Melatonin mitigates the adverse effect of hypoxia during myocardial differentiation in mouse embryonic stem cells

BackgroundHypoxia causes oxidative stress and affects cardiovascular function and the programming of cardiovascular disease. Melatonin promotes antioxidant enzymes such as superoxide dismutase, glutathione reductase, glutathione peroxidase, and catalase.ObjectivesThis study aims to investigate the correlation between melatonin and hypoxia induction in cardiomyocytes differentiation.MethodsMouse embryonic stem cells (mESCs) were induced to myocardial differentiation. To demonstrate the influence of melatonin under hypoxia, mESC was pretreated with melatonin and then cultured in hypoxic condition. The cardiac beating ratio of the mESC-derived cardiomyocytes, mRNA and protein expression levels were investigated.ResultsUnder hypoxic condition, the mRNA expression of cardiac-lineage markers (Brachyury, Tbx20, and cTn1) and melatonin receptor (Mtnr1a) was reduced. The mRNA expression of cTn1 and the beating ratio of mESCs increased when melatonin was treated simultaneously with hypoxia, compared to when only exposed to hypoxia. Hypoxia-inducible factor (HIF)-1α protein decreased with melatonin treatment under hypoxia, and Mtnr1a mRNA expression increased. When the cells were exposed to hypoxia with melatonin treatment, the protein expressions of phospho-extracellular signal-related kinase (p-ERK) and Bcl-2-associated X proteins (Bax) decreased, however, the levels of phospho-protein kinase B (p-Akt), phosphatidylinositol 3-kinase (PI3K), B-cell lymphoma 2 (Bcl-2) proteins, and antioxidant enzymes including Cu/Zn-SOD, Mn-SOD, and catalase were increased. Competitive melatonin receptor antagonist luzindole blocked the melatonin-induced effects.ConclusionsThis study demonstrates that hypoxia inhibits cardiomyocytes differentiation and melatonin partially mitigates the adverse effect of hypoxia in myocardial differentiation by regulating apoptosis and oxidative stress through the p-AKT and PI3K pathway.     

Endless variety for bovine virus diarrhea viruses: new members of a novel subgroup into Pestivirus A from Turkey

Tropical Animal Health and Production - As ubiquitous pathogens, bovine virus diarrhea viruses (BVDVs) in cattle have been reported several times in Turkey. Over time, the frequency and importance...     

Two different types of malignant fibrous histiocytomas from pet dogs

We describe 2 cases of malignant fibrous histiocytomas (MFHs) that spontaneously developed in young pet dogs. To classify these tumors, we applied a panel of antibodies (vimentin, desmin, α-SMA, and ED1) and Azan staining for collagen. The MFHs were most consistent with osteoclast-like giant and inflammatory cell types. The first case had positive staining for ED1 and vimentin, and given the osteoclast-like giant cells, calcification sites accompanying peripheral giant cell infiltrates. The latter case, the inflammatory cell type, exhibited a storiform-pleomorphic variant of neoplastic cells, including an ossifying matrix. MFHs are among the most highly aggressive tumors occurring in soft tissue sarcomas in elderly dogs; however, MFHs have been poorly studied from a diagnostic point of view. Herein, we describe the histologic and immunohistologic features of MFHs in detail, thus classifying the subtypes of these tumors.     

Development of gates to measure the immature platelet fraction in C57BL/6J mice using the Sysmex XN-V series multispecies hematology analyzer

The immature platelet fraction (IPF) is a measure of newly released platelets, which has been used as a marker of platelet production in multiple human studies but is not widely available in multispecies analyzers. We developed gates to measure the IPF in diluted and undiluted murine blood samples on the Sysmex XN-1000V multispecies hematology analyzer. IPF gates were created using undiluted and diluted (1/10) blood samples obtained from adult and newborn (postnatal day 10, P10) C57BL/6J wild-type (WT) mice, and from 3 murine models of thrombocytopenia: c-MPL^−/− mice, which lack the thrombopoietin receptor (hyporegenerative); antibody-mediated thrombocytopenia; and acute inflammation-induced thrombocytopenia. P10 mice were chosen because, at their size, we could consistently obtain (by terminal phlebotomy) the blood volume needed to run an undiluted sample. The undiluted blood IPF gate successfully differentiated between mechanisms of thrombocytopenia in both adult and P10 mice. For diluted samples, 2 IPF gates were generated: a thrombocytopenic (T) gate, which performed well in samples with platelet counts (PCs) <800 × 10⁹/L in adult mice and <500 × 10⁹/L in newborn mice, and a non-thrombocytopenic (NT) gate, which performed well in samples with PCs above these thresholds. PCs and IPFs measured in diluted blood using these gates agreed well with those measured in undiluted blood and had good reproducibility. These diluted gates allow for the accurate measurement of PCs and IPFs in small (10 µL) blood volumes, which can be obtained easily from adult and newborn mice as small as P1 to assess platelet production serially.     

Stiffness of normal, opposite, and tension poplar wood determined using micro-samples in the three material directions

Mechanical tests on micro-samples were performed in the three material directions in normal, opposite, and tension wood collected from a poplar tree. Two custom micro-devices were designed and built in the laboratory to test samples under pure tension in the transverse direction and under 4-point bending conditions in the longitudinal direction. Both devices were designed to handle samples with a small transverse section (a few square mm), which allowed to select zones with homogenous anatomical features. The results indicate a very high longitudinal stiffness in tension wood (up to 35 GPa compared to an average of 18 GPa for normal wood). Considering wood density, the value represents a specific modulus that is nearly 70 % crystalline cellulose. However, tension wood is slightly less stiff in the tangential and radial directions (1,150 vs. 1,500 MPa for normal wood in the radial direction and 430 vs. 530 MPa in the tangential direction).     

End-capped polyethylene terephthalate having superior carbonaceous char yield

End-capping of PET with triphenyl phosphate (TPP), bisphenol A bis(diphenyl phosphate) (BDP), or resorcinol bis(diphenyl phosphate) (RDP) as phosphorous containing flame retardants was performed by using benzoyl peroxide as initiator. ³¹P NMR and FT-IR spectroscopy were used to evaluate end-capping of PET and structural changes of specimens. From thermal characteristics evaluated by differential scanning calorimetry and thermogravimetric analysis, BDP and RDP end-capped PETs show flame retarding mechanisms similar to those of blended counterparts. However, TPP end-capped PET presented superior carbonaceous char yields of up to 46.7 %, which was attributed to the condensed-phase action of TPP anchored at the ends of PET.     

Use of canine small intestinal submucosa allograft for treating perineal hernias in two dogs

Here, we describe two dogs in which canine small intestinal submucosa (SIS) was implanted as a biomaterial scaffold during perineal herniorrhaphy. Both dogs had developed severe muscle weakness, unilaterally herniated rectal protrusions, and heart problems with potential anesthetic risks. Areas affected by the perineal hernia (PH) located between the internal obturator and external anal sphincter muscles were reconstructed with naïve canine SIS sheets. In 12 months, post-operative complications such as wound infections, sciatic paralysis, rectal prolapse, or recurrence of the hernia were not observed. Symptoms of defecatory tenesmus also improved. Neither case showed any signs of rejection or specific immune responses as determined by complete and differential cell counts. Our findings demonstrate that canine SIS can be used as a biomaterial scaffold for PH repair in dogs.     

Profiles of Fatty Acids in Different Bone Structures of Growing Chicks

This study was concerned with the dynamics of fatty acids in tissues involved in the growth and mineralization of the femur of Starbro chicks from 1 to 50 days of age. Four chickens that died of suffocation were obtained each day. Altogether, 400 femoral bones were studied in five age groups (I–V). Compact bone, spongy bone and articular cartilage were sampled. Lipids were extracted according to Folch and fatty acids were separated using gas chromatography. The fatty acid profile was found to change with age. Fatty acids with the highest content in bone were C_18:1, C_16:0, C_18:2 and C_18:0. The highest content of fatty acids was found in spongy bone and the lowest in articular cartilage. Several correlations were revealed between individual fatty acids. The following conclusions were drawn. (1) Fatty acid profiles in compact bone, spongy bone and articular cartilage change with age of chicks. (2) Oleic (C_18:1), palmitic (C_16:0), linoleic (C_18:2), stearic (C_18:0) and arachidonic (C_20:4) acids accounted for most of the fatty acid pool. (3) Correlations in the content of fatty acids were noted between bone structures.     

Pathogenicity of Vietnamese enterotoxigenic Escherichia coli strains in colostrum-deprived one-day-old piglets

Preweaning colibacillosis is a major cause of economic loss to the swine industry in Vietnam. The aim of this study was to examine the enteropathogenicity of representative enterotoxigenic Escherichia coli (ETEC) strains obtained during an earlier epidemiologic survey conducted in five provinces in North Vietnam. This included isolates belonging to serotype O8 that produced heat-stable and heat-labile enterotoxins but did not produce any of the recognized fimbriae (F4, F5, F6, F41, F18). In vitro hemagglutination (unique mannose-resistant hemagglutination activity with guinea pig, sheep, human, and chicken red blood cells at 37 degrees C, but not at 18 degrees C) and enterocyte brush border attachment assays suggested that the F- ETEC strains produced an unidentified colonization factor that promoted adherence to the intestinal epithelium. Colostrum-deprived 1-day-old piglets challenged with an F- strain (1-2 x 10(9) bacteria) developed acute watery diarrhea within 4 hours of inoculation and suffered up to 20% weight loss, with comparable severity to piglets challenged with conventional F4 and F5 strains. At necropsy, viable counts and histopathologic examination of intestinal sections demonstrated colonization of the duodenum, jejunum, and ileum by F4-positive strains. In comparison, the F- and F5-positive strains attached exclusively to the ileum. Transmission electron micrographs of negatively stained F- cells grown at 37 degrees C demonstrated the presence of fimbriae. These results confirm the presence of a potentially new pathogenic ETEC fimbrial type in piggeries in Vietnam, with a unique hemagglutination property and attachment characteristics similar to ETEC bearing F5 fimbriae.