Anti-oxidative effect of Klotho on endothelial cells through cAMP activation

Klotho, a regulatory factor implicated in countering the aging process, has been reported to ameliorate endothelial dysfunction in vivo. To clarify whether Klotho protein directly affects endothelial cell function, we studied the effects of membrane-form Klotho on manganese superoxide dismutase (Mn-SOD) expression and nitric oxide production in human umbilical vein endothelial cells (HUVEC). We incubated HUVEC with conditioned medium from COS-1 cells transfected with expression vector, pCAGGS-klotho (Klotho-CM) or a recombinant, purified 6His-tagged Klotho protein. Both Klotho-CM and 6His-tagged Klotho protein enhanced Mn-SOD expression by approximately two-fold, partially via activation of the cAMP signaling pathway. Furthermore, Klotho-CM increased nitric oxide production, which also contributed to the up-regulation of Mn-SOD. Using the oxidation-sensitive dye dihydroethidium, we found that Klotho inhibited angiotensin II-induced reactive oxygen species production in HUVEC. These findings provide new insights into the mechanisms of Klotho action and support the therapeutic potential of membrane-form Klotho to regulate endothelial function. Hiromi Rakugi and Naomichi Matsukawa contributed equally to this work.     

Intravenous immunoglobulin therapy for Miller Fisher syndrome

We analyzed clinical recovery of 92 patients with Miller Fisher syndrome who had been treated with IV immunoglobulin (IVIg; n = 28), plasmapheresis (n = 23), and no immune treatment (n = 41). IVIg slightly hastened the amelioration of ophthalmoplegia and ataxia, but the times of the disappearances of those symptoms were similar among three groups. In Miller Fisher syndrome, IVIg and plasmapheresis seem not to have influenced patients' outcomes, presumably because of good natural recovery.     

沙生蜡菊花中黄酮类成分的分离与鉴定

目的研究沙生蜡菊(Helichrysum arenarium(L.)Moench)花的化学成分。方法采用硅胶柱色谱?ODS柱色谱和HPLC柱色谱分离纯化,依据理化性质、波谱数据分析进行结构鉴定。结果从沙生蜡菊花的甲醇提取物中分离得到8个黄酮类化合物,分别鉴定为山奈酚3-O-β-D-吡喃葡萄糖苷(kaempferol 3-O-β-D-glucopyranoside,1)、木犀草素3′-O-β-D-吡喃葡萄糖苷(luteolin3′-O-β-D-glucopyranoside,2)、木犀草素7-O-β-D-吡喃葡萄糖苷(luteolin7-O-β-D-glucopyranoside,3)、木犀草素6-羟基-7-O-β-D-吡喃葡萄糖苷(luteolin6-hydroxy-7-O-β-D-glucopyranoside,4)、木犀草素3′-甲氧基-6-羟基-7-O-β-D-吡喃葡萄糖苷(luteolin3′-methoxyl-6-hydroxy-7-O-β-D-glucopyranoside,5)、黄芩素7-O-β-D-吡喃葡萄糖苷(scutellarein7-O-β-D-glucopyranoside,6)、山柰酚3-O-β-D-龙胆二糖苷(kaempferol3-gentiobioside,7)、山柰酚3-O-(3-β-D-吡喃葡萄糖基)-β-D-吡喃葡萄糖苷(kaempferol3-O-(3-β-D-glucopyranosyl)-β-D-glucopyranoside,8)。结论化合物2、4~8为首次从蜡菊属植物中分离得到。     

Effects of ethanol on excitatory and inhibitory synaptic transmission in rat cortical neurons

BACKGROUND: The gamma-aminobutyric acid-A (GABA(A)) receptor and glutamate receptors are among the most important target sites for the behavioral effects of ethanol. However, data in the literature concerning the ethanol modulation of the GABA(A) and glutamate receptors have been controversial. The activity of the neuronal nicotinic acetylcholine (ACh) receptors (nAChRs) has recently been reported to be potently augmented by ethanol. The activation of nAChRs is also known to cause the release of various neurotransmitters including GABA and glutamate. Thus, ethanol potentiation of nAChRs is expected to stimulate the GABAergic and glutamatergic systems. METHODS: Whole-cell patch clamp experiments were performed using rat cortical neurons in primary culture to record spontaneous miniature inhibitory postsynaptic currents (mIPSCs) and spontaneous miniature excitatory postsynaptic currents (mEPSCs). RESULTS: Two types of neurons were distinguished: bipolar neurons possessed alpha4beta2 nAChRs generating a steady current in response to 30 nM ACh, and multipolar neurons that did not generate a current by ACh application. Acetylcholine greatly increased the frequency of mEPSCs and mIPSCs in bipolar neurons but not in multipolar neurons. The amplitude of neither type of neuron was affected by ACh. Ethanol at 10 to 100 mM suppressed the amplitude of mEPSCs while augmenