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We examined the effects of treatment with pulsed electromagnetic fields (PEMFs) on cumulus cells and buffalo somatic cell nuclear transfer (SCNT) embryos. PEMF treatment (30 μT for 3 hours) of cumulus cells increased (p < 0.05) the relative cell viability and cell proliferation and the expression level of OCT4, NANOG, SOX2, P53, CCNB1, and GPX, but decreased (p < 0.05) that of DNMT1, DNMT3a, GSK3b, and BAX, whereas the expression level of DNMT3b, GLUT1, BCL2, CASPASE3, SOD1, and CATALASE was not affected. PEMF treatment of SCNT embryos at the beginning of in vitro culture increased (p < 0.05) the blastocyst rate (51.4% ± 1.36% vs. 42.8% ± 1.29%) and decreased (p < 0.01) the apoptotic index to the level in in vitro fertilization blastocysts, but did not significantly alter the total cell number and the inner cell mass:trophectoderm cell number ratio of blastocysts compared to the controls. PEMF treatment increased the expression level of NANOG, SOX2, CDX2, GLUT1, P53, and BCL2 and decreased that of BAX, CASPASE3, GSK3b, and HSP70, but not OCT4, DNMT1, DNMT3a, DNMT3b, HDAC1, and CCNB1 in blastocysts. It increased (p < 0.001) the global level of H3K27me3 but not H3K18ac. These results suggest that PEMF treatment of SCNT embryos improves their developmental competence, reduces the level of apoptosis, and alters the expression level of several important genes related to pluripotency, apoptosis, metabolism, and stress.

Skeletal muscle atrophy is debilitating consequence of a large number of chronic disease states, aging, and disuse conditions. Skeletal muscle mass is regulated through coordinated activation of a number of signaling cascades. Transforming growth factor‐β activated kinase 1 (TAK1) is a central kinase that mediates the activation of multiple signaling pathways in response to various growth factors, cytokines, and microbial products. Accumulating evidence suggests that TAK1 promotes skeletal muscle growth and essential for the maintenance of muscle mass in adults. Targeted inactivation of TAK1 leads to severe muscle wasting and kyphosis in mice. However, the mechanisms by which TAK1 prevents loss of muscle mass remain poorly understood. Through generation of inducible skeletal muscle‐specific Tak1‐knockout mice, we demonstrate that targeted ablation of TAK1 disrupts redox signaling leading to the accumulation of reactive oxygen species and loss of skeletal muscle mass and contractile function. Suppression of oxidative stress using Trolox improves muscle contractile function and inhibits the activation of catabolic signaling pathways in Tak1‐deficient muscle. Moreover, Trolox inhibits the activation of ubiquitin‐proteasome system and autophagy markers in skeletal muscle of Tak1‐deficient mice. Furthermore, inhibition of oxidative stress using Trolox prevents the slow‐to‐fast type fiber transition and improves mitochondrial respiration in skeletal muscle of Tak1‐deficient mice. Overall, our results demonstrate that TAK1 maintains skeletal muscle mass and health through redox homeostasis.

Skeletal muscle wasting causes both morbidity and mortality of cancer patients. Accumulating evidence suggests that the markers of endoplasmic reticulum (ER) stress and unfolded protein response (UPR) pathways are increased in skeletal muscle under multiple catabolic conditions, including cancer. However, the signaling mechanisms and the role of individual arms of the UPR in the regulation of skeletal muscle mass remain largely unknown. In the present study, we demonstrated that gene expression of Toll-like receptors (TLRs) and myeloid differentiation primary response gene 88 (MyD88) was increased in skeletal muscle in a Lewis lung carcinoma (LLC) model of cancer cachexia. Targeted ablation of MyD88 inhibits the loss of skeletal muscle mass and strength in LLC tumor-bearing mice. Inhibition of MyD88 attenuates the LLC-induced activation of the UPR in skeletal muscle of mice. Moreover, muscle-specific deletion of X-box binding protein 1 (XBP1), a major downstream target of IRE1α arm of the UPR, ameliorates muscle wasting in LLC tumor-bearing mice. Our results also demonstrate that overexpression of an active form of XBP1 caused atrophy in cultured myotubes. In contrast, knockdown of XBP1 inhibits myotube atrophy in response to LLC or C26 adenocarcinoma cell conditioned medium. Collectively, our results demonstrate that TLR/MyD88-mediated activation of XBP1 causes skeletal muscle wasting in LLC tumor-bearing mice.

The purpose of this study was to determine the effect of oviduct specific proteins as a media supplement for in vitro embryo development in cattle. The proteins were extracted from oviducts of cows and precipitated by ammonium sulfate (30%, 40%, 50% and 60%) followed by dialysis in 50 mM Tris–HCl (pH 7.0) buffer. The dialyzed proteins were fractionated into acidic, basic and neutral fractions using SP sephadex cation exchange and DEAE sephadex anion exchange column chromatography respectively. Cow oviduct specific proteins (cOSPs) constituting all the extracted proteins were used as media supplement in three different concentrations (10, 50 and 100 μg/ml) for in vitro maturation, fertilization and culture (IVMFC) of cow oocytes. Acidic, basic and neutral (unbound) fractions were also used as media supplement in three different concentrations (10, 30 and 50 μg/ml) for IVMFC. Cumulus oocytes complexes were collected from slaughterhouse ovaries, washed thoroughly and cultured in maturation media for 24 h in 5% CO 2 at 38.5 °C with maximum humidity. In vitro matured oocytes were co-incubated with in vitro capacitated sperm in Fert-BO media at 38.5 °C for 18 h in 5% CO 2 . The fertilized oocytes were washed and cultured in embryo development media for cleavage. After 40–42 h cleavage was observed and embryos were put in the replacement media for further development. The cleavage rates (%) for cOSPs were observed as 68.24±2.46, 69.28±2.05, 61.77±0.93 and 42.62±1.31 at concentrations of 0, 10, 50 and 100 μg/ml respectively. Rates of blastocyst stage development were 14.49±3.61, 21.17±2.77, 14.66±1.06 and 11.98±1.84. These results indicate that addition of cOSP at10 μg/ml increased blastocyst formation as compared to other concentrations (0, 50 and 100 μg/ml). Although acidic, basic and neutral fractions seemed to have no major effect on cleavage rate, but both acidic and neutral fraction of oviduct specific proteins improved the cleavage rate at 30 μg/ml concentration and basic fraction improved the blastocyst formation at 10 μg/ml concentration.

