Neuromedin B stimulates proliferation of mouse chondrogenic cell line ATDC5

Neuromedin B (NMB), which was originally isolated from porcine spinal cord, is a mammalian bombesin-related peptide that exerts various physiological effects. Previously, we observed expression of NMB in rib cartilage from chicken. Here, we report the initial attempt to elucidate the role of NMB in cartilage. We used RT-PCR to measure the expression of NMB and its receptor (NMB-R) in mouse chondrogenic cell line ATDC5. During chondrogenic differentiation of ATDC5 cells, NMB mRNA transiently increased on day 4 and then decreased on day 14, whereas NMB-R mRNA decreased on days 7 and 14. We also characterized immunoreactive NMB in ATDC5 culture medium using a combination of specific radioimmunoassay (RIA) and reverse phase-high performance liquid chromatography (RP-HPLC). Furthermore, using the WST-8 assay, we demonstrated that NMB significantly induced ATDC5 proliferation; this was inhibited by NMB-R antagonist, BIM 23127. These results implicate that NMB is involved in cartilage development, either in an autocrine or paracrine manner.     

Quercetin offers cardioprotection against progression of experimental autoimmune myocarditis by suppression of oxidative and endoplasmic reticulum stress via endothelin-1/MAPK signalling

In order to test the hypothesis that treatment with quercetin at a dose of 10 mg/kg protects from the progression of experimental autoimmune myocarditis (EAM) to dilated cardiomyopathy (DCM), we have used the rat model of EAM induced by porcine cardiac myosin. Our results identified that the post-myocarditis rats suffered from elevated endoplasmic reticulum (ER) stress and adverse cardiac remodelling in the form of myocardial fibrosis, whereas the rats treated with quercetin have been protected from these changes as evidenced by the decreased myocardial levels of ER stress and fibrosis markers when compared with the vehicle-treated DCM rats. In addition, the myocardial dimensions and cardiac function were preserved significantly in the quercetin-treated rats in comparison with the DCM rats treated with vehicle alone. Interestingly, the rats treated with quercetin showed significant suppression of the myocardial endothelin-1 and also the mitogen activated protein kinases (MAPK) suggesting that the protection offered by quercetin treatment against progression of EAM involves the modulation of MAPK signalling cascade. Collectively, the present study provides data to support the role of quercetin in protecting the hearts of the rats with post myocarditis DCM.     

Index finger position fluctuations reflect multi-muscle coordination

We hypothesized that movement fluctuations in the index finger reflect the integrated result of the coordination of multiple muscles because index finger movements are determined by the cooperation of multiple muscles spanning the metacarpophalangeal (MCP) joint. To evaluate this hypothesis, the aim of the present study was to examine the fluctuations of the index finger in abduction-adduction and extension-flexion directions during a position-holding task using two laser displacement sensors. Eleven healthy men maintained their index finger position while supporting a load at 5% of the maximal voluntary contraction force. To maintain the position of the index finger, displacement of the index finger in the abduction-adduction and extension-flexion directions was measured from a distance with two laser displacement sensors that were positioned to the lateral side of and above the index finger. The index finger movements fluctuated around the target position in not only the abduction-adduction direction but also the extension-flexion direction. The path length of finger displacement and the standard deviation of finger acceleration were significantly greater in the extension-flexion direction than in the abduction-adduction direction. These results suggest that the index finger movements quantified by two laser displacement sensors reflect the coordination of multiple muscles spanning the MCP joint.     

Primary culture of human fallopian tube epithelial cells and co-culture of early mouse pre-embryos.

We have established a monolayer culture system for human fallopian tube epithelial cells. The cells were isolated from tubes using collagenase digestion, and were cultured in Ham's F-10 supplemented with 15% fetal calf serum. The epithelial cells derived from culture were characterized using immunocytochemical staining and electron microscopy. These cells were stained with antikeratin and anti-epithelial membrane antigen, but showed no staining after treatment with antivimentin. Electron microscopy showed many microvilli on the cell surface and tight junctions or desmosomes at areas of cell-cell contact. Cell proliferation was enhanced by epidermal growth factor, but not by fibroblast growth factor, insulin, transferrin, estradiol-17 beta, or progesterone. The 2-cell ICR mouse pre-embryos were co-cultured for 4 days with tubal epithelial cells (A) (n = 98), in cell-conditioned medium (B) (n = 83), or in medium alone (C) (n = 72). During the first 24 h in culture, for groups A and B, the rates of cleavage to the 4-cell stage were 90.9% and 81.9%, respectively. Cleavage rates in these two groups were significantly higher (P = 0.0012, P less than 0.00001) than in group C (56.9%). After 72 h in culture, the rates of development to the blastocyst stage were significantly higher for groups A and B compared to group C (89.6% and 73.5% vs. 54.5%, P less than 0.00001, P = 0.0002). These results suggest that factor(s) from tubal epithelial cells may facilitate the development of mouse pre-embryos throughout the pre-implantation stages.     

