posture-training

‘Tech Neck’ Can Turn Your Good Posture into Bad Posture

Ever found yourself trying to force your body into a ‘perfect’ posture? Your body is now in perfect alignment and your spine can maintain its natural curvature, unstrained, right? Just so you know, a good posture always starts from the centre of the body. Good posture is affected by and affects your breathing, whereby there’s more capacity inside your chest for your lungs in a good open posture. You see, what actually happens is that when your lungs don’t have enough space, it affects your breathing and when your posture is good, your back is straight, and there is no pressure on your bones and ribs which then it is but very obvious that you will be able to breathe easily. Our affordable personal training package retrains your body to assume the correct posture. You might remember your parents telling you to “stop slouching” and “sit up straight!”. Well, we don’t do that, we activate the muscles that have weakened over time from neglect. Bad posture can be dangerous and can give you back problems. Having a bad posture means increased body pain especially in the bones and joints and muscle groups feel the strain. Most of the people complaining about lower back pain and some even suffer from which again is a result of a bad posture. When your body isn’t aligned properly, it starts putting pressure on your lower back and all the other joints in your body. On the other hand, properly aligned body or in other words a body with a good posture puts no stress on shoulders and places like the lower back which ultimately results in decreased body pain.

Research has shown that time spent sitting at a desk has a positive correlation with the strength of lower back pain and neck-shoulder pain. Awkwardly, back and neck misalignment can be caused by spending a lengthy time sitting at a desk. You have probably heard the claim that bad posture causes back pain, or that you should try to improve your posture while sitting at work to avoid lower back pain. New research from the British Chiropractic Association (BCA) has revealed that our posture while at work has been highlighted as the biggest culprit. So you’ll be pleased to hear that there are some movements that you can take to maintain a good posture when you’re at work. Even if you’re not experiencing problems now, improving your posture is something that you should consider to prevent issues from arising in the future.
When you are sitting down for a prolonged period, what should you be doing? Place both of your feet firmly on the floor and sit back resting against your seat so that your feet do not dangle. Try and sit back in the chair with a neural spine and head. Offer your back some support with shoulders relaxed. If you head is tilted forward, the weight on the spine puts pressure on the spinal cords, causing back discomfort and neck pain also now referred to as ‘tech neck’.

Improve Your Mood with Good Posture

Sleeping, sitting, standing in a slumped posture is one of the first visual signs of a bad mood. You feel comfortable in that position and even when you are meditating you will attain the same position thinking it is correct. You’ll still be thinking negatively because of the slumped posture. We don’t want you to miss out on having a good posture and a happy future. Good posture has been linked to higher self-esteem, more social engagement and less anxiety. People overlook the importance of a good posture, slouching may possibly be the reason behind peoples afternoon headaches.. An overly-curved back, tilted head and hunched shoulders – affects your mood by reducing the amount of oxygen you’re breathing in. So, posture can have a knock-on effect on your mental health.

How to Improve Bad Posture

The best thing you can do is some daily posture exercises with your personal trainer. Core strength exercises reduce back pain. Your core muscles are essential to support your posture, so the more you can work them, the better your posture will be. Excessive watching or texting on mobile-phones for a long time which can lead to stress injuries and pain in the neck and lower-back severe shoulder pain.
These are some of the main benefits of a good posture – your body looks slimmer, smarter and aligned. A healthy body means a good posture and we don’t want you to miss out on this. The bottom line is that good body posture is considered very important.

smoothies

Healthy Breakfast Smoothies for Health and Weight Loss

Do you need more greens in your breakfast? I bet you can you squeeze some delicious shakes into your breakfast. I think you want a positive mentality, weight loss, and health boost in one drink too.
Green smoothies are healthier than fruit or vegetable juices. Juices are extracted from fruit or vegetables, the fibre portion of the plant is removed. Green smoothies contain a large amount of water, as well as the fibre of the fruit pulp, which helps boost your metabolism.
Your hunger will be satisfied at breakfast by consuming fruits and berries, juice and smoothie to satisfy your nutritional needs, cancer-fighting, blood pressure beating, bone strengthening, cardiovascular helping, brain boosting, skin smoothing, vision improving, inflammation reducing, energy increasing, and more.

A green smoothie recipe is a yummy serving of blended fruits and veg or a combination of both. They are an easy way to flush your body of toxins, get lots of healthy nutrients, and lose weight fast.


Why green smoothies?


Green smoothies are becoming incredibly popular despite the look and taste and may not exactly look like the most delicious meal. The reason for the rise in popularity is that they are great for weight loss and detox. Find your weight loss and detox goals are easier to achieve. Full of phytonutrients, protein, vitamins, antioxidants
The thought of granola and yogurt may not be appealing due to the fat and sugar content. After eating a healthy breakfast smoothie in the morning, I don’t feel heavy or bloated compared to an “old-fashioned” breakfast with eggs, meats, and slices of bread/croissants.
Green smoothies are often green or bright green and, you’ll love your simple green smoothies you’ll feel great and lose weight as well.

What do green smoothies usually contain? Basically anything healthy and green in colour will do:
Raw leafy greens
Young or mature spinach leaves, lettuce heads, mint leaves, Kale, chard (silver beet), Cabbage, Dandelion Greens, Carrot Tops
Green fruit and vegetables
Green apples, avocado, kiwi, Celery, Green peppers, Cucumber, juice/zest/flesh of lime
Green powders
Chlorella and Spirulina

Get More Green Nutrients


The nutrient filled the juice of greens are nourishment powerhouses, full of fibre from the edible slices of the plant, packed with vitamin B12 vitamins A, C, E, K, and folate; minerals like iron and calcium; carotenoids; antioxidants; omega-3s; and phytochemicals, and also insoluble fibre, which is like a miraculous sponge, helping us regularly eliminate pounds of toxins. This absorption of toxins from the liver is transported out of the body and gives your metabolism a great kick start.
When you eat fruits and vegetables in the raw state you get double the nutrients, because when you cook food it loses half of the vitamins and nutrients.
Green smoothies are homemade at low cost and you’re getting the freshest fruit and vegetables possible. Ooo Lala!
If you’re not a fan of drinking something “green”, try starting out with baby spinach in all your green smoothies. A spinach smoothie recipe has a very mild flavour and will naturally add vitamins and boost your metabolism. You won’t even know it’s in your smoothie, all you will taste is the fruit. Over time, try swapping half of the spinach for mint or Dandelion.

Enter the protein smoothie

Plan ahead to have a protein smoothie you whip up a protein smoothie in under 5 minutes. A protein smoothie can be made with just protein powder and water. As you age, you actually need to add more protein to your diet, getting 20 grams of body awakening protein to start your day.
Green healthy smoothies are easy to make. It’s pretty easy to start the smooth blending process with these directions:
Step 1 – add all ingredients to blender
Step 2 – blend until smooth
Step 3 – enjoy

split-squat

The Split-Squat

Leg exercises can build muscles using bodyweight, dumbbells or barbell. For an overall ripped, lean body it’s important to know what muscles they work and how placing your hand on a stationary object can assist with balance.
The split squat is frequently put into training programs to add strength and power to the lower body for sports. The split squat is a squat using a staggered stance, supporting one leg up on a bench or ball (lessening stability) usually as an additional exercise to gain more once the back squat has to be used and mastered. With the back foot on a bench, ~85% of the load is held on the front foot.
Splitting your squat into a one-sided movement is the equivalent to progressing from barbell bench press to dumbbell bench press. Squatting may be easier to perform but the split squat displays greater hamstrings, external oblique, and gluteus medius muscle activity but less quadriceps muscle activity than the back squat. Remarkably, it exhibits similar rectus abdominis and erector spinae muscle activity to the back squat. Make sure to keep your back leg stable, as to not fall forward or have your foot inadvertently slip off the bench pad or stability ball.
Keep torso upright during the split-squat and squatting deeply will target your stabilizer muscles and allow for further depth in the squat and it is crucial to remember that split squats can burn your stabilizers out. You must be wary of where and how you decide to perform them.
The split squat more hip-dominant than the back squat Flexible hip flexors are important. Although it is called a single-leg exercise, it still uses both legs. The further your front leg is extended out, the deeper you can sit into the squat, the greater hip extension in the movement.
Step to The Split Squat:
• Stand facing away from bench.
• Extend leg back and place top of foot on bench.
• Squat down by flexing knee and hip of front leg
• Rear knee almost contact with floor.
• Return to original standing position by extending hip and knee of forward leg

squat

The Squat Movement

The goal of the squat exercise is to develop lower-body strength to build muscle or lose weight. Is the squat one of the most popular, practical and established lower-body exercises worldwide in the gym today? It can be one of the most difficult exercises to do correctly, except if you actually know what you’re doing. Every day, as a personal trainer in Kensington, I see people doing squats and estimate that 90% of gym goers do them incorrectly.
The optimal squat technique is:
• a wide stance (shoulder width +) with natural foot placing,
• unrestricted movement of the knees,
• full-depth while the curve of the lumbar spine is sustained
• forward or upward gaze.
Personal Trainers should cue you to, “sit back into the squat,” as a way to teach proper squatting technique.
The squat movement accomplishes various tasks associated with activities of daily living, sitting on our office chairs, jumping up, reaching down or sliding into our cars from the side.

