Tips To Maintain Good Hair Hygiene

http://www.boldsky.com/beauty/hair-care/2013/maintain-hair-hygiene-032717.html

Do you want a stylish hair? Then get a new hair style. Try straightening, curling or colouring you hair for a makeover. But if you want to have healthy hair, then you should give prime importance to hygiene. Sebum produced by sebaceous glands in the hair follicles are beneficial as it keeps the hair healthy. However, excess production or accumulation of sebum is not desirable as it cause hair loss. 

We all worry about various hair problems, but with a habit of good hygiene, most of this can be solved. Healthy hair definitely has an important role in keeping your confidence level high. No matter how appealing you are, a bad odour from your hair is strong enough to make you feel embarrassed. Now stop worrying about your hair problems, and follow these simple steps to maintain a good hair hygiene. 





Washing your hair: It is the first and foremost need to keep your hair and scalp clean. Washing your hair and massaging your scalp will resolve most of your hair complaints. But it is important to select a mild shampoo which is formulated for your hair type. 

Use conditioners: Frequent drying, colouring, straightening, and usage of chemical products can affect your hair health. Hair conditioners are usually helpful to maintain proper hair hygiene, as they will form a light layer over the hair and keep it protected. 

Clean your scalp: Massaging your hair with warm oil will help to clean your scalp. Massaging will help remove dandruff or dirt from the scalp. Oil can act as a conditioner and will protect your hair from getting rough and dry. 

Remove excess oil: While applying oil is healthy for hair, keeping excess oil on hair without washing will affect the health of your hair. It will accelerate the accumulation of dirt on the scalp. Always remember to wash off the excess oil with a mild shampoo. 

Clean your hair accessories and tools: Washing your comb is very important to maintain hair hygiene. Do not share your comb. Clean your hair accessories like hair clips or hair bands. 

Do not use expired hair products: The composition of these can change over time which will eventually result in hair fall. To keep your hair and scalp hygienic and clean, use quality products only. 

Treat dandruff: Dandruff occurs mainly due to poor hygiene of your hair, which can further cause hair fall. Take necessary measures to resolve dandruff and treat hair fall. You can try natural remedies and if it is not working, consult a doctor. 

Wash off sweat: Washing your hair whenever sweaty is important to remove the dirt and dead skin of scalp which are accumulated due to sweat. Excessive sweating will cause bad odour as well. Keep hair clean and neat to maintain hygiene and reduce hair problems like hair fall and itching. 

Not loose, not tight: Keeping hair loose will settle dirt and dust. Loosened hair is more prone to hair splitting. Tying hair too tight may cause excess sweating. So, do tie your hair too tight or loose. 

During travel: Always keep a mini pack of oil, shampoo and conditioner with you if you are travelling for several days. Try to keep your hair oil free while travelling. Do not let sweat stay on scalp for a long time. Follow these tips to maintain hair hygiene.


Read more at: http://www.boldsky.com/beauty/hair-care/2013/maintain-hair-hygiene-032717.html

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CROSS STITCHING...In the Shade of Love...

CROSS STITCHING...In the Shade of Love...

This is a cross stitching done by me. 

Cross-stitch is a popular form of counted-thread embroidery in which X-shaped stitches in a tiled, raster-like pattern are used to form a picture. 

Cross-stitch is often executed on easily countable evenweave fabric called aida cloth. But, here I used a frame. 

We have to count the threads in each direction so that the stitches are of uniform size and appearance. 

This form of cross-stitch is also called counted cross-stitch in order to distinguish it from other forms of cross-stitch.

To customize it I just left one portion of the thread free instead of stitching. In final outcome it gave a 3D result.

Try it. It really needs patience  and composure to a great extent. But, once you complete one, you will really love it. 

Since my wall is ivory in color, I gave a black frame for it.  

Try doing one and enjoy the fun. 

Framed and hanged...

UNDERSTAND AND OVERCOME DEPRESSION

My article in Ethnic Health Court http://ethnichealthcourt.com/2013/02/25/understand-and-overcome-depression/

UNDERSTAND AND OVERCOME DEPRESSION

My article in Ethnic Health Court http://ethnichealthcourt.com/2013/02/25/understand-and-overcome-depression/

Most of us feel gloomy and sad one time or the other in our life. But don’t be depressed thinking that you have depression. True clinical depression is a mood disorder in which these negative feelings interfere with everyday life for weeks or longer. According to World Health Organization, depression is a mental disorder, characterized by sadness, loss of interest or pleasure, feelings of guilt or low self-worth, disturbed sleep or appetite, feelings of tiredness and poor concentration.

