. You can find it in liver, beef kidney, egg yolk, oysters, cereals, raw clams, and red meat. A supplement of 10 to 18 mg per day is sufficient. Check your iron supplement to see if it contains "Ferrous Sulfate" an inorganic iron which can destroy vitamin E in your system. Try to avoid this preparation. For women, whose bodies use much more iron than men's, a supplement is most likely needed.
  - Magnesium - it aids the cardiovascular system, aids nerve and muscle functions, and is needed for calcium and vitamin C metabolism. It is found in figs, apples, grapefruits, lemons, seeds, nuts, yellow corn and dark green vegetables. A necessary amount of supplement is 300 to 400 mg. daily. If you live in area with hard drinking water, or consume large amounts of nuts, seeds and green vegetables, you're probably getting sufficient amounts of magnesium.
  - Phosphorus - it aids growth and provides energy. The best sources for it are fish, poultry, eggs, meats, nuts, seeds and whole grains. As for supplements, RDA is 800 to 1200 mg per day for adults. Most diets already include adequate amounts of phosphorus or "phosphates" in them, so check before taking a supplement.
  - Other important minerals are:
    - Potassium - Sources include: citrus fruits, watercress, bananas, potatoes and green vegetables.
    - Sodium - Sources include: salt, shellfish, carrots, beets, bacon and kidney.
    - Sulfur - Sources include: lean beef, fish eggs, cabbage and dried beans.
    - Zinc - Sources include: steak, eggs, lamb chops, brewer's yeast and pumpkin seeds.
- Proteins- everyone's protein requirements are different, depending upon your size, age and physical condition. A large, young person will require more protein than a small, older person. Proteins have different functions, and work in different areas of the body. There are basically two types of protein: complete and incomplete protein.
  - Complete Protein: provides the proper balance of eight necessary amino acids that build tissues, and are found in foods of animal organ such as meats, poultry, seafoods, eggs, milk and cheese.
  - Incomplete Protein: lacks certain essential amino acids and is not used efficiently when eaten alone. However, when it is combined with small amounts of animal?source proteins, it becomes complete. It is found in seeds, nuts, peas, grains and beans. Mixing complete and incomplete proteins can give you better nutritional value than either one alone.
- Water- water is the basic solvent for all the products of digestion. It is essential for removing waste from our bodies and 6 to 8 glasses daily is considered healthy.
Eat a steady amount of food. That's why dieting is not recommended as long as you're still growing. Not eating a good quantity of food will deprive your body of the substances detailed above, therefore stunting your growth. If you want to lose weight, decrease the amount of fats, sugar and carbohydrates from your alimentation, but eat the rest of foods in good, moderate amounts. Also try losing weight through sports. After all, you can still lose weight later, but you must make the most out of your adolescence if you want to grow taller.
Schedule your meals. 4-5 hours is the time needed for insulin to disappear from your bloodstream so that growth hormone can work on building your tissue. This is the period of time that you should wait between meals. Proteins, which are absorbed slowly, should keep you from becoming hungry. Your last meal should be 3-4 hours before you go to sleep. Small amounts of protein shake are allowed right before your bedtime.

Exercising

Tickle Massage.

This works best if somebody else does it for you: Massage your knee for 5-10 minutes. This should be very ticklish to some others. Massaging your knees with your hands allows your knees to release growth chemicals that helps you grow your bones.

Stretch. Stretching is the most effective form of exercising when it comes to height increase. Doing intense stretching exercises can add a few extra inches to your height, even short after your growth has stopped. For best results, you should do these exercises two times a day, after you wake up and before you go to bed. Start with easy exercises, then gradually move on to more difficult ones. Here is a list of stretching exercises, divided in three categories, from the easiest to the most difficult:
- Preliminary exercises - These exercises will play the role of a warm-up, in order to prepare the body for the more difficult ones. These preliminary exercises require very little effort and should not result in tiredness or fatigue. You will also notice that the side benefits include good posture development, straightening of the spine, and stretching the body. All of these will contribute to height increase. It is recommended that you continue these exercises during the complete time you are performing the other exercises in the following chapters. Perform all of the exercises twice a day, for 7 days, before you start the regular exercises. You should be able to complete all of the preliminary exercises within 15 minutes.
  - Before getting out of bed each morning (and before going to sleep each evening), stretch your arms and legs to their limit. Point your toes towards the foot of the bed, point your outstretched arms towards the head of the bed, and stretch your body to its limits. Twist and turn your body in every possible direction, stretching every joint and muscle in your body simultaneously.
  - Still in bed, lying flat on your back, place your hands on your hips and lift your legs and lower torso into the air so that your weight is resting on your elbows and upper back. In this position, with your feet straight up, rotate your legs in the same manner as if you were riding a bicycle. Continue the pedaling motion for 60 seconds.
  - In the sitting position, while still in bed, allow your head to droop forward with your chin as close to your chest as possible. Rotate your head to the left, then backward, to the right, and then forward. Repeat this circular rotation of the head several times, and then rotate the head in the opposite direction several times. Extend the head as far to the left and right, and as close to the chest and back as possible. Loosen up those neck joints!
  - In the standing position, arms stretched out horizontally away from the body, rotate the arms in a circle approximately 2 feet in diameter. Keep the arms extended to the side, and do not bend the elbows. Rotate the arms from the shoulder joints. After several rotations, rotate the arms in the opposite direction several more times. Extend the arms as far backwards as possible during each rotation.
  - Stand away from all walls or other objects with your feet about 18 inches apart. Allow your head to fall loosely backwards as far as it will go without straining your neck. Raise both arms sideways, away from the body, and stretch them outward as far as they will go. Hold them there, level with your shoulders. Start the exercise by swinging your torso all the way to the left, and then all the way to the right. Keep your arms stiff and straight. Keep performing this swinging movement, to the left, to the right, then left, then right, left, right. Do it naturally and smoothly. During the motion, extend your arms as far outward as you can, and twist the body as far as you can in each direction. Repeat this exercise for 60 seconds, and then clasp your hands behind your neck, and perform the same swinging movements, left, and right, for another 60 seconds.
  - In the standing position, facing a wall, with your stomach and toes touching the wall, raise your left hand and reach as high up on the wall as you possibly can. Do not lift your heels. Let your fingers touch the wall as high as possible, and try to move your fingers up the wall a little further. When you have reached as far as possible, hold that position for several seconds, and slowly lower your arm to your side. Repeat the same procedure with each hand, a total of 3 times. Turn your left side to the wall, and perform the exercise with your left hand, 3 times. Turn your right side to the wall, and perform the exercise with your right hand, 3 times. Turn your back to the wall, clasp your hands together, behind your head, and raise both hands as high up on the wall as possible, without unclasping your hands, and without lifting your heels. Perform this 3 times.
- Regular exercises - These exercises are designed to not only help in the continued straightening of the spine but, also in stretching the body and strength of the muscles involved. This is how you're going to accomplish your ultimate goal of more height. You will continue to perform the preliminary exercises every day in addition to the regular exercises. To avoid over exerting yourself, we advise doing only 5 out of the 15 regular exercises each day and rotating them each day. For example you would perform the first 5 on the first day, the second 5 on the second day, and the last 5 on the third day. Repeat this cycle 6 more times, for a total period of 21 days doing both preliminary and regular Exercises.
  - Stand erect behind a chair, feet together with hands gripping the back of the chair. Your feet should be about 12 inches away from the chair. Lift your left leg back and up as far as possible, maintaining your grip on the chair for support. Bring your leg down, and repeat the same procedure with your right leg. Perform the leg lifts slowly and stretch them out as far as possible. Repeat the leg lifts 10 times for each leg.
  - Lie flat on your back on a firm surface. Lift your left leg up, bending it at the knee, and touch your chin to your knee. Grasp your leg with both hands below the knee, and pull your knee to your neck. You may lift your head off the floor, towards your knee, but do not lift; your shoulders off the floor. When your knee touches your neck, hold that position for a few seconds, and then return to the original starting position. Perform the exercise with the other knee. Alternating legs, repeat the exercise 10 times with each leg. When bringing knee into chest remember to inhale so lungs are filled with oxygen. Holding that position, then exhale while returning to starting position.
  - Stand erect, knees and heels together with arms relaxed at your sides. Raise your arms outward and up until they meet at the highest point over your head with the knuckles of each hand facing and touching each other. As you raise your arms, lift your heels so that all your weight is on your toes. Stretch your arms and body up as far as you can. As the arms are raised, inhale and fill your lungs to full capacity. Lower your arms in the same course as you raised them, exhale slowly, and lower your heels until they touch the ground. Repeat this exercise for 1 minute the first time, 2 minutes the second time, and 3 minutes every time thereafter.
  - Lie down on your stomach with your hands behind your back, clasp your hands together and interlock your fingers. Arch your body so that your head, shoulders and legs are raised off the ground and maintain this position. Rock your body forward and then backward several times, and then relax. Repeat this exercise 5 times. While still on your stomach, stretch both arms out in front of you and rest them on the floor. Commence raising and stretching your legs upwards, alternating legs, without bending your knees. Do this 5 times with each leg. As in all exercises, if at any time you become very tired, stop and rest before continuing.
  - Stand erect with your arms high over your head and your thumbs interlocked. Stretch your body upwards vigorously without lifting your heels, and then bend far to the right. Return to the position with your arms over your head and bend far to the left. Repeat this movement to the right and to the left alternately for 10 times. Rest 1 minute, and repeat it another 10 times. Do it slowly, but keep stretching your arms and torso all during the exercise.
  - Lie flat on your back with your hands below your buttocks. Raise both legs off the ground, straight up. Bend your knees and lower your toes so that they touch the floor. Lift your hips off the floor, supporting your body with your hands on the floor. Arch your body so that your weight lies only on your shoulders and your toes. Lower your hips so they rest on your hands, lift your toes, straighten your legs, and lower them to the floor so that you are in the starting position. Repeat this procedure 10 times, each time trying to arch your body as much as possible when you touch the floor with your toes.
  - Seated in a large arm chair, stretch your feet straight out and stiffen your body so that the only points of contact between your body and the chair are at the top of the chair and at the front of the seat. Your buttocks should be resting on the front of the seat. Lean slightly forward, and at the same time bring your knees up to your chest. Use your arms to help bring your knees closer to your chest by wrapping your arms around your bent legs and pulling them towards you. Breathe normally during this exercise. Release your legs and return to the original position. Repeat this exercise 5 times.
  - In the standing position, feet about 18 inches apart, place your palms on the back of your thighs. Without bending your knees, slide both hands down the sides of your legs as far as you can reach. You will have to bend forward to perform this exercise, but be sure to maintain contact between your palms and your legs. The further you reach down, the more excessive the strain on the back of your knees. Do not bend them. Vary this exercise by placing your hands on your buttocks and moving them down the back of your legs, bending your body backwards to enable you to perform this movement. Perform each of these exercises, slowly, 5 times.
  - Stand erect with your back to the wall, feet flat on the floor. Your feet should be about 24 inches away from the wall on the first time you perform this exercise. On following days, you will increase your distance from the wall 3 additional inches each time. Stretch the arms forward, upward, and then backwards over your head until your fingers touch the wall behind you. Do not allow your body to touch the wall. If you find it very easy to touch the wall, move a few more inches away from the wall. You should have to stretch your body to enable you to touch the wall. After your fingers achieve contact with the wall, return to the original positions by bringing the arms back over the head and then down. Repeat this exercise 7 times. Keep a careful record of the distance from the wall the last time you performed this exercise so you can increase this distance by three inches each different day you perform it.
  - In the standing position, place your left leg far out in front of your right leg, and distribute your weight evenly on both legs. Without lifting your right leg, shift your weight forward to your left leg, bending the left knee and placing your left hand on your left knee to maintain your balance. Lean forward as far as you can, placing most, of your weight on your left foot. You may lift your right heel, but do not allow your right toes to lift off the floor. Stretch your body forward as far as possible, hold this position for 3 seconds, and return to the starting position. Switch positions of your legs, and perform the exercise by shifting your weight forward to your right leg, and maintain balance with your right hand. Perform the exercise 5 times in each position.
  - In the standing position, feet slightly apart, hands on hips, body straight and erect, head up. Slowly bend the knees while keeping the rest of your body straight and erect, and come to a squatting position with the knees straight ahead (not spread apart). As you slowly bend your knees, extend your arms straight ahead, stretching the arms and fingers to their fullest extent. Hold this position for a few seconds before slowly returning to the original position. Repeat this exercise 10 times.
  - Sit on the floor with your legs extended straight out in front. Place your feet under an article of furniture (chair, table, or sofa) to keep them from moving during the exercise. Interlock your fingers behind your neck, and start the exercise. Slowly revolve your trunk in a large circle, bringing your upper torso forward, to the right, backward and to the left, as far as possible. Rotate your body in as wide a circle as possible, and then reverse directions. Repeat this exercise for 30 seconds, rest 15 seconds, repeat 30 seconds, rest 15 seconds, etc., for a period of six 30 second units.
  - In the standing position, with your legs spread widely apart, raise both hands straight overhead and overlap your hands. Bend over forward and touch the floor between your legs, then return to the original position. Perform this exercise 10 times. You will notice that the wider your legs are spread apart, the easier it will be to touch the floor. As you progress in this and the other exercises, you should bring your feet closer together so that it will be more difficult to touch the floor with your fingers. If you can touch the floor with your feet close together, you should then strive to touch the floor with your palms.
  - In the prone position, lying flat on your back with your arms by your side, raise your arms and bring them straight back over your head until they are stretched out on the floor, pointing away from your head. Resting all of your weight on your outstretched arms, your shoulders, and on your heels, slowly raises the back, hips, torso, and upper legs off the floor. Stretch your body up as high as you can. Hold that position for a few seconds, and return to the original position. Perform this exercise 5 times.
  - In the standing position, feet spread slightly apart, hands behind the neck with your fingers interlocked. Bend the body forward from the waist, without bending the knees, and try to bring your head down between your legs. Of course, it will be impossible to bring your head all the way down, but stretch your body as far as possible without straining yourself. Do not remove your hands from behind your head. Use your hands to help push your head down further. When you have reached as far down as possible, return to the starting position. You will perform this exercise with more ease after one or two weeks. Perform this exercise 5 times.
- Advanced exercises
Swim.
Raise your seat while cycling.
Play sports like basketball or volleyball, which involve jumping a lot.
Hang off a bar while wearing ankle weights.
Sprint.
Jump rope.
Perform kicks. Repeated kicking helps lengthen the shin and thigh bone. Ever noticed how kick-boxers have longer than average legs? It's the same principle as far as baseball pitchers too. Pitchers usually have a pitching arm that is 1-2 inches (2-5 cm) longer than their other arm. Same principle. Repetitive stress forces the bones to lengthen due to the stress. There are only 2 kicks you will have to do in this program. Perform 3 sets of both kicks each day. Shoes are optional.
- Front Snap Kick - the first kick is the basic front snap kick. It's very simple. Keep both feet facing forward in a comfortable position, with a slight bend to the knees for balance. Raise your knee until the thigh is parallel to the floor. While doing this, the ankle should be flexed downwards and your toe should be pointed towards the floor. Next, quickly snap your foot outward with your toe pointing outward. Once that is done, quickly return your toe to its previous downward pointing position and lower the foot onto the ground. That is one repetition. The kick will be quick. It doesn't matter how high off the ground you kick - our focus here is simply on executing the quick snapping motion of the lower leg. Each kick will last approximately than 1 second. Perform 20 kicks per leg then switch to the other leg and repeat. That is 1 set. Rest for 1 minute between sets then repeat the process again for a total of 3 sets.
- Straight Leg Kick - the second kick is a straight leg kick. The best way to explain this is to pretend that you are punting a ball. Keep both feet facing forward in a comfortable position, with a slight bend to the knees for balance. Unlike the snap kick, you will NOT bend your knees while performing this kick. Keeping your leg straight, raise your leg quickly as if you were punting an imaginary ball. Raise your leg as high as possible with your toe pointing outward and upward. I find it helpful to imagine that there is a target in front of me at about eye level that I am trying to kick with my foot. Quickly lower your leg to starting position and place your foot on the floor. That is one repetition. Once again, the kick will be quick. Your goal is to keep your leg perfectly straight and kick as high as possible. Each kick will last approximately 1.5 seconds. Perform 10 kicks per leg then switch to the other leg and repeat. Rest for 1 minute between sets then repeat the process again for a total of 3 sets.

Sleeping

Sleep 8 to 10 hours a day. Most of your growing takes place while sleeping, because that's when most HGH is released. Not getting enough hours of sleep will stunt your growth.
Keep your body as straight as possible while sleeping. your spine must be as straight as possible. Lie on you back with your arms and legs stretched toward the foot of the bed. Do not exert any effort or pressure to stretch your limbs. Allow your body to be completely relaxed. You may let your head turn to the right or left and bend your arms if it is more comfortable to you. The important thing is to keep your body (torso and legs) as straight as possible. This position may prove to be uncomfortable for the first few nights, but your body will soon become accustomed to this manner of sleeping and before long you will discover not only extra inches but, also a. more comfortable sleep.
- Kick the 'pillow habit'. This is a very common mistake made by most of us because we are led to believe that a pillow allows for a more comfortable night's sleep so, through habit, we become attached and generally accept this as the most comfortable way to sleep. However, nothing could be further from the truth. The use of a pillow is an incorrect form of sleeping and should be avoided. While lying on your back with your head resting on a pillow, your neck is bent forward in a very unnatural position. In this position, your head is being pushed forward and your back is arched, also a very unnatural position. If you suffer from frequent neck or back pains, in the majority of cases you can probably blame it on your pillow or mattress.
Make sure your mattress is firm and capable of giving your body full support. This is to aid in keeping your spine as straight as possible while sleeping. A soft or sagging mattress will tend to bend the spine and curve the torso in a sinking effect, which must be avoided. A good mattress will support the whole body, which will keep it in a straight posture setting - a must for obtaining greater height.

Posture

Correct your posture. Many people rob themselves of extra height because they fail to realize that a good posture is essential for maximum height increase. Correct posture involves more than just standing straight and erect. You must train each part of your body to maintain its proper position. You must learn how to hold your head, your pelvis, your legs, sit correctly, walk correctly, plus numerous other do's and don'ts to assure you of achieving every possible inch of height.

Tips

You're taller in the morning than in the evening by up to a full inch. This is due to decompression of spine discs, after you lay in horizontal position for many hours (like when you sleep). After standing up for a few hours, you will lose that height. So, when measuring yourself, it's recommended to count your evening height as your 'official' height, because measuring in the morning can be deceiving. Also, try to measure yourself at the same time of the day.
Also recommended as preliminary exercises are physical recreation activities such as walking, jogging, tennis, swimming, bicycling, handball, baseball, soccer and any type of activity involving muscle exertion. Your body is a machine, and if you do not use it regularly, the working parts will become "rusty" and inoperable. To look your best, to feel your best, and to be able to do your best, you must exercise regularly. That is man's nature, and modern technology can't change it.

Warnings

There are no magic formulas or secret compounds that will help you grow. Don't buy eBooks which will offer no more information that you can read here for free.
Your growth is determined largely by genetic factors. If your parents are both small, there isn't much chance of being tall (unless you have another relative who is tall). However, the advice in this article will still help you reach your full height.

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How To Grow Taller on Howcast

Things to do or not to do after blood donation (post donation precautions)

2010-03-12T23:38:00.001-08:00

Do not drive for half an hour. Due to blood donation there is slight drop in blood pressure. That may cause dizziness ,light headiness and sometime nervousness. So wait for half an hour in this time blood pressure comes to normal and body adjusts to blood loss.
Do not smoke for half an hour. Because of blood loss, blood volume is less in circulation and heart tries to compensate this loss by pumping more blood in your body. And you know tobacco products contain nicotine which causes elevation in blood pressure and it can cause angina and even heart failure.
Do not go in direct sunlight for longer time. As i stated above due to blood loss there is tachycardia (increase in heart beat) due to blood loss and heart is compensating to blood loss. if you go out in direct sunlight there will be sweating and evaporation of water, it will leads to more tachycardia and more prone to heart attack.
Take enough liquid for next 24 hour. Due to blood loss the first thing required to manage in body is water and remaining things like protein, minerals and cells loosed in blood donation is less important. As theories body compensate this loss of water from extra vascular space but there is imbalance in homeostasis. So drink as much water you can drink for next 24 hours. At least 4 glass within 6 hrs.
Do not take any alcoholic product for 24 hrs. Alcoholic products causes dehydration and pooling a lot of water from body to urine. And you should avoid to loosing water so early so please avoid alcoholic drink for next day.
Do not go for heavy physical activity for next 24 hour. As you have already understand that water is losses from your body and you have to prevent this water, and you also knows heavy exercise cause sweating so better to avoid heavy exercise, cycling, football playing etc.
Take your normal diet. Take your normal diet because you have not loosed anything special in blood donation. Lets find out what you have loosed, protein, minerals, cells, water and some clotting factors. Protein is manufactured in liver from amino acid and body have a lot of amino acid reserve. Second thing is minerals, kidney is enough capable to preserve enough minerals from loss in urine, even our diet supply that much. Cells like RBC, WBC and platelet is manufactured in bone marrow from cell division so you don’t need to worry for that just take some more iron content in your diet because loss of RBC needs iron to form hemoglobin.
If you feel dizziness. Dizziness is caused when blood is not approaching in brain. Due to blood donation, blood volume is low and this amount is not sufficient to provide enough oxygen in brain so you feel dizziness. This is most common problem in blood bank and just solved by relaxing for some time. If you feel this problem then just sit down or lay down for some time.
If you feel light headed. The problem is same like dizziness so you need to lay down and elevate you foot above your head level. The problem is usually solved in five to ten minutes under expert supervision but if you are alone then just wait until you feel to pee.
Bleeding is not stopping or brushing. This is also a common problem in blood bank. If your bleeding is not stopping apply pressure for 5- 10 min and still it is bleeding consult to doctor. Bruising is causes by blood that collects under skin that causes metabolism of hemoglobin and formation of blue/black colored pigment under you skin. To remove this apply ice pack frequently for next 24 hours.(commonly 4hrly gap).

If you have a question please leave comment i will glad to answer this.

How to Do Pranayam

2010-03-10T20:04:00.001-08:00

Pranayam (also spelled Pranayama) is an ancient practice concerned with breath control. Research has shown that that practicing Pranayama can relieve symptoms of asthma.It is also beneficial in treating stress related disorders, such as anxiety and depression.There are a total of six types of Pranayam practice, all of which are detailed here.

Bhastrika Pranayam

Sit in comfortable position, you may sit even on a chair.
Breathe in through your nostrils until the lungs are full. Feel the diaphragm move down to allow the lungs to expand and forcing the abdomen out, followed by the mid section of your chest expanding and finally your collar bone rising.
Breath out forcefully and uniformly, again through your nostrils. Again feel the collar bone dropping, chest deflating and the diaphragm moving up as the lungs collapse allowing the abdomen to be sucked in. This process of exhaling should be much faster than the process of inhaling.
Repeat the process. When correctly done, your chest will expand when you breathe in and deflate when you breathe out. Continue doing this for 5 minutes.

Kapalbhati Pranayam

Sit erect.
Inhale through your nostrils a little and exhale through both nostrils forcefully.
Inhale again a little and follow with another forceful exhalation passively and effortlessly.
Continue these cycles. The frequency should be about 60 strokes/minute.
Continue doing this for 15 minutes. You may take a minutes rest after every five minutes.

Anulom Vilom Pranayam

Sit comfortably.
Close your eyes.
Close the right nostril with the right thumb.
Inhale slowly through the left nostril and fill your lungs with air.
Close your left nostril with the ring and middle fingers of the right hand and open the right nostril.
Exhale slowly and completely with the right nostril.
Again inhale through the right nostril and fill your lungs.
Close the right nostril by pressing it with the right thumb.
Open the left nostril, breathe out slowly. (This process is one round of Anulom Vilom Pranayam.)
Continue for 15 minutes. You may take a minute's rest after every five minutes of exercise.

Bahya Pranayam

Breathe air out, touch chin to chest, squeeze stomach completely and hold for a while.
Release chin, breathe in slowly.
Repeat 3 to 5 times.

Bhramari Pranayam

Close ears with thumb, index finger on forehead, and rest three on base of nose touching eyes.
Breathe in. And now breathe out through nose while humming like a bee.
Do this three times.

Udgeeth Pranayam

Breathe in deeply, and chant OOOOOOm (long O and small m).
Do this 3 times.

Video

Watch this video to understand how to do

Tips

Do Pranayam preferably in the morning with an empty stomach.
If you prefer to do Pranayam in the evening, do it on an empty stomach and keep a gap of at least 5 hours between your meals and Pranayam.
Those who cannot sit in Asanas can sit on the chair and do Pranayam.
Never hold your breath while doing Pranayam.

Warnings

Pregnant woman and persons having a fever should consult a doctor before doing Pranayam.
Children above 5 years should do Bhastrika Pranayam for only 2 minutes, Kapalbhati and Anulom Vilom Pranayam each for 5 minutes.
Even after consulting doctors, it is important to remember that Kapalbhati can be risky in those who have abdominal wounds, surgical operations, hernia, peritonitis, appendicitis, prolapse of rectum or uterus, hiatus hernia and recently delivered women.
Some of the pranayam is not recommended for people with High Blood Pressure.

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Creatinine

2010-03-08T01:01:00.001-08:00

Creatinine is synthesizes from creatine in muscles by creatinine kinase enzyme. The conversion is fairly constant beside some muscle degeneration disorders, extensive exercise and some other problems.

Formation of creatine

Creatine is synthesizes in liver, kidney and pancreases by two enzymatic reaction, in first reaction transmidation reaction of arginine and glycine forms guadinoacetic acid and in second reaction methylation of guadinoacetic acid occurs with S-adenomethionine as the methyl donor occurs.

Then this creatine stored in muscles, brain and high oxygen demanding tissues. Where it phosphorylated to phosphocreatine a high energy compound like ATP.

Function of creatine

Supplement of energy: When oxygen is not available in muscle and body needs energy creatine phosphate donates phosphate to compensate energy crisis.
It also stimulates muscle growth by accumulating water molecules due to presence of phosphate.
Increases protein synthesis.

Clinical significance

Creatinine is measured in medical science as indicator of renal damage.

Analytical method for creatinine

There are various creatine measurement procedure are available chemical and enzymatic. I am going to explain few of them and continue to update when new technology comes out.

Jaffe reaction: this method was first described by Jaffe in 1886. In this method the alkaline picrate reacts with creatinine to produce a orange-red complex. The mechanism of this reaction still not clear but this is the best method for routine laboratory in term of economical, nearby to reference method and more reproductive. The produced color is then compared with chemical standard in same environment of test and read in spectral measurement devices. Because Jaffe reaction is not specific for creatinine and we not know the theory of chemical reaction a lot of chemical like protein, glucose , ascorbic acid, ketone bodies, pyruvate, guanidine and cephalosporines are shows interference either positive or negative. To reduce this now this Jaffe reaction is modified from endpoint reaction to kinetic reaction to reduce these interferences. In kinetic reaction we takes two reading of mixture at 20 second and second on 80 sec to reduce protein and bilirubin interference.
Creatinase method : In this method creatinine reacts with creatinase to produce creatine.then creatine reacts with creatine kinase in the presenace of ATP to form creatine kinase and ADP. The resulted ADP reacts with pyruvate kinase in the presence of phosphoenol pyruvate to produce pyruvate and again ATP.Then pyruvate reacts with lactate dehydrogenase in presence of NADH to produce lactate and NAD. The conversion of NADH to NAD is monitored at wavelength of 340 nm.

