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Notes: Background: The IgA1 molecule, which is predominantly deposited in glomeruli of IgA nephropathy, has the characteristic O-glycan side chain in its hinge region. Several lines of evidence have demonstrated that underglycosylated IgA1 acquires self-aggregation1 and binding affinities to the extra-cellular matrix,1,2 resulting in deposition in the glomeruli.3,4 However, there is little information concerning actual biological activities of the underglycosylated IgA1 deposited in glomeruli, which lead to mesangial cell proliferation and/or matrix expansion.Aim: To clarify the influence of O-glycan side chain in the hinge of IgA1 on its biological activity in mesangial cells, we performed a comprehensive gene expression profiling analysis of human cultured mesangial cells stimulated by enzymatically underglycosylated IgA1 using cDNA array.Methods: IgA1 was purified by affinity chromatography using anti IgA column followed by Jacalin column. Asialo/agalacto IgA1 was obtained by enzyme digestion using sialidase and β-galactosidase. Heat aggregated IgA1 was obtained by incubation at 63°C for 150 min. Cultured human mesangial cells were stimulated for 3 h by non-treated IgA1, heat aggregated IgA1 or asialo/agalacto IgA1, respectively, and total RNA was obtained. Only the enzyme stimulation was performed as a negative control for asialo/agalacto IgA1. After DNase treatment, isotope labelled probes were prepared by reverse transcription and hybridized to the Atlas Human 1.2 Array (CLONTECH, Palo Alto, CA, USA) according to the manufacturer’s protocol. A total of 1176 arrayed genes were quantitatively evaluated using BAS.Results: Expression profiles of 1176 genes were obtained. In all experiments, none of the three negative controls on the cDNA array was detected. The expression profiles of mesangial cells by each IgA1 stimulation resembled one another, but they were widely different from that of mononuclear cells. On the whole, 10% of the genes expressed in the mesangial cells were up-regulated by asialo/agalacto IgA1 stimulation compared to that by enzyme stimulation. The results suggested that under-O-glycosylated IgA1 has a biological function in human mesangial cells in vitro. In good agreement with previous studies,5 heat aggregated IgA1 stimulation up-regulated IL-6 and TNF-α expressions in mesangial cells compared to by non-treated IgA1. Asialo/agalacto IgA1 stimulation also up-regulated these molecules, suggesting its pathogenetic role in IgA nephropathy. However, since a similar tendency was revealed by enzyme stimulation alone, further study may be necessary to establish these observations.Conclusion. In the present study, we investigated the comprehensive expression profiles of mesangial cells stimulated by various types of IgA1 using cDNA array. This approach may serve as a useful database not only to evaluate the biological activities of under-O-glycosylated IgA1 but also to elucidate the pathogenesis of IgA nephropathy.

Notes: Polymerization of C60 clusters epitaxially grown on Si(111)-(7×7) substrates was found to be induced by electron injection from the probe tips of scanning tunneling microscopes (STM) as the sample bias was increased from +4.0 to +5.5 V, exhibiting an evolution behavior characterized by an incubation, a linear growth, and a saturation. The incubation time and the growth rate are dependent greatly on the sample site, which is explained by a model taking into account the pre-existing stress as the driving force of the polymerization and the internal stress built up as a consequence of polymerization producing a stress for backward reactions. © 2000 American Institute of Physics.

Notes: Melanin-concentrating hormone (MCH) has been reported to be involved in the regulation of feeding behaviour in rats and mice. Because many neuropeptides that influence ingestive behaviour also regulate reproductive function, the present study was designed to determine if central administration of MCH changes pulsatile secretion of luteinizing hormone (LH) in the rats. Wistar-Imamichi strain female rats were ovariectomized and implanted with oestradiol to produce a moderate inhibitory feedback effect on LH release. The effects of i.c.v. injections of MCH on LH release were examined in freely moving animals. Blood samples were collected every 6 min for 3 h through an indwelling cannula. After 1 h of sampling, MCH (0.1, 1 or 10 μg/animal) or vehicle (saline) was injected into the third cerebroventricle. Because MCH is also reported to affect the hypothalamo-pituitary-adrenal (HPA) axis, which in turn, can influence reproductive function, plasma corticosterone concentrations were determined in the same animals at 30-min intervals during the first and last hours and every 12 min during the second hour of the 3-h sampling period. When expressed as per cent changes, mean plasma LH concentrations after MCH administration were significantly lower in the animals injected with all doses of the peptide compared with vehicle-treated animals; LH pulse frequency was significantly lowered by 1 μg of MCH. Per cent changes in mean plasma corticosterone levels were not significantly affected by MCH administration. These results in oestradiol-treated ovariectomized rats indicate that central MCH is capable of inhibiting pulsatile LH secretion. We have previously shown that 48-h fasting suppresses pulsatile LH release in the presence of oestrogen. Take together, these results raise the possibility that MCH could play a role in mediating the suppression of LH secretion during periods of reduced nutrition.

Notes: Background:  Using the sera from buckwheat (BW)-allergic patients, several putative causative molecules were reported. However, few molecules were determined on the molecular structure. We demonstrated in 2000 that the major allergen with 24 kDa (BW24KD) is a legumin-like storage protein.Objective:  The aim of this study was to isolate and characterize further a major allergen with 10 kDa by molecular cloning.Methods and results:  Buckwheat allergens were identified by immunoblotting analysis using sera from 14 allergic and two nonallergic individuals. We identified a protein with 10 kDa (BW10KD) that reacted with immunoglobulin E (IgE) more strongly than with IgG and IgA in 57% of the allergic patients but not with IgE in nonallergic individuals. Analyses were performed by N-terminal amino acid sequencing and molecular cloning. Physiological significance was assessed by an immunoblotting experiment showing that the reactivity of an allergic patient's serum IgE to BW10KD was competitively inhibited by natural BW extracts.Conclusion:  Molecular cloning experiments indicated that BW10KD as a BW allergen was a member of the 2S-albumin multigene family.