Don’t think that depression is associated with sad conditions only. Major good events like marriage, new job, professional achievement or even any important responsibility can cause anxiety and depression.

 CAUSES

  There is no single cause for depression. It can be due to various triggers.

·         Alcohol or drug abuse

·         Many central nervous system illnesses and injuries

·         Certain medical conditions, including under-active thyroid, cancer

·         Certain medications such as steroids, beta-blockers or reserpine

·         Sleeping problems

·         Stressful life events such as death of dear ones, conflict, relation problems, job stress, social isolation etc.

·         Past physical, sexual, or emotional abuse

·         Genetics

·         Delivery (postpartum depression)

SYMPTOMS

·         Restlessness or irritability

·         Difficulty in concentrating

·         Continuous low mood or sadness

·         Change in appetite, often with weight gain or loss

·         Fatigue

·         Loss of self esteem

·         Feeling anxious or worried

·         Feelings of guilt

·         Loss of interest in activities that were once enjoyed

·         Thoughts of death or suicide and thoughts of harming yourself

·         Psychotic symptoms, such as hallucinations and delusions in severe cases.

DIAGNOSIS

Diagnosis is mainly based on clinical presentation. No specific blood test or scan is available to confirm depression. Butat the same time some tests are used to rule out other medical conditions that may cause similar symptoms. That can be blood tests, CT, MRI or EEG.

TREATMENT

In general, treatments for depression include:

·         Antidepressants

·         Psychotherapy

Cases with mild depression need only one of these treatments. Severe depression usually need a combination of both treatments. Don’t expect a sudden cure from depression, but wait for a slow improvement.

If the patient has suicidal or self harming tendency, help of a trained and qualified councilor is needed.

MEDICATIONS

Drugs used to treat depression are called antidepressants. Common types of antidepressants include:

·         Selective serotonin re-uptake inhibitors (SSRIs), including fluoxetine (Prozac), sertraline (Zoloft), paroxetine (Paxil), fluvoxamine (Luvox), citalopram (Celexa), and escitalopram (Lexapro).

·         Serotonin norepinephrine reuptake inhibitors (SNRIs), including desvenlafaxine (Pristiq), venlafaxine (Effexor), and duloxetine (Cymbalta).

Other medicines used to treat depression include:

·         Tricyclic antidepressants, Bupropion (Wellbutrin), Monoamine oxidase inhibitors

Lithium (or other mood stabilizers) and thyroid hormone supplements also maybe added to help the antidepressants work better.

COMPLICATIONS

·         Increased risk of associated health problems

·        Suicide

MANAGEMENT

Specialized treatment is needed only in advanced cases. Most cases can be successfully treated by a primary health care professional. Help of a councilor will improve the confidence of the patient.

TIPS TO PREVENT DEPRESSION FROM RULING YOUR  LIFE

Be confident: Low self-esteem will definitely put you down in every phase of your life. Be confident in your physical appearance and mental strength.

Find out what is the problem: In most cases what we hear people telling is’ I don’t know the reason, but am not feeling good’. Remember if you don’t have any problems, then you will never get worried. The fact is, you know what it is, but not ready to accept it. If you want to be happy, then take steps to solve your problem.

Connect with others: Involving in group activity is very important. If you find you are unhappy, don’t try to isolate yourself from others. Look forward for social connections that can uplift you.

Enjoy a hobby: Focusing on activities that keep you busy and entertained is very important. Engaging in something meaningful will make you energetic. It will help you dream about tomorrow.

Maintain a good sleeping habit:  Let your body take rest and see how things will change with magical effect. Relaxing your mind and body is needed for a perfect healthy life.

Share your problem: Something you think difficult to solve will be simpler than anything else for another person. So don’t hesitate to share your problems with anyone you find caring for you.  That can be a friend, partner, parents, teachers, siblings or relatives. But what if your problem itself is not having anybody to care you? Don’t worry. Contact a qualified and trained counselor.

Be positive: When you have all the freedom to think positive or negative about a situation, why not opting for positive thoughts? The situation is not what will change for you. The attitude that you have towards a situation should change. Accept the problem as it is and think positively to resolve it.