HDL: unknown roles

2010-02-28T20:48:00.001-08:00

Emerging evidence suggests that HDL function is not always accurately predicted by HDL cholesterol levels. The functions of HDL include reverse cholesterol transport and modulation of inflammation. These functions appear to have evolved as part of the innate immune system. In healthy individuals, in the absence of systemic oxidative stress and inflammation, HDL is anti-inflammatory. Now i will tell you different unknown facts about HDL.

HDL can modulate LDL oxidation. Lipoproteins evolved to facilitate the extracellular transport of lipids in multi cellular organisms. The major protein in LDL is apolipo protein (apo)-B. This protein contains a binding domain that causes LDL to be deposited in the extra cellular matrix of many tissues, particularly in arteries that are predisposed to atherosclerosis. Unlike LDL, HDLs do not normally bind to extra cellular matrix molecules, and the concentration of the main protein in HDL (apoA-I) in the sub endothelial space of normal arteries is only one-fifth the concentration found in plasma. Addition of normal HDL abolished this process, indicating that normal HDL is capable of
preventing LDL oxidation and the inflammatory response induced by LDL.
The acute-phase response changes HDL’s ability to inhibit LDL oxidation. HDL loses its ability to inhibit LDL oxidation during the acute-phase response. HDL from the same rabbits or humans, isolated at the peak of an acute-phase response, was less effective in inhibiting LDL oxidation and actually increased LDL-induced MCP-1 production.
The chronic acute-phase response. By this definition, diseases characterized by persistent elevations of acute-phase reactants such as C-reactive protein (CRP) are examples of a chronic acute-phase response. The important clinical implications of the persistent elevations of these plasma biomarkers in chronic disease states have been extensively studied.
HDL in diabetes, the metabolic syndrome, obesity, and familial hypercholesterolemia Hedrick et al. found that glycation of HDL by incubation under hyperglycemic conditions caused the HDL to lose its ability to inhibit monocyte adhesion to human aortic endothelial cells exposed to oxidized LDL. Glycation of the HDL associated enzyme PON1 also prevented its ability to inhibit monocyte adhesion to the endothelial cells.
HDL in rheumatic diseases Systemic lupus erythematosus (SLE) is an autoimmune disease with chronic inflammation and episodes of acute inflammation. Pro inflammatory HDL was found in 44.7% of women with SLE, 20.1% of women with rheumatoid arthritis, and only 4.1% of healthy women, even though all three groups had normal HDL cholesterol levels.
HDL in leprosy. it is reported that normal HDL strongly promoted the conversion of monocytes into CD1b dendritic cells. It is found that HDL from leprosy patients was
proinflammatory (i.e., HII 1.0) and was also less effective in promoting the conversion of monocytes into CD1bdendritic cells than anti-inflammatory HDL from control
subjects.
HDL as a potential therapy and therapeutic target. In animal models, HDL and apoA-I have been highly efficacious in the treatment of atherosclerosis . In pilot studies in humans, apoA-I has also shown promise, including in patients with diabetes.

Written by M. NAVAB AND ASSOCIATES in article

HDL as a Biomarker, Potential Therapeutic Target, and
Therapy

Breast cancer: causes

2010-02-27T21:45:00.001-08:00

Breast cancer affects one in eight women during their lives. Breast cancer kills more women in the United States than any cancer except lung cancer. No one knows why some women get breast cancer, but there are a number of risk factors.

Certain changes in DNA can cause normal breast cells to become cancerous. DNA is the chemical in each of our cells that makes up our genes -- the instructions for how our cells work. Some inherited DNA changes can increase the risk for developing cancer and are responsible for the cancers that run in some families. But most breast cancer DNA changes happen in single breast cells during a woman's life rather than having been inherited. These are called acquired changes, and most breast cancers have several of these acquired gene mutations. But so far, the causes of most acquired mutations that could lead to breast cancer remain unknown.

While we do not yet know exactly what causes breast cancer, we do know that certain risk factors are linked to the disease. A risk factor is anything that affects a person's chance of getting a disease such as cancer. Different cancers have different risk factors. Some risk factors, such as smoking, drinking, and diet are linked to things a person does. Others, like a person's age, race, or family history, can't be changed. But risk factors don't tell us everything. Having a risk factor, or even several, doesn't mean that a person will get the disease. Some women who have one or more risk factors never get breast cancer. And most women who do get breast cancer don't have any risk factors. While all women are at risk for breast cancer, the factors listed below can increase a woman's chances of having the disease.

Although many risk factors may increase your chance of developing breast cancer, it is not yet known exactly how some of these risk factors cause cells to become cancerous. Hormones seem to play a role in many cases of breast cancer, but just how this happens is not fully understood.

Risk factors you cannot change

Gender: Simply being a woman is the main risk for breast cancer. While men also get the disease, it is about 100 times more common in women than in men.

Age: The chance of getting breast cancer goes up as a woman gets older. About 2 out of 3 women with invasive breast cancer are age 55 or older when the cancer is found.

Genetic risk factors: About 5% to 10% of breast cancers are thought to be linked to inherited changes (mutations) in certain genes. The most common gene changes are those of the BRCA1 and BRCA2 genes. Women with these gene changes have up to an 80% chance of getting breast cancer during their lifetimes. Other gene changes may raise breast cancer risk as well.

Family history: Breast cancer risk is higher among women whose close blood relatives have this disease. The relatives can be from either the mother's or father's side of the family. Having a mother, sister, or daughter with breast cancer about doubles a woman's risk. (It's important to note that 70% to 80% of women who get breast cancer do not have a family history of this disease.)

Personal history of breast cancer: A woman with cancer in one breast has a greater chance of getting a new cancer in the other breast or in another part of the same breast. This is different from a return of the first cancer (which is called recurrence).

Race: White women are slightly more likely to get breast cancer than are African-American women. But African American women are more likely to die of this cancer. At least part of the reason seems to be because African-American women have faster growing tumors. Asian, Hispanic, and American Indian women have a lower risk of getting breast cancer.

Dense breast tissue: Dense breast tissue means there is more glandular tissue and less fatty tissue. Women with denser breast tissue have a higher risk of breast cancer. Dense breast tissue can also make it harder for doctors to spot problems on mammograms.

Menstrual periods: Women who began having periods early (before age 12) or who went through the change of life (menopause) after the age of 55 have a slightly increased risk of breast cancer. They have had more menstrual periods and as a result have been exposed to more of the hormones estrogen and progesterone.

Earlier breast radiation: Women who have had radiation treatment to the chest area (as treatment for another cancer) earlier in life have a greatly increased risk of breast cancer.

Treatment with DES: In the past, some pregnant women were given the drug DES (diethylstilbestrol) because it was thought to lower their chances of losing the baby (miscarriage). Recent studies have shown that these women (and their daughters who were exposed to DES while in the womb), have a slightly increased risk of getting breast cancer.

Breast cancer risk and lifestyle choices

Not having children or having them later in life: Women who have had not had children, or who had their first child after age 30, have a slightly higher risk of breast cancer. Being pregnant more than once and at an early age reduces breast cancer risk. Pregnancy reduces a woman's total number of lifetime menstrual cycles, which may be the reason for this effect.

Recent use of birth control pills: Studies have found that women who are using birth control pills have a slightly greater risk of breast cancer than women who have never used them. Women who stopped using the pill more than 10 years ago do not seem to have any increased risk.

Postmenopausal hormone therapy (PHT): Postmenopausal hormone therapy (also known as hormone replacement therapy or HRT), has been used for many years to help relieve symptoms of menopause and to help prevent thinning of the bones (osteoporosis). There are 2 main types of PHT. For women who still have a womb (uterus), estrogen and progesterone (known as combined PHT) are generally prescribed. Estrogen alone can increase the risk of cancer of the uterus, so progesterone is added to help prevent this. For women who no longer have a uterus (those who've had a hysterectomy), estrogen alone can be prescribed. This is commonly known as estrogen replacement therapy (ERT).

Combined PHT: It has become clear that long-term use (several years or more) of combined PHT increases the risk of breast cancer and may increase the chances of dying of breast cancer. The breast cancer may also be found at a more advanced stage, perhaps because PHT seems to reduce the effectiveness of mammograms. Five years after stopping PHT, the breast cancer risk seems to drop back to normal.

ERT: The use of estrogen alone does not seem to increase the risk of developing breast cancer much, if at all. But when used long-term (for more than 10 years), some studies have found that ERT increases the risk of ovarian and breast cancer.

Not breast-feeding: Some studies have shown that breast-feeding slightly lowers breast cancer risk, especially if the breast-feeding lasts 1½ to 2 years. This could be because breast-feeding lowers a woman's total number of menstrual periods, as does pregnancy.

Alcohol: Use of alcohol is clearly linked to an increased risk of getting breast cancer. Women who have one drink a day have a very small increased risk. Those who have 2 to 5 drinks daily have about 1½ times the risk of women who drink no alcohol. The American Cancer Society suggests limiting the amount you drink to one drink a day.

Being overweight or obese: Being overweight or obese is linked to a higher risk of breast cancer, especially for women after change of life and if the weight gain took place during adulthood. Also, the risk seems to be higher if the extra fat is in the waist area. But the link between weight and breast cancer risk is complex, and studies of fat in the diet as it relates to breast cancer risk have often given conflicting results. The American Cancer Society recommends you maintain a healthy weight throughout your life and avoid gaining too much weight.

Lack of exercise: Studies show that exercise reduces breast cancer risk. The only question is how much exercise is needed. One study found that as little as 1 hour and 15 minutes to 2½ hours of brisk walking per week reduced the risk by 18%. Walking 10 hours a week reduced the risk a little more. The American Cancer Society suggests that you exercise for 45 to 60 minutes 5 or more days a week.

Uncertain risk factors

High fat diets: Studies of fat in the diet have not clearly shown that this is a breast cancer risk factor. Most studies found that breast cancer is less common in countries where the typical diet is low in fat. On the other hand, many studies of women in the United States have not found breast cancer risk to be linked to how much fat they ate. Researchers are still not sure how to explain this difference. More research is needed to better understand the effect of the types of fat eaten and body weight on breast cancer risk.
The American Cancer Society recommends eating a healthy diet that includes 5 or more servings of vegetables and fruits each day, choosing whole grains over processed (refined) grains, and limiting the amount of processed and red meats.

Antiperspirants and bras: Internet e-mail rumors have suggested that underarm antiperspirants can cause breast cancer. There is very little evidence to support this idea. Also, there is no evidence to support the idea that under wire bras cause breast cancer.
Abortions: Several studies show that induced abortions do not increase the risk of breast cancer. Also, there is no evidence to show a direct link between miscarriages and breast cancer.

Breast implants: Silicone breast implants can cause scar tissue to form in the breast. But several studies have found that this does not increase breast cancer risk. If you have breast implants, you might need special x-ray pictures during mammograms.

Pollution: A lot of research is being done to learn how the environment might affect breast cancer risk. At this time, research does not show a clear link between breast cancer risk and environmental pollutants such as pesticides and PCBs.

Tobacco Smoke: Most studies have found no link between active cigarette smoking and breast cancer. An issue that continues to be a focus of research is whether secondhand smoke (smoke from another person's cigarette) may increase the risk of breast cancer. But the evidence about secondhand smoke and breast cancer risk in human studies is not clear. In any case, a possible link to breast cancer is yet another reason to avoid being around secondhand smoke.

Night Work: A few studies have suggested that women who work at night (nurses on the night shift, for example) have a higher risk of breast cancer. This is a fairly recent finding, and more studies are being done to look at this issue.

By slivi

Appendicitis

2010-02-27T21:31:00.001-08:00

The appendix is a small outgrowth of tissue forming a tube-shaped sac attached to the lower end of the large intestine. Inflammation of the appendix presents itself in acute and chronic forms and affects both the sexes equally. This disease accounts for about half the acute abdominal emergencies occurring between the ages of ten and thirty.

Appendicitis symptoms

Pain in centre of abdomen, discomfort in abdomen
Appendicitis usually begins with a sudden pain in the centre of the abdomen. The pain may be preceded by general discomfort in the abdomen, indigestion, diarrhoea, or constipation. Gradually, the pain shifts to the lower right side, and is usually accompanied by a fever varying from 38 oC to 39 oC.
Nausea and Vomitting
Nausea is common and the patient may vomit once or twice. In the chronic state of appendicitis, the patient may suffer from recurrent pain in the right lower abdomen, constipation, loss of appetite, and mild nausea.

Causes of Appendicitis

Excessive amount of poisonous waste material in caecum
Appendicitis is initiated by the presence of an excessive amount of poisonous waste material in the caecum. As a result, the appendix gets irritated and inflamed. Inflammation and infection are caused by certain germs which are usually present in the intestinal tract.