Aging Process Slower with a Healthy Lifestyle 1

Aging Process Slower with a Healthy Lifestyle

Exercise may be the best anti-ageing pill
You can slather on all the vitamin C and D serums and night creams, but a beautiful body and feeling younger stats can be obtained with a little change in your lifestyle. Summing it up, getting quality sleep, mindful eating and right exercise are critical elements to aging gracefully. If you are a health junky, you likely have an idea about the importance of exercise to keep your body healthy and preclude symptoms of aging.
How does exercise slacken the aging process?
There are numerous outward elements linked with aging, from obese to skinny figure and from fresh to wrinkly skin. Younger-looking skin is associated with the proteins (myokines) secreted by working muscles. Myokines is utilized by our body to stay young. Aging starts in your body at the age of 35 when you gain fat and lose body mass. Exercise keeps your metabolism high, weight on target and manages lean body mass. Exercise accentuates the muscles, which in reaction create a protein which movies around the blood-brain barrier and perks up the brain.
How does exercise affect inside?
When it is about preventing a specific disease, exercise plays a vital role – from strengthening bones to precluding osteoporosis to minimizing the risk of cardiovascular disease to diabetes, it’s difficult to notice a disorder that exercise can help with. Exercise improves the quality of sleep, baffles thousands of genes and squinches the white fat which amends the danger of heart disease and diabetes. Exercise also enhances circulation, efficiency and functionality of our heart and reduces the risk of heart disease. Plus, exercise provokes the hippocampus which means better memory which we all want till our old age.
Walk, resistance training, jumping and cardio are some useful exercises which are recommended as anti-aging.

wonder-women

Gal Gadot: Wonder Woman 1984

Introduction to The Wonder Women Movie


Wonder Woman 1984 (WW84) is an upcoming theatrical superhero film scheduled to be released by Warner Brothers Pictures next year.
In August, shooting on location in London: St. Andrew’s Place, Regent’s Park and the Royal College of Physicians. Production again came back to the UK in October, with filming on location at the Imperial War Museum in Duxford, and Hyde Park and the Savoy Hotel in Central London.
The movie is directed by Patty Jenkins and written by Jenkins, Geoff Johns, and David Callaham, from a story written by Johns and Jenkins.
It stars shining light, Gal Gadot, in the leading role. Former beauty pageant contestant and signed model, she is naturally slim and playing a warrior woman meant she had to put on muscle to satisfy us with a superhero look. A strong, powerful body, independent, talented, and even relatable woman in a prominent role, effortlessly battling it out on screen.

Gal Gadot: Wonder Woman 1984 2

That’s our kind of gal!

Gal Gadot’s Training: WW84 Movie


Gal Gadot and the cast of Amazonians all had tailored fitness plans personally suited their needs by a personal trainer.
The healthy heroines trained for six months to play Wonder Woman in Arguably the fiercest display of strong females. The routine included an hour of strength and conditioning both in mind set and muscles, topped off with two hours of stunt training. Gadot team need to be fit, agile and powerful for the stunts. With an amazing feat of sword-fighting, that uses core strength, flexibility and agility.

The 45-minute full-body wonder women inspired workout made from simple exercises taken from some of the toughest disciplines on the planet!

kickboxing-london

Exercise 1: Warm Up Rowing Machine – low-impact
a. 5 min low intensity for upper body warm up and pulse raiser.
b. Intervals – 30-second intense “sprint- row,” and a 60-second easy pace.

Exercise 2: Animal Inspired Movements.
a. Bear Crawl 40-metres.
b. Crab Walks 25 metres.


Exercise 3: Army Inspired Exercises
a. Burpee Broad Jumps
b. Pull Ups

Exercise 4: Olympic Gymnastics Inspires Exercise
a. Ring Push Ups.
b. Hanging leg raises

weight-loss-love

Weight Loss Ideas: Surprise your Lover this Valentine’s Day!

Weight loss ideas surprise your love on this Valentine’s Day!
Are you worried about the upcoming Valentine’s Day? Don’t want to look shapeless or obese in front of your prince charming? Keep reading the article for healthy weight loss ideas to surprise your love!
Although there are endless supplements, diet tips and meal replacement strategies affirming rapid weight loss they still lack scientific pieces of evidence. However, there are some schemes supported by science that can strike on weight management. These schemes involve a careful observing of calorie intake, exercise, minimizing the number of carbohydrates in your diet and intermittent fasting. I have listed here three incredible methods for weight loss.

  1. Intermittent Fasting
    Intermittent fasting is an irregular pattern of eating which includes short-term fasts and intake of a meal within a smaller period. It has been pointed in several studies that a 24 weeks intermittent fasting can tether weight loss in overweight people. The most effective intermittent fasting involves these methods;
    • Fast every second day and prefer eating a veg meal on the non-fasting day.
    • Fast on two days per week and only eat 500-600 calories on a fasting day.
    • Fast for 16 hours and eat healthy during an 8-hour window.
  2. Track your diet and exercise
    If you are interested in losing obesity, you must be aware of everything you drink or eat every day. The best way to do it is to write everything you consume either in an online food tracker or in a journal. More than 3.7 billion health apps were launched in 2018 to track diet, weight loss and physical activity and using an app to check weight loss progress or following physical activity can be useful in managing weight.
  3. Eat carefully
    Eating in a careful, deliberate manner is a healthy practice to observe how and where you have eaten food. This practice enables people to cherish the food they consume and manage a healthy weight. Every one of us is living a busy life, we, most of the time; tend to eat fast being barely aware of the food intake. Strategies for mindful eating are;
    a. Pay attention to your meal and enjoy every bite.
    b. Don’t get distracted by TV, phone or laptop while you’re eating.
    c. Properly chew and savor the food. This strategy helps your brain recognize the signal that the food intake is enough for now.
    d. Choose nurturing foods that can satisfy your hunger for hours.
    Bottom line
    It is critical to keep in mind that when you are struggling to reduce weight, there are no quick fixes. The ideal way to reach a healthy weight is, eat a healthy, balanced and nourishing diet including whole grains, protein-rich food, fruits and vegetables. Also, Do exercise regularly for 30-minutes at least.
refresh-eyes

3 of the Safest Ways to Refresh Your Eyes

In a normal day, you face long deskbound hours staring at your computer monitor. There are daily commutes and other stressful chores causing your eyes to scream for a relaxing bath. Many times we need a break to relax. In summer the side-effects of heat include body dehydration, dry eyes, and worse skin. Moreover, people who wear contact lens don’t find it a very cheery season as the eye’s natural lubricant dries because of the climate. The outcome can be itchy, dry eyes just because they are sunbaked.
Similarly, some people experience heavy eyelids in the morning just after waking up. At initial stages, you may experience low productivity, physical tiredness, and more work errors and eye strains. Later, the eye strain can cause red eyes, eye twitching, blurry vision, and headache. Here are top 3 ways to soothe your strained, dried and tired eyes.

  1. Wash your eyes with cold water
    Begin with sprinkling cold water on your face; it results in constriction of facial arteries and low blood flow to the face. This decrease in blood flow activates the nervous system reflex to set off and fight against this condition. Reduced blood flow to the eyes decreases eye inflammation, reduces dryness, natural tears are made by increasing eye closure and spreading tear film.
    (*splash the water three times after testing the temperature of water. It must not be freezing.)
  2. Try deluging your face in cold water and follow with a warming massage
    Deepen your cold water wake-up by submerging your face in a tub or bowl of cold water for 30 seconds. Take out your face of the water when you need to breathe. If you feel pain or any other symptom appears – consult your doctor.
  3. Use lubricating eye drops
    There are numerous eye drop varieties which aid in soothing eye strain. Lubricating eye drops soothe tired eyes. Natural tear element is also added by these drops to hydrate eyes. Before using eye drops read instructions on the packaging. Note: If you feel chronic pain, go for an eye check-up to your eye doctor for a correct diagnosis of your eye problem.
    You can also do simple exercises to refresh your eyes and have a break from the monotony of job so that you can get back to it with an energy boost!
lung-cancer-hpv

Human Papilloma Virus in Lung Cancer Patients

Question: What steps would you need to take to develop an immunohistochemically method to identify Human Papilloma Virus (HPV) in lung cancer tissue removed from a patient? 


ABSTRACT
Human Papilloma Virus (HPV) is a virus which infects the skin, and particularly moist membranes lining the body. Evidence that HPV may be correlated with the development of lung squamous cell carcinoma came from Syrjane et al 1972, as a result of examining histological samples of oesophageal and cervical cancers, and finding a similarity between the examinations.
Immunohistochemistry (IHC) methods are discussed in this paper, to identify HPV in a lung cancer tissue removed from a patient. The tissue is subjected to multiple processing stages, to ensure the cell morphology, tissue architecture as well as the antigenicity of the HPV epiptopes are maintained and retrieved. Monoclonal mouse primary antibody is used to target the HPV antigen and the signal is amplified via the Labelled Streptavidin Biotin (LSAB) detection system, for visualisation under bright field microscopy. IHC has revolutionised the field of clinical diagnostics as well as medical research, due to being a highly sensitive and specific method, capable of detecting a variety of antigens.

INTRODUCTION
It is estimated that the Human Papilloma Virus (HPV) is directly related to 15-20% of all human cancers (Giuliani, Favalli et al. 2007). With the virus consistently proven to be, not just related to, but the leading cause of carcinomas within the uterus, particularly cervical cancer, where it displays a causality rate approaching 100% (Nour 2009). In other vulnerable areas of the body, more recent scientific research has begun to establish that HPV is implicated in the formation of squamous cell carcinomas of the lungs (Giuliani, Favalli et al. 2007). The first evidence of an association between HPV and lung sqamous cell carcinoma was demonstrated by Syrjane in the early 1980’s where he reported a similarity between the histological characteristics of invasive oesophageal SCC and cervical cancer, suggesting the possibility of HPV aetiology (Syrjanen, Syrjanen et al. 1982).
Further studies have shown the virus capable of interfering with the cell cycle and interrupting tumour suppressive functions, resulting in cell oncogenesis. Oncoproteins E6 and E7, have been strongly linked to interference with the tumour suppressive, p53 and Rb proteins (Prabhu, Jayalekshmi et al. 2012).
With mounting research establishing HPV to be an exceptionally versatile biomarker, and increasing incidences of HPV association with lung cancers, it has become a growing necessity to develop a test for routine HPV virus within lung cancer cells (de Freitas, Gurgel et al. 2016). To develop and implement such a standardised method, a technique that is both highly accurate and cost effective, while also being easily transferable to existing pathology laboratories is required.
Immunohistochemistry (IHC) has become a recognised and adaptable methodology for investigation and diagnostic purposes, utilising antigen-antibody reactions within specific cells or tissues. Once the antigen-antibody binding occurs, the immunohistochemical reaction is demonstrated by the production of a coloured product, which is visible under bright field microscopy (Montero 2003).
This helps to create a technique highly valuable in diagnostic situations, specifically where clinical data and observed morphology under a microscope have proved insufficient to produce a clinical diagnosis (Duraiyan, Govindarajan et al. 2012). Therefore, due to its proven success at identification, through the localisation of tissue antigens, IHC must be considered a highly useful diagnostic tool, when used alongside scientific research. There is no doubt that it would form a solid base for developing a new method to identify HPV within lung cancer tissue.

METHOD

Tissue Processing
Sample Preparation
Initially, the fresh lung carcinoma tissue will be fixed on the tissue slide by applying a fixative such as 10% neutral buffered formalin (Werner, Chott et al. 2000). Next the tissue would need to be embedded in medium such as paraffin, and sections of 4-5um in thickness are cut on a microtome instrument (Grizzle 2009). Fixation is essential for tissue and antigen preservation. The tissue sections must then be mounted on charged slides and dried overnight. This ensures that the section of the tissue with adhere to the slide (Chen, Cho et al. 2010)

Deparaffinization and Rehydration of Tissue Section
To perform antibody staining, paraffin wax must be removed from the tissue sample and the sample must be rehydrated.
• To remove the paraffin wax, immerse the sections in a clearing agent such as xylene for a period of 5 minutes (Williams, Mepham et al. 1997).
• Xylene must be removed by graded washes of xylene and alcohol and the sections are rehydrated, via repeated washes of ethanol with water. For example, subject the tissue sections in 100% ethanol for 10 minutes, and then place them in 95% alcohol for another 10 minutes. In order to complete the rehydration process was the slides in distilled water (dH2O) for a period of 5 minutes (Paulsen, Dimke et al. 2015).
Antigen Retrieval
The next step is to be able to retrieve the antigen epiptopes which have been masked by cross-linking of “methyl bridges” between the proteins in the tissue, due to the fixative, formaldehyde. (Leong, Leong 2007). This can lead to a greater improvement in the detection of antibody staining.
The most efficient epiptope retrieval method, which can be used to retrieve the HPV antigen, is known as the Heat Induced Epiptope Retrieval (HIER) method. This method utilises the heat, which can come from a variety of sources including pressure cooker, autoclaves, or microwaves to retrieve the antigens. Figure 1 demonstrates the process of how antigen retrieval can be performed by using the heat mediated method.
• For example the tissue sections can be subjected to a heat-mediated solution such Citrate buffer in a high heat environment such as in a microwaveable vessel for 20 minutes (Shi, Shi et al. 2011), (Yemelyanova, Gravitt et al. 2013).
Blocking of non-specific background staining
Endogenous peroxidase activity is found naturally occurring in tissues and cells, which can react with the HRP conjugated antibody (Buchwalow, Samoilova et al. 2011). This can result in high non-specific background staining when view under the microscope. Therefore, the tissue must be treated with a blocking agent such as Hydrogen Peroxidase (H2O2), which significantly reduces non-specidinc background staining, resulting to a less likelihood of a false positive outcome (Radulescu, Boenisch 2007).

