Treatment for influenza

Because it takes a lot of eggs to make the influenza vaccine each year, I wonder if this is a feasible solution for the epidemic:
At the beginning of any flu season, identify the first individual who recovers from that year’s virus infection, take blood sample and isolate individual B-cells into several sets of 96-well plates. Extract RNA from these cells and prepare recombinant antibodies and screen the ones that react with the virus. Once identified, we will have an unlimited supply of “monoclonal” antibodies that can be used for passive transfer to highly susceptible individuals. While the idea may be labor intensive, there are probably companies that are already spending huge sums of money to address the possible epidemic.
Along these lines, I saw a recent paper in which they obtained human monoclonals from a vaccinated individual.
http://www.ncbi.nlm.nih.gov/pubmed/20096915

DO CRP AND FIBRONGEN HAVE A DIRECT RELATIONSHIP WITH BODY FAT MASS??

In the paper reviewed in class called, “The relation of body fat mass and distribution to markers of chronic inflammation” the authors do an excellent job explaining the relationship between CRP and fibrinogen to body fat mass and see what effects insulin resistance produced on this relationship. Since most of us have background in science we know that CRP (C-reactive protein) produced by the liver and released by adipocytes plays a critical role in chronic inflammation. Also, we know that fibrinogen which is also synthesized by the liver is a glycoprotein that is converted by thrombin into fibrin during the blood coagulation.

The study included over 1500 participants and an array different variable such as ethnicity, gender, BMI, height, etc were used to measure their relationship to body mass. The results indeed showed that there was a correlation between the two and specified how these relationships were split up according to gender. Overall the results showed that women had higher levels of CRP and fibrinogen along with increased BMI, adipose body mass; whereas men had increased in height, weight, and waist-hip ratio. In conclusion, there was a relationship between increased levels of CRP and fibrinogen and body fat mass.

Festa, A., R. D’Agostino, K. Williams, AJ Karter, EJ Mayer-Davis, RP Tracy, and SM Haffner. "The Relation of Body Fat Mass and Distribution to Markers of Chronic Inflammation." International Journal of Obesity (2001): 1407-415. Print.

NKTs? (Natural Killer T cells)

Here's a practical question. I want to clone out human CD8+ T cells, starting with frozen PBMCs. Before I stimulated them with CMV peptide (and an artificial APC), I used magnetic beads to produce a somewhat more purified population. I used anti-CD56 beads to remove NK cells, and anti-CD19 beads to remove B cells. After a few rounds of stimulation/proliferation followed by rest, I sorted only the CD8+ cells also expressing early activation markers (CD69 and CD137). I then cultured these through several more cycles of stimulation and rest.

I then stained the cells with certain markers I was interested in, and performed flow cytometry. I found 14% of the population of one of the cultures to be CD8+, CD4+, CD3+ triple positive cells. Unfortunately, I did not stain with anti-CD56. My guess is that these peripheral double positive T cells are NKT cells. Does anyone have any ideas?

Macrophages of Subcutaneous Fat and Visceral Fat: Amount vs Proximity?

We hear about the additional dangers that come with visceral fat since it surrounds our internal organs. However, does this make subcutaneous fat less dangerous? In an article about adipose tissue macrophages, Wentworth et al show that the macrophage profile differs between subcutaneous (under the skin) and omental (a part of the visceral tissue) tissue in samples taken from women. In subcutaneous fat, they showed a more dense population of adipose tissue macrophages with CD11c+ than in visceral fat. These macrophages have pro-inflammatory entities, one example is a protein related to reactive oxygen species release (CD11c+). Reactive oxygen species (ROS), which are related to stress, can lead to further inflammation and immune response. In regards to obesity, the group of scientists suggested that subcutaneous tissue may play a greater role in inflammation and insulin resistance than omental tissue. The inflammation may result from CD11c+ macrophages containing increased mitochondria concentration and mitochondria activity as well as fat concentration, entities that would increase ROS production.

