Biosynthesis and Characterization of an Anticoagulant Chitinase from Fermented Wheat Bran & Shrimp Shells’ Substratum

Solid state fermentation (SSF) of wheat bran coupled with shrimp shellfish waste Citrobacter freundii str. nov. haritD11 was optimized conventionally (112.43 U/gds) and statistically (124.73 U/gds). Chitinase was purified 4.24-fold with 31% yield and specific activity of 64.87 U/mg protein. The purified chitinase had a specific activity of 64.87 U/mg with optimal activity at pH 9 and temperature 45 ºC. The enzyme was stable at 8.0–9.5 pH range with 90% stability and between 45 °C – 60 °C for 1 hour. The Km value of the Citrobacter freundii haritD11 purified chitinase with swollen chitin (substrate) is 7.53 mg/mL with a Vmax of 2.27 mmol h−1mL−1. The purified chitinase was halotolerant showing maximum activity and stability up to 9% Sodium chloride, it also possessed potential antifungal and anticoagulant activity. This is the first report to date elucidating the production of halotolerant chitinase from wheat bran supplemented with shrimp shellfish waste using Citrobacter freundii haritD11 with notable tolerance to heavy metal ions, its application as an antifungal and anticoagulant agent.HIGHLIGHTS•Optimization of solid-state fermentation of Citrobacter freundii str. nov. haritD11.•The halotolerant chitinase was produced from wheat bran supplemented with shrimp shellfish waste.•The enzyme had notable tolerance to heavy metal ions.•Chitinase was purified 4.24-fold with 31% yield and specific activity of 64.87 U/mg.•The enzyme can be applied as an antifungal and anticoagulant agent

Pyrroloquinoline quinone increases the expression and activity of Sirt1 and -3 genes in HepG2 cells.

Sirtuin (Sirt) 1 and Sirt 3 are nicotinamide adenine dinucleotide ((+))-dependent protein deacetylases that are important to a number of mitochondrial-related functions; thus, identification of sirtuin activators is important. Herein, we hypothesize that pyrroloquinoline quinone (PQQ) can act as a Sirt1/Sirt3 activator. In HepG2 cell cultures, PQQ increased the expression of Sirt1 and Sirt3 gene, protein, and activity levels (P < .05). We also observed a significant increase in nicotinamide phosphoribosyltransferase gene expression (as early as 18 hours) and increased NAD(+) activity at 24 hours. In addition, targets of Sirt1 and Sirt3 (peroxisome proliferator-activated receptor γ coactivator 1α, nuclear respiratory factor 1 and 2, and mitochondrial transcription factor A) were increased at 48 hours. This is the first report that demonstrates PQQ as an activator of Sirt1 and Sirt3 expression and activity, making it an attractive therapeutic agent for the treatment of metabolic diseases and for healthy aging. Based on our study and the available data in vivo, PQQ has the potential to serve as a therapeutic nutraceutical, when enhancing mitochondrial function

Sales Management Portal

The use of the Web (World Wide Web) has had many positive effects on E commerce. It overcomes time and space limitations from traditional marketing. Consumer and customer are now using the Web to access vast amounts of information and resources in the cyberspace. Also, marketing via the Web enables both synchronous and asynchronous communication. Despite of many benefits of the Web. The model can also enhance the business of GSU Corporation is a world-renown provider of communication solutions. We implement our model and show that it can be applied for scales management as an instance. The Sales Management portal is web based application where the users will be able to make purchases of different sales based products available at the store. The web based application will be having three levels of access where each user will be having different menu options available for performing various levels tasks based on the level of authorization. The web based application will be dealing with the selling of the on-line based products and generating the sales information to create plans and deals for the future to gain more profits. The importance of the project lies in the generation of the sales data by which the vendors or the sellers can gain knowledge about the products and the likeliness to buy the product

Mono-(2-ethylhexyl) Phthalate Increases Oxidative Stress Responsive miRNAs in First Trimester Placental Cell Line HTR8/SVneo

Phthalates, an endocrine disruptor group, cause oxidative stress (OS) in the placenta. However, no studies have reported OS-related miRNAs induced by phthalates. In the present study, we demonstrate that mono-(2-ethylhexyl) phthalate (MEHP) induces OS responsive miR-17-5p, miR-155-5p, and miR-126-3p in HTR8/SVneo in a dose- and time-dependent manner. Furthermore, MEHP altered the expression of phosphoinositide-3-kinase regulatory subunit 1α, phosphatase and tensin homolog, CDKN2A interacting protein, superoxide dismutase 2, and 3β-hydroxysterol-D24 reductase, which are involved in OS and predicted to be regulated by these miRNAs. Our results suggest that placental exposure to MEHP may result in aberrant miRNA expression leading to pregnancy complications

Pyrroloquinoline quinone increases the expression and activity of Sirt1 and -3 genes in HepG2 cells.