Success will not come in one day: Don’t think that you are useless in failures. It is just a part of game. Tomorrow is open for you to have your next trial. Learning from your past mistakes will help you make your success more colorful.

Don’t conclude before the beginning: Taking a negative conclusion before starting a task will never take you to success. This will lead you to depression and anxiety. Feel free to face life as it comes.

Study the situation:  Don’t try to assume anything based on a particular situation. Wait and study what happened. Never think ‘I don’t deserve anything better’.  Tell yourself that you are strong enough to handle the situation.

Leave your past and live today:  Never allow mistakes that happened in the past to rule your present and the future. Learn from mistakes, take measures to correct it, leave it and be happy.

Regular exercise and yoga: Maintaining a healthy mind and body will not only help you beat depression, but increase inner peace and joy.

Be yourself:  If you want to be confident, then you should stop worrying about what others will think about you. Realize nobody other than your family is too concerned about you. This itself will give you all the courage to face the life with confidence.

Be realistic: Don’t expect that miracles are about to happen in your life. Being in a dream world is more likely to take you to depression. Focus on realistic expectations and work hard for it.

My article in oneindia.com

http://www.boldsky.com/pregnancy-parenting/kids/2013/tips-for-effective-communication-with-kids-032416.html

TIPS FOR EFFECTIVE COMMUNICATION WITH CHILDREN

Happy family is the best gift that anyone can ever get. But in this modern age, family bonds are getting loosened due to various reasons. Each member of a family has a different personality and age. The best way to bring them all close to each other is effective communication. Kids brought up in a happy family will always come out as a responsible and efficient social beings. Expressing your love and discussing any problem will result in a happy family. 

Here are some tips for communicating with kids. 

1. Listen to your children. Let them know that you are interested in knowing about them. Ask them what happened in school and share how your day was. Don't turn to television, newspaper or telephone when they start talking. 

2. Talk to them. Stop debating in social networks and talk to your child about his favorite game, cartoon or book. Be surprised to see how talkative they will be. 

3. Give them some time to respond. Children may take some time to process and understand what you have said. 

4. Computers and internet has become inevitable part of our daily life. Children are growing up watching and copying their parents spending free time with internet. Give them your time when your kids want it the most and they will like it more than playing video games. 

5. Make your communication with kids simple and clear. Don't make children confused between your attitude and words. Gestures are as important as words for children. 

6. Appreciate them generously and loudly for any achievement. Also, make sure you comfort them in failures. 

7. Communication should always be something that motivates your child. Make them feel secure. Give them full freedom that they can run to you at any time. 

8. While giving instructions, remember to keep it positive. For example, instead to telling "John, don't take that book" you can try "John, keep that book in place." 

9. Respond with patience. Think with a kid's mind after listening to them. Take some time to think about what your child want to convey and respond in a gentle and caring way. 

10. Never use abusing words in front of children. Don't discuss family matters in front of children if they are not good enough to take it positively. At the same time, consider their opinion also when taking any family related decision. 

Communication is the best way to express your feelings for your child. Let them know that they are blessed with the best family in this world.

Read more at: http://www.boldsky.com/pregnancy-parenting/kids/2013/tips-for-effective-communication-with-kids-032416.html

FLOWER MAKING... Here it is...Some together...

These are some flowers and painted vases which I made. Please go through individual posts for more details about what materials are used and how to make. Hope it will help you to try one.

GLASS PAINTING - FAIRY

GLASS PAINTING-CHINESE KOI FISH

Acrylic painting

This is my all time favorite as this is my first sold out acrylic painting.

Started with red and moved along to yellow for the horizon.

And how could a tree branch be lively with out some birds, especiaally two love birds..... so they are here...

Acrylic Painting - TULIPS

Work of Tulips in progress...

Work in progress..

I preffered painting the minute details first after giving a light shade in the background.This helped me to assume how it will and how it should look after completing it.

Arylic painting...Tulips

Added the needed green and loved using a light brown shade for the tree and just brushed it for the background also. Play with colors and try experimenting.

Asha Das

YARN WORK - God's Eye wall hanging

YARN WORK - God's Eye wall hanging

The Ojo de Dios or God's eye is a ritual or cultural symbol of Huichol peoples of western Mexico.The God's Eye is considered as a symbolic representation of the power of seeing and understanding that which is unknown and unknowable,The four points represent the elemental processes: earth, fire, air, and water.

