Adam Wiss and Will Judson Biographies on the CU website!

Two Biology/Biochemistry majors have been highlighted on Cumberland University’s website this week.

Read about Will Judson, a biology/biochemistry double major, student athlete, and aspiring doctor, here.

Read about Adam Wiss, a biology major, a student athlete, and aspiring pharmacist, here.

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Journal Club: Mg(HSO4)2/SiO2 as a Highly Efficient Catalyst for the Green Preparation of 2-Aryl-1,3-Dioxalanes/Dioxanes and Linear Acetals

This blog post was written by William Greene.  His article is titled “Mg(HSO4)2/SiO2 as a Highly Efficient Catalyst for the Green Preparation of 2-Aryl-1,3-Dioxalanes/Dioxanes and Linear Acetals” by Jafari, et al., (Organic Chemistry International Volume 2012, Article ID 475301, 5 pages doi:10.1155/2012/475301). 

This article is about scientist trying to figure out how to conduct a green preparation of 2-Aryl-1,3-Dioxalanes/Dioxanes and Linear Acetals  that is efficient as possible. The reaction process needs to have high yields with low costs and low hazard risks. To get the reactions started either Magnesium Sulfate or SiO2 catalyst needs be used for the acetylization reaction. There are many amazing compounds like fragrances for example that can be made by these processes. But at what cost does the manufacturing of these chemicals have? Current manufacturing methods have high pollution rates with relatively low product yields.

The reactions conducted in this experiment are solvent free because the addition of a  catalyst induces the reaction without the need of a strong chemical solvent. This method greatly reduces the amount of pollution and harmful byproducts from the reaction. Magnesium Sulfate is considered a green compound because as a catalyst it emits low pollution levels. Despite being highly recyclable, magnesium sulfate maintains high reactivity levels making it an ideal catalyst.

Fariba Jafari and Saeed Khodabakhshi first needed to calculate the ideal concentration of the magnesium sulfate catalyst for their solvent free reaction. In order to do this they added different amounts of the catalyst to a reaction with benzaldehyde with ethandiol. The greatest percent yield and the lowest reaction time was when 0.2 grams of magnesium sulfate was added to the reaction. So the determined ideal catalyst concentration is 0.2 grams. A condensation reaction with many different organic compounds was conducted all for ten minutes and with 0.2 grams of the Silika supported Magnesium Sulfate catalyst solution added. The percent yields were then recorded.

The syntheses of fifteen different products were documented and the results seemed overwhelmingly positive that the Magnesium Sulfate solution-less catalyst reactions are quite efficient. The lowest percent yield was 87%. Jafari and Khodabakhshi also gave a means of comparison to previous (current manufacturing) methods of synthesizing these compounds some of which had percent yields as low as 58%.

What this means is that there are more efficient means to manufacture products than the ones currently used today. As the human population increases along with increases in product demands it is becomes more and more important to be efficient as possible to cut back on pollution and prolong the use of nonrenewable resources. The solution-less green reaction approach will also lead to lower manufacturing costs which means savings to the consumer. It will take time to implement new manufacturing techniques but the payoff for business and consumers will be great.

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Journal Club: Hepatotoxic effects of acetaminophen. protective properties of tryptophan derivatives

This post was written by Victoria Bennett. 

Hepatoxic Effects of Acetaminophen. Protective Properties of Tryptophan Derivatives.

Introduction

Many people use acetaminophen daily to help with aches, pains, fevers, arthritis, and symptoms that come with common colds. Users of acetaminophen may exceed the recommended dosage instruction not knowing the possible side effects of this medicine. The purpose of this study was to show the effects of taking too much acetaminophen has on the liver and what may possibly help reduce these risks. These studies are usually conducted on rats, so scientists are able to have control groups, control their surroundings easily, and control dosages administered to the animals.