Amplify the Signal
The staining process that will be discussed in this method is the Labelled Streptavidin Biotin (LSAB) method. The first step would be to inoculate the tissue sections with a primary monoclonal antibody which is specific to the HPV antigen epiptope, such as mouse anti-HPV, followed by the addition of a secondary HRP conjugated Ab, i.e horse anti-HPV (Cardiff, Miller et al. 2014)

An example of such a method is demonstrated below:
• The slides are inoculated serially diluted Anti-HPV, which is a monoclonal mouse anti-HPV protein. Then incubate the slides for 60 minutes at 37oC.
• The next step would be to apply serially diluted horse biotinylated anti-mouse IgG linked Ab to the slides, ensuring that the tissue slice is coated thoroughly. Again the sections must be incubated at room temperature for 30 minutes (Sano, Oyama et al. 1998).

Detection of bound secondary antibodies
In order to detect the secondary Biotinylated anti-mouse IgG, an enzyme conjugated streptavidin can be added to the slides.
• For example apply diluted Streptavidin-Horseradish Peroxidase (HRP) antibody conjugate to the tissue within the wax sections, and then incubate for at least 20 minutes to ensure that all biotin-binding sites on the enzyme conjugated complex are filled (Hofman 2002).

Staining and detection
For HRP-conjugated antibodies, the substrate known as, 3,3’-diaminobenzidine (DAB), can be used, which yields a brown insoluble product.
• For instance a few drops of DAB can be added to the slides and incubated for a period 5 minutes.
• Counter-staining must then is performed using Haematoxylin Mayer, which is a nuclear contrast stain, capable of specifically staining the nuclear chromatin without staining the cytoplasmic organelles, allowing the pathologist to identify to morphology of the cells. The standard factor for a positive reaction is dark brown nucleus of the HPV (Fischer, Jacobson et al. 2008).

Controls
The validity of the IHC results interpretations relies upon appropriate usage of positive and negative controls. Positive controls demonstrate that the antibodies and reagents are functioning properly, whereas negative controls are incorporated in the procedure, to demonstrate that the reaction observed is due to the antigen-antibody complex and not to other exogenous factors, ensuring that results are a true positive (Hewitt, Baskin et al. 2014).

• The positive control would consist of a lung cancer tissue slide treated with the primary mouse anti-HPV, to show specificity of the Ab to the HPV detection. Another tissue slide would be treated with the secondary horse anti-HPV, to demonstrate that the label is specific to the primary mouse anti-HPV.
• The negative control would be a lung cancer tissue slide not incubated with the primary mouse anti-HPV.
• It’s also essential to incorporate a labelling control during the IHC procedure, in order to show that the labelling of the result slides is due to the label added and not to other factors such as endogenous labelling or reaction to other chemical products. A sample of a tissue slide which has undergone all the preparations in detergents and buffers, but has not been exposed to the antibodies, enzyme or the dye would be appropriate as the label control (Burry 2011).

DISCUSSION

An immunohistochemically method was discussed, with the purpose of being able to detect HPV in tissue of a lung cancer patient. The different IHC stages were discussed, including the tissue processing steps, antigen-antibody detection processes, and the staining technique.
Whilst immunohistochemistry is considered a routine procedure, several interdependent factors can affect the outcome result of the immunostaining. It is strongly affected by formalin based fixation, as well as the steps that follow tissue processing in addition to antigen retrieval. Furthermore, the quality as well as specificity of the primary antibody is essential to the final result, as well as the technique of the amplification of the secondary antibody (Otali, Stockard et al. 2009).
Formalin is the gold standard for of fixative used in routine histology, and was the fixative used in the method above. It yields good morphological details of the cells, as well as a consistency in staining. Additionally, it’s highly effective at preventing growth of microorganisms, and being able to stabilise and preserve the tissue for decades (Grizzle 2009).
Antigen retrieval, performed by the direct heat method used in the method above has revolutionised the detection an antigens in tissue which have been subjected to fixatives such as formalin. It can unmask epiptopes of antigens by hydrolysis of methylene cross-links. However a disadvantage of this method is that it frequently results in the damage or even loss of tissue (Shi, Shi et al. 2011).

