Cardiomyopathy (CM) is a primary degenerative disease of myocardium and is traditionally categorized into hypertrophic and dilated CMs (HCM and DCM) according to its gross appearance. Cardiomyopathic hamster (CM hamster), a representative model of human hereditary CM, has HCM and DCM inbred sublines, both of which descend from the same ancestor. Herein we show that both HCM and DCM hamsters share a common defect in a gene for ␦-sarcoglycan (␦-SG), the functional role of which is yet to be characterized. A breakpoint causing genomic deletion was found to be located at 6.1 kb 5 upstream of the second exon of ␦-SG gene, and its 5 upstream region of more than 27.4 kb, including the authentic first exon of ␦-SG gene, was deleted. This deletion included the major transcription initiation site, resulting in a deficiency of ␦-SG transcripts with the consequent loss of ␦-SG protein in all the CM hamsters, despite the fact that the protein coding region of ␦-SG starting from the second exon was conserved in all the CM hamsters. We elucidated the molecular interaction of dystrophin-associated glycoproteins including ␦-SG, by using an in vitro pull-down study and ligand overlay assay, which indicates the functional role of ␦-SG in stabilizing sarcolemma. The present study not only identifies CM hamster as a valuable animal model for studying the function of ␦-SG in vivo but also provides a genetic target for diagnosis and treatment of human CM.Cardiomyopathy (CM) manifests dyspnea, cardiac failure, or sudden death, causing serious morbidity and mortality. Clinical features and molecular genetic studies of CM demonstrate a wide variety of possible genetic causes of this disease but the causative genes and pathogenesis are poorly understood (1-3). Medical treatment for this progressive disease are only palliative with poor prognosis. Syrian hamsters with CM are known to inherit both CM and muscular dystrophy as an autosomal recessive trait but the genetic cause still remains to be elucidated (4-6). Recent studies on muscular dystrophy revealed the genetic importance of sarcoglycans (SGs), a subcomplex of dystrophin-associated glycoprotein complex (DAGC), in this disease (7-10).Distinct sublines of Syrian hamster manifesting hypertrophic CM (HCM; BIO 14.6 and its descendant UMX7.1) or dilated CM (DCM; TO-2) have been established from the original line BIO1.50 (5, 6). We have reported to the DDBJ (DNA Data Base of Japan) that no mutation exists in the coding regions of cDNAs of BIO14.6 for ␣-, ␤-, or ␥-SGs, all of which are lost in cardiac and skeletal muscles of this animal, where dystrophin is normally expressed (11). Our latest study revealed that these SGs are also deficient in UMX7.1 and TO-2 (vide infra), suggesting a hypothesis that both HCM and DCM share the loss of SG subcomplex as a common causative feature in hamster. In addition, ␦-SG, which was identified recently, seemed to constitute DAGC together with ␣-, ␤-, and ␥-SGs (12). These facts prompted us to identify the causative gene common to HCM and DCM wit...

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A novel protein-encapsulation technique using sol-gels was developed for the preparation of monolithic capillary columns for capillary electrochromatography. Two chiral compounds, bovine serum albumin (BSA) and ovomucoid (OVM) from chicken egg white, were encapsulated in tetramethoxysilane-based hydrogel and their chiral selectivity was evaluated for the separation of some selected enantiomers (tryptophan, benzoin, eperisone, chlorpheniramine). The protein encapsulation was carried out within a capillary in a single step under mild conditions. The resultant monolithic columns showed adequate chromatographic performance, including mechanical strength, penetration of pressurized flow, and chiral separation. Two different proteins, BSA and OVM, were successfully encapsulated into the gel matrixes by changing the alkoxysilane compositions of the gel. Run-to-run repeatability was quite satisfactory. The consecutive analysis of the neutral compound, benzoin, by the OVM-encapsulated column showed good repeatability in the retention time (RSD = 1.23% for the first peak, N = 10). Under optimized conditions, the theoretical plate number for the first peak of benzoin reached 72,000 plates/m.

A novel trypsin-encapsulation technique using the sol-gel method was developed for the preparation of an on-line enzyme reactor integrated into capillary electrophoresis. Trypsin was encapsulated in tetramethoxysilane-based hydrogel, and its enzymatic activity was evaluated using alpha-N-benzoyl-L-arginine ethyl ester and two peptides (bradykinin and [Ter8]-bradykinin). The enzyme encapsulation was carried out in a single step under mild conditions within a capillary, and 1.5-cm gel was formed at the inlet of the capillary. The resultant monolithic reactor showed excellent enzymatic activity, which was approximately 700 times higher than that in free solution, without stopping the flow. Separation of the unreacted substrates and products in the same capillary also showed high selectivity, and sample size in this system decreased 3 orders of magnitude from conventional tryptic reaction schemes. The encapsulated trypsin maintains its substrate specificity even in a sol-gel matrix. Furthermore, the encapsulated trypsin exhibits increased stability even after continuous use compared to that in free solution.

Trypsin-encapsulated sol-gel was fabricated in situ onto a plastic microchip to form an on-chip bioreactor that integrates tryptic digestion, separation, and detection. Trypsin-encapsulated sol-gel, which is derived from alkoxysilane, was fabricated within a sample reservoir (SR) of the chip. Fluorescently labeled ArgOEt and bradykinin were digested within the SR followed by electrophoretic separation on the same chip. The plastic microchip, which is made from poly(methyl methacrylate), generated enough electroosmotic flow that substrates and products could be satisfactorily separated. The sol-gel in the SR did not alter the separation efficiency of each peak. With the present device, the analytical time was significantly shortened compared to conventional tryptic reaction schemes. This on-chip microreactor was applicable to the digestion of protein with multiple cleavage sites and separation of digest fragments. Furthermore, the encapsulated trypsin exhibits increased stability, even after continuous use, compared with that in free solution.

Chronic subdural hematoma can be treated with tranexamic acid without concomitant surgery. Tranexamic acid might simultaneously inhibit the fibrinolytic and inflammatory (kinin-kallikrein) systems, which might consequently resolve CSDH. This medical therapy could prevent the early stages of CSDH that can occur after head trauma and the recurrence of CSDH after surgery.

Alzheimer's disease (AD) is an irreversible, progressive brain disease and can be definitively diagnosed after death through an examination of senile plaques and neurofibrillary tangles in several brain regions. It is to be expected that changes in the concentration and/or localization of low-molecular-weight molecules are linked to the pathological changes that occur in AD, and determining their identity would provide valuable information regarding AD processes. Here, we propose definitive brain metabolic profiling using ultra-performance liquid chromatography coupled with electrospray time-of-flight mass spectrometry analysis. The acquired data were subjected to principal components analysis to differentiate the frontal and parietal lobes of the AD/Control groups. Significant differences in the levels of spermine and spermidine were identified using S-plot, mass spectra, databases and standards. Based on the investigation of the polyamine metabolite pathway, these data establish that the downstream metabolites of ornithine are increased, potentially implicating ornithine decarboxylase activity in AD pathology.

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