Summary and Procedure

In this study, rats were given 500-1500 milligrams of acetaminophen (APAP) per kilogram of body weight intrgastrically. The scientists wanted to find ways to prevent liver damage, so they hypothesized that tryptophan derivatives, melatonin, and N-acetyl-nitrosotryptophan (NNT) might help stop the harmful side effects. These are believed to be hepatoprotectors, which are substances that help prevent damage to the liver. Acetaminophen was used because it is one of the most commonly used drugs to conduct chemically driven liver damage. Forty male Wister rats were used in this study. The rats were exposed to light for twelve hours and followed by a twelve hour period of darkness for one week. The animals were divided into five groups which were control, APAP, APAP+NNT (N-acetyl-nitrosotryptophan), APAP+Melatonin, and NNT. Each rat in the groups APAP, APAP+NNT, and APAP+Melatonin was given 1500 mg/kg of body weight of acetaminophen intragastrically. After 4 and 6 hours the groups APAP+NNT and NNT were given intraperitoneal injections of NNT at 2 mg/ kg of body weight. The group APAP+Melatonin also received intaoperitoneal injections of melatonin at 10 mg/kg of body weight. Approximately 24 hours after the injections and dosages were administered, the rats were decapitated and blood and liver tissue samples were prepared for analysis and study.

Tables and Trends

The general trend of Figure 1 shows that as Acetaminophen dosage increases, the mitochondrial protein drastically decreases by two-thirds. The second part of the chart shows that as APAP is increased, the postmitochondrial fraction decreases by almost half. The second chart shows the alanine aminotransferase (ALT), aspartate aminotransferase (AST), and bilirubin in blood plasma. The control group showed the lowest levels of ALT, AST, and bilirubin. The only section that has more ALT than AST is the APAP group. In the APAP+NNT and APAP+Melatonin groups, the AST and ALT are equivalent. The NNT group has the second lowest amount of AST, ALT, and bilirubin. Table 1 shows that there was not a decrease in the activity of liver enzymes in the mitochondria, but there was an increase in succinate dehydrogenase. Table 2 shows that there was not a large decrease in protein SH groups in the postmitochondrial fraction, but the glutathione (GSH) decreased by almost 45%. GSH, a main antioxidant in the body, helps the body detoxify itself. Once GSH is decreased dramatically, it causes the person or animal’s immune system to weaken, therefore increasing the risk of infection.

Conclusion

In conclusion, studies from the table show that N-acetyl-nitrosotryptophan was best in stopping elevated levels of ALT, AST, and bilirubin. By decreasing those three, there is less damage to the liver. When a liver blood test is done, elevated levels in ALT and AST signify that there is evidence of liver damage. When bilirubin levels were increased in the blood stream, it indicated that medictions, such as acetaminophen, had been taken causing drainage ducts in the liver to be blocked and leak bilirubin. There may be better ways to take acetaminophen, so that it may bypass the liver causing less harmful side effects. Research would need to be conducted to make sure it would not cause harm to other organs. Acetaminophen has many positive uses when taken properly and in the right dosage. People need to be aware of the harmful side effects when APAP is taken in too high of a dosage and too frequently. Studies need to be done with different other possible hepatoprotectors, such as combining melatonin, NNT, and N-acetyl-cysteine. Studies have been done showing that some of these reduce APAP hepatoxicity, so if combined, these effects may be decreased. The experiment was valid, and each step of the process was recorded precisely in measurement. The only possible problems might include the drugs used were not left in the rats for a long enough time to see other possible side effects. Other studies derived from this experiment could test the side effects that happen to the other organs in the body.

Works Cited

Dremza, I. Cheshchevik, VT. Zabrodskaya, SV. Maksimchik, YuZ. Lapshina, EA. Zavodnik, IB.  (2009, January 20). Hepatotoxic effects of acetaminophen. protective properties of tryptophan derivatives. Retrieved from http:/search.proquest.com.biologyjourndocview/748033150/139C843929931202A9D/63?accountid=26997

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Journal Club: Perinatal Exposure to Bisphenol A Alters Early Adipogenesis in the Rat

This post was written by Katie Jones. 

Bisphenol A (BPA) is a chemical compound found in many plastics. It has been identified as an environmental pollutant and can be a potential danger to human health as a disruptor of the endocrine system. When buying plastic baby bottles, cups, and food containers, many will have a BPA-free stamp somewhere on the container for this reason. As someone who worked in a dental office as a sterilization technician for two years, BPA has continued to fascinate me because many of the instruments I worked with were made of plastic. My father has also been diagosed with non-Hodgkins lymphoma within the last year, which his oncologists have said may have a BPA link.

With the increasing prevalence of childhood obesity, scientists at the University of Geneva in Switzerland decided to research the effects of BPA exposure on perigonadal adipose tissue in rats from early in gestation to three weeks after birth. Their research is summarized in a paper titled Perinatal Exposure to Bisphenol A Alters Early Adipogenesis in the Rat. It was published by Emmanuel Somm et al. and appeared in the October 2009 issue of Environmental Health Perspectives from pages 1549-1555.