Notes: Flaking type failure in rolling-contact processes is usually attributed to fatigue-induced subsurface shearing stress caused by the contact loading. Assuming such crack growth is due to mode II loading and that mode I growth is suppressed due to the compressive stress field arising from the contact stress, we developed a new testing apparatus for mode II fatigue crack growth. Although the apparatus is, as a former apparatus was, based on the principle that the static KI mode and the compressive stress parallel to the pre-crack are superimposed on the mode II loading system, we employ direct loading in the new apparatus. Instead of the simple four-point-shear-loading system used in the former apparatus, a new device for the application of a compressive stress parallel to the pre-crack has been developed. Due to these alterations, mode II cyclic loading tests for hard steels have become possible for arbitrary stress ratios, including fully reversed loading (R=−1); which is the case of rolling-contact fatigue. The test results obtained using the newly developed apparatus on specimens made from bearing steel SUJ2 and also a 0.75% carbon steel, are shown.

Notes: Porphyromonas gingivalis has been shown to attack host defense systems through proteolytic cleavage of a wide variety of members of the systems. In this study, we examined the ability of P. gingivalis culture supernatant to alter the expression of human T cell surface proteins. As judged by flow cytometric analysis, detection of CD4 expression was completely eliminated by the supernatant, but CD8 was less sensitive. When the culture supernatant was added with reducing agents, proteolytic activity was enhanced, resulting in the cleavage of CD8. Mitogenic response of T cells to phytohemagglutinin or concanavalin A was decreased by the treatment of the cells with the culture supernatant of P. gingivalis. The three forms of gingipains (high molecular mass arginine-specific gingipain, arginine-specific gingipain 2 and lysine-specific gingipain) purified from the culture supernatant of P. gingivalis actively cleaved CD4 and CD8 on human T cells, indicating that proteolytic activity of the culture supernatant was due to gingipains. These results suggest that cysteine proteinases like gingipains released from P. gingivalis cleave T cell surface proteins and impede T cell function.

Notes: Abstract Background. 5-Methyl-1-phenyl-2-(1H)-pyridone (pirfenidone [PD]) has anti-fibrotic and anti-inflammatory effects. Previous reports have indicated that PD can prevent the progression of chronic kidney fibrosis in some animal models. It has not yet been reported, however, whether PD is capable of controlling acute inflammation of the kidney. Methods. The present study was designed to observe the effect of PD on an acute phase of anti-Thy-1 nephritis, a well known model for acute kidney inflammation. Male standard Wistar rats (specific pathogen-free [SPF] level), 7 weeks old (day −10, 141.52 ± 2.60 g) were divided into two groups. In group C (n = 7), 0.7 ml of anti-Thy-1 antibody was injected intravenously on day 0. In group P (n = 6), anti-Thy-1 antibody was injected similarly, and PD (500 mg/kg BW per day) was given daily by dietary intake starting 1 day before the first injection and continuing throughout the study. All rats were killed on day 7 and subjected to light microscopic and serum biochemical examinations. The degree of glomerular extracellular matrix (ECM) expansion was determined as a matrix score, using a semiquantitative method. Differences between the two group scores were analyzed by a non-parametric test. Results. The mean matrix score for group C was 87.6 ± 11.8, against 71.5 ± 12.5 (P 〈 0.05) for group P. The mean cell count for the 140 glomeruli in group C was 75.3 ± 6.1, and for the 120 glomeruli in group P it was 78.2 ± 11.6 (no significant difference). However, the possibility could not be ruled out that the degree of initial mesangiolysis in group P had been smaller than that in group C. Serum creatinine and blood urea nitrogen (BUN) levels were within the normal range in both groups. Conclusions. It is possible that PD is capable of partially controlling acute phases of anti-Thy-1 nephritis. More detailed studies in the future will establish the validity of the short-term effects of PD on this animal model.

Notes: Abstract: We have reported that a cell population obtained from fetal rat mandible with neutral protease (Pro I) has a unique differentiation sequence in which the elevation of alkaline phosphatase (ALPase), calcium accumulation, and collagen synthesis occurs simultaneously. In this report, we further characterized Pro I-released population of cells by studying the effect of dexamethasone (Dex) or β-glycerophosphate (β-GP) on the formation of bone nodules. The formation of bone nodules in Pro I-released population of cells (ProIRPC) was augmented by the addition of Dex (10−7 M) from days 3 to 14, suggesting that Pro IRPC contained osteoprogenitor (OP) cells. A 24-hour pulse treatment of ProIRPC released population of cells with Dex on days 9 and 12 resulted in an increase in the number of nodules but treatment on days 3, 6, or 15 did not. The number of bone nodules formed in Pro IRPC pulse treated with Dex on day 9 was comparable with that in Pro IRPC treated with Dex from days 3 to 14. Dex caused an earlier elevation of ALPase, in which maximal expression was observed on day 10. β-GP caused a prolonged elevation of ALPase, but did not affect the formation of bone nodules. Unlike Pro I-released population of cells, rat calvarial cells did not form mineralized nodules without β-GP, and showed that a Dex-responsive period on bone nodule formation in rat calvarial cells was at preconfluency (days 0 and 1). Thus, it appeared that the Dex-induced differentiation of early OP cells in Pro IRPCs occurred during the limited period from day 9 to day 12. Pro IRPC was found to have an unique characteristic that bone nodule formation was not affected by β-GP.