Biochemical characterization and tissue distribution of hamster complement C1s

Several mAb (PG11, NG7, and ED4) against hamster complement C1s were obtained. PG11 and NG7 were shown to cross-react with human and rat C1s. By using an immunohistochemical method, we examined localization of C1s in tissues of hamsters and rats. Present results revealed a widespread yet specific staining of hamster C1s which is associated with endoderm-, mesoderm-, and neuroectoderm-derived cells. For example, chondrocyte of hyaline cartilage and surface epithelium of the stomach were strongly positive. Intestinal epithelium, muscle cells, pia mater and epithelium of the choroid plexus of the ventricle, and hepatocytes were also stained. The synthesis of hamster C1s in these organs was confirmed by RNA blot hybridization. Secretion of C1s into the culture medium was revealed by immunoblot analysis in cell lines of hepatocytes, kidney cells, and myoblasts of rat or hamster.     

A gene structure of testosterone 6 beta-hydroxylase (P450IIIA)

Genomic clones of a rat testosterone 6 beta-hydroxylase have been isolated and characterized as the first gene (P450/6 beta A) among P450IIIA subfamily. This gene spans about 25Kb and consists of 13 exons, which is the largest number of exons among cytochrome P-450 genes reported previously. The nucleotide sequence of the exon region showed high similarity to those of P450PCN2 and P450PCN1 cDNA (Gonzalez, F.J. et al. (1987) Mol. Cell. Biol. 2969-2974), but several replacements and deletions of nucleotide were found between the P450/6 beta A gene and both cDNAs, indicating the existence of multiple P450IIIA genes in rats.     

Disrupted-in-schizophrenia 1 (DISC1) Regulates Dysbindin Function by Enhancing Its Stability

Dysbindin and DISC1 are schizophrenia susceptibility factors playing roles in neuronal development. Here we show that the physical interaction between dysbindin and DISC1 is critical for the stability of dysbindin and for the process of neurite outgrowth. We found that DISC1 forms a complex with dysbindin and increases its stability in association with a reduction in ubiquitylation. Furthermore, knockdown of DISC1 or expression of a deletion mutant, DISC1 lacking amino acid residues 403–504 of DISC1 (DISC1^Δ403–504), effectively decreased levels of endogenous dysbindin. Finally, the neurite outgrowth defect induced by knockdown of DISC1 was partially reversed by coexpression of dysbindin. Taken together, these results indicate that dysbindin and DISC1 form a physiologically functional complex that is essential for normal neurite outgrowth.     

Early intervention to promote oral feeding in patients with intracerebral hemorrhage: a retrospective cohort study

Background  

Stroke is a major cause of dysphagia, but little is known about when and how dysphagic patients should be fed and treated after an acute stroke. The purpose of this study is to establish the feasibility, risks and clinical outcomes of early intensive oral care and a new speech and language therapist/nurse led structured policy for oral feeding in patients with an acute intracerebral hemorrhage (ICH).     

Administration of D-aspartate increases D-aspartate oxidase activity in mouse liver

Oral administration of D-aspartate to mice for 2 weeks by addition of the amino acid to drinking water produced a nearly 4-fold increase in liver D-aspartate oxidase (EC 1.4.3.1) activity, whereas no increase was induced by L-aspartate administered in the same way. Administration of D-aspartate also produced a small significant increase in the kidney enzyme activity, but L-aspartate administration increased the activity as well. The enzyme activity in the brain and muscle was not affected by administration of either D- or L-aspartate. Intraperitoneal administration of D-aspartate increased the enzyme activity only in the liver, and other compounds tested, including D-glutamate and D-alanine, could not replace D-aspartate. The results indicate a specific relationship between D-aspartate and D-aspartate oxidase and suggest that the amino acid is, in fact, a physiological substrate of the enzyme.     

Ab initio molecular orbital study of the interaction of Li, Na and K with the pore components of ion channels: Consideration of the size, structure and selectivity of the pore of the channels

Ab initio molecular orbital calculations were made for the various types of structures of the pore of the ion channel and the results were applied to the permeability model by Hille, an extension of the Eyring rate theory. In Hille's model, ion passage through the channel is regarded as a kinetic process. Accordingly, it is thought that the interaction energy between cation and ligand, the easier the passage is, due to the lower activation energy. The calculated interaction energy was in the order Li+ greater than Na+ greater than K+ for all models. The optimum size of the pore determined from the interaction energy depends on the structure of the filter. The size for the pentagon was largest, followed by the hexagon and tetragon. On the other hand, the size depends hardly at all on the kind of ligand molecules. In the case of the tetragon, the sizes for the Na and K channels were nearly the same as those estimated from the model building and inhibitor-blocking experiment. The interaction energy between the ionized carboxyl group and the cation was extremely large, clearly reflecting the experimental fact that the carboxyl group in the pore has an important role in making the passage of the cation through the channel easier by dehydrating the water molecules. By analysis of the interaction energy, it was revealed that the contribution of the electrostatic energy was predominant, although the contributions of the other effects might not be negligible. Among these effects, the value of the charge transfer energy is largest, and this is noteworthy in connection with the selective transmission of cations through the overlap of orbitals. It is concluded that the quantum-chemical indices such as interaction energy and the electronic charge calculated by the sophisticated ab initio method help to shed light on the nature of the pore of the ion channel.