Asha Das

Tissue Paper Flowers...

Tissue Paper Flowers...

How to make tissue paper flowers?

You will need:

• Tissue paper
• Scissors
• 2 paper clips
• Floral wire
• Optional:floral tape,wire cutters

Tissue Paper Flowers...a closer view..

1.  Take 5-6 tissue papers one over the other.Fold it half way and then again to a half. Draw a 

     circle on the top side. You can use a bottle cap with required diameter. I feel that a circle of 

     around 7 cm diameter will give a realistic size.

2.   Now cut the circle out. Clip the paper for keeping them altogether. 

3.   Make a small hole at center of the circle and insert a floral wire through it.Tie it firmly.

4.   For making the petals, separate the first layer of tissue paper and crumble it upwards.

5.   Now crumble each layer one by one.

6.   Use the floral tape to wrap the stem all the way to bottom which will give a more realistic look.

 Try it out. I feel it simple and interesting. 
 What about you?



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ഞാന്‍ ഒരു  മഞ്ഞുതുള്ളിയും 

നീ ജ്വലിക്കുന്ന  സൂര്യനുമായാല്‍

ചിലപ്പോള്‍ നീ എന്റെ തിളക്കം കൂട്ടും 

അല്ലെങ്കില്‍ ഞാനുരുകി ഇല്ലാതാകും 

പാതി മുറിഞ്ഞൊരു സുന്ദര സ്വപ്നത്തിന്‍
ആലസ്യമിപ്പോഴും വിട്ടോഴിഞ്ഞില്ലതിന്‍ 
പൊട്ടിയ കഷണങ്ങള്‍  ചേര്‍ത്തു വെച്ചിന്നൊരു 
പുതിയ സ്വപ്നത്തിന്‍ പണിപ്പുരയിലാണ് ഞാന്‍  

പൂനിലാവിന്‍ കരതലത്തില്‍ തല ചായ്ച്ചു
ഉറങ്ങാന്‍ കിടന്നൊരു തൈമുല്ല മൊട്ടു പോല്‍
നിന്‍ പ്രണയനിലാവില്‍ മുഖം ചേര്‍ത്ത് 
പുതിയൊരു പുലരിയിലേക്ക് തിരിക്കവെ

എത്തി ഞാന്‍  ഇതുവരെ കാണാത്ത നിന്‍ 
ഹൃദയ കൊട്ടാര മുറ്റത്തെ പൂവാടിയില്‍  
ചാമരം വീശുന്നു വെണ്‍ മേഘജാലങ്ങള്‍ 
സുഗന്ധം പരത്തുന്നു പനിനീര്‍ പൂവുകള്‍

ഒഴുകിയെത്തും ഇളം കാറ്റിനിരുന്നാടാന്‍ 
ഊഞ്ഞാല് തീര്‍ക്കുന്നു വര്‍ണ മഴവില്ല്
മിന്നി തിളങ്ങുന്ന മഞ്ഞുത്തുള്ളികള്‍ 
മണിമാല ചാര്‍ത്തുന്ന പുല്‍തലപ്പുകള്‍

പ്രകൃതി തന്‍ കൈ തട്ടി ചിതറി തെറിച്ച 
നിറങ്ങള്‍ മിഴിവേകുന്ന ചിത്രശലഭങ്ങള്‍ 
കണ്ണിലും കരളിലും പുതിയൊരു നിര്‍വൃതി
ചേര്‍ത്ത് വെച്ചിന്നവയ്ക്കൊപ്പം പറന്നു ഞാന്‍

നിന്നു നിന്‍ ഹൃദയ കവാടത്തിന്‍ മുന്നില്‍ 
ഒരു മാത്ര എന്‍ ഹൃദയം എന്തിനെന്നറിയാതെ 
കൈനീട്ടി ആ വാതില്‍ തുറക്കുന്നതിന്‍ മുന്‍പേ
കാത്തിരിപ്പിന്‍ മഞ്ഞുരുകുന്നതിന്‍ മുന്‍പേ

എത്തി ഞാന്‍ പുതിയൊരു പുലരിതന്‍ തീരത്ത്
അവിടെ നീയില്ല പൂവില്ല പൂമ്പാറ്റകളില്ല 
പൊന്‍ പ്രഭാതതിന്‍ തൂവേളിച്ചതിലാ 
 മാസ്മര കാഴ്ചകളെല്ലാം മറഞ്ഞു പോയി