Introduction:  Based on previous studies, it has been found that BPA is in the serum of pregnant women and umbilical cord serum collected at birth.1 It has also been indicated that BPA has a presence in amniotic fluid and placenta.1 When rats were exposed to BPA in utero, females showed accelerated puberty and increased body weight,2 changes to the mammary glands,3 and abnormal development of the genitals.1 Males showed abnormal changes in structure and function of the prostate.4,5

 In the body, BPA has been known to disrupt normal endocrine functioning by binding to estrogen receptors alpha and beta.6,7 It also acts as a thyroid hormone antagonist and targets protein disulfide isomerase, which is involved in the folding, assembly, and shedding of many proteins in the cells.8,9 BPA also interferes with glucose homeostasis10 and increases the presence of adipose tissue cells in the body with increased enzyme levels and fatty acid binding proteins.11

 At the time, there was a lack of research concerning BPA exposure in vivo, which also accounted for the unexplained trends of obesity in industrialized countries. Because of this, Somm et al. studied the effects of BPA on adipose tissue and its gene expression in Sprague-Dawley rats three weeks after birth. They also monitored the body weight and food intake of these same rats if they were fed a standard diet or high-fat diet after the three-week mark.

Methods:  Animal Care, Diets, and BPA Exposure before Weaning: The male and female rats were given a rodent experimental diet low in phytoestrogens 10 days before mating until the end of the gestation period. Pregnant female rats were exposed to BPA at a concentration of 1 milligram per liter in the water supply from day six of the gestation period until three weeks after the birth of the offspring. Pregnant females in the control group were given 1% ethanol instead of BPA. All plastics used during the research were BPA-free to avoid contamination. Daily food intake, volume of water consumed, and body weight were measured daily in the mother rats before the three-week post-birth mark.

Animal Care and Diets after Weaning: At the three-week mark, a group of rats that were from both the BPA-exposed group and the ethanol control group were anesthetized and decapitated. Epididymal white adipose tissue (eWAT) was removed from males and parametrial white adipose tissue (pWAT) was removed from females. Brown adipose tissue was also collected near the shoulder region from both males and females. These fat pads were weighed and then frozen in liquid nitrogen for further analysis.

After the three-week mark, a second group of rats that were from both BPA-exposed and ethanol control groups started to get normal drinking water and were fed with either a standard rat diet or a high-fat diet. Their body weight was measured from ages four weeks to fourteen weeks. Their food intake was also measured by weighing the solid pellets on grids atop the cages.

Histologic Examination of Adipose Tissue: The pWAT that was removed from the first group of rats was fixed in a paraformaldehyde solution for 24 hours and then set in paraffin. Small sections (5 micrometers) were cut and stained.

RNA Preparation and mRNA Quantification: RNA was extracted from the adipose tissue and liver samples from the rats using Trizol reagent. 5 micrograms total of RNA were reverse transcribed using a leukemia virus reverse transcriptase, RNAsin ribonuclease inhibitor, primers, dNTP (deoxyribonucleotide triphosphate) and DTT (dithiothreitol). Polymerase chain reaction (PCR) determined the coding of CAAT enhancer binding protein alpha (C/EBP-α), peroxisome proliferator-activated receptor gamma (PPAR-ɣ), sterol regulatory element binding protein 1-C (SREBP-1C), GATA binding protein 2 (GATA-2), preadipocyte factor 1 (Pref-1), lipoprotein lipase (LPL), acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), peroxisome proliferator-activated receptor alpha (PPAR-α), peroxisome proliferator-activated receptor-gamma coactivator (PGC-1α), stearoyl-CoA desaturase 1 (SCD-1) and glucose transporter 4 (GLUT4). The primer sequences for each were listed in this same section.

Plasma Measurements: Plasma glucose, noesterified fatty acid and triglyceride levels, and cholesterol levels were measured.

Glucose Tolerance Test: 14-week-old male rats from both the BPA and control groups had been fed a high-fat diet and fasted from the previous day. They were injected with 1.5 milligrams of glucose per gram of body weight. Blood samples were then collected through the tail at certain time intervals (0, 15, 30, 60, and 120 minutes) following the administration of glucose.