Home Remedies for Appendicitis

Appendicitis treatment using Green Gram
Green gram is a proven home remedy for acute appendicitis. An infusion of green gram is an excellent medicine for treating this condition. It can be taken in a small quantity of one tablespoon three times a day.
Appendicitis treatment using Fenugreek Seeds
Regular use of tea made from fenugreek seeds has proved helpful in preventing the appendix from becoming a dumping ground for excess mucus and intestinal waste. This tea is prepared by putting one tablespoon of the seeds in a litre of cold water and allowing it to simmer for half an hour over a low flame and then strained it. It should be allowed to cool a little before being drunk.
Appendicitis treatment using Vegetable Juices
Certain vegetable juices have been found valuable in appendicitis. A particularly good combination is that of 100 ml each of beet and cucumber juices mixed with 300 ml of carrot juice. This combined juice can be taken twice daily.
Appendicitis treatment using Buttermilk
Buttermilk is beneficial in the treatment of chronic form of appendicitis. One litre of buttermilk may be taken daily for this purpose.
Appendicitis treatment using Whole Wheat
The consumption of whole wheat, which includes bran and wheat germ, has been found beneficial in preventing several digestive disorders, including appendicitis. The bran of wheat can be sterilised by baking after thorough cleaning. This sterilised bran can be added to wheat flour in the proportion of one to six by weight. Two or three chapatis mane from this flour can be eaten daily for preventing this disease.
Appendicitis diet
Fasting and nothing except water
At the first symptoms of severe pain, vomiting, and fever, the patient should resort to fasting and nothing except water should enter the system.
Fruit juices and All-fruit diet
Fruit juices may be given from the third day onwards for the next three days. Thereafter the patient may adopt an all-fruit diet for a further four or five days.
Well-balanced diet
After this tightly regulated regimen, he should adopt a well-balanced diet, consisting of seeds, nuts, grains, vegetables, and fruits.

Other Appendicitis treatments

Half litre of Warm-water enema
When the first symptoms of pain, vomiting, and fever occur, the patient must be put to bed immediately, as rest is of the utmost importance. A low enema, containing about half a litre of warm water, should be administered once every day for the first three days to cleanse the lower bowel if it can be tolerated with comfort.
Hot compresses and abdominal packs of wet sheet strip
Hot compresses may be placed over the painful area several times daily. Abdominal packs, made of a strip of wet sheet and covered by a dry flannel cloth bound tightly around the abdomen, should be applied continuously until all acute symptoms subside.
Three litres of warm-water enema
When the acute symptoms subside by about the third day, the patient should be given a full enema containing about three litres of warm water, and this should be repeated daily until all inflammationand pain have subsided.
Avoid constipation
In other words, the patient of appendicitis should adopt all measures to eradicate constipation. Once the waste matter in the caecum has moved into the colon and is then eliminated, the irritation and inflammation in the appendix will subside and surgical removal of the appendix may not be necessary.

Make your mind sharp 10 great ways

2010-02-18T22:24:00.001-08:00

We expect the prowess of our joints and lungs to slowly decline as we age, but the thought of our minds doing the same is intolerable. Here are some top prevention tips worth their weight in wits, plus a few to forget.

1. TEASE YOUR BRAIN
Whether crossword puzzles, sudokus and other brain teasers actually keep your brain in shape, has not been well-established. However, lack of education is a strong predictor of cognitive decline. The more you've tried to learn, the better you'll be at mental sit-ups in old age. The key may be tackling something new; the challenge of the unknown is likely more beneficial than putting together the same jigsaw puzzle over and over again.

2. SKIP THE SUPPLEMENTS
Supplements have been getting a bad rap recently, with even the familiar multivitamin now looking like a waste of money -- or worse. Brain pills, such as ginkgo and melatonin, likely belong in the trash as well. Despite their "natural" origins, they are not free of potential side effects, such as high blood pressure, digestion trouble, fertility problems and depression. And among healthy individuals, ginkgo offers no brain benefits beyond that of a placebo. (In some cases, the placebo worked better.)

3. CHILL OUT
Stress takes a toll on the brain by washing harmful chemicals over the hippocampus and other brain areas involved in memory. Some scientists suspect that living a balanced lifestyle and pursuing relaxing activities such as yoga, socializing and crafting may delay memory impairment by reducing stress.

4. EAT FISH
Some theories credit the introduction of fish into the human diet with the evolution of our tremendous cognitive prowess. Essential fatty acids, such as Omega 3s, are critical to brain function and are proving beneficial for treating such brain-sapping ailments as depression. Studies on the efficacy of Omega 3 supplements, however, have had mixed results, so get doses from food sources, such as flax seeds, fatty fish and grass-fed animals.

5. ENJOY YOUR COFFEE
Growing evidence suggests a caffeine habit may protect the brain. According to large longitudinal studies, two to four perk-me-ups a day may stave off normal cognitive decline and decrease the incidence of Alzheimer's by 30 to 60 percent. It is unclear whether the benefits come from caffeine or the antioxidants found in coffee and tea, but that latte may improve cognition this afternoon and several decades from now.

6. GET YOUR BEAUTY REST
When we rest and dream, memories are sifted through, some discarded, others consolidated and saved. When we don't sleep, a recent study found, proteins build up on synapses, possibly making it hard to think and learn new things. Furthermore, chronically sleeping poorly (in contrast to not enough) is linked to cognitive decline in old age, although the relationship may not be causal.

7. TAKE CARE OF YOUR BODY
Largely preventable diseases -- such as Type II diabetes, obesity and hypertension -- all affect your brain, too. System-wide health concerns have been linked to an increased risk of cognitive decline and memory impairments. Keeping your circulatory system in working order, by, say, avoiding cigarettes and saturated fat, lessens the onslaught of age-related damage to the brain.

8. WATCH THAT DIET
While overindulging can make the brain sluggish and lead to long-term detriments to your brain, too few calories can also impair brain function. Extreme dieting can cause some diehards to feel stretches of calm -- a feeling that may underlie the addiction of anorexia -- but many studies have also linked dieting with distraction, confusion and memory impairment.

9. EAT, EAT, EAT
Too much or too little energy throws a kink in the brain's delicate machinery. A low glycemic diet -- high fiber, with moderate amounts of fat and protein -- is broken down more slowly in the body than high glycemic foods, such as sweets and white starches. A steady pace of digestion in the gut gives a more reliable flow of energy to the brain, likely optimizing the organ's long-term health and performance.

10. DO SOMETHING!
Scientists are starting to think that regular aerobic exercise may be the single most important thing you can do for the long-term health of your brain. While the heart and lungs respond loudly to a sprint on the treadmill, the brain is quietly getting fitter with each step, too. For mental fitness, aim for at least 30 minutes of physical activity every other day.

Written by Sheivi on medicalgeek.

Ten ways to birth control from ancient to future

2010-02-18T20:45:00.001-08:00

Birth control is commonly used as part of family planning.
The history of birth control began with the discovery of the connection between coitus and pregnancy.
Ever since humans first figured out that Tab A + Slot B = Baby, hard attempts have been made to defy that equation -- and there are still more innovations to come.
The following list gives a short summary of the various methods which were used previously, which are being used actually and which will be used in the future for birth control.

1) Wood Block Pessary
The word "pessary" covers a wide variety of objects that women have used over the centuries to block or kill sperm inside their vaginas. One of the more cringe-inducing options was the wood block pessary, a door-stop-sized, 6-sided block with divots carved on each side. The idea was that one of the divots would cover the cervix. The hope was that nobody would get a splinter.

2) Crocodile Dung
Recipes for pessaries found in ancient Egyptian papyrus texts may have been surprisingly effective. Some would have blocked the cervix, much like a modern diaphragm. Others probably changed the acidity of the vagina, making it an unfriendly environment for sperm. Honey, medicinal plants, and other not-especially-horrific-sounding substances were also recommended, but it's the crocodile dung that really stands out for some reason.

3) Booze and Beaver Balls
This creative combination was the favored folk remedy among early European settlers in New Brunswick, Canada. Frontier women who didn't want to get pregnant drank a concoction of white lightning infused with a powder made from dried beaver testicle. Needless to say, this method was pretty much useless...unless, of course, you were trying to get drunk.

4) Sponges
Yesterday's sponge was literally just that: The dried body of a sea sponge. Mentioned in the Talmud, this was the favored method of birth control in ancient Jewish communities. The sponge was wrapped in silk and had a string attached for easy removal. Women often soaked the sponge in vinegar or lemon juice before putting it in, creating a one-two punch of sperm-killing acidity and semen-absorbing sponge.

5) Condoms
Originally made out of animal guts or linen, the condoms staying power is a testament to its effectiveness and reliability. But one of the oldest birth control methods still in use today is probably even older than you realize. Remains of actual condoms, dating to the 1600s, have been found in England. Artistic representations, on the other hand, are positively ancient, including an Egyptian drawing more than 3,000 years old and 12,000-year-old French cave paintings.

6) Career Pill
In the future, women may be able to combine normal birth control with a factor that helps them stay fertile longer, effectively pushing menopause back by decades. Every month, dozens of eggs begin to grow in a woman's ovary. Usually, only one fully matures. The others simply die off. According to fertility expert Dr. Roger Gosden, a career pill would stop this waste, leaving more eggs available as a woman reaches her 30s and 40s.

7) Adjudin
While some women's products are in development, the real future of birth control is men. Contraceptives for guys could help women avoid the side-effects of hormonal pills and give men more control over their fertility. One possible option, currently being researched by the Population Council, is Adjudin, a chemical that prevents sperm from maturing, so they can't fertilize eggs. Ten years or more away from being ready, Adjudin would probably come in a patch or implant, rather than a pill.

8) Phthalates
Phthalates are chemicals added to plastic to make it more flexible, showing up in everything from children's toys to shampoo bottles. They're also a major health concern, with mounting evidence that they can affect puberty and the human reproductive system. But at the Population Council, researchers are working on a way to turn this straw into gold. Because they can change testosterone levels and lower sperm count, phthalates may hold clues to developing new male birth control methods.

9) Reversible Inhibition of Sperm Under Guidance (RISUG)
RISUG is one of the most promising male contraceptives in development. All it takes it one injection to the vas deferens, the tube that carries sperm from the testes to the urethra. A chemical compound partially blocks up the tube and any sperm that make it past the blockade come out damaged, unable to fertilize an egg. One application of RISUG can last up to 10 years and it appears to have none of the nasty side effects associated with hormone-based birth control. Better yet, it's already in Phase III clinical trials in India, which means there's a good chance of it hitting the market before you're too old to care.

10) Dry Orgasm Pill
Doctors have known since the 1950s that a couple of medications, usually taken for high A and schizophrenia, could also make male patients sterile. In November 2006, researchers figured out why: Both medications freeze the muscle contractions that propel sperm through the male reproductive tract. The result: A normal orgasm, sans sperm. Work is underway to turn this discovery into a marketable pill, but it's still a long way off.

What you think about your diet, myths

2010-02-17T21:51:00.001-08:00

Here are some common myths about diet..

■If you eat late at night, the food turns straight into fat.
Not true.If your overall calories are appropriate for weight loss, you certainly can eat something after dinner. Late night calories will ultimately get used the next day (and even while you sleep).
However, for the sake of energy, it is always better to eat your calories during the day when your body needs the fuel. Plus, eating consistently throughout the day will stabilize blood sugar levels -- so you’ll feel energized and experience fewer cravings. If you are going to snack after dinner, I suggest choosing something 250 calories or less.

■Fresh fruits and vegetables are more nutritious than frozen.
Not necessarily. Frozen can be a great produce option (just avoid varieties with added salt, sugar, and sauce). Frozen foods are picked in the peak of ripeness, then frozen. You can eat them as you need them -- and most of the nutrients are locked in. On the other hand, fresh fruit and vegetables are typically harvested before they ripen, and can have nutrient variability. Also, the longer fresh produce sits around in your fridge, the less nutrients it will contain.
Bottom line: Buy both fresh and frozen and eat as much as you can.

■Cravings are your body's way of telling you it needs something.
This has never been proven. You normally crave what you like to eat (or smell or see someone else eating). Also, hormonal changes are sometimes responsible for food cravings. Ice cream and pickles anyone?

■Any type of water is always better than soda.
No. There are a few caloric waters with sexy marketing ploys. In fact, some brands have quite a bit of sugar. Always check labels.

■Certain foods, like grapefruit, celery, or cabbage soup can burn fat and make you lose weight.
These are anecdotal stories that have no scientific back up. It’s true these foods are low in calories, but they do NOT actually burn fat.

■Exercise in the morning burns more calories.
Studies show that people who exercise in the morning tend to be more consistent with their daily workouts. However, exercising in the morning does not actually burn more calories than exercising later in the day.

■Dairy is bloating.
Dairy is only bloating for people with lactose intolerance… and in some instances, for people with irritable bowel syndrome (IBS). For people without lactose intolerance or dairy specific IBS, low-fat milk, yogurt and cheese should not cause bloating.

■You can “save” calories by skipping breakfast.
Studies report that breakfast eaters weigh less than breakfast skippers (obviously, it has to be a healthy breakfast). Plus, breakfast skippers tend to overeat after dinner.

■Colonics help you lose weight.
Colonics simply dehydrate you -- you may weigh less, but it’s mainly water weight NOT fat weight. Lost water weight typically comes right back on after a few glasses of fluid.

■Weight lifting makes you bulky.
Appropriate weight lifting will not make you bulky (unless that’s your goal and your program takes this into consideration). Light weight lifting helps to increase lean body mass, which helps you burn more calories 24-7. In the end it will help you lose weight and enable you to eat more.

■The scale is your absolute best indicator of weight loss.
Checking your weight loss progress on a scale is certainly simple and encouraging (depending upon the outcome!). However, there are other effective ways to track your progress: take body measurements with a tape measure, test body fat, compare personal photos, and assess the fit of your clothing.

Its time for male contraceptive injection !

2010-02-17T19:31:00.001-08:00

If you are in committed relationship and positive to each other then you may try this injection which reduces sperm count for shorter time and when you want to back to normal you can do it.

Professor Richard Anderson, from the University of Edinburgh, who is heading this research found that when a mixture of testosterone and female hormone progesterone injected twice a month that leads to temporary lowering in sperm count and when discontinued it come back to normal level. Normally it stimulates brain to lowers those hormones that involves in maturation and secretion of sperms.

This method is only useful in committed relationship and can’t prevent to sexually transmitted disease so it is not an alternative to physical contraceptive devices and will use in those people which is minimal risk of diseases and in which female, no other contraceptive drug is useful.

This artical is written on

Now birth control shot men

How to check your skin cancer

2010-02-10T18:34:00.001-08:00

World wide there are more than 10 million skin cancer patient from them about 1000 died every year. Skin cancer is a malignant growth of skin from the layers like basal cell, squamous cell layer, melanoma cells. Skin cancer are always visible on the skin because they originates on superficial layer of skin. This is most common detected cancer surpassing lungs, breast, colon and prostate cancer. The basic stimulus is too much absorption of sun ultraviolet light.

Signs and symptoms

There are variety of sign like changes in skin that do not heal, ulcer in skin, discoloration of skin and changes in existing mole.

Basal cell carcinoma- usually looks like raised, smooth, pear like bump on sun exposed skin. Crushing and bleeding always occurs.
Squamous cell carcinoma- it is commonly red , scaling, thickened patch on sun exposed skin If it not treated it develops in large mass, so it is dangerous.
Malenoma - they are brown/black looking lesion.
Markel cell carcinoma- they are very rapidly developing that dont produces red or any visible lesion on the skin. Even they are not itchy or painful.

Causes

Studies says that smoking can doubles risk of cancer.
Overexposure to ultraviolet light causes skin cancer via direct DNA damage or via indirect DNA damage.

Check your cancer

Schedule a examination plan. Make a monthly schedule to examine your body once in a month by standing against the mirror. Also make a annual plan for skin doctor visit.
Don’t get panic just by pimples or bruise or anything like that first check your symptoms .
Get familiar with signs and act fast before get delay make some time to go doctor if any no healing or chronic bleeding ulcer seen.
Abcd of melanoma

A - Asymmetry, one half of a mole or birthmark does not match the other.
Normal mole Asymmetrical mole
B - Border is irregular, ragged, notched, or blurred.
Normal mole Irregular border
C - Color varies (brown, black, red, white blue).
Normal moleVarying color
D - Diameter is larger than 6 millimeters across (about 1/4 inch -- the size of a pencil eraser).
Normal moleEnlarged mole

Watch a video how to check your skin cancer

Self-Check

How To Check For Skin Cancer

Now you should act fast to cure yourself.

This posting is modified by wikipedia and associated sites.

10 ways to stay away from stress

2010-01-26T20:29:00.001-08:00

Stress is a biological condition in which our body fails respond appropriately to emotional or physical threads whatever it is actual or imagined. The symptom of stress is commonly include a state of alarm and adrenaline production , short term resistance as coping management , exhaustion,, irritability, muscular tension, inability to concentrate, headache and increased heart rate.
Signs : It may be cognitive, emotional or behavioral. Sings include poor judgment , a negative outlook, excessive worrying, moodiness, irritability, agitation, inability to relax, feeling lonely, isolated or depressed , aches and pains, diarrhea or constipation, nausea, dizziness, chest pain, rapid heart beating, eating too much or nothing,sleeping too much or not enough, social withdraws, procrastination or neglect of responsibility, increased alcohol, drugs and nicotine consumption and nail biting.
Relationship with disease:

Stress directly acts on mental, physiological and immunological system of body. if immunological system is compromised it will made our body venerable to infections, it can increase chances to viral, fungal and bacterial outbreak.
Chronic stress leads to developmental growth impairment. This leads to decreased pituitary hormones ( growth hormone) so leads to impaired body growth. Chronic stress may be originated from bad home environment, parental alcoholism, and child abuse.
It also increases visceral fat deposition and disarranged metabolic status of body.

Stress Management

Exercise It seems to be very effective just due to it involves physiological and muscular system. And also increase blood flow to every organs and so in brain. It controls secretion of stress hormone(epinephrine and nor epinephrine).
Cognitive therapy : You should start meeting with positive minded peoples, find your stressor and try to control it yourself. Don’t compare yourself to another people neither economically nor socially.
Deep breathing : It is also a very effective therapy when you start deep breathing a cyclic pattern originates in your mind and blood so it also controls hormone release.
Get a hobby : Hobby keeps you busy in interesting work rather than being bore and more stressed. Start whatever you like, you feel enjoys doing this adopt a hobby.
Autogenic training : In this training person told to repeat a similar kind of visual, exercise or something other thing in repeated steps usually three time a day.It should be exercised in morning, noon and evening and the exercise is laying down, sitting in a posture or sitting like a doll.
Meditation : Meditation increases hormonal balance by acting directly on pituitary gland so it balance all the hormone related problems.
Spending time in nature: nature feels so artistic. so spend time in nature.
Break during work: Fractional break during work also seems to very effective.
Social involvement : Be social and start working with social organizations which involves in improvement in socially and physically challenged peoples.
Donate blood : Blood donation leads to secreation of a hormone call endorphin which is a stress busting hormone so make a habit to donate blood after certain intervals.

Algal Toxins: It is a poison ?

2009-09-26T09:52:00.001-07:00

Amnesic Shellfish Poisoning (ASP) It is caused by domoic acid produced by several species of Pseudonitzschia diatoms. The main contamination problems include mussels, clams, and crabs of the Pacific Northwest of the United States and Canada.

Paralytic Shellfish Poisoning (PSP) It is caused by the saxitoxin family (saxitoxin + 18 related compounds) produced by several species of Alexandrium dinoflagellates. The main contamination problems include mussels, clams, crabs, and fish of the Pacific Northwest and Northeast Atlantic.

Neurotoxic Shellfish Poisoning (NSP) It causes sickness in humans lasting several days. NSP is not fatal to humans; however, it is known to kill fish, invertebrates, seabirds, and marine mammals (e.g., manatees). It is caused by the brevetoxin family (brevetoxin + 10 related compounds produced by the dinoflagellate Karenia brevis a.k.a. Gymnodinium breve.

Diarrheic Shellfish Poisoning (DSP). It is caused by chemicals of the okadaic acid family (okadaic acid + 4 related compounds) produced by several species of Dinophysis dinoflagellates.

Ciguatera Fish Poisoning (CFP) Caused by consumption of reef fishes (e.g., grouper, snapper), sickness in humans lasts several days to weeks, but the human fatality rate is low. It is caused by the ciguatoxin family (ciguatoxin + 3 or more related compounds) and produced by several species of dinoflagellates including Gambierdiscus, Prorocentrum,
and Ostreopsis.

Cyanobacterial (Blue-Green Bacteria) Toxins. It is caused by a variety of biotoxins and cytotoxins, including anatoxin, microcystin, and nodularin produced by several species of cyanobacteria, including Anabaena, Aphanizomenon, Nodularia, Oscillatoria, and Microcystis. The main contamination problems include all eutrophic freshwater rivers, lakes, and streams.

Ambush Predator (Pfiesteria piscicida and Toxic Pfiesteria Complex) Toxins.Produced by several dinoflagellate species including, Pfiesteria piscicida, Pfiesteria shumwayae, and perhaps several other unidentified, un-named dinoflagellates belonging to the potentially toxic Pfiesteria complex. Main problems include major fish kills in North Carolina and
Maryland and potential human health problems.

Yes it is

Anti-tumor vaccine: A immunological approach by Antigen Presenting Cells

2009-09-26T06:24:00.001-07:00

In order for cells involved in the immune response to react to foreign organisms or cells, the non-self antigens in these organisms or cells must be ‘‘presented’’ in a way that the immune cells can ‘‘see’’ them. This requires that the foreign antigens be processed into smaller bits of information and be presented to immune cells as part of a complex with cell surface MHC molecules. There are two types of MHC molecules: class I, expressed on all cells, and class II, expressed on macrophages, dendritic cells, B cells, and occasionally on other cell types. All three of these latter cell types can present antigens to T lymphocytes. However, there are two ways that antigen loading onto MHC occurs. Intracellular antigens such as viral or cytosolic tumor peptides are complexed with MHC class I via intracellular processing pathways and presented to CD8þ (cell surface marker) T lymphocytes
(cytotoxic T cells); however exogenous antigens such as those from pathogens are processed via a different pathway and presented with MHC class II molecules to CD4 þ
T cells (helper cells). Antigen-derived peptides are generated in antigen presenting cells (APCs) by one of two routes. Antigenic peptides present in the cytosol, for example, from viral-infected cells or from engulfed tumor-associated antigens, are derived by proteolytic degradation and transported into the endoplasmic reticulum where they bind to nascent MHC class I molecules. In this way, T lymphocytes bearing the CD8 cell surface marker (CD8þ cells) become stimulated to become cytotoxic T cells (CTLs). Antigenic peptides
generated during endocytosis of engulfed antigenic molecules can bind to MHC class II molecules targeted to this cellular compartment on their way to be cycled back to the cell surface. Antigens presented in conjunction with MHC class II stimulate CD4þ T cells, which become activated T-helper cells. Macrophages scavenge dead and dying cells, engulf and kill many types of bacteria, present antigens to T cells, and are themselves effectors cells in cell-mediated immune reactions and when activated can kill tumor cells.Phagocytic dendritic cells are present in many tissues where they can pick up and process antigens and then migrate to lymphoid tissue. Those that differentiate in lymphoid tissues acquire the ability to present antigens to T cells, and they are highly effective in activating CD4þ cells. They do not appear to have a cytotoxic effector function themselves. B cells have also been implicated as ‘‘presenters’’ of soluble protein antigens to CD4 þ cells. Although intact antigenic proteins need to be processed to generate antigenic peptides and co-presented with MHC class I or II molecules, soluble peptides can also bind directly to empty class I or class II molecules present on the cell surface of APC cells. Such empty MHC molecules are potentially important targets for synthetic immune system–stimulatory peptides that could be anti-tumor vaccines

Does cell phone causes cancer ?

2009-09-26T05:57:00.001-07:00

There is a lot of myth about the relationship between cancer and cell phone. Is it is true that a cell phone causes cancer? First of all you should know about what frequency uses a cell phone and how much heat or energy it produces. Any typical cell phone uses or emit radiofrequency between 800 to 2000 MHz that is in microwave range. Radio-frequency can produces small amount of heat even when it puts on higher capacity. Any typical cell phone which uses this frequency can increase a temperature up to 0.18 C. And at this temperature till now we have not absorbed any biological effect on cell‘s morphological or biochemical level. And even radiofrequency does not posses sufficient energy that can causes any removal of electron or chemical changes thus it does not causes any ionization change in DNA so it does not causes cancer. Because of the weak thermal and ionizing potential of RF from cell phones, it seems highly unlikely that this RF would cause cancer.
But some studies also shows in mouse, using this range of radiofrequency some mouse developed some type of cancer. But in human even with experimental or occupational workers data shows no evidence of cancer development.
SO, STILL IT IS SAFE TO USE CELL PHONE.

Mechanisms of Tumor Initiation

2009-09-25T10:37:00.001-07:00

Initiation of malignant transformation of normal cells by a carcinogenic agent involves a permanent, heritable change in the gene expression of the transformed cell. This could come about by either direct genotoxic or mutational events, in which a carcinogenic agent reacts directly with DNA, or by indirect or ‘‘epigenetic’’ events that modulate gene expression without directly reacting with the base sequence of DNA. Most investigators favor the mutational theory of carcinogenesis—that is, that the initiating events involve a direct action on the genome. The mutational theory depends on three kinds of evidence:

1. Agents that damage DNA are frequently carcinogenic. As discussed previously, chemical carcinogens are usually activated to form electrophilic agents that form specific
reaction products with DNA. The extent of formation of some of these reaction products, for example, alkyl-O6-guanine, has been shown to correlate with mutagenicity and carcinogenicity of certain chemical agents. Ultraviolet and ionizing radiation also interact with DNA at doses that are carcinogenic.

2. Most carcinogenic agents are mutagens. A number of in vitro test systems using mutational events in microorganisms have been developed to rapidly screen themutagenic
potential of various chemical agents. One of the best known of these, the Ames test, is based on certain characteristics of specially developed strains of the bacterium Salmonella typhimurium. The tester strain, amutant line that requires exogenous histidine for its growth (hisauxotroph), has a poor excision repair mechanism and an increased
permeability to exogenously added chemicals. Using this system, together with a liver microsomal fraction that has the capacity to activate most chemical carcinogens metabolically, Ames and colleagues have shown that about 90% of all carcinogens tested are also mutagenic.36 Moreover, few noncarcinogens show significant mutagenicity in this test system. Malignant transformation can be induced in a variety of cultured mammalian cells by agents that are mutagenic for the same cells. For example, carcinogenic polycyclic hydrocarbons cause mutations, as measured by induction of resistance to 8-azaguanine, ouabain, or elevated temperature, in Chinese hamster V79 cells if the cells are
cocultured with lethally irradiated rodent cells that can metabolize the hydrocarbons to their electrophilic, activemetabolite.37,38 In these studies,mutagenicity was obtained with the carcinogenic hydrocarbons 7,12- dimethylbenz(a)anthracene, benzo(a)pyrene, and 3-methylcholanthrene. There was no mutagenicity with a noncarcinogenic hydrocarbon, and the degree of mutagenicity was related to the degree of carcinogenicity of the chemicals in vivo.

