77 research outputs found

    Occurrence of gangliosides in the common squid and pacific octopus among protostomia

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    AbstractAcidic lipids from tissues of the common squid Todarodes pacificus and the pacific octopus Octopus vulgaris were characterized. Hepatopancreatic tissues of both animals had complex compositions of resorcinol-positive acidic lipids, many of which became reactive with cholera toxin B subunit and anti-GM1 antibody after in situ treatment with sialidase on TLC. One of the major acidic lipids in squid tissue was isolated and examined for its structure. This acidic lipid was identified to be the ganglioside GD1a based upon the susceptibility to sialidases of different substrate specificity, characterization of reaction products, and electrospray ionization-mass spectrometry of the lipid. Hepatopancreatic tissues of squid and octopus also contained acidic lipids that reacted with A2B5, a monoclonal antibody specific to c-series gangliosides. Cerebral ganglia of both animals expressed resorcinol-positive acidic lipids, though their compositional patterns differed from the hepatopancreatic tissues. N-Acetylneuraminic acid was identified as the main species in lipid-bound sialic acid in both tissues. The contents of lipid-bound sialic acid in cerebral ganglia were significantly lower than those of hepatopancreatic tissues in both animals. The present study presents the first evidence for the occurrence of gangliosides in protostomia

    Effects of Tablet Size and Head Posture on Drug Swallowing: A Preliminary Examination Using Endoscopy in Healthy Subjects

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    Tablet size and head posture have been reported to affect swallowing of medications, but no previous studies have evaluated their effects in detail. Our aim was to investigate for the first time the effect of tablet size and head posture on drug swallowing by endoscopic evaluation in healthy subjects. Round tablets (7 , 10 , 12, and 14 mm in diameter) were swallowed by 15 healthy adults with an endoscope inserted in the neutral, head flex-ion, and head extension positions. Evaluation of swallowing difficulty using a numeric rating scale (NRS), presence or absence of pharyngeal residue and its location, and tablet oral transit time (TOTT) were recorded. In the neutral position, the NRS score was higher with the 14 mm tablets than with the 7 mm tablets. The TOTT with the 7 mm tablets was significantly shorter in the head extension than in the neutral position. Swallowing difficulty increased when the tablet diameter was more than 7 mm. Residues were found in the epi-glottis, pyriform sinus, and tongue base. These findings suggest that head extension shortens the TOTT and assists oral-pharyngeal transport

    Cerebellar Ataxia and Overactive Bladder after Encephalitis Affecting the Cerebellum

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    The cerebellum is one of the regions that contribute to urinary dysfunction in humans. A 43-year-old woman at age 35 had an acute onset of encephalitis that led to fever, generalized convulsion and coma. Six months after the disease onset, she regained consciousness and developed generalized myoclonus, cerebellar ataxia and overactive bladder, e.g., urinary urgency, daytime urinary frequency, and urinary incontinence. Eight years after the disease onset, she was revealed to have cerebellar atrophy on MRI, cerebellar hypoperfusion on SPECT, and detrusor overactivity on urodynamic study. Selective inflammation in the cerebellum seemed to produce cerebellar ataxia and overactive bladder in our case

    LAMPhimerus: a novel LAMP assay for detecting Amphimerus sp. DNA in human stool samples

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    [EN]Amphimeriasis, a fish-borne zoonotic disease caused by the liver fluke Amphimerus spp., is a highly prevalent parasitic infection affecting an indigenous Amerindian group, the Chachi, living in rural and remote tropical areas along the RõÂo Cayapas and its tributaries in the north-western coastal rainforest of Ecuador. Very little is known about the clinical course and treatment of this disease, and the only method for diagnosing it is the parasitological microscopic detection of eggs from Amphimerus spp. in patients' stool samples. This method lacks sensitivity, and the morphology of the eggs may be confounded with other liver and intestinal flukes. New diagnostic tools that can improve the sensitivity and specificity for diagnosing Amphimerus spp. infection would be desirable. At present, LAMP technology shows all the characteristics required of a real-time assay with simple operation for potential use in the clinical diagnosis of infectious diseases, particularly in the field conditions in developing countries for most neglected tropical diseases. In this study, we developed and successfully evaluated a LAMP assay for detecting Amphimerus sp. in human stool samples. After further validation, our LAMP assay (LAMPhimerus) could be readily adapted for effective field diagnosis and disease surveillance in amphimeriasis- endemic areas

    Four-electron reduction of dioxygen by a multicopper oxidase, CueO, and roles of Asp112 and Glu506 located adjacent to the trinuclear copper center

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    金沢大学理工研究域物質化学系The mechanism of the four-electron reduction of dioxygen by a multicopper oxidase, CueO, was studied based on reactions of single and double mutants with Cys500, a type I copper ligand, and the noncoordinating Asp112 and Glu506, which form hydrogen bonds with the trinuclear copper center directly and indirectly via a water molecule. The reaction of C500S containing a vacant type I copper center produced intermediate I in an EPR-silent peroxide-bound form. The formation of intermediate I from C500S/D112N was restricted due to a reduction in the affinity of the trinuclear copper center for dioxygen. The state of intermediate I was realized to be the resting form of C500S/E506Q and C500S of the truncated mutant Δα5-7CueO, in which the 50 amino acids covering the substrate-binding site were removed. Reactions of the recombinant CueO and E506Q afforded intermediate II, a fully oxidized form different from the resting one, with a very broad EPR signal, g < 2, detectable only at cryogenic temperatures and unsaturated with high power microwaves. The lifetime of intermediate II was prolonged by the mutation at Glu506 involved in the donation of protons. The structure of intermediates I and II and the mechanism of the four-electron reduction of dioxygen driven by Asp112 and Glu506 are discussed. © 2009 by The American Society for Biochemistry and Molecular Biology, Inc