The antigen staining detection method discussed above was the indirect method, known as labelled streptavidin-biotin method (LSAB). This compromises of three steps, whereby the unlabelled primary antibody against the target antigen is bound with biotinylated secondary antibody. Then this is followed by the addition of biotin-avidin complex, with the HRP enzyme, bound via the biotin. This in turn produces the substrate product DAB, which is a brown colour precipitation (Hofman 2002). Below is a diagram representation of the LSAB

The advantages of the using this method is that it’s more sensitive than the direct method, which compromises of only the primary antibody and the detective marker. This is because in the LSAB method the detection signal is amplified, due to increased number of binding molecules of the secondary antibody to the primary antibody (Cardiff, Miller et al. 2014).

Peroxidase enzyme, is the preferred staining marker among the immunohistochemistry field. Its unique properties with regards to its stability as well as effective staining intensity related to the DAB substrate make it a standard choice for immunostaining (Bussolati, Radulescu 2011). The expectant result of the lung carcinoma tissue would be a brown nuclear staining of the HPV antigen, as demonstrated in figure 3 below.

Fig 3. An illustration of the HPV immunostain positive, in lung cancer tissue (100x magnification) (van Boerdonk, Daniels et al. 2013)

Overall immunohistochemistry biomarkers, such as the HPV antigen can used to assist in the early detection of lung carcinoma, as well as in diagnosis. It can also prove valuable for prognosis of lung carcinoma. IHC has proven as a highly valuable tool for both routine diagnostics as well as in the field of scientific research, and can be a highly effective method to help identify HPV in lung cancer

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References
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BURRY, R.W., 2011. Controls for immunocytochemistry: an update. The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 59(1), pp. 6-12.
BUSSOLATI, G. and RADULESCU, R.T., 2011. Blocking endogenous peroxidases in immunohistochemistry: a mandatory, yet also subtle measure. Applied immunohistochemistry & molecular morphology : AIMM, 19(5), pp. 484.
CARDIFF, R.D., MILLER, C.H. and MUNN, R.J., 2014. Manual immunohistochemistry staining of mouse tissues using the avidin-biotin complex (ABC) technique. Cold Spring Harbor protocols, 2014(6), pp. 659-662.
CHEN, X., CHO, D.B. and YANG, P.C., 2010. Double staining immunohistochemistry. North American journal of medical sciences, 2(5), pp. 241-245.
DE FREITAS, A.C., GURGEL, A.P., DE LIMA, E.G., DE FRANCA SAO MARCOS, B. and DO AMARAL, C.M., 2016. Human papillomavirus and lung cancinogenesis: an overview. Journal of cancer research and clinical oncology, 142(12), pp. 2415-2427.
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FISCHER, A.H., JACOBSON, K.A., ROSE, J. and ZELLER, R., 2008. Hematoxylin and eosin staining of tissue and cell sections. CSH protocols, 2008, pp. pdb.prot4986.
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stress

Health and Stress

Introduction: Health and Stress


It was interesting to learn was how stress can have a negative impact of the body, and how it evolved because of flight-fight response. Our bodies are intended to respond to our condition with an end goal to safeguard homeostasis. However it was new to me to learn that the Autonomic Nervous System (ANS) and the Hypothalamic-Pituitary-Adrenal (HPA) axis play a major role in responding to stress. Chronic stress can have detrimental effects on the body including decreased immune functions, atherosclerosis as well as bone demineralisation etc. We also learn that due to trauma such as cell damage or infection a cascade of processes take place which result to inflammation. Inflammation is essential because it is responsible to eliminate the tissue damage or pathogenic microbes, which attracts immune cells into cells and which results to granulation, and leads to tissue repair.

Question: How does chronic stress lead to compromised health?


Chronic stress is maladaptive and detrimental to organism’s health, if it is repeatedly activated. Chronic stimulation of the Sympathetic Nervous System (SNS) due to chronic stress, results to a stimulation of cardiovascular system. This leads to an increased blood pressure and vascular hypertrophy. The muscles responsible for constricting the vasculature thicken, resulting to elevated rested blood pressure. Chronic high blood pressure consequently forces the heart to function harder, and results to hypertrophy of the left ventricle. The end result leads to damaged arteries and plaque formation [1]. Chronic stress can also result to a significantly elevated stress hormone, such as cortisol and glucacartinoids which suppress immunity and particularly affect the distribution of cytokine presence. Cytokines are communication molecules, which mainly produced by immune cells. T helper 1 (Th1) and T helper 2 (Th2) which are involved in cellular and humoral immunity respectively become primarily dysregulated during exposure to chronic stress. This results to slower wound healing, and slower recovery from surgery. In addition chronic stress can be particularly detrimental to the elderly. There is a steady loss of immune function due to ageing, and when this is coupled with chronic stress it can lead to elderly being less able to produce antibodies to vaccinations, and their immune system in less effective at responding to a viral or bacterial infection [2].