3. Incidence of cancer in patients with DNArepair deficiencies is increased. In individuals with certain recessively inherited disorders, the prevalence of cancer is significantly higher than in the general population. 39 The connecting link between these disorders is the inability to repair certain kinds of physical or chemical damage to DNA. The high incidence of cancer in these diseases constitutes the best available evidence for a casual relationship between mutagenicity and carcinogenicity
in humans. One example of xeroderma pigmentosum (XP) is characterized by extreme sensitivity of the skin to sunlight and is the most widely studied of the repair-deficient human diseases. Virtually 100% of affected individuals will eventually develop some form of skin cancer. In addition, heterozygotes who carry the XP gene but do not have the disease appear to have a higher incidence of nonmelanoma skin cancer.40 All individuals with XP are defective in repair of ultraviolet damage to DNA, and most of them have a defect in the excision repair pathway (see below). The repair defect ranges from 50% to 90% repair efficiency in cells from different patients, and there is good correlation between the severity of the molecular defect and the extent of the disease. The defect in most patients appears to be at the nicking or incision step of excision repair, although
patients in one complementation group have normal excision repair and are defective in postreplication repair. The XP cells are also less efficient at repairing
chemically induced damage to their DNA. Other examples of enhanced susceptibility to cancer in individuals with DNA repair deficiencies are ataxia telangiectasia (AT), Fanconi’s anemia (FA), and Bloom’s syndrome (BS). Cell lines derived from AT patients are defective in repair replication following exposure to irradiation or carcinogenic chemicals. Patients with AT are more prone to develop leukemia and some other cancers. FA cells have a defect in the repair of cross-linked bases in DNA; they also appear to have a slight deficiency in the repair of g-ray- or ultraviolet-induced damage, and patients with FA are at increased risk to develop cancer. Lymphoblastoid cell lines from patients with a type of BS characterized by a high rate of sister chromatid exchange (SCE) aremore highly tumorigenic after brief exposure in vitro to 4-nitroguinoline-N-oxide andN-methyl-N0- nitro-N-nitrosoguanidine (MNNG) in the nude mouse assay than are lymphoblastoid cells from normal individuals

QUALITY ASSURANCE (QA) PROCEDURES

2009-09-24T09:00:00.001-07:00

QUALITY ASSURANCE (QA) PROCEDURES

Over the last 20 years the reliability of data produced by analytical laboratories has increased dramatically. Strict requirements have ensured that the data were produced
under defined standards of quality with a stated level of confidence. The routine day-to-day activities (e.g., matrix fortifications) to control, assess, and ensure the quality of generated data are the quality controls associated with analytical processes. The management of the system that ensures that these processes are in place and
functional is the quality assurance portion of the laboratory program to produce reliable data.
Quality assurance (QA) is an essential part of analytical protocols. Each laboratory is required to detect and correct problems in analytical processes and to reduce errors to agreed-upon limits. To produce data that have acceptable quality, all laboratory members must follow established guidelines and protocols. Some of the essential elements that must be included in a QA program are as follows:
1. Laboratory practices (e.g., glass washing protocols) must be developed, reviewed, and updated with the staff’s participation on a scheduled basis and followed strictly by all laboratory members;
2. Operating procedures (e.g., SOPs monitoring freezer temperatures daily) must be standardized, documented, and supplied to each member of the laboratory staff and updated on a set schedule;
3. Monitoring programs (e.g., surface water monitoring of supplies furnishing public drinking water) must be carefully designed.
4. Maintenance of equipment and instruments must be documented in LIMS or appropriate maintenance books kept with the equipment.
5. Expiration dates of analytical standards, chemicals, and solvents must be observed and replacements made prior to their expiration date.
6. Good laboratory practices (GLPs) must be implemented as needed.
7. Audits must be performed on a scheduled basis to verify that all aspects of the QA program are operating sufficiently.

QUALITY CONTROL (QC) PROCEDURES
Quality control (QC) concerns procedures that maintain a measurement system in a state of statistical control. This does not mean that statistics control the analytical procedures but that statistical evidence is used to ensure that the procedure is working under the conditions set by protocol. The accuracy of an analytical method depends on statistical control being conducted prior to determining any other parameter. How well the basic method will work with the sample matrix being evaluated will depend on the way the QC samples are examined. A comprehensive QC analytical procedure would include the following:
1. Replicated environmental samples to test the precision of the sampling or analytical procedures.
2. Replicated analyses conducted on the same sample multiple times in order to determine analytical precision.
3. Trip blanks to determine if contaminants are introduced the processes of collecting, shipping, or storing of samples.
4. Matrix-fortified laboratory blanks consisting of solvent and reagent blanks to determine levels of bias due to matrix effects or analytical method problems.
5. Sample blanks (a sample matrix that does not contain the toxicant, although this is sometimes difficult to obtain) to ensure no extraneous or interfering peaks; the peaks indicate where a problem might exist in the method used.
6. Fortified field blanks to determine the effects that the matrix might have on analyte recovery.

Laboratory Information Management System (LIMS)

2009-09-24T08:55:00.001-07:00

As computers and software became more sophisticated, it was only a matter of time before attention was turned to laboratory management. Through large databases, applications
were designed to manage and store information associated with a laboratory, and to include many unique features. These laboratory information management systems
(LIMS) have found widespread use in regulatory laboratories. A typical LIMS contains the following functions:

1. A sample tracking module, using a unique computer-generated identification number, that allows sample tracking from the time the sample enters the laboratory
until it is analyzed.
2. A sample scheduling module that automatically logs in the sample, prints barcoded labels and assigns analyses for routine projects.
3. A personnel and equipment module that maintains employee training records, tracks instrument calibration, repairs, costs, and so on.
4. A data entry module that allows the chemist to enter results into the LIMS, assign QC runs, and prepare reports.
5. A QA/QC module that allows generation of control charts and produces graphs that detail trends.
6. An electronic data transfer module that automatically transfers data generated from the electronics of the analytical instrument into the LIMS, thus reducing transcription errors.
7. A chemical and reagent inventory module that manages the purchase and use of laboratory supplies, keeps track of lot number purchases, shelf lives, and costs.
8. A maintenance module that allows laboratory managers to manage the database, monitor studies, methods, priorities of projects, and so on.

LIMS have become more sophisticated and recently have been combined with two newly developed systems, the chemical information management system (CIMS), which searches for, manipulates, and stores chemical structures, reactions, and associated data, and the analytical information management system (AIMS), which acts as a central repository for instrumental data (e.g., chromatograms).

Quality assurance in clinical lab

2009-09-23T09:05:00.001-07:00

To produce a reliable report we should run Periodic quality control . the reagent , manpower, and instrument should go through quality assurance program.Here are few topic i am going to define that will help you to understand the organization of quality assurance.

Quality control: this is the common method that is used to decide that the method in use is enough reliable to produce an accurate result or the test reagent or performance must have to reject. This is usually done by running control sera with each lot of test.
Control sera: Control sera is a sample which concentration is known and is used to run with sample in identical manner. It should be identically same as sample in physical or chemical manner.
Control charts: In manner to apply the originated control data on mathematical manner There are few chart are available called as levy-jenning chart, KUSUM chart,

Parathion

2009-09-23T08:15:00.001-07:00

Parathion is one of several organophosphorus insecticides that has had great economic importance worldwide for several decades. Organophosphate toxicity is the result of excessive stimulation of cholinergic nerves, which is dependent on their ability to inhibit acetylcholinesterases. Interestingly the parent organophosphates are relatively poor inhibitors of acetylcholinesterases, requiring metabolic conversion of a P=S bond to a P=O bond for acetylcholinesterase inhibition.In vitro studies of rat and human liver have demonstrated that CYP is inactivated by the electrophilic sulfur
atom released during oxidation of parathion to paraoxon. Some have shown that the specific isoforms responsible for the metabolic activation of parathion are destroyed in the process. For example, preincubations of NADPH-supplemented human liver microsomes with parathion resulted in the inhibition of some isoform-specific metabolites including testosterone (CYP3A4), tolbutamide (CYP2C9), and 7-ethylresorufin (CYP1A2) but not aniline (CYP2E1). These losses of metabolic activity were also associated with the loss of CYP content as measured by the CO-difference spectra. These
results suggest that parathion acts as a suicide substrate, in that its metabolism results in the destruction of the particular isoforms involved in its metabolism. This becomes particularly important because the principal CYP involved in parathion metabolism is CYP3A4, which is the dominant CYP in humans; accounting for between 30–50% of the total liver CYP. Because of this enzyme’s importance in drug metabolism, the strong potential for inhibition by organophosphate compounds may have serious consequences in individuals undergoing drug therapy.

Causes of cancer(part 2)

2009-09-23T07:56:00.001-07:00

ONCOGENE
An oncogene is a special type of gene that is capable of transforming host cells and triggering carcinogenesis. The name is derived from the Greek onkos, meaning bulk, or mass, because of the ability to cause tumor growth. Oncogenes were first discovered in retroviruses (viruses containing the enzyme reverse transcriptase, and RNA, rather than DNA) that were found to cause cancer in many animals (e.g., feline leukemia virus, simian sarcoma virus). Although this is a relatively common mechanism of oncogenesis in animals, very few oncogene-carrying viruses have been identified in man. The ones that are known include the papilloma virus HPV16 that is associated with cervical cancer, HTLV-1 and HTLV-2 associated with T-cell leukemia, and HIV-1 associated with Kaposi sarcoma. Studies of humans led to the discovery of related genes called proto-oncogenes that exist naturally in the human genome. These genes have DNA sequences that are similar to oncogenes, but under normal conditions, the proto-oncogenes do not cause cancer. However, specific mutations in these genes can transform them to an oncogenic form that may lead to carcinogenesis. So, in humans, there are two unique ways in which oncogenesis occurs, by true viral infection and by mutation of proto-oncogenes that already exist in human cells.

Causes of cancer (part 1)

2009-09-23T06:55:00.001-07:00

There are a list of cancer producing agents (carcinogen) that causes cancer.

IRRADIATION CARCINOGENESIS : Both x ray and ultraviolet rays cause cancer by inducing DNA damage that leads to activate erroneous repair mechanism that results in mutation.

When cell are exposed to ultraviolet rays in the 240-300 range, the bases acquired the excitation state, producing photochemical reaction between the nucleotides. The final resultant product in ultraviolet excitation is cyclobutane formed between the adjacent pyramidine diamer. This product leads to carcinogenic activity.
Heavy exposure to sunlight also induces same reaction.
Cancer due to ionizing radiation is caused by x ray, gamma rays and beta radiation. The resultant cancer involves blood cancer(leukemia), lymphomas, skin cancer, brain cancer etc. the carcinogenic activity of this ionizing radiation is due to the damage to DNA, and hence its mutagenic and carcinogenic effect, is due to the generation of free radicals as the radiation passes through tissues. The amount of radical formation and ensuing DNA damage depend on the energy of the radiation. In general, X-rays and gamma rays have a low rate of
linear energy transfer, generate ions sparsely along their tracks, and penetrate deeply into tissue. This profile contrasts with that of charged particles, such as protons and a particles, which have a high linear energy transfer, generate many more radical ions locally, and have low penetration through tissues. The damage to DNA can include single- and double-strand breaks, point mutations due to misrepair deletions, and chromosomal translocations.

Diagnosing Cystic Fibrosis

2009-09-22T08:30:00.001-07:00

Introduction: It is a autosomal recessive disorder. It is a multisystem disorder that affect pulmonary, gastrointestinal, and reproductive organ. the phenomenal expression of this disorder that’s severity is hertogenous, ranging from mild pulmonary symptoms to sever mucunium ileus and sever respiratory disease and even sometime no evidence of disease. Mortality of this disease is basically due to accumulation of mucus and infection with unusual pathogen like Pseudomonas ariginosa.

The gene is mapped to 7q31(long arm of chromosome 7, bending region 31) in 1985d that was cloned subsequently. The Cystic fibrosis gene was extreamly large containing 27 exons and the product is 6.5 kb long.It codes for transmembrain conductance regulatory protein of 1480 amino acid. The molecule is located within lipid bylayer predominantally at the apical membrain of seceretory epithelial cells where it serves as cyclic adenosine monophosphate(cAMP) ctivated clorine ion chanel to regulate chloride ion conductance.

Diagnosis

The routine sweet chloride test- In this test we measure the concentration of chloride in sweat. First of all you should tale an measured peace of filter paper and put on the body surface of patient and weight again. the difference of weight will tell you the presence of chloride. more than 60m mol in children and more than 40m mol in adult is considered as positive.
Pcr testing: there are two panel of mutants for this test. They are

Mutation	Frequency
AF508	72.42%
G542X	2.28
G55ID	2.25
621+1G>T	*1.57
W1282X	1.50
N1303K	1.72
R553X	0.88
^1507	0.87
R117H	0.70
R117H	0.70
3849+10kbC>T	0.58
2789+5G>A	0.48
1717-1G>A	0.48
R347P	0.45
711+1G>T	0.43
R560T	0.38
3659delC	0.34
A455E	0.34
G85E	0.29
R1162X	0.23
2184delA	0.17
1898+1G>A	0.16
R334W	0.14
1148T	0.09
3120+1G>A	0.80
1078delT	0.02
TOTAL	88.40

Quality Control in PCR

2009-09-22T06:26:00.001-07:00

1. Introduction
Polymerase chain reaction (PCR), like any laboratory procedure, can be subject to a range of experimental or procedural error. A clear consideration of where such potential errors may occur is essential to minimize their impact. Careful quality control of equipment and reagents is essential.

2. Equipment
In an ideal world, and any diagnostic or commercial laboratory, each piece of equipment should be serviced and calibrated on a regular basis, with careful records being kept of this maintenance. Unfortunately, not every laboratory has funds for full-service contracts on all equipment. There are a few fundamental procedures, however, which will reduce errors from equipment problems.

• Be consistent in the equipment used for any given PCR. If it works on Monday but not Tuesday, this may simply be to the result of using a different PCR block. Even the most
modern and expensive thermal cyclers deteriorate with age.
• Check pipetting devices on a regular basis (weekly is not excessive) to ensure they pipette the correct volume. This is easily performed by pipetting and weighing water. Most
manufacturers produce inexpensive service packs for their pipettors.