Statistics: The results were expressed as the mean plus or minus the standard error of measurement (SEM). The analysis of variance (ANOVA) method was used. When taking into account birth weight and the weight at three weeks between the BPA and control groups, a regression method was used.

Results:  Maternal Physiology during Gestation: During the pregnancies of the female rats, the weight gain throughout the gestational period was not significantly different in those exposed to BPA (135 ± 7 g) versus the control group (140 ± 6 g). Food intake was also similar between the two. The daily amount of water consumed was not changed by the presence or absence of BPA. By the end of gestation, the scientists estimated that the total exposure of BPA to each rat was approximately 70 micrograms of BPA per kilogram of body weight each day once gestation had ended. Bottle leakage, spills, and evaporation were not taken into account. I saw this as problematic because leakage, spills, and evaporation could have accounted for a large portion of the measured intake and dramatically changed the results.

Body Weight of Offspring at Birth, Sex Ratio, and Litter Size: At birth, the body weight of male rats born to BPA-exposed mothers was significantly higher (7.33 ± 0.12 g) than those of the control group (6.91 ± 0.15 g). The same applied to the female rats born to BPA-exposed mothers (7.03 ± 0.11 g) versus the control group (6.47 ± 0.12 g). This trend is seen in Figure 1A-B. Litter size was determined to not affect the results. The sex ratio of the newborn rats was not affected by prenatal BPA exposure.

Body Weight and Fat Pad Weights at Weaning: Body weight measurements were repeated at age three weeks, which can be seen in Figure 1C-D. The body weight of BPA-exposed male rats (53.36 ± 1.02 g) was not much different from the control group (52.24 ± 1.11 g). However, the BPA-exposed females still had a higher body weight on day 21 (53.73 ± 0.65 g) versus the control group (47.79 ± 1.44 g).

To study the fat pads, a few of these rats were sacrificed in the same manner as those mentioned earlier. The BAT and eWAT/pWAT were removed and weighed. In males, the BAT weight remained unchanged but the eWAT mass was higher in the BPA group (56 ± 6 mg) compared to the control group (39 ± 6 mg). In the pWAT of females, the mass was significantly higher in the BPA group (95 ± 9 mg) than the control group (33 ± 6 mg). In contrast to the males, the BAT was also higher in the BPA group (178 ± 13 mg) than the control group (116 ± 11 mg). These results can be seen in Figure 2A-B. The results also suggest a positive correlation between the increase in body weight of the BPA-exposed female rats and the increase in fat pad weight. This is shown in Figure 2C.

Overall, the results showed that females are more sensitive to weight gain when exposed to BPA. With BPA binding to estrogen receptors, which are seen more in females, this seems to make sense.

Expression of Genes Involved in Adipogenesis and Lipogenesis in Adipose Tissue At Weaning: Histologic sections of the females’ pWAT tissue were taken. In the control group, the cells appeared normal, but in the BPA group, the adipose cells were much larger. This is seen in Figure 2D.

The mRNA levels of the preadipogenic transcription factors C/EBP-α, PPAR-ɣ, and SREBP-1C were significantly increased by about double each in the BPA female rats’ pWAT. The levels of the inhibitors GATA-2 and Pref-1 remained unchanged. The levels of lipogenic enzymes LPL, FAS, and SCD-1 also increased in the pWAT of the BPA females. Levels of GLUT4 also increased. These results are shown in Figure 2E-F. These results compared with the histologic examinations of the pWAT tissue show that the exposure to BPA enhances adipogenesis by increasing gene transcription. This explains why the adipose cells are much larger and the pWATs were heavier.

Expression of Genes Involved in Lipogenesis in Liver at Weaning: In BPA-exposed females, there were significant increases in SREBP-1C, ACC, and FAS, but no changes to PPAR-α or PPAR-ɣ (Figure 3A). There were no significant differences in circulating triglycerides, which could have altered gene expression (Figure 3B-E).

Postweaning Body Weight: The BPA-exposed rats were weighed weekly from ages four to fourteen weeks. There was no statistical difference in the body weight of BPA-exposed males versus the control males on the standard rat diet, but the BPA-exposed males on a high-fat diet were much heavier than the controls (Figure 4A-B) In females, the BPA-exposed were heavier than the controls on both standard rat and high-fat diets (Figure 4C-D). These results show a sex-specific difference in weight gain in BPA-exposed rats.