ഞാന്‍ ആ മണിവാതില്‍ തള്ളി തുറന്നുവോ
നീ തുറന്നെന്നെ അകത്തേക്ക് ക്ഷണിച്ചുവോ
മാറോട്‌ ചേര്‍ത്തെന്നെ  പുല്‍കി നീ എന്‍ കാതില്‍
കാത്തിരിക്കയായിരുന്നെന്നു മന്ത്രിച്ചുവോ

പാതിവഴിയില്‍ മുറിഞ്ഞൊരു സ്വപ്നത്തിന്‍ 
കൊഴിഞ്ഞ നിമിഷങ്ങള്‍ താലോലിക്കുവാന്‍
സങ്കല്‍പ്പദളങ്ങള്‍ ചേര്‍ത്തുവച്ചിന്നൊരു 
പുതിയ സ്വപ്നത്തിന്‍ പണിപ്പുരയിലാണ് ഞാന്‍ 

എന്‍റെ പൊന്നോമനയ്ക്ക് ...

ഓമന കുഞ്ഞേ നീ ചായുറങ്ങിടുകെന്‍
താമര പൂവേ നീ ഉറങ്ങുറങ്ങ്
ജീവന്റെ ജീവനെ നിന്നെ ഉറക്കുവാന്‍ 
അച്ഛനും അമ്മയും ചാരെയുണ്ട്

പൊന്മാനും പൂമീനും വെണ്‍പ്രാവും
പൊന്നൂഞ്ഞാലാടുന്ന ചേലില്‍ നിന്‍ പൂന്തൊട്ടിലില്‍
ചാഞ്ചാടിയാടി  ഉറങ്ങിടുകെന്‍  മുത്തേ
സുന്ദര സ്വപ്‌നങ്ങള്‍ കണ്ടുറങ്ങു

തുമ്പയും തുമ്പിയും തുള്ളിക്കളിക്കുന്ന

പൂമുറ്റത്തോടി കളിചീടണ്ടെ

കൈ വളരുണ്ണി നിന്‍ കാല്‍ വളരുണ്ണി നിന്‍

കണ്ണും കാതും നന്മയ്ക്കായി വളര്

നല്ലത് ചെയ്യുവാന്‍ നല്ലത് ചൊല്ലുവാന്‍ 

നല്ലത് മാത്രമേ  ചിന്തിച്ചീടാന്‍ 

അച്ഛനും അമ്മയ്ക്കും പോന്നുണ്ണിയായി നീ 

നാളെ വളര്‍ന്നീടാന്‍ ഇന്നുറങ്ങ് 

Dear friend, come lets go back

Over the fields of memories

Leaving the present behind

To the long ago you and me...

We laughed and we cried..

We played for hours

Sat quietly by each others side

Hold in the hands of innocence

As safe as it can be,,

Sharing palm for friendship

Sharing heart for love

Sharing words for care

The long ago you and me

Sharing you... 

Sharing me..

നിന്നുടെ പാതി ഞാന്‍ 

എന്നുടെ പാതി നീ

നമ്മുടെ പാതികള്‍ ചേര്‍ന്ന് ജനിക്കുന്നു 

നാം തന്നെയാകുമൊരു പൊന്നോമന 

ഭാവനകള്‍ വരകളിലാക്കുവാന്‍ ചുവരുകള്‍ ഇല്ലാതിരുന്ന  ഒരു  കാലത്ത് നടവഴിയിലെ പൂഴി മണലില്‍ ഞാന്‍ എന്റെ ചിത്രങ്ങള്‍ കോറിയിട്ടു. നോക്കിയും നോക്കാതെയും , കണ്ടും കാണാതെയും പലരും അത് ചവിട്ടി  കടന്നു പോയി. മാഞ്ഞു പോകുന്ന വരകള്‍ ചേര്‍ത്ത് വെച്ച് വീണ്ടും ഞാന്‍ കാത്തിരുന്നു. ഒരിക്കല്‍ നീ മാത്രം ഒരു നിമിഷം നോക്കിയ ശേഷം എന്റെ വര്‍ണങ്ങളില്ലാത്ത വരകളില്‍ ചവിട്ടാതെ കടന്നു പോയപ്പോള്‍ , അതായിരുന്നു എനിക്ക് ലഭിച്ച ആദ്യത്തെ അംഗീകാരം