Articles
• Inflammatory response and chronic disease
MURAKAMI M, & HIRANO T. (2012). The molecular mechanisms of chronic inflammation development. Frontiers in Immunology. 3.

This article reviews the inflammatory the mechanisms and pathogenesis associated with inflammation. Inflammation acts as a host defence against infectious agents as well as injury, however it can result to pathophysiology of many chronic diseases. Interactions between cells of the innate and adaptive immune system, in addition to inflammatory mediators can contribute to acute and chronic inflammation. Common effectors mechanisms of the inflammatory system, such as the complement system and phagocytosis result to tissue injury, angiogenesis, oxidative stress and fibrosis of tissue. The article mentions atherosclerosis as an example of a disease which is mediated by various inflammatory mediators involving both the innate and adaptive immune system. Overall I believe this article makes a significant impact in the field of cellular pathology, as a greater understanding and knowledge of the inflammatory response will help scientists devise new strategies to predict disease susceptibility. Also this base of knowledge could aid in the development of new approaches to prevent and treat chronic diseases such as atherosclerosis [3].

• Pathophysiology of the inflammatory response
BRØCHNER, A. C., & TOFT, P. (2009). Pathophysiology of the systemic inflammatory response after major accidental trauma. Scandinavian Journal of Trauma, Resuscitation and Emergency Medicine. 17, 1-10.
The article explores the pathophysiology of the inflammatory response to trauma. Local inflammatory response happens due to trauma of tissue. Local mediators, such as kinis and arachidonic acid metabolites, as well as histamine released from mast cells are all activated during the response. They have a short half life, however their effects are long lasting. Following trauma, the complement system is also activated. The article explains that the system is activated by three pathways, including the antigen-antibody complex, bacterial cell wall components as well as mannan binding lectin pathway. The activation results to components of the cascade system that are able to oponise pathogens, lyse bacteria attract neutophils as well as attract platelets at the site of trauma [4]. I believe that overall this article produces an excellent literature with regards to how the inflammatory response reacts to trauma. It explains well the complex mechanisms involved during the inflammatory response, and I therefore believe that it makes a significant contribution to the clinical field of the inflammation response.

• Mechanisms of Phagocytosis
ADEREM A. (2003). Phagocytosis and the inflammatory response. The Journal of Infectious Diseases. 187, 340-5.
The article explores the mechanism pathways involved in phagocytosis. Phagocytosis is the first line of defence step in the triggering of the host defence, in addition to inflammation. It’s essential for the removal of vast amounts of senescent cells that that are no longer necessary, as well in tissue remodelling and embryonic development. Phagocytosis activated by Pathogen-Associated Molecular Patterns (PAMPS), which results to the activation NF-kB, which is responsible for the control of DNA, cytokine production and overall survival of the cell. In addition oposonins such as antibodies and C3b can aid phagocytosis by acting as attachment positions. The article further explains that engulfment of foreign bodies is supported by the actin-myosin contractile system [5].
Overall, I believe that this article has made a vital contribution in the understanding on phagocytosis, given its crucial role in the regulation of the immune response.

References

  1. HENRY JP, STEPHENS PM, & SANTISTEBAN GA. (1975). A model of psychosocial hypertension showing reversibility and progression of cardiovascular complications. Circulation Research. 36, 156-64.
  2. SCHNEIDERMAN, N., IRONSON, G. AND SIEGEL, S. D. (2005). Stress and Health: Psychological, Behavioral, and Biological Determinants. Annual Review of Clinical Psychology, 1(1), pp.607-628.
  3. MURAKAMI M, & HIRANO T. (2012). The molecular mechanisms of chronic inflammation development. Frontiers in Immunology. 3.
  4. BRØCHNER, A. C., & TOFT, P. (2009). Pathophysiology of the systemic inflammatory response after major accidental trauma. Scandinavian Journal of Trauma, Resuscitation and Emergency Medicine. 17, 1-10.
  5. ADEREM A. (2003). Phagocytosis and the inflammatory response. The Journal of Infectious Diseases. 187, 340-5.