Sirtuin (Sirt) 1 and Sirt 3 are nicotinamide adenine dinucleotide ((+))-dependent protein deacetylases that are important to a number of mitochondrial-related functions; thus, identification of sirtuin activators is important. Herein, we hypothesize that pyrroloquinoline quinone (PQQ) can act as a Sirt1/Sirt3 activator. In HepG2 cell cultures, PQQ increased the expression of Sirt1 and Sirt3 gene, protein, and activity levels (P < .05). We also observed a significant increase in nicotinamide phosphoribosyltransferase gene expression (as early as 18 hours) and increased NAD(+) activity at 24 hours. In addition, targets of Sirt1 and Sirt3 (peroxisome proliferator-activated receptor γ coactivator 1α, nuclear respiratory factor 1 and 2, and mitochondrial transcription factor A) were increased at 48 hours. This is the first report that demonstrates PQQ as an activator of Sirt1 and Sirt3 expression and activity, making it an attractive therapeutic agent for the treatment of metabolic diseases and for healthy aging. Based on our study and the available data in vivo, PQQ has the potential to serve as a therapeutic nutraceutical, when enhancing mitochondrial function

Effect of Chronic Western Diets on Non-Alcoholic Fatty Liver of Male Mice Modifying the PPAR-γ Pathway via miR-27b-5p Regulation

Western diets contribute to metabolic diseases. However, the effects of various diets and epigenetic mechanisms are mostly unknown. Here, six week-old C57BL/6J male and female mice were fed with a low-fat diet (LFD), high-fat diet (HFD), and high-fat high-fructose diet (HFD-HF) for 20 weeks. We determined that HFD-HF or HFD mice experienced significant metabolic dysregulation compared to the LFD. HFD-HF and HFD-fed male mice showed significantly increased body weight, liver size, and fasting glucose levels with downregulated PPARγ, SCD1, and FAS protein expression. In contrast, female mice were less affected by HFD and HFD-HF. As miR-27b contains a seed sequence in PPARγ, it was discovered that these changes are accompanied by male-specific upregulation of miR-27b-5p, which is even more pronounced in the HFD-HF group (p < 0.01 vs. LFD) compared to the HFD group (p < 0.05 vs. LFD). Other miR-27 subtypes were increased but not significantly. HFD-HF showed insignificant changes in fibrosis markers when compared to LFD. Interestingly, fat ballooning in hepatocytes was increased in HFD-fed mice compared to HFD-HF fed mice, however, the HFD-HF liver showed an increase in the number of small cells. Here, we concluded that chronic Western diet-composition administered for 20 weeks may surpass the non-alcoholic fatty liver (NAFL) stage but may be at an intermediate stage between fatty liver and fibrosis via miR-27b-5p-induced PPARγ downregulation

Regulation of Adipocyte Differentiation by the Zinc Finger Protein ZNF638*

Zinc finger proteins constitute the largest family of transcription regulators in eukaryotes. These factors are involved in diverse processes in many tissues, including development and differentiation. We report here the characterization of the zinc finger protein ZNF638 as a novel regulator of adipogenesis. ZNF638 is induced early during adipocyte differentiation. Ectopic expression of ZNF638 increases adipogenesis in vitro, whereas its knockdown inhibits differentiation and decreases the expression of adipocyte-specific genes. ZNF638 physically interacts and transcriptionally cooperates with CCAAT/enhancer-binding protein (C/EBP) β and C/EBPδ. This interaction leads to the expression of peroxisome proliferator-activated receptor γ, which is the key regulator of adipocyte differentiation. In summary, ZNF638 is a novel and early regulator of adipogenesis that works as a transcription cofactor of C/EBPs