Protein phosphatases play vital roles in phosphorylation-mediated cellular signaling. Although there are 11 serine/threonine protein kinases in Mycobacterium tuberculosis, only one serine/threonine phosphatase, PstP, has been identified. Although PstP has been biochemically characterized and multiple in vitro substrates have been identified, its physiological role has not yet been elucidated. In this study, we have investigated the impact of PstP on cell growth and survival of the pathogen in the host. Overexpression of PstP led to elongated cells and partially compromised survival. We find that depletion of PstP is detrimental to cell survival, eventually leading to cell death. PstP depletion results in elongated multiseptate cells, suggesting a role for PstP in regulating cell division events. Complementation experiments performed with PstP deletion mutants revealed marginally compromised survival, suggesting that all of the domains, including the extracellular domain, are necessary for complete rescue. On the other hand, the catalytic activity of PstP is absolutely essential for the in vitro growth. Mice infection experiments establish a definitive role for PstP in pathogen survival within the host. Depletion of PstP from established infections causes pathogen clearance, indicating that the continued presence of PstP is necessary for pathogen survival. Taken together, our data suggest an important role for PstP in establishing and maintaining infection, possibly via the modulation of cell division events.

Polyhydroxyalkonoates (PHAs) are biological in origin, organic polyesters comprising the industrial and biomedical interest. This chapter summarizes the current advances, applications, limitations, and challenges of biopolymers in medicine. Biopolymers not only substitute the existing polymers, but novel combinations of diverse PHAs broaden the applicability and utility. The PHA-based implants are new dimensions of future in biomedical engineering.

Biofilms are bacteria’s way of behaving like a multi-cellular organism. Bacteria have been constantly evolving in the face of myriad natural challenges since the first life form appeared. Over the centuries, they have survived under extreme conditions by virtue of their ability to form biofilms. Biofilms have proved to be an immensely strong collaborative effort of bacteria, and this interaction is administered via their quorum-sensing mechanism. A biofilm plays a crucial role in survival, dispersal, transfer of resistance genes, and generation of diversity among bacteria. It can also act as a pathogenic factor for virulent bacteria and biofilms are often listed as the major cause of many diseases, such as endocarditis, cystic fibrosis, etc. Bacterial biofilms can be formed on almost every surface and, thus, have deleterious effects on many indwelling medical devices and industrial equipment. Many methods—from antibiotics to ultraviolet (UV) radiation—are currently being used to eliminate or reduce biofilm. However, the most effective and eco-friendly measure involves the targeting of the root phenomenon, quorum sensing. Biofilm formation and dispersal mechanisms are being studied to increase the efficiency of biofilm elimination. Despite the many harms they can pose, these biofilms have been efficiently manipulated to be used for various purposes such as wastewater treatment, microbial fuel cells, drug delivery, nanobiotechnology, etc. Biofilms bring about a very detailed level of complexity that helps for better persistence of the bacterial population and at the same time, provides us a valuable tool to address several important environmental issues. Thus, it will be appropriate to term bacterial biofilms as remarkably proficient assemblies of the life forms.

The objective of this study was to see the effect of purified heparin binding oviduct specific proteins (OSP) as media supplement on in vitro embryo developmental competence in cattle. The oviduct specific proteins were isolated from abattoir cattle oviducts and precipitated, dialyzed and at the end purified by high performance liquid chromatography system. The SDS-PAGE profile of eluted heparin binding protein (HBP) fraction showed bands between approximately 66-approximately 97 kDa, while heparin unbinding protein (HUBP) fraction showed two bands at approximately 66 kDa and in total protein (TP) bands were approximately 60-approximately 95 kDa. Collected all three OSP fractions were used as a media supplement in three different concentrations (0, 5 and 20 microg/mL) for in vitro maturation of immature oocytes, in vitro fertilization and culture of presumptive embryos at 38.5 degrees C in 5% CO2 incubator with maximum humidity. The highest cleavage rate (73.40 +/- 2.36%) was observed at 5 microg/mL concentration level and lowest cleavage rate (27.63 +/- 1.89%) was obtained in 20 microg/mL total protein (TP) fraction. The highest blastocyst formation (26.47 +/- 1.47%) also occurred in 5 microg/mL concentration of total protein (TP) fraction and the lowest blastocyst rate (3.60 +/- 1.80%) was achieved at 20 microg/mL HBP fraction. The highest cleavage rate in the control group was 60.45 +/- 2.66% in TP fraction and blastocyst formation was 11.66 +/- 2.54% in HUBP fraction which was not significantly differ from HBP fraction. These results indicate that at 5 microg/mL of total OSP fraction (TP) and HBP used as media supplement increased the cleavage rate significantly as compared to HUBP fraction, and total OSP fraction (TP) increased blastocyst formation significantly (P0.05) as compared to HBPHUBP fraction.