Galactosylated chitosan as a synthetic extracellular matrix for hepatocytes attachment

Galactose moiety as the hepatocyte anchorage was covalently coupled with chitosan for the development of synthetic extracellular matrix. Hepatocytes adhesion to galactosylated chitosan (GC)-coated polystyrene (PS) dish became as high as 94.7% after 2 h incubation whereas the hepatocytes adhesion to chitosan-coated PS dish was 69.1%, indication of galactose-specific recognition between GC molecules and asialoglycoprotein receptors of hepatocytes. The DNA synthesis of the hepatocytes adhered to GC-coated dish was increased in the presence of epidermal growth factor (EGF) at low concentration of GC (0.05 microg/ml) whereas the DNA synthesis of the hepatocytes adhered to GC-coated dish was decreased in the presence of EGF at high concentration of GC (5 microg/ml). The spreading shapes of the hepatocytes adhered to the surface in the presence of EGF at low concentration of GC (0.05 microg/ml) were enhanced than in the absence of EGF. The hepatocytes adhered to the surface at high concentration of GC (5 microg/ml) showed round shapes and exhibited many spheroid formation after 24 h in the presence of EGF.     

Galactosylated PVDF membrane promotes hepatocyte attachment and functional maintenance

One of the major challenges in BLAD design is to develop functional substrates suitable for hepatocyte attachment and functional maintenance. In the present study, we designed a poly(vinylidene difluoride) (PVDF) surface coated with galactose-tethered Pluronic polymer. The galactose-derived Pluronic F68 (F68-Gal) was adsorbed on PVDF membrane through hydrophobic-hydrophobic interaction between PVDF and the polypropylene oxide segment in Pluronic. The galactose density on the modified PVDF surface increased with the concentration of the F68-Gal solution, reaching 15.4 nmol galactosyl groups per cm2 when a 1 mg/ml of F68-Gal solution was used. The adsorbed F68-Gal remained relatively stable in culture medium. Rat hepatocytes attachment efficiency on F68-Gal modified PVDF membrane was similar to that on collagen-coated surface. The attached hepatocytes on PVDF/F68-Gal membrane self-assembled into multi-cellular spheroids after 1 day of culture. These attached hepatocytes in spheroids exhibited higher cell functions such as albumin synthesis and P450 1A1 detoxification function compared to unmodified PVDF membrane and collagen-coated surface. These results suggest the potential of this galactose-immobilized PVDF membrane as a suitable substrate for hepatocyte culture.     

太空发育鸡胚的前庭感受器细胞形态学研究

为了探讨太空微重力对鸡胚前庭感觉上皮细胞的形态发育的影响 ,选取在航天飞机 (STS-2 9)发育鸡胚和地面发育鸡胚各两只 ,利用计算机显微测量技术分别测量椭圆囊和球囊的毛细胞、支持细胞核的切面面积、周长、形状系数。太空发育鸡胚的球囊支持细胞核的切面面积、周长显著大于地面组 ,形状系数无差异 ;太空发育鸡胚的椭圆囊支持细胞核的切面面积、周长、形状系数以及椭圆囊和球囊毛细胞的切面面积、周长与地面发育鸡胚相比无明显差异。微重力可能对球囊支持细胞核的体积发育有影响 ,对椭圆囊和球囊的毛细胞以及椭圆囊支持细胞核的形态发育无影响。     

Myelin Degeneration in Multiple System Atrophy Detected by Unique Antibodies

A rabbit antiserum (anti-EP), induced against a synthetic peptide corresponding to residues 68 to 86 of guinea pig myelin basic protein, powerfully immunostained abnormal-appearing oligodendrocytic processes and cell bodies in demyelinating areas associated with multiple system atrophy (MSA). However, as we reported previously, the antiserum, which is highly specific for the sequence QDENPVV corresponding to human myelin basic protein residues 82 to 88, failed to recognize any structures in normal human brain. QD-9, a mouse monoclonal antibody raised against human myelin basic protein residues 69 to 88, which also recognizes specifically the epitope QDENPVV, gave the same results as did anti-EP. The unusual epitope recognized by anti-EP/QD-9 antibodies appears to be accessible in areas of myelin degeneration, and the antibodies have been shown to detect such areas in multiple sclerosis and infarcted brains. These antibodies detect myelin degeneration more widely than previous conventional methods. The present study emphasizes the importance of myelin degeneration in the pathogenesis of multiple system atrophy.     

Myoblasts fail to stimulate T cells but induce tolerance

Recent interest in myoblast transfer and in the use of myoblastsas vehicles in gene therapy has made it important to understandthe potential immunogenicity of allogeneic or neoantlgen-expresslngmyoblasts. Given the problems of producing a pure populationof myoblasts, In this study we used a tumour-derived musclecell line (TE671), with phenotyplc features of myoblasts, whichwe transfected to express HLA-DR1. However, this cell line wasunable to stimulate either established HLA-DR1-specific alloreactlveT cell clones or a primary alloresponse. Nor could it presenthaemagglutlnln peptide HA 306–324 to DR1-restricted, HA306–324-speciflc T cell clones or lines. Indeed, prelncubatlonwith DR1-expressing TE671 and HA 306–324 rendered suchT cells tolerant as Judged by their subsequent inability toproliferate in response to a DR1⁺ B cell line plus peptide HA306–324. These results imply that myoblasts do not providecostlmulatory signals, and are therefore unlikely to stimulateallospeclfic T cells following myoblasts transplantation orto initiate neoantlgen-speclfic immune responses following Invivo transfection.     