Erythrocyte Glutathione-S-Transferase Activity in Diabetics and its Association with HBA1c

http://www.webmedcentral.com/article_view/2004

Erythrocyte Glutathione-S-Transferase Activity in Diabetics and its Association with HBA1c

Abstract

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The generation of reactive oxygen species is increased in both types of diabetes and is closely associated with oxidative stress. Erythrocytes are equipped with a highly effective anti-oxidant defence system. Oxidative denaturation of hemoglobin leads to the release of hemin into the RBC membrane and the released hemin is capable of oxidizing membrane proteins via a thiyl radical intermediate. GST can bind free hemin that is released during Hb oxidation presumably reducing damage to RBC membrane. We hypothesized that as oxidative stress and hyperglycemia are major etiologic and pathologic factors of diabetes mellitus, then the antioxidant enzyme, Glutathione – S – Transferase (GST) in red blood cells, are likely to have a role in the glycation of Hb in diabetic patients. Hence we designed this study to evaluate the activity of red cell Glutathione-S-Transferase and correlate its association with HbA_1c. This is a cross – sectional study, conducted in subjects attending Kasturba Hospital, Manipal. All patients who were advised to do a blood test for glycated haemoglobin were included in this study. Hemolysed blood samples were excluded. Erythrocyte GST activity was determined using the method of Habig et al and glycated Hb (HbA_1c) was estimated using the turbidimetric inhibition immunoassay (TINIA) for hemolyzed whole blood by Roche Cobas Integra 400 auto analyzer. 68 patients who were confirmed to have type 2 diabetes mellitus were included in diabetic group whereas rest 32 patients without diabetes were included in non-diabetic group. The data were analysed using Mann Whitney U test and Pearson correlation coefficient test by SPSS (v. 14.0) software. From this study we could found out that there is no significant difference in erythrocyte GST activity between diabetics and non-diabetics (p value = 0.08). However a positive correlation between erythrocyte GST activity and HbA_1c concentration was observed in diabetic patients (r = 0.239, p = 0.089 ). The chronicity of the disease along with treatment modalities might have played a significant role in the outcome of our study as the subjects included in diabetic group were selected without considering their glycemic control status.

Introduction

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It is well known that oxidative stress plays an important role in the pathophysiology of diabetes mellitus by impairing various cellular functions. Abnormally high levels of free radicals and the simultaneous decline of antioxidant defense mechanisms can lead to damage of cellular organelles and enzymes, increased lipid peroxidation, and development of insulin resistance leading to diabetic complications [1].
The easy accessibility, finite life span and relative simplicity of erythrocytes make them an attractive model to study the oxidative stress status of the body in diabetes mellitus. Erythrocytes, which are equipped with a highly effective anti oxidant defence system, can scavenge free radicals by the action of their antioxidant enzymes [2,3].
In erythrocytes, glutathione detoxifies free radicals through GST conjunction. GST also possesses peroxidase activity and can directly attack the peroxides. Interaction of free radicals with hemoglobin can denature Hb. Oxidative denaturation of hemoglobin leads to the release of hemin into the RBC membrane and the released hemin is capable of oxidizing membrane proteins via a thiyl radical intermediate. GST can bind free hemin that is released during Hb oxidation presumably reducing damage to RBC membrane [4-6].
Considering all these factors we hypothesized that if oxidative stress and hyperglycemia are major etiologic and pathologic factors contributing towards diabetes mellitus and its complications, then Glutathione – S – Transferase in red blood cells, are likely to have a role in the glycation of Hb in diabetic patients. Hence we designed this study to evaluate the activity of red cell Glutathione-S-Transferase and correlate its association with Glycated Hemoglobin (HbA1c).