The interesting article caught my eye in its focus on subcutaneous tissue over omental tissue regarding obesity and inflammation. There is no doubt that visceral fat should be kept in check. However, the comparative amount of subcutaneous tissue usually out weights the amount of visceral fat in any one person. The pure amount of pro-inflammatory CD11c⁺ cells could be the reason why they have such an effect on insulin resistance. More specifically, it could play a part in how individuals continue to experience states of chronic inflammation. Sine this data was taken from female samples, it would be interesting to test the article’s hypotheses in men, children and across cultures to see if the results are consistent.

Looking at obesity through an inflammation perspective brings up the fascinating topic of adipocytes functioning as an immune organ. Exploring adipocytes and adipose tissue macrophages the past two weeks has shown the possibility of fat specific immune cells leading to inflammation. Especially through this article, it provides a reason to care about subcutaneous fat. Even though the imminent threat of visceral tissue is present, the mere amount of subcutaneous fat can also pose a serious problem to an individual’s health.

Wentworth, John, et al. Pro-Inflammatory CD11c⁺ CD206^+‑ Adipose tissue Macrophages Are associated with Insulin resistance in Human Obesity. Diabetes 2010; 59:1648-1656.

Type 2 Diabetes, Fatty Acids, and Genetics:

In the brief report by Pisabarro et. al, the authors examine a potential connection between a diet high in trans and saturated fatty acids and individuals with the Ala allele for peroxisome proliferator-activated receptor (PPAR) γ2 and incidience of type 2 diabetes. PPAR functions as a transcription factor and plays a role in the regulation of cell differentiation, development, and metabolism, and the Ala allele is a common polymorphism. Aside from the fact that the authors never suggest a mechanism for why carriers of the Ala allele are more likely to develop type 2 diabetes as a result of a diet high in trans and saturated fatty acids, their statistics to argue that a connection exists are even weaker.

According to their results, only 50% of the individuals found to have this polymorphism had impaired glucose metabolism, and while that may sound encouraging, this equates to 6 individuals. Furthermore, the data that suggests that type 2 diabetes occurs at a younger age, lower BMI, lower insulin levels, and lower waist circumference was not found to be significant. In general, it makes sense that a diet high in trans and saturated fatty acids could be suspected to be correlated to incidence of type 2 diabetes, yet the results presented in this paper and the lack of a mechanism raise significant questions about the validity of the authors’ claims. I am curious as to what other work has been done in examining a connection between diets high in trans and/or saturated fatty acids, genetic polymorphisms, and incidence of type 2 diabetes, but as of this point, I remain skeptical of a direction relationship between the Ala allele and such diets based on this evidence (or lack thereof) alone.

Incubation time

Once a human is infected by a pathogen, the dendritic cells present the antigen to the T cells and the whole cascade results in the adaptive immune response. Why is it that in some virus infections the "incubation time" is longer than others before the clinical manifestation is seen?

Type 2 Diabetes and Breastfeeding

An article in the LA Times discussed a study where women who breastfed their infants were less likely to develop Type 2 diabetes. Of the women who did not breastfeed, 27% developed diabetes, compared to the 16% of women who did breastfeed. Two theories behind why breastfeeding reduces the risk of diabetes are that breastfeeding helps women lose weight, and lactation may help with glucose metabolism.

Interestingly, I found a couple articles claiming that breastfeeding reduces an infant’s chances of developing Type 2 diabetes later in life, and that non-diabetic people who were breastfed as infants have marginally lower insulin concentrations than non-diabetics that were formula-fed. There are many chemicals in breast milk that make it superior to formula, and breast milk has been shown to have some anti-inflammatory effects, namely due to the presence of interleukins. However it is still unknown if this or other factors are what protect against diabetes.

Other things to consider are mothers of lower socio-economic status or obese mothers are more likely to formula feed, and thus their children have an increased risk for diabetes and obesity due to environmental factors besides being formula-fed.

There are different possible reasons why breastfeeding may decrease the risk of Type 2 diabetes (and I suspect the real reason is still unknown), but it is interesting that it seems to have the same health benefit for both the mothers and infants.

Shari Roan. "Study: Women who breastfeed even one month reduce their diabetes risk." Los Angeles Times, Aug 31 2010

Undurti N Das. "Breastfeeding prevents type 2 diabetes mellitus: but, how and why?" Am. J. Clinical Nutrition, May 2007; 85: 1436-1437

Christopher G Owen. "Does breastfeeding influence risk of type 2 diabetes in later life? A quantitative analysis of published evidence." Am. J. Clinical Nutrition, Nov 2006; 84: 1043-1054

ACE Inhibitors and NSAIDS: More Pieces to the Diabetes Puzzle?