3. Reagents
As with all laboratory procedures, it generally pays dividends to use high-quality reagents from reputable suppliers. You may well know someone who brews their own Taq polymerase in a vat in the garage, but do they control for batch-to-batch variability? It is important to remember, however, that these are single-stranded DNA molecules and are therefore relatively labile. Repeated freeze/thawing will cause degradation to shorter products, which will either not anneal, or if the annealing temperature is low enough, will anneal promiscuously, yielding multiple products. Simple aliquoting primers into manageable volumes will reduce both the scope for contamination and degradation. This practice
should also be adopted for dNTP stocks for the same reason. which will either not anneal, or if the annealing temperature is low enough, will anneal promiscuously, yielding multiple products. Simple aliquoting primers into manageable volumes will reduce both the scope for contamination and degradation. This practice should also be adopted for dNTP stocks for the same reason.

4. Operator Errors
• Use appropriate pipetting devices. A pipettor designed for the 20- to 200-μL range will not accurately dispense 10 μL.
• The use of master mixes not only reduces the dependence on accurately pipetting small volumes but also improves the control over reaction contents.
• Practice with the same reaction until consistent results are obtained.

5. PCR-Specific Difficulties
Although much of the above could apply to any analytical laboratory technique, PCR also is subject to the confounding problem of contamination. Cross contamination
of samples is of concern in any discipline, and good laboratory practice, such as careful pipetting and the constant changing of disposable pipet tips, will minimize the opportunity of this occurring. Where PCR differs from most other procedure is in the production of vast quantities of the analyte during the procedure. The presence of billions of copies of potential template can create severe problems. These problems can be minimized by physically separating the pre- and postamplification processes (ideally in different rooms with different pipettors, etc.); however, they should constantly be monitored by the inclusion of appropriate controls.

6. Controls
• No DNA. Although it can seem extravagant to constantly set up reactions without template, this is the best way to monitor for contamination. A separate “no DNA” control should be set up for each master mix or each individual reaction. If contamination is discovered, the pipettor should be decontaminated (as per manufacturers guidelines), and the reagent aliquot should be rechecked or discarded.
• Positive control. PCR is often used simply to detect the presence of specific sequence. In such circumstances, it is essential to include at least one reaction with a template known to contain the sequence.
• Internal control. Even when master mixes have been used to ensure consistency of reaction components, and a positive control is used, there is the possibility that template may be omitted from individual tubes. This can be addressed by the inclusion within each reaction tube of primers, which will amplify a target known to consistently be present in the test DNA.

7. Regional Quality-Assurance Programs
In addition to the in-house precautions detailed above, there are a growing number of specialist quality-assurance programs that have been developed for most diagnostic PCR. These programs distribute test material to laboratories, who then report their results centrally. Results from all participating centers are compared and confidential reports issued to each center. If such a program exists in your field, details will probably be available on the Internet: join it.

What Are the Strengths and Limitations of Contemporary DNA Purification Methods?

2009-09-21T06:44:00.001-07:00

Salting out and DNA Precipitation
Mechanism
Some of the first DNA isolation methods were based on the use of chaotropes and cosmotropes to separate cellular components based on solubility differences (Harrison, 1971; Lang, 1969). A chaotrope increases the solubility of molecules (“salting-in”) by changing the structure of water, and as the name suggests, the driving force is an increase in entropy. A cosmotrope is a structure-maker; it will decrease the solubility of a molecule (“salting-out”). Guanidium salts are common chaotropes applied in DNA purification. Guanidinium isothiocyanate is the most potent because both cation and anion components are chaotropic. Typical lyotropes used for salting out proteins are ammonium and
potassium sulfate or acetate. An all solution based nucleic acid purification can be performed by differentially precipitating contaminants and nucleic acids. Cells are lysed with a gentle enzyme- or detergent-based buffer (often SDS/proteinase K). A cosmotrope such as potassium acetate is added to salt out protein, SDS, and lipids but not the
bulk of nucleic acids. The white precipitate is then removed by centrifugation. The remaining nucleic acid solution is too dilute and in a buffer incompatible with most downstream applications, so the DNA is next precipitated as described above.

Features
Protocols and commercial products differ mainly in lysis buffer composition.Yields are generally good, provided that sample lysis was complete and DNA precipitation was thorough.These procedures apply little mechanical stress, so shearing is generally not a problem.
Limitations
If phenolic contaminants (i.e., from plants) are a problem, adding 1% polyvinylpyrrolidine to your extraction buffer can
absorb them (John, 1992; Pich and Schubert, 1993; Kim et al., 1997). Alternatively, add a CTAB precipitation step to remove polysaccharides (Ausubel et al., 1998).

Extraction with Organic Solvents, Chaotropes, and DNA Precipitation
Mechanism
Chaotropic guanidinium salts lyse cells and denature proteins, and reducing agents (b-mercaptoethanol, dithiothreitol) prevent oxidative damage of nucleic acids. Phenol, which solubilizes and extracts proteins and lipids to the organic phase, sequestering them away from nucleic acids, can be added directly to the lysis buffer, or a phenol step could be included after lysis with either GTC- or SDS-based buffers as above. GTC/phenol buffers often require vortexing or vigorous mixing. The affinity of nucleic acids for this two-phase extraction system is pH dependent. Acidic phenol is applied in RNA extractions because DNA is more soluble in acidic phenol; smaller DNA molecules (<50 kb) will be found in the organic phase and larger DNA molecules (>50 kb) in the interphase. When purifying RNA via this procedure, it is essential to shear the DNA to ensure a light interphase. Phenol titrated to a pH of 8 is used to separate DNA from proteins and lipids, since DNA is insoluble in basic phenol.Whether protocols call for a GTC/phenol, a GTC, or an SDS based step followed by phenol, it is best to follow a phenol extraction with chloroform in order to extract residual phenol from the aqueous phase. Phenol is highly soluble in chloroform, and chloroform is not water soluble. Remaining lipids may also be removed by this step. Phenol extractions are followed by nucleic acid precipitation steps as described above.
Features
Though caustic and toxic, this strategy still has wide use because yield, purity, and speed are good, and convenient for working with small numbers of samples.
Limitations
If lysis is incomplete, the interphase between organic and aqueous layers becomes very heavy and difficult to manipulate, and may trap DNA. Phenol is not completely insoluble in water, so if chloroform steps are skipped, residual phenol can remain and interfere with downstream applications. High salt concentrations can also lead to phase inversion, where the aqueous phase is no longer on top (problematic if colorless phenol is used). Diluting the aqueous phase and increasing the amount of phenol will correct this inversion. When working with GTC/phenol-based extraction buffers, cross-contamination of RNA with DNA, and vice versa, is frequent.

Glass Milk/Silica Resin-Based Strategies
Mechanism
Nucleic acids bind to glass milk and silica resin under denaturing conditions in the presence of salts (Vogelstein and Gillespie, 1979). Recent findings indicate that binding of some nucleic acids might even be feasible under nondenaturing conditions (Neudecker and Grimm, 2000). The strong, hydrophobic interaction created in the presence of chaotropic substances can be easily disrupted by removal of salt. The adsorption is followed by wash steps, usually with salt/ethanol which will not interfere with the strong binding of nucleic acids but will wash away remaining impurities and excess chaotrope. Depending on the protocol, this can be followed by a low salt/ethanol wash step that can lead to a reduction in yield. Finally nucleic acids are eluted from the glass in a salt or TE buffer. Nucleic acids are then ready
for use. Most methods create a denaturing adsorption environment by using guanidium salts for one-step lysis and binding. The strength of the binding depends on the cation used to shield the negative charges of the phosphate backbone and the pH (Romanowski et al., 1991). Slightly acidic pH and divalent cations, preferably magnesium, seem to work best. Differences between glass milk, silica resin, and powdered glass consist mainly in capacity and adsorption strength, a function of impurities present in the binding resins. Diatomaceous earths seem to have an especially high binding capacity (http://www.nwfsc.noaa.gov/protocols/dna-prep.html). Pure silica oxide has the lowest affinity to nucleic acids (Boom et al., 1990), but this can improve recovery even though initial binding capacity is lower. Glass milk is silica presuspended in chaotropic buffer, whereas the silica resin is a solid, predispensed matrix usually found in spin or vacuum flow-through format. Glass milk gives more flexibility for scale of prep, predispensed resin is more convenient for highthroughput applications. Glass milk or silica-based kits are available from numerous vendors, and even though the basic principle is the same, there can be significant differences in efficiency, purity, and yield.
Features
DNA purification based on hydrophobic adsorption to glass or silica is fast, simple, straightforward, and scalable. No additional time-consuming and yield-reducing precipitation steps are required. Depending on binding and wash buffer composition, very good yield and purity values are obtained. This purification approach can also allow restriction digestion/ligation reactions directly on the glass surface, improving transformation efficiency of complex ligation mixtures (Maitra and Thakur, 1994).

Limitations
One of the dangers of silica-based strategies is underloading the sample. Even though yields are good, there is always sample loss due to some remaining material on the resin or filter.The smaller the DNA fragment, the tighter is the interaction. Oligonucleotide primers are actually removed because binding to the glass becomes virtually irreversible. Underloading can become a critical issue when working with small samples and large volumes of glass milk or silica filter. Some of the older methods utilized unstable buffer components, such as NaI, that tended to oxidize over time, leading to very poor recoveries. Some procedures required the addition of reagents to produce functional wash or elution buffers. If the concentrations were incorrect, or if volatile reagents (i.e., ethanol) were added and the buffers stored long term, these buffers lost their effectiveness. Incomplete sample lysis can be problematic because intact cells may also bind to silica and lyse under low- or no-salt elution conditions, leading to degradation of nucleic acids. Incomplete ethanol removal after wash steps will cause the problems described earlier for ethanol precipitation (discussed below under the question What Are The Fundamental Steps Of DNA Purification?). Ethanol must be completely removed from the samples after wash steps to avoid problems such as diffusion out of agarose gel wells (“unloadable” DNA/RNA) or undigestable DNA. Overdrying will lead to irreversible binding of nucleic acids to the resin severely impairing yields.

Anion Exchange (AIX) Based Strategies
Mechanism
Nucleic acids are very large anions with a charge of -1/base and -2/bp; hence they will bind to positively charged purification resins (commonly referred to as anion exchangers). After washes in low-salt buffers, the DNA is eluted in a high-salt buffer. AIX strategies are applied to purify genomic and plasmid DNA. Logic might suggest that the greater the strength of the anion exchanger, the more DNA it would bind (and more tightly), which would make for superior DNA purification. In practice, however, if an anion exchanger is too strong, most DNA is never recovered. This is especially problematic when working with small samples and with spun column formats. Forcing liquid through porous chromatography resins via centrifugation does not allow for even flow rates, hence resolution is poor. For this reason some spun column plasmid purification procedures advise the recovery of only a portion of the potential total material to avoid contamination by genomic DNA. Procedures where the buffer flows through columns packed with AIX resins under the force of gravity (as in standard column chromatography) can overcome this problem, but are slower. Gravity flow-based columns can clog if lysis is incomplete or if removal of protein or lipid is incomplete. Resolution
is very much flow rate dependent, and tight control of linear flow rates on HPLC or FPLC™ systems are superior to gravity flow and/or spun column formats when it comes to resolution and scale-up.
Features
These methods can produce very pure DNA, but the yields in small-scale applications tend to be low, especially in spun column formats.
Limitations
Not the most robust method, and recoveries tend to be lower, and the final elution step of AIX protocols involves high-salt buffers. The 0.7 to 2M sodium chloride eluate needs to be desalted, usually by a precipitation step, which decreases recovery and increases the overall procedure time. The binding capacities tend to be low (0.25–2 mg/ml of resin), increase with pH, and decrease with increasing size of the DNA. The amount of RNA present in the sample will also affect binding capacity because RNA will compete with DNA for binding.

Hydroxyapatite (HA) Based Strategies
Mechanism
Nucleic acids bind to crystalline calcium phosphate through the interaction of calcium ions on the hydroxyapatite and the phosphate groups of the nucleic acids. An increase in competing free phosphate ions from 0.12 to 0.4M will elute nucleic acids, with single-stranded nucleic acids eluting before double-stranded DNA. The entire experiment needs to be run at 60°C for thermal elution (Martinson and Wagenaar, 1974) or in the presence of formamide at room temperature (Goodman et al., 1973). Sodium phosphate buffers are most commonly used; the phosphate salt affects the selectivity of the resin (Martinson and Wagenaar, 1974). Nucleic acids may also be eluted by increasing the temperature until nucleic acid strands melt and elute from the column.

Features
Excellent separation of single-stranded from double-stranded DNA molecules.
Limitations
The quality and performance of hydroxyapatite can vary from batch to batch and between manufacturers. Thermal elution procedures require reliable temperature control, but fluctuations occur because of lack of heat-regulated chromatography equipment. These elevated temperatures can also produce bubbles in the buffer that can interfere with the separation. Hydroxyapatite has poor mechanical stability. Hydroxyapatite procedures often employ high-salt buffers and lead to sample dilution, requiring an additional precipitation step. For these reasons hydroxyapatite is not extensively referenced. It is mostly limited to subtractive cDNA cloning (Ausubel et al., 1998), removal of single-stranded molecules, and DNA reassociation analysis (Britten, Graham, and Newfeld, 1974).

ISOLATING DNA FROM CELLS AND TISSUES

2009-09-21T06:26:00.001-07:00

What Are the Fundamental Steps of DNA Purification? The fundamental processes of DNA purification from cells and
tissues are sample lysis and the segregation of the nucleic acid away from contaminants.While DNA is more or less universal to all species, the contaminants and their relative amounts will differ considerably.The composition of fat cells differs significantly from muscle cells. Plants have to sustain high pressure, contain chloroplasts packed with chromophores, and often have a very rigid outer cell wall. Bacteria contain lipopolysaccharides that can interfere with purification and cause toxicity problems when present in downstream applications. Fibrous tissues such as heart and skeletal muscle are tough to homogenize. These variations have to be taken into consideration when developing or selecting a lysis method.