    Minilaparotomy Approach for Giant Mutinous Cystadenoma of the Ovary in Children: Report of Two Cases

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    Mucinous cystadenomas (MCAs) are rare benign neoplasms in children. To the best of our knowledge, only 22 cases in children have been described. MCAs may reach huge sizes, and thus are not readily amenable to laparoscopic treatment due to the risk of rupture and the limited working space. We report two cases of giant MCA of the ovary treated by minilaparotomy. In case 1, a 12-year-old girl was admitted with abdominal pain and vomiting. Diagnostic imaging showed a large polycystic mass occupying nearly the whole abdominal cavity. With a provisional diagnosis of ovarian cyst, surgery was performed. The cyst was punctured under direct vision though a small subumbilical incision. After aspiration of 2,000ml of mucinous fluid, laparoscopic examination revealed a tumor originating from the left ovary. Left oophorectomy was performed through an 8-cm incision in the left lower abdomen. The histopathological diagnosis was MCA. In case 2, a 15-year-old girl presented with slowly increasing abdominal distension over 5 months. A polycystic mass measuring 36 × 21 × 14cm was evident on imaging. After drainage of 9,500ml of clear mucinous fluid, right oophorectomy was performed through a small (5cm) midline incision. The final pathology revealed a benign MCA. No recurrence has been detected for 2 years postoperatively in case 1 and for 6 years postoperatively in case 2

    Exportin 4 Interacts with Sox9 through the HMG Box and Inhibits the DNA Binding of Sox9

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    Sox9 is a transcription factor that is required for tissue development in mammals. In general, such transcription factors require co-regulators for precise temporal and spatial control of the activation and inactivation of the numerous genes necessary for precise development during embryogenesis. Here we identify a new Sox9 co-regulator: Using affinity chromatography with immobilized Sox9 protein, we identified exportin 4 (Exp4) as an interacting protein of Sox9 in human cultured cells. Interaction between endogenous Exp4 and Sox9 proteins was confirmed in the human osteosarcoma U2OS cells by immunoprecipitation experiments using anti-Sox9 antibody. siRNA depletion of Exp4 enhanced transcription of Sox9 target genes in U2OS cells, but did not affect nuclear localization of Sox9. These results suggest that Exp4 regulates Sox9 activity in the nucleus. Furthermore we found that the HMG box of Sox9 was responsible for binding to Exp4, and the HMG box was required for suppression of Sox9-mediated transcription. This contrasts with the known Sox9 co-regulators which bind to its transcriptional activation domain. Chromatin immunoprecipitation analyses revealed that Exp4 prevents Sox9 binding to the enhancers of its target genes. These results demonstrate that Exp4 acts as a Sox9 co-regulator that directly regulates binding of Sox9 to its target genes

    Genome-Wide Association Study Confirming Association of HLA-DP with Protection against Chronic Hepatitis B and Viral Clearance in Japanese and Korean

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    Hepatitis B virus (HBV) infection can lead to serious liver diseases, including liver cirrhosis (LC) and hepatocellular carcinoma (HCC); however, about 85–90% of infected individuals become inactive carriers with sustained biochemical remission and very low risk of LC or HCC. To identify host genetic factors contributing to HBV clearance, we conducted genome-wide association studies (GWAS) and replication analysis using samples from HBV carriers and spontaneously HBV-resolved Japanese and Korean individuals. Association analysis in the Japanese and Korean data identified the HLA-DPA1 and HLA-DPB1 genes with Pmeta = 1.89×10−12 for rs3077 and Pmeta = 9.69×10−10 for rs9277542. We also found that the HLA-DPA1 and HLA-DPB1 genes were significantly associated with protective effects against chronic hepatitis B (CHB) in Japanese, Korean and other Asian populations, including Chinese and Thai individuals (Pmeta = 4.40×10−19 for rs3077 and Pmeta = 1.28×10−15 for rs9277542). These results suggest that the associations between the HLA-DP locus and the protective effects against persistent HBV infection and with clearance of HBV were replicated widely in East Asian populations; however, there are no reports of GWAS in Caucasian or African populations. Based on the GWAS in this study, there were no significant SNPs associated with HCC development. To clarify the pathogenesis of CHB and the mechanisms of HBV clearance, further studies are necessary, including functional analyses of the HLA-DP molecule

    Visual Properties of Transgenic Rats Harboring the Channelrhodopsin-2 Gene Regulated by the Thy-1.2 Promoter

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    Channelrhodopsin-2 (ChR2), one of the archea-type rhodopsins from green algae, is a potentially useful optogenetic tool for restoring vision in patients with photoreceptor degeneration, such as retinitis pigmentosa. If the ChR2 gene is transferred to retinal ganglion cells (RGCs), which send visual information to the brain, the RGCs may be repurposed to act as photoreceptors. In this study, by using a transgenic rat expressing ChR2 specifically in the RGCs under the regulation of a Thy-1.2 promoter, we tested the possibility that direct photoactivation of RGCs could restore effective vision. Although the contrast sensitivities of the optomotor responses of transgenic rats were similar to those observed in the wild-type rats, they were enhanced for visual stimuli of low-spatial frequency after the degeneration of native photoreceptors. This result suggests that the visual signals derived from the ChR2-expressing RGCs were reinterpreted by the brain to form behavior-related vision
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