Type 2 diabetes, commonly referred to as adult onset, is becoming a larger and larger public health issue. It is especially important because people are beginning to experiencing the disease at younger ages. Left untreated, type 2 diabetes can lead to stroke and cause damage to the kidneys and retina of the eye. Identifying early signs of insulin resistance, may help identify potential type 2 diabetics. As cited in “Inflammation and the etiology of type 2 diabetes,” almost 90% of type 2 diabetes patients show signs of reduced insulin sensitivity before type 2 diabetes symptoms became apparent (Sjöholm 2005)

As discussed in the previous blogs, studies have indicated that a low level chronic inflammation (inflammation lasting for long periods of time) may be a factor in type 2 diabetes. In a review article cited above, Sjöholm and Nyström indicate that when compared to a placebo group, ACE inhibitors and angiotensin receptor antagonists which affect the renin-angiogensin system showed lower proportions of newly diagnosed diabetes patients. Another interesting drug they note is the possibility of NSAIDs (i.e. aspirin) to improve glucose tolerance and the affect of insulin. When compared, ACE inhibitors, angiotensin receptor antagonists and the NSAIDs are vasoactive substances. They work to improve blood flow. Interestingly enough, increased blood flow and delivery of nutrients to active tissue reduces stress and reduces inflammation. This data provides us with a potential direction to further explore the relationship between chronic inflammation and type 2 diabetes. The scientists of the review hope to identify potential drug targets and a more complete biological profile can help treat and identify pre-diabetic individuals.

Although pharmacological advances will play a decent role in reducing the rate of diabetes, it is imperative that physicians stress the importance of physical activity and nutritious diet as a natural preventative measure. It is especially important at this time since treatment/management of type 2 is hardly straightforward due to the difference among patients. Moving people away from the “quick fix” attitude, where they can make bad decisions and “start over,” will hopefully cause individuals to realize the importance of exhibiting and maintaining a health life style. Learning this knowledge and passing it down generations by example to children and the community is where I believe we can make the greatest leap in battling type 2 diabetes.

Sjöholm, Åke and Thomas Nyström. Inflammation and the etiology of type 2 diabetes. Eiabetes/Metabolism Research and Reviews 2005; 22: 4-10.

LAP-BAND surgery: YEA or NEA?

According to the recent article discussed in class, “Surgery may cure diabetes in overweight” first time readers may be given a false first impression that the article actually has found a “cure” for diabetes. It isn’t only until you read the entire article that you notice something misleading and that no cure was found just an expensive quick alternative. Let me first begin my mentioning that the study was funded by Allergan Health and was provided laparoscopic adjustable gastric bands without charge from the manufactures. Therefore a bias is noted from the beginning in favor of the $20,000 LAP-BAND surgery (coincidence??). The study presented in the article involved 55 randomized patients that were obese and have had a history of type 2 diabetes. These patients were divided into two different groups; first group focused on treating the patients with standard diabetic care and the second group focused on treating with the LAP-BAND surgery. The results presented in this article showed that 22 out of the 29 patients in the LAP-BAND surgery group showed diabetes remission while 4 out of 26 patients with the standard diabetic care showed diabetes remission. With such staggering results one might overestimate the accuracy of this study, at least at first glance. Even though not explained in this article, the study published in the journal of the American Medical Association mentioned that the definition of diabetes remission used was a fasting plasma glucose level of less than 126mg/dL. Therefore not even an oral glucose tolerance was necessary to determine diabetes remission. Not only was this study limited because of size, it is also limited because the results did not show an accurate way to determine whether diabetic remission was achieved.

My personal opinion about the LAP-BAND surgery according to this article is NEA! I think that there is more to “curing” diabetes than just a quick fix surgery. Patients have to make lifestyle changes. They have to change their sedentary lifestyle and start involving diet and exercise into their daily activities. Yes, the surgery will provide temporary weight loss, but is it the pounds that matters or the eventual remission of diabetes? Overall, there are more factors than obesity that influence the onset and progression of type 2 diabetes and LAP-BAND surgery doesn’t show a permanent cure for the disease.