Lysis
Detergents are used to solubilize the cell membranes. Popular choices are SDS, Triton X-100, and CTAB(hexadecyltrimethyl ammonium bromide). CTAB can precipitate genomic DNA, and it is also popular because of its ability to remove polysaccharides from bacterial and plant preparations (Ausubel et al., 1998). Enzymes attacking cell surface components and/or components of the cytosol are often added to detergent-based lysis buffers. Lysozyme digests cell wall components of gram-positive bacteria. Zymolase, and murienase aid in protoplast production from yeast cells. Proteinase K cleaves glycoproteins and inactivates (to some extent) RNase/DNase in 0.5 to 1% SDS solutions. Heat is
also applied to enhance lysis. Denaturants such as urea, guanidinium salts, and other chaotropes are applied to lyse cells and inactivate enzymes, but extended use beyond what is recommended in a procedure can lead to a reduction in quality and yield. Sonication, grinding in liquid nitrogen, shredding devices such as rigid spheres or beads, and mechanical stress such as filtration have been used to lyse difficult samples prior to or in conjunction with lysis solutions. Disruption methods are discussed at http://www.thescientist.com/yr1998/nov/profile2_981109.html.

Segregation of DNA from Contaminants
The separation of nucleic acid from contaminants are discussed below within the question, What Are The Strengths and Limitations of Contemporary Purification Methods? DNA Precipitation To concentrate nucleic acids for resuspension in a more suitable buffer, solvents such as ethanol (75–80%) or isopropanol (final concentration of 40–50%) are commonly used in the presence of salt to precipitate nucleic acids (Sambrook, Fritsch, and Maniatis 1989; Ausubel et al., 1998). If volume is not an issue, ethanol is preferred because less salt will coprecipitate and the pellet is more easily dried. Polyethylene glycol (PEG) selectively precipitates high molecular weight DNA, but it is also more difficult to dry and can interfere with downstream applications (Hillen, Klein, and Wells, 1981). Trichloroacetic acid (TCA) precipitates even low MW polymers down to (5kDa) (http://biotechserver.biotech.ubc.ca/biotech/bisc437/lecture/e-na-isoln/na-isoln3.html), but nucleic acids cannot be recovered in a functional form after precipitation. Salt is essential for DNA precipitation because its cations counteract the repulsion caused by the negative charges of the phosphate backbone. Ammonium acetate is useful because it is volatile and easily removed, and at high concentration it selectively precipitates high molecular weight molecules. Lithium chloride is often used for RNA because Li+ does not precipitate double-stranded DNA, proteins, or carbohydrates, although the single-stranded nucleic acids must be above 300 nucleotides. To efficiently precipitate nucleic acids, incubation at low temperatures (preferably £-20°C) for at least 10 minutes is required, followed by centrifugation at 12,000 ¥ g for at least five minutes. Temperature and time are crucial for nucleic acids at low concentrations, but above 0.25 mg/ml, precipitation may be carried out at room temperature. Additional washing steps with 70% ethanol will remove residual salt from pelleted DNA. Pellets are dried in a speed vac or on the bench and are resuspended in water or TE (10mM Tris, 1mM EDTA). Do not attempt to precipitate nucleic acids below a concentration of 20 ng/ml unless carrier such as RNA, DNA, or a high molecular weight co-precipitant like glycogen is added. In the range from 20 ng/ml to 10mg/ml, either add carrier or extend precipitation time, and add more ethanol.
Polyethylene glycol (PEG) precipitation is even more concentration dependent and will only work at DNA concentrations above 10mg/ml (Lis and Schleif, 1975). Pellets will dissolve better in lowsalt buffers (water or TE) and at concentrations below 1 mg/ml. Gentle heating can also help to redissolve nucleic acids

What Practices Will Maximize the Quality of DNA Purification?

2009-09-21T06:18:00.001-07:00

The success of DNA purification is dependent on the initial quality of the sample and its preparation. It would be nice to have a simple, straightforward formula that applies to all samples, but some specimens have inherent limitations.The list below will help guide your selection and provide remedies to nonideal situations:

1. Ideally start with fresh sample. Old and necrotic samples complicate purification. In the case of plasmid preparations, cell death sets in after active growth has ceased, which can produce an increase in unwanted by-products such as endotoxins that interfere with purification or downstream application. The best growth phase of bacterial cultures for plasmid preparations may be strain dependent. During the log phase of bacterial culture, actively replicating plasmids are present that are “nicked” during replication rather than being supercoiled. Still some researchers prefer mid to late log phase due to the high ratio of DNA to protein and low numbers of dead cells. Others only work with plasmids that have grown just out of log phase to avoid co-purification of nicked plasmid. If old samples can’t be avoided, scaling up the purification can compensate for losses due to degradation. PCR or dot blotting is strongly recommended to document the integrity of the DNA.

2. Process your sample as quickly as possible. There are few exceptions to this rule, one being virus purification. When samples can’t be immediately purified, snap freeze the intact sample in liquid nitrogen or hexane on dry ice (Franken and Luyten, 1976; Narang and Seawright, 1990) or store the lysed extract at -80°C. Commercial products, such as those from Ambion, Inc., can also protect samples from degradation prior to nucleic acid purification. Samples can also be freeze-dried.

3. Thorough, rapid homogenization is crucial. Review the literature to determine if your sample requires any special physical or mechanical means to generate the lysate.

4. Load the appropriate amount of sample. Nothing will impair the quality and yield of a purification strategy more than
overloading the system.Too much sample can cause an increase in the viscosity of the DNA preparation and lead to shearing of genomic DNA. If you do not know the exact amount of starting material, use 60 to 70% of your estimate

What Interferes with Nucleic Acid Purification?

2009-09-21T06:10:00.001-07:00

Nuclease
One of the major concerns of nucleic acid purification is the ubiquity of nucleases.The minute a cell dies, the isolation of DNA turns into a race against internal degradation. Samples must be lysed fast and completely and lysis buffers must inactivate nucleases to prevent nuclease degradation. Most lysis buffers contain protein-denaturing and enzyme inhibiting components. DNases are much easier to inactivate than RNases, but care should be taken not to reintroduce them during or after purification. All materials should be autoclaved or baked four hours at 300°F to inactivate DNases and RNases, or you should use disposable materials. Use only enzymes and materials guaranteed to be free of contaminating nucleases. Where appropriate, work on ice or in the cold to slow down potential nuclease activity. Smears and lack of signal, or smeared signal alone, and failure to amplify by PCR are indicative of nuclease contamination. The presence of nuclease can be verified by incubating a small aliquot of your sample at 37°C for a few hours or overnight, followed by evaluation by electrophoresis or hybridization. If nuclease contamination is minor, consider repurifying the sample with a procedure that removes protein.

Shearing

Large DNA molecules (genomic DNA, bacterial artificial chromosomes, yeast artificial chromosomes) can be easily sheared during purification. Avoid vortexing, repeated pipetting (especially through low-volume pipette tips), and any other form of mechanical stress when the isolate is destined for applications that require high molecular weight DNA.

Chemical Contaminants
Materials that interfere with nucleic acid isolation or downstream applications involving the purified DNA can originate from the sample. Plants, molds, and fungi can present a challenge because of their rigid cell wall and the presence of polyphenolic components, which can react irreversibly with nucleic acids to create an unusable final product. The reagents of a DNA purification method can also contribute contaminants to the isolated DNA. Reagents that lyse and solubilize samples, such as guanidinium isothiocyanate, can inhibit some enzymes when present in trace amounts. Ethanol precipitation of the DNA and subsequent ethanol washes eliminate such a contaminant. Phenol can also be problematic. If you experience problems with DNA purified by a phenol-based strategy, apply chloroform to extract away the phenol. Phenol oxidation products may also damage nucleic acids; hence re-distilled phenol is recommended for purification procedures. A mixture of chloroform and phenol is often employed to maximize the yield of isolated DNA; the chloroform reduces the amount of the DNA-containing aqueous layer at the phenol interphase. Similar to phenol, residual chloroform can be problematic, and should be removed by thorough drying. Drying is also employed to remove residual ethanol. Overdried DNA can be difficult to dissolve, so drying should be stopped shortly after the liquid can no longer be observed. Detailed procedures for the above extraction, precipitation and washing steps can be found in Sambrook, Fritsch, and Maniatis (1989) and Ausubel et al. (1998). Ammonium ions inhibit T4 polynucleotide kinase, and chloride can poison translation reactions (Ausubel et al., 1998). The common electrophoresis buffer, TBE (Tris, borate, EDTA) can inhibit enzymes (Ausubel et al., 1998) and interfere with transformation due to the increased salt concentration (Woods, 1994). Phosphate buffers may also inhibit some enzymes, namely T4 Polynucleotide kinase (Sambrook et al., 1989), alkaline phosphatase (Fernley, 1971), Taq DNA polymerase (Johnson et al., 1995), and Poly A polymerase from E. coli (Sippel, 1973).Agarose can also be a problem but some enzyme activity can be recovered by adding BSA to 500mg/ml final concentration (Ausubel et al., 1998). EDTA can protect against nuclease and heavy metal damage, but could interfere with a downstream application. The anticoagulant heparin can contaminate nucleic acids isolated from blood, and should be avoided if possible (Grimberg et al., 1989). Taq DNA polymerase is inhibited by heparin, which can be resolved by the addition of heparinase (Farnert et al., 1999). Heparin also interacts with chromatin leading to release of denatured/nicked DNA molecules (Strzelecka, Spitkovsky, and Paponov, 1983). Narayanan (1996) reviews the effects of anticoagulants.

Inherited disease(Top ten genetic disorder)

2009-09-21T00:27:00.001-07:00

There is a list of inherited disease and i will explain each disease in detail latter

Cystic fibrosis: CF is a multi organ disease that causes failure of different organ and effects mainly in pulmonary, gastrointestinal and reproductive organ.
Heredity homochromatosis: It is a autosomal recessive disorder of iron regulation that causes excessive storage of iron in tissues.
Carbomylase phosphate synthetase I deficiency: The effect newborn appears normal but within 24-72 hours develops vomiting, lethargy, and blood ammonia level continue to increase, coma and death is imminent unless treatment started.
Achrondoplastia: This is common type of human dwarfism.
Charcot-Marie tooth disease : Sometime refers as heredity motor and sensory neuropathy that characterized chronic motor and sensory neuropathy.
Huntington’s disease : This is autosomal disorder that couses late onset neurodegenerative disorder.
Hemophilia A : This is X linlked ressesive bleeding disored coused by deficiency of coagulation factor VIII.
Duchenne’s muscular dystrophy: That is a X linked disorder characterized by progressive skeletal muscle wasting syndrome.
Fragile X syndrome: This is a most common inherited disease that causes mental retardation.
Inherited cancers: Breast cancer, colon cancer and many more cancer are there which is inherited.

Customized medicine

2009-09-20T23:54:00.001-07:00

Are you fighting with cancer, taking chemotherapy, radiotherapy or some antibody that antagonists the cell development? Or a diabetic, arthritis or certain type of genetic disorder. That cause an intense side effect on your whole system or taking any nonspecific drug that causes an adverse reaction.

Then why you don't try your customized medicine. In future many companies are looking forward in costamised medicine system in which every person have their genotypic and phenotypic profile created by drug companies or private lab which is used by the medicine companies to prepare the customized drugs. That have little or no side effect and made for you specially using our own genetic substance like DNA, or protein from your own cell that is very specific. The genotype and phenotyping is already got in streamline in developed countries and being used for many care. In this system their are two type of testing is available one is known as genotyping an which your genetic profile about a specific protein or enzyme is detected. The second method is rapid in which your physically expressed protein or enzyme is detected in laboratory by reacting your proposed medicine and its efficiency and dose is defined.

The whole approach is called as Pharmacogenetics. The common feature of pharmacogenetics is proven useful in cancer chemotherapy, immunosuppressant, antidepressant, antipsychotics, anticoagulants, Antihypertensive, cardiac medication, and lipid lowering therapies.

Basic clinically related pharmacogenetics targets are

Thiopurine-S-methyletransferase TPMT is a metabolic enzyme that catalysesthe inactivation of 6 methylepurine thus preventing it from formation of thioguanine nucleotide. This is done in acute lymphocytic leukemia.
Cytochrome P4502D6 – It has a name called debrisoquine hydroxilase that participate in more than 100 of drug metabolism. This is used in antidepressant medication, angina and hypertension treatmment.
Cytocrome P450 2C19
N-Acetyle transferase NAT1 or NAT2

More in research just wait and your customized medicine is on your doorstep.

Target neurons and forget addiction

2009-09-20T23:04:00.001-07:00

Every drugs or addictive substance activates a certain area of mind and activates neurons that in turn causes activated secretion of substance either activator or depressant which causes pleasure.The activated brain or group of neurons always want to stay in the same state that causes addiction.

What if we target that group of neurons or part of brain and modified the response either depress or stimulate or even antagonize the whole response.

Let your heart heal themself

2009-07-26T07:22:00.000-07:00

How would you feel if your heart repaire themself after a heart attack or if the prone of being heart patient could be reduced to 0%. Your cardiomyocyte prolifrates(heart cells) at the rate as liver cell does. How would you feel if your heart disese could cured just thought some protein ingections. Is it not a great alternative to bipass surgery or high risk surgery or expensive procedures. But how??

It may be possibe very soon if every things planned by Bernhard kuhn and his associates goes smother. He has shown that a protein given to mice not only induced cardiomyoctes prolifration but also decrease cardiac cell damage after heart attack.

But how it is possibe? That is by injecting a well studies protein called neuregulin 1 that is responsible for cardiac cell prolifration in feutus. Kuhn team created artificial heart attack by removing its major arterey that couse heart attack in mouse then some of mice given the protein that couse speedy scare dispared and their heart pumps more blood in their artery thereafter.

But their are some criticism also following this research that it will turn the cardiac cell in to cancerous cell. Lets watch that when our heart start healing himself.

Blog Rolls

2009-05-06T01:09:00.000-07:00