Why is the diverse U letter relationship between embarrassment and psychological distance observed? In view of Schlenker and Leary (1982)'s self-presentation model of social anxiety

The aim of the present study is to explain why there is "the diverse U letter relationship" between embarrassment and psychological distance empirically, in view of Schlenker and Leary (1982)'s self-presentation model of social anxiety. In Study 1, those relationships were observed in three different situations. In Study 2, the main effect of motivation for avoiding rejection and the damage of self-image, and their interaction with embarrassment was examined by hierarchical regression analysis. In the first step, the psychological distance, in the second step, the main effect of both, and in the third step, the interaction was entered. As a result, in the second step, both main effects were significant, but in the third step, only interaction was significant, and both main effects were not significant. This showed that the interaction could predict embarrassment, although the psychological distance was controlled. Finally, this result was discussed in view of social norm.     

<Emphasis Type="Italic">GPC5</Emphasis> is a possible target for the 13q31-q32 amplification detected in lymphoma cell lines

Comparative genomic hybridization (CGH) analyses have detected gains of copy number on 13q, especially at 13q31-q32, in cell lines and primary cases of various types of lymphoma. Since amplification of chromosomal DNA is one of the mechanisms that can activate tumor-associated genes, and because 13q amplification had been reported in various other types of tumors as well, we attempted to define by fluorescence in situ hybridization (FISH) a common region at 13q31-q32 in which to explore genes that might be targets for the amplification events. Although the commonly amplified region we defined was relatively large (approximately 4 Mb), only one true gene, GPC5, was found there. GPC5 was over-expressed in lymphoma cell lines that had shown amplification, in comparison with those that had not. Our findings suggest that GPC5 is a likely target for amplification, and that over-expression of this gene may contribute to development and/or progression of lymphomas and other tumors.     

Melanocytes and melanosis of the oesophagus in Japanese subjects — analysis of factors effecting their increase

Summary Normal oesophagus specimens taken from 65 autopsy cases and surgical specimens from 127 oesophageal carcinoma cases were examined histopathologically to determine melanocyte incidence and distribution. Melanocytes were found in the epithelio-stromal junction in 7.7% of normal oesophagus specimens examined at autopsy, and in 29.9% of surgical cases with oesophageal carcinoma. Positive specimens in the latter groups, especially from pre-operatively irradiated individuals, showed a more remarkable increase of melanocytes than was evident in any of the normal oesophageal samples. There were no significant differences in incidence between males and females, or between age groups. In cases where the cancer invaded into deeper stroma, the melanocytes were mainly observed in the normal epithelium around the carcinomas. Epithelial and stromal elements of the melanotic mucosa commonly showed hyperplastic changes such as acanthosis or basal cell hyperplasia, and chronic oesophagitis. Melanocytes were observed most commonly in the lower part of the oesophagus, the site where malignant melanoma of the oesophagus, most often originates. These results strongly suggest that the melanocyte increase observed in areas of hyperplastic epithelium and chronic oesophagitis may play an important role as a precursor lesion for malignant melanoma in the oesophagus.     

Investigation of optimal drug in moderate bronchial asthma]

BACKGROUND: Many drugs and the combinations of drugs are recommended for each treatment step in bronchial asthma. However, there are few issues examined about the optimal drug and combination of drugs in a long term prognosis. In this study, we investigated the optimal drugs and combinations of drugs from a point of view of prognosis. METHODS: One hundred and ninety four patients who visited our hospital for treatment from November, 2003 to October, 2004 and were managed according to GINA guideline were surveyed retrospectively. We compared the rate of step up and the frequency of urgent visit and urgent hospitalization in one year between drug groups in each treatment step. RESULTS: The rate of step up was significantly higher in leukotriene receptor antagonist (LTRA) group than in inhalation corticosteroid (ICS) group and theophylline group in Step 2. The frequency of urgent visit and urgent hospitalization was significantly higher in ICS+LTRA group than in ICS+theophylline group and ICS+long-acting beta 2-agonist (LABA) group in Step 3. CONCLUSION: There is a possibility that the prognosis becomes bad when we use LTRA in the practical treatment according to GINA guideline.     

维生素D3受体对hsp90β基因表达的调控作用

鉴于热休克蛋白９０β（ｈｓｐ９０β）基因内含子中含有维生素Ｄ３受体（ＶＤＲ）结合位点，为探讨作为核受体家族成员的ＶＤＲ是否对核受体特异分子伴侣的ｈｓｐ９０β基因的表达具有调控作用，我们开展了本项研究。分别将野生型ＶＤＲ、含Ｎ端（１～１３３氨基酸残基）及Ｃ端（２８１～４２７氨基酸残基）片段的ＶＤＲ突变体真核表达质粒与人ｈｓｐ９０β基因调控片段（－１０３９／＋１５３１）介导的氯霉素乙酰基转移酶（ＣＡＴ）报告基因质粒共转染Ｊｕｒｋａｔ细胞，检测正常及经热休克（４２℃，１ｈ）处理后细胞裂解液中ＣＡＴ活性。结果表明ＶＤＲＮ端增强、而Ｃ端抑制ｈｓｐ９０β的组成性表达；在热诱导条件下野生型ＶＤＲ对ｈｓｐ９０β的表达有一定的抑制作用，而其Ｃ端片段的抑制较强。为进一步研究ＶＤＲ对细胞内源性热休克基因表达的影响，我们用ＲＴＰＣＲ方法研究了ＶＤＲ的对细胞内ｈｓｐ９０β基因ｍＲＮＡ水平的影响，发现ＶＤＲ过表达对ｈｓｐ９０β的热诱导表达明显抑制。结果提示ＶＤＲ对ｈｓｐ９０β基因的组成性和热诱导表达的调控机制不同。