Methods

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This study was designed as a cross – sectional study, conducted in subjects attending Kasturba Hospital, Manipal. All patients who were advised to do a blood test for HbA1c were included in this study. Hemolysed blood samples were excluded.
Specimen Collection: Na2EDTA-Fluoride anticoagulated venous blood was used for both GST activity and HbA1c estimations. The blood sample was centrifuged at 3000 rpm for 20 min to separate the packed red cells. The separated red cells were washed 3 times with normal saline.
Erythrocyte GST activity
Erythrocyte GST activity was determined using the method of Habig et al (1974). This assay is based on the principle that GST catalyzes the conjugation of L-glutathione with CDNB through the thiol group of the glutathione to form GS-DNB Conjugate, which has maximum absorbance at 340 nm. The rate of increase in the absorption is directly proportional to the GST activity in the sample[7].
The hemolysate was diluted 1 in 5 with phosphate buffer and used for the assay. To the reaction tube, 0.1 ml of 30 mM GSH, 0.1 ml of CDNB and 2.7 ml of 100 mM KPO4 buffer (pH 6.5) were added . The reaction was initiated by adding 0.1ml of hemolysate, mixed thoroughly, and absorbance was recorded at 340 nm every minute for 5min period. The average change in absorbance ΔA(avg) per minute was calculated.
Estimation of Glycated hemoglobin (HbA1c)
The concentration of HbA1c was estimated using the turbidimetric inhibition immunoassay (TINIA) for hemolyzed whole blood by Roche Cobas Integra 400 auto analyzer. In this method the polyhaptens react with excess anti-HbA1c antibodies to form an insoluble antibody polyhapten complex which can be determined turbidimetrically. Here the hemoglobin concentration has been determined in a second channel. Liberated hemoglobin in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum which is measured bichromatically.
Statistical analysis
Erythrocyte GST activity in diabetics and non-diabetics were compared with Mann Whitney U test. The Pearson correlation coefficient test was applied to know the association of erythrocyte GST with HbA1c. A p value of ≤ 0.05 was considered statistically significant. The analysis was performed using SPSS (v. 14.0) software.

Results

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After the eligibility criteria, a total of 100 subjects were included in this study. The subjects were divided into “diabetic” and “non-diabetic” based on clinical diagnosis. 68 patients who were confirmed to have type 2 diabetes mellitus were included in diabetic group whereas rest 32 patients without diabetes were included in non-diabetic group [Table 1.].
Erythrocyte GST activity in diabetic patients
The erythrocyte GST activity in the diabetic patients was assessed by comparing with that in non-diabetics. Data were expressed as median and interquartile ranges. The GST activity in the diabetic group was 83.3(41.6-145.8)IU/L and that of non-diabetic group was 83.4 (47-153.6)IU/L .The Mann Whitney U test revealed that there is no statistically significant difference in the distribution of erythrocyte GST between diabetic patients and non-diabetic patients (p value = 0.08).
Association between erythrocyte GST and HbA1c
The association of erythrocyte GST and HbA1c was studied considering the possible effect of glycemic control on erythrocyte GST. The Pearson’s correlation study yielded a mild positive correlation (r = 0.239, p = 0.089 ) between erythrocyte GST and HbA1c among the diabetic patients [Fig. 1.] and no association in non-diabetic patients [Fig. 2.].

Discussion

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Increased hyperglycaemia induced oxidative stress is known to be a component of molecular and cellular tissue damage, associated with complications of diabetes. Impaired antioxidant status is known to be an indicator of oxidative stress..
Glutathione metabolism and GST distribution in the tissues may play an important role in the etiology, pathology and prevention of diabetes . Low levels of GSH in diabetics are found to potentiate the effects of the increased reactive oxygen species. Changes in GSH dependent enzyme activities, such as glutathione peroxidase, γ-glutamyl transpeptidase, and glutathione S-transferase(GST) are also noted in diabetes.
Evidences from few studies suggest that erythrocyte GST tend to decrease in diabetic patients. But there are studies which show increased GST activity in diabetics independent of their glycemic status. According to our study, there is no significant difference in erythrocyte GST activity between diabetics and non-diabetics. Decreased red blood cell GSH as well as increased serum total GST levels may be due to a compensatory mechanism of the antioxidants to combat the oxidative stress in diabetic conditions with or without complications . The regulation of GST is subject to a complex set of endogenous and exogenous parameters. These include developmental, gender, and tissue-specific factors, as well as a large number of xenobiotic-inducing agents.

Conclusion

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The association of erythrocyte GST with HbA_1c was tested since HbA_1c is an established marker of glycemic control. The hypothesis underlying this study was that there will be a correlation between erythrocyte GST and HbA_1c. Our study shows no significant difference in erythrocyte GST activity among diabetic and non diabetic patients. However a positive correlation between erythrocyte GST activity and HbA_1c concentration was observed in diabetic patients. The chronicity of the disease along with treatment modalities might have played a significant role in the outcome of our study as the subjects included in diabetic group were enrolled without considering their glycemic control status.