This was a lay article obtained from azcentral.com.

Johnson, Carla. "Surgery May Cure Diabetes in Overweight." Azcentral.com. Web. .

Main Culprit in Obesity and Related Diseases: Inflammation!

Obesity is obviously a growing concern within the United States. With the lack of desire to lead an active lifestyle, to the convenience of fast-food, Americans are adding inches to to their waistlines while increasing their risks for obtaining different obesity-related diseases such as hypertension and diabetes. What many Americans don't know is the role that inflammation plays in their weight gain, and in turn how this process can lead to other obesity comorbidities.

Within Robert W. O'Rourke's article, "Inflammation in Obesity-related Diseases", the biological process of inflammation within the body, particularly in adipose tissue, is explained in detail when related to obesity. Obesity is associated with a low-grade, systemic inflammation, and it is this biological response that is the fundamental tie between most obesity-related diseases. It primarily takes place within the adipose tissue of the body where adipocytes have the ability to transdifferentiate into macrophages (which are crucial to the regulation of immune responses). The article takes this a step further by then labeling adipose tissue as a potential "immune organ" because of the large amounts of adipose tissue-associated macrophages, or ATMs, residing there. Inflammation within obesity is also related to an increase in the levels of adipocytokines such as TNFα and IL-6 and a decrease in those that downregulate inflammation, such as adiponectin. The increased release of adipocytokines also affects other organ systems within the body via various hormonal pathways. Adipocyte enlargement can also take place within the adipose tissue, and once the adipocyte reaches a size that doesn't allow the most effective amount of oxygen to diffuse into the cell, adipocyte hypoxia occurs. The deprivation of oxygen to the adipocytes can then cause them to become necrotic, which then signals ATMs and other inflammatory markers to the site, causing (you guessed it) inflammation!

Most people hear the word "inflammation" and associate it with cuts and colds rather than conditions such as obesity and diabetes. Increasing awareness of how inflammation can affect a person chronically, with the possibility of transforming into something worse than just gaining a few pounds, may be the insight needed to encourage people change their lifestyles for the better.

Inflammation, Diabetes, and Obesity: What's to Blame?

Recently a large amount of evidence has linked type 2 diabetes with obesity. However, type 2 diabetes has also been closely associated with chronic systemic inflammation. As the review article by Kolb and Mandrup-Poulsen entitled "An Immune Origin of Type 2 Diabetes?" suggests, obesity (or a high calorie diet) is not enough to induce the insulin resistance that is characteristic of type 2 diabetes. In contrast, the research appears to indicate that several proinflammatory genes and infiltration of adipocytes by macrophages in response to inflammatory cytokines ultimately cause insulin resistance that results in type 2 diabetes. This evidence suggests that if an individual has obesity as well as a genetic predisposition (activated proinflammatory genes) then the infiltration of adipocytes by macrophages, insulin resistance, and type 2 diabetes would be likely to result.
In particular, cytokines such as TNF-α and IL-6 have been implicated in this process as they are produced by the activated macrophages. As the article states, disruption of genes for several TNF- α receptors can actually prevent insulin resistance in animal models. However, such drastic measures are obviously not desirable in humans. Moreover, the result of creating such a gene defect is likely to differ given different genetic backgrounds (i.e. humans and mice). There is also indication that IL-6 may also be beneficial in lowering blood glucose levels as a result of muscle-derived IL-6 released during exercise. This conflicting evidence for the role of IL-6 as a component of inflammation connected with type 2 diabetes helps to illustrate just how complicated this disorder is. Ultimately, the question becomes what method is best to treat this low-level inflammation in order to reduce the risk of developing insulin resistance. Is it really beneficial to treat patients with NSAIDS and other anti-inflammatory drugs? What are the long-term risks of suppressing or, at least, decreasing levels of inflammation within the body?

Childhood Obesity and Insulin Resistance

There is no denying that obesity in adolescents is on the rise and increasing greatly past previous decade's statistics. One in every three children ages 2-19 is overweight or obese. The devastating consequences of this epidemic create a desperate need for action. With the major changes that have taken place in the American lifestyle in the past several decades, its no wonder we are seeing this trend. Coupling processed foods and unhealthy fast-food diets with virtually no physical activity, the outcome is a more overweight generation than ever seen in the past. With this rise of obesity in children comes the risk of future health problems such as high cholesterol, high blood pressure, heart disease, insulin resistance, and eventually type II diabetes.

With in the study "Young adults obese subjects with and without insulin resistance: what is the role of chronic inflammation and how to weigh it non-invasiveley?", the goal was to obtain the differences between young obese individuals with and without insulin resistance, the role that visceral and subcutaneous abdominal adipose tissue have on hepatic steatosis, and determining spleen size and which inflamation markers correspond with insulin resistance and hepatic steatosis. The results showed that CRP and fibrinogen as well as spleen diameter were increased in the obese young with insulin resistance compared to non-insulin resistance young adults. Insulin resistance was significantly related to spleen size, hepatic steatosis, and the inflammation marker CRP. Metabolic syndrome was much more prevelant in obese young adults with insulin resistance, being that it is associated with a proinflammatory state that contributes to insulin resistance.

Raising awareness in prevention of childhood obesity and switching to a healthier lifestyle is crucial if we want to see a decline in this life-threatening trend. Switching from the research article to recent events, First Lady Michelle Obama has become an extremely influential advocate in this particular issue. In February, she launched a nationwide campaign, "Let's Move", in hopes to solve the growing challenge of childhood obesity. Her goals are to empower parents and caregivers with the knowledge needed to make changes in their children's lives, provide healthier food in schools, improve access to healthy, affordable food, and motivate children to become more physically active. The threat of childhood obesity is one that will not go away instantly, but the steps in Michelle Obama's campaign are key components to changing this generation's lifestyle for the better.

Inflammation, processed sugars and mysterious sources of inflammation

Americans, and now the larger industrialized world, are getting fatter and suffering from it. Type II diabetes is rampant (the ADA stated in 2007 that 10-11% of adults 20 and over are affected), and inflammation caused by a “western lifestyle” has been linked to a diverse array of maladies that lie somewhere between autoimmunity and inappropriate inflammation. From heart disease to gout we now suspect faulty and personally neglected signaling pathways related to insulin. The events leading up to diabetes actually may cause the most damage.

According to an article published by the ADA there is good evidence for the existence of a syndrome linking increased levels of insulin to increased triglycerides and hypertension^[1]. Most of the manifestations of this syndrome seem to relate to stress placed on muscle tissue in the presence of a high carbohydrate diet and lack of that fleeting pass-time, exercise. See, negative feedback is a wonderful thing, until you break a part of the cycle. In this case, muscle tissue fails to dispose of dietary blood glucose appropriately (or at least fails to do so in a timely manner) and the pancreas picks up the slack by stimulating the systemic uptake of glucose with insulin. You may have guessed by now that this creates a nasty little cycle, with the muscle becoming less stimulated and the pancreas secreting more insulin.

Unfortunately while the insulin-caused-insulin-resistance view of things supports the development of an insulin resistant state, it doesn't produce a culprit or even a biased, that-guy-looks -shady suspect for other inflammation-associated diseases. Diseases like stroke, certain cancers, asthma and others too numerous to list here, need another explanation. This is tricky, we can't point at something like peanuts and claim, “that causes inflammation.” The medical community tried that with the cholesterol hypothesis, and statin treatment; of which the most comprehensive studies are particularly lukewarm ^[2]. What we can point to, are the rise of the sedentary lifestyle and a trend towards highly processed foods as the norm. Facets of western culture that, if abandoned, might alleviate the suffering without significant medical intervention.

As far as the miracle of Bromelain discussed in “revisiting inflammation”, I would tend to be very skeptical of an enzyme based oral medication. Seeing as the enzyme is the meat tenderizer found at your local butcher, and your digestive tract releases a good deal of similar enzymes, it probably won't hurt^[3]. Although the literature is still out on the actual medical benefits ^[4]. After the cholesterol/statin hullabaloo, I would expect the community to drag its feet regarding broad statements on the matter.