Postweaning Food Intake and Glucose Tolerance at Adulthood: There was no impact of perinatal BPA exposure on daily food intake or energy intake in males or females on either diet. Glucose tolerance tests were performed in the same manner in both BPA-exposed males and control males. No disturbances in glucose tolerance were found.

Discussion:  Based on this study, it is evident that exposure to bisphenol A perinatally affects the generation of fatty tissue. The body weight of the rats exposed to BPA was higher than that of the control groups. The genes, proteins, and enzymes responsible for formation of adipose cells were also overexpressed, resulting in hypertrophied adipose cells in rats exposed to BPA. It was also indicated that being exposed to BPA as well as a high-calorie diet makes one more susceptible to obesity.

Some explanations for the increased weight gain in the BPA-exposed rats can be examined by looking at the mechanisms of BPA. By binding to estrogen receptors, BPA can directly act on the hypothalamus and control appetite. This happens because BPA can cross the blood-brain barrier. This could explain why those rats exposed to BPA had an increased food intake. It could also explain why the females consistently had higher body weights than the controls, as there is a higher presence of estrogen in females than in males.

In conclusion, the direct exposure to BPA through the parent (placenta and milk) in vivo increases adipose tissue creation and storage in a sex-specific manner. Using these results, BPA exposure could be related to the increase in childhood obesity, but further studies are needed to confirm this.

Final Thoughts:

 Because this study is one of very few of its kind, I would say it is too soon to accept or dismiss these results. Given the circumstances, I do believe this is an interesting and exciting idea in the application to childhood obesity. Knowing the dangers of BPA and our exposure to plastics on a normal basis, this experiment could be very valid in determining the relation. On another note, I did feel this experiment could have had completely different results in a few areas. For example, the amount of  BPA consumed by the pregnant rat during the gestational period was 70 micrograms of BPA per gram of body weight each day. This BPA was in the water bottle, and the scientists did not take into account spills, leaks, or evaporation, and these were stated as an error. I understand that those variables were very hard to measure, but they could have dramatically altered the results had they been attainable.

This study also had multiple variables that were tested, including body weight, RNA analysis, histological analysis, glucose testing, and the effect on the liver. These variables may have been necessary to fully explain the results, but at times the article became muddled and confusing to me because there were so many different variables. If I had performed this same experiment, I would have focused on less variables, probably excluding the glucose tolerance and effect on the liver since they did not yield the significant results that the others did.

Overall, I enjoyed this aspect of relating BPA to obesity. It will be interesting to see the grey areas be answered in the future, such as, if BPA and a high-calorie diet increases susceptibility to obesity, would the same result be seen if plastics were regulated to be BPA-free and exposure was much lower? That experiment would require a longitudinal study (and probably several decades), but this study opened up opportunities for questions such as that one to be answered.

References:

  1. Ikezuki, Y, Tsutsumi, O, Takai, Y, Kamei, Y, Taketani, Y. 2002. Determination of bisphenol A concentrations in human biological fluds reveals significant early prenatal exposure. Hum Reprod 17(11): 2839-2841.
  2. Takahashi, O, Oishi, S. 2000. Disposition of orally administered 2,2bis(4-hydroxyphenyl) propane (bisphenol A) in pregnant rats and the placental transfer to fetuses. Environ Health Perspect 108: 931-935.
  3. Honma, S, Suzuki, A, Buchanan, DL, Katsu, Y, Watanabe, H, Iguchi T. 2002. Low dose effect of in utero exposure to bisphenol A and diethylstilbestrol on female mouse reproduction. Reprod Toxicol 16(2): 117-122
  4. Markey, CM, Luque, EH, Munoz De Toro, M, Sonnenschein, C, Soto, AM. 2001. In utero exposure to bisphenol A alters the development and tissue organization of the mouse mammary gland. Biol Reprod 65(4): 1215-1223.
  5. Ramos, JG, Varayoud, J, Sonnenschein, C, Soto, AM, Munoz De Toro, M, Luque, EH. 2001. Prenatal exposure to low doses of bisphenol A alters the periductal stroma and glandular cell function in the rat ventral prostate. Biol Reprod 65(4): 1271-1277.
  6. Hiroi, H, Tsutsumi, O, Momoeda, M, Takai, Y, Osuga, Y, Taketani, Y. 1999. Differential interactions of bisphenol A and 17beta-estradiol with estrogen receptor alpha (ER alpha)  and ER beta. Endocr J 46(6): 773-778.
  7. Kurosawa, T, Hiroi, H, Tsutsumi, O, Ishikawa, T, Osuga, Y, Fujiwara, T, et al. 2002. The activity of bisphenol A depends on both the estrogen receptor subtype and the cell type. Endocr J 49(4):465-471.
  8. Hiroi, T, Okada, K, Imaoka, S, Osada, M, Funae, Y. 2006. Bisphenol A binds to protein disulfide isomerase and inhibits its enzymatic and hormone-binding activities. Endocrinology 147(6): 2773-2780.
  9. Okada, K, Hiroi, T, Imaoka, S, Funae, Y. 2005. Inhibitory effects of environmental chemicals on protein disulfide isomerase in vitro. Osaka City Med J 51(2): 51-63.
  10. Sakurai, K, Kawazuma, M, Adachi, T, Hariyaga, T, Saito, Y, Hashimoto, N, et al. 2004. Bisphenol A affects glucose transport in mouse 3T3-F442A adipocytes. Br J Pharmacol 141(2): 209-214.
  11. Masuno, H, Iwanami, J, Kidani, T, Sakayama, K, Honda, K. 2005. Bisphenol A accelerates terminal differentiation of 3T3-L1 cells into adipocytes through the phosphatidylinositol 3-kinase pathway. Toxicol Sci 84(2): 319-327.
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Journal Club: Crystal Growth Inhibitors for the Prevention of l-Cystine Kidney Stones Through Molecular Design

This post was written by Simone Ryan. The article is entitled “Crystal Growth Inhibitors for the Prevention of l-Cystine Kidney Stones Through Molecular Design” by Jeffrey D. Rimer et al. (Science 330, 337, 2010). 

Introduction
The kidney acts as a garbage man collecting trash and wastes that comes in the body. Eventually the kidney wants to excrete these wastes and get rid of them by urinating. Sometimes these wastes causes a build up in the kidney commonly referred to as kidney stones. There are different types of kidney stones. The type of kidney stone discussed in this research article is known as L- Cystine stones. They are relatively larger than the more common type of kidney stones found in the body and are more likely to cause chronic kidney disease. L- Cystine build up is caused by mutations in one of two different genes (type A cystinuria or type B cystinuria). There are a few different types of treatments for this problem, one is the ingestion thiol drugs and another is high fluid intake increasing the urine pH diluting the stones. The purpose of this experiment was to see if alternative approaches to minimizing L- Cystine will be more effective in crystal growth inhibition or more effective in previous processes.

Methods
The binding of tailor Growth Inhibitors L- CDME (L- Cystine dimethylester) and L- CME (L- Cystine methylester are used for the prevention of L- Cystine kidney stones. The synthesis of L- CME and L- CDME were done. The preparation of the hexagonal L- Cystine crystals were performed and bulk crystallization in the presence of additives were done. The research paper goes into details showing how this was done using figures. Figure 1 shows the molecular structure of L- Cystine and the inhibitors L- CME and L- CDME. Figure 2. shows the human stones and helical make-up of these stones as well. This figure is important and showing how the whole process will work. Other figures in the paper further explain and shows what was done and results of the experiment.

Results
L- CME binds at ledge sites and the difference of L- CME and L- CDME shows a outstanding difference of molecular recognition  between tailor inhibitors and crystal surfaces. L- CDME was extremely effective in the reduction of other kidney stones. The molecular additives were less effective.  L- CDME behavior was viewed to possibly be therapeutic to preventing L- cystine kidney stones, thus being more effective than the other treatments such as thiol drugs and high fluid intake which could have a lot of side effects.
Discussion
The article shows an effective and alternative way in the reduction of L- cystine kidney stones. The different treatments used in decreasing kidney stones or the reduction of them may be ineffective and  problematic. Some treatments even have a bunch of side effects that can be annoying. It is good that the experiment tests different ways in minimizing these problems by finding an alternative way to prevent L- Cystine stones.

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Journal Club: Amino Acid Supplements and Recovery From High-Intensity Resistance Training

This blog post was written by Taylor Watson.  The article is titled “Amino Acid Supplements and Recovery From High-Intensity Resistance Training” by Sharp, et al., (Journal of Strength and Conditioning Research; Apr 2010; 24, 4; ProQuest Science Journals pg. 1125). 

In this paper a study was done on 8 men to show whether a branched amino acid supplement would help with recovery from high-intensity resistance training.  These men were put into two groups in which the one group took the amino acid supplement and the other took a placebo for four weeks.  On the fourth week both groups were subjected to high-intensity training.  During this week blood was drawn four times and tested for the levels of testosterone, cortisol, and creatine kinase in the serum.  The results were then compared to the level of each before the study began.

In this study a group of 8 men of around the same age, height, and weight with previous resistance training were chosen.  All of these men were tested for their 1-rep max on 8 different exercises and also had blood drawn to record their original levels of testoerone, cortisol, and creatine kinase.  These men were separated into two different groups.  One group was given a branched amino acid supplement (Nutri-Build II) while the other group was given lactose and used as a control.  Neither group knew what they were taking so as to not skew the study.  The groups took either the supplement or placebo for 3 weeks with no resistance training during that period.  Then the groups continued to take either supplements or placebo for a forth week but in this week four training sessions were also done on four separate days.  The groups blood was taken at the beginning of the week, 1 hour after training days 2 and 4, and 36 hours after the final training session.  The levels for testosterone, cortisol, and creatine kinase were compared to the first sample taken.  A five-week period was then given to allow all of the supplements to be excreted by the body.  After this period the groups were then switched the placebo group was given the supplements and vice-versa.

The results of this study showed there to be decrease in levels of cortisol, which will tear muscles down, in the serum after taking the branched amino acid supplement.  With the levels of  cortisol being lowered with the use of branched amino acid it shows that recovery can happen quicker because there are less stress signals being sent from the muscles.  The level of creatine kinase rose in both groups but the levels rose significantly less in the branched amino acid group.  Testosterone levels were significantly greater after taking the branched amino acid supplement.  This helps the muscles recover and causes protein sythesis.

Figure 1 in the article shows the times blood was drawn and when resistance training was done during this study.  Figure 2 shows the average of the cortisol levels of both the branched amino acid group and the placebo group in the final week in which the resistance training was done.  It shows that the average level of corisol in the placebo group went up and the average level of cortisol in the branched amino acid group went down.  Figure 3 shows the percent change of corisol levels in the both groups from the beginning of the study to the end between the two groups.  Again the levels of corisol in the placebo group rose and the level in the branched amino acid supplement went down.  Figure 4 shows the percent change of serum testosterone levels between both groups.  Testosterone levels in the placebo group decreased and in the branched amino acid group the levels rose.  Figure 5 showed the ratio between testosterone and cortisol over the study.  In the placebo group the ratio was lowered and in the branched amino acid group the ratio was increased.  Figure 6 shows the average levels of serum creatine kinase over the study.  In both groups creatine kinase levels rose but the level of rise was much lower in the branched amino acid group.

I found this article to be very useful because being a football player my body depends on working out to develop my body.  This article showed me a practical application of a supplement that really does help my body in several ways while working out. I also think this article could be used to show people an alternate safe way other than illegal ways of gaining strength such as steroids. I feel like 2 groups of females should have also been included to see if the results were alike between the different sexes.

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Journal Club: Synthesis of new pyrazolyl-2, 4-thiazolidinediones as antibacterial and antifungal agents

This post was written by Nilu Nurinova.

Passing by Wal-Marts’ personal hygiene sections or cleaning supplies sections we see numerous and a variety of antimicrobial products. Many of them aimed at different types of microbes. Lysol, antimicrobial hand soap, disinfecting wipes all of them have several chemical compounds which target particular bacteria or fungus. But, why we need so many of them? Is one type of antimicrobial product not enough? Well, apparently not. Microbes evolve with their resistant properties. Overtime, the product that we usually use in the household will not kill the germs we want anymore. On a biochemical level, microbes and germs incorporated the chemicals into their system and develop immunity, just like humans towards certain diseases.

Many scientists develop new antimicrobial chemical products daily and it is becoming a challenging task for researchers to develop fastest and easiest methods to derive them.

The paper by Aneja et al. “Synthesis of new pyrazolyl-2, 4-thiazolidinediones as antibacterial and antifungal agents” published in Organic and Medicinal Chemistry Letters 2011 shows another shorter and modified method to synthesize new derivatives of antimicrobial chemical compounds.

Here, I present a brief summarization of the research.

