Serum levels of inflammatory biomarkers, glycaemic control indices and leptin receptors expression in adult male Wistar rats exposed to Pyrethroids
TL;DR: It is self-evident that pyrethroid chemicals exposure may disrupt carbohydrate and lipid metabolism by raising the risk of obesity among the exposed groups.
Abstract: BACKGROUND: Due to widespread use of pesticides, an individual’s lifetime exposure to pesticide mixtures is unavoidable, resulting in potentiation of toxic effects such as changing metabolic signaling temporarily or permanently due to the formation of free radicals among exposed individuals. OBJECTIVE: This study then looked at the expression of leptin and insulin receptors in Wistar rats exposed to pyrethroid and its effects on adipokines and glycemic indices. METHODS: 84 Wistar rats were randomly selected and divided into two groups. 51 rats were exposed to 1.2 % w/v pyrethroid insecticides, while the remaining 33 rats were grouped as non-exposed. The groups were divided into three different groups, each with 7, 21, and 41 days. The animals were anesthetically sacrificed and samples of blood were collected after days of exposure. The levels of total antioxidant status (TAS), malondialdehyde (MDA), glutathione (GSH), hydrogen peroxide (H2O2), nitric oxide (NO), glucose and lipids were determined. Blood activities of superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT) were also determined. ELISA was used to assess leptin, adiponectin, insulin, CRP, IL-6, and TNFα. Glycaemic indices were calculated appropriately. Leptin and Insulin expression was determined using the reverse transcriptase-polymerase chain reaction (RT-PCR) method. Data obtained were statistically compared, and P≤0.05 was considered significant. RESULTS: The mean TChol, TAG, and HDL-C were significantly different (p < 0.05) among the exposed groups. The serum mean levels of SOD, GPx, CAT, GSH, TAS, QUICKI, TNFα, IL6, and CRP were significantly reduced (p < 0.05), while mean levels of MDA, H2O2, NO, insulin, HOMA-IR, FIRI, leptin, and adiponectin were significantly elevated (p < 0.05) in exposed groups. The relative expression of insulin and leptin genes in exposed and non-exposed groups was also shown by column. CONCLUSION: It is self-evident that pyrethroid chemicals exposure may disrupt carbohydrate and lipid metabolism by raising the risk of obesity among the exposed groups.
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TL;DR: This protocol provides an overview of the comparative CT method for quantitative gene expression studies and various examples to present quantitative gene Expression data using this method.
Abstract: Two different methods of presenting quantitative gene expression exist: absolute and relative quantification. Absolute quantification calculates the copy number of the gene usually by relating the PCR signal to a standard curve. Relative gene expression presents the data of the gene of interest relative to some calibrator or internal control gene. A widely used method to present relative gene expression is the comparative C(T) method also referred to as the 2 (-DeltaDeltaC(T)) method. This protocol provides an overview of the comparative C(T) method for quantitative gene expression studies. Also presented here are various examples to present quantitative gene expression data using this method.
20,580 citations
TL;DR: Control of portion size, consumption of a diet low in fat and energy density, and regular physical activity are behaviors that protect against obesity, but it is becoming difficult to adopt and maintain these behaviors in the current environment.
Abstract: The current epidemic of obesity is caused largely by an environment that promotes excessive food intake and discourages physical activity. Although humans have evolved excellent physiological mechanisms to defend against body weight loss, they have only weak physiological mechanisms to defend against body weight gain when food is abundant. Control of portion size, consumption of a diet low in fat and energy density, and regular physical activity are behaviors that protect against obesity, but it is becoming difficult to adopt and maintain these behaviors in the current environment. Because obesity is difficult to treat, public health efforts need to be directed toward prevention.
2,302 citations
TL;DR: Biochemical and physiological characterization of catalases from many different organisms has revealed a surprisingly wide range ofCatalatic efficiencies, despite similar sequences.
Abstract: More than 300 catalase sequences are now available, divided among monofunctional catalases (> 225), bifunctional catalase-peroxidases (> 50) and manganese-containing catalases (> 25). When combined with the recent appearance of crystal structures from at least two representatives from each of these groups (nine from the monofunctional catalases), valuable insights into the catalatic reaction mechanism in its various forms and into catalase evolution have been gained. The structures have revealed an unusually large number of modifications unique to catalases, a result of interacting with reactive oxygen species. Biochemical and physiological characterization of catalases from many different organisms has revealed a surprisingly wide range of catalatic efficiencies, despite similar sequences. Catalase gene expression in micro-organisms generally is controlled either by sensors of reactive oxygen species or by growth phase regulons, although the detailed mechanisms vary considerably.
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TL;DR: A first systematic analysis of microbiota changes in the ileum and colon using multiple diets and investigating both fecal and mucosal samples demonstrates correlations between the microbiota and dysfunctions of gut, adipose tissue, and liver, independent of a specific disease-inducing diet.
Abstract: Development of non-alcoholic fatty liver disease (NAFLD) is linked to obesity, adipose tissue inflammation, and gut dysfunction, all of which depend on diet. So far, studies have mainly focused on diet-related fecal microbiota changes, but other compartments may be more informative on host health. We present a first systematic analysis of microbiota changes in the ileum and colon using multiple diets and investigating both fecal and mucosal samples. Ldlr−/−.Leiden mice received one of three different energy-dense (ED)-diets (n = 15/group) for 15 weeks. All of the ED diets induced obesity and metabolic risk factors, altered short-chain fatty acids (SCFA), and increased gut permeability and NAFLD to various extents. ED diets reduced the diversity of high-abundant bacteria and increased the diversity of low-abundant bacteria in all of the gut compartments. The ED groups showed highly variable, partially overlapping microbiota compositions that differed significantly from chow. Correlation analyses demonstrated that (1) specific groups of bacteria correlate with metabolic risk factors, organ dysfunction, and NAFLD endpoints, (2) colon mucosa had greater predictive value than other compartments, (3) correlating bacteria differed per compartment, and (4) some bacteria correlated with plasma SCFA levels. In conclusion, this comprehensive microbiota analysis demonstrates correlations between the microbiota and dysfunctions of gut, adipose tissue, and liver, independent of a specific disease-inducing diet.
1,097 citations
TL;DR: TRIzol solubilization and extraction is a relatively recently developed general method for deproteinizing RNA, which is particularly advantageous in situations where cells or tissues are enriched for endogenous RNases or when separation of cytoplasmic RNA from nuclear RNA is impractical.
Abstract: TRIzol solubilization and extraction is a relatively recently developed general method for deproteinizing RNA. This method is particularly advantageous in situations where cells or tissues are enriched for endogenous RNases or when separation of cytoplasmic RNA from nuclear RNA is impractical. TRIzol (or TRI Reagent) is a monophasic solution of phenol and guanidinium isothiocyanate that simultaneously solubilizes biological material and denatures protein. After solubilization, the addition of chloroform causes phase separation (much like extraction with phenol:chloroform:isoamyl alcohol), where protein is extracted to the organic phase, DNA resolves at the interface, and RNA remains in the aqueous phase. Therefore, RNA, DNA, and protein can be purified from a single sample (hence, the name TRIzol). TRIzol extraction is also an effective method for isolating small RNAs, such as microRNAs, piwi-associated RNAs, or endogeneous, small interfering RNAs. However, TRIzol is expensive and RNA pellets can be difficult to resuspend. Thus, the use of TRIzol is not recommend when regular phenol extraction is practical.
990 citations