Introduction:  The following research experimented and investigated biochemical properties of newly synthesized organic molecules which aid in antibacterial and antifungal properties of today’s different cleaning products. A group of researchers from the Department of Microbiology and Pharmaceutical Sciences at Kurukshetra University in India, experimented better methods to synthesize newer derivatives of antimicrobial compounds in order to combat the rising resistance of microbes towards contemporary germicidal agents. Here, the research group Aneja et al. presents the synthesis of antimicrobial compound pyrazole’s derivatives which contain thiazolidindiones (TZD).  TZDs are heterocyclic compounds and along with other heterocyclic compounds like rhodanines became famous in pharmaceutical industry due to their diverse biological activities and pharmaceutical applications. TZDs in particular, have long been known for their bactericidal and fungicidal effects, especially.

While researching for TZDs antimicrobial efficacy, continuous modification of these heterocyclic compounds i.e. fused or replaced with variety of substituents on certain carbons in the cyclic chain became main focus during this synthesis. TZDs are more effective biologically and can be applied for variety of medical purposes, if they are fused or incorporated into other biologically active compounds. Hence, TZDs, could serve more than antimicrobial agents, as an anticonvulsant or as other anti inflammatory agents according to the authors. In this research, TZD is fused with the pyrozole compound and the newly synthesized molecule pyrazolyl-2, 4 –TZD was tested for its antimicrobial property. However, synthesizing pyrazolyl-2, 4 –TZDs with the desired modifications, could challenged scientists  and the group of Aneja, provided that Knoevenagel condensation was the best method for the compound synthesis. (Knoevenagel condensation is the reaction where aldehyde or ketones, which don’t normally contain an a hydrogen, perform a condensation reaction with compounds of the form Z-CH2-Z’ or Z-CHR-Z’. The Z groups are electron withdrawing groups, such as CHO, COR, COOH, COOR, CN, NO2, SOR, SO2R, SO2OR or similar groups.)

Experimental Methods and Data:  The research includes chemical and physical data. The chemical data shows the synthesis of the desired compound pyrazolyl-2, 4 –TZDs,  via Knoevenagel condensation* and acid hydrolysis, and Vilsmeier – Haack reaction  involving many other multistep processes. To establish the structures, synthesized compounds were subjected to H NMR and IR spectra, which showed the locations of carbonyl, ester, cyclic, amine and TZD groups on the synthesized compound. The analytical data  and the H NMR analysis showed that all the derivatives in this study were obtained in Z isomer form because Z forms of pyrazolyl-2, 4 –TZDs were thermodynamically favored.

Results:  According to the physical data table, the reactions showed the average of 94-95% yield of all compounds synthesized including very similar melting points. Similar melting points are indicative of similarly formed compounds.

The biological data shows that all synthesized compounds were tested for anti fungal and antibacterial properties. For antifungal test, Aspergillus niger and Apergillus flavus were chosen and tested in vitro. Flucanozole, a standard over counter antifungal drug was used for comparison. For antibacterial test, two gram positive bacteria, Staphylococcus Aureus and Bacillus subtillis, including two gram negative bacteria, E-Coli and Pseudomonas aeruginosa were chosen. Ciprofloxacin antibiotic was used for comparison, too.

Results for antifungal test showed a comparable myecelial growth inhibition, comparable to flucanozole. Results for antibacterial test showed no susceptibility to gram negative bacteria but variable susceptibility to gram positive bacteria, and S.Aureus showed the highest susceptibility to pyrazolyl-2,4-TZDs.

Discussion:  Development of antimicrobial agents is only beneficial as alternatives. Considering that antimicrobial chemicals over used by today’s society, more intense modification of these chemical are required. Bacteria and fungus have different chemical properties on cellular level, yet developing compounds such as pyrazolyl-2,4-TZDs which can target eukaryotic and prokaryotic organisms at the same time is quite an achievement.

Having such great antifungal results at hand, provide alternatives to contemporary antifungal drugs like flucanozole. Although, the antibacterial activity showed great results in terms of combating gram positive bacteria, maybe developing or using another synthetic methods will leave hope to modify the pyrazolyl-2,4-TZDs so it would inhibit gram negative bacteria, too.

Final note:  This article awoke my interest due its synthetic methods applied in development of new derivatives of existing compounds. Modification of chemical compounds on molecular level, especially which have antimicrobial properties always presents a giant challenge especially with today’s evolving resistant microbiota. When something is newly synthesized resulting in effective and diverse biological activity, brings hope that science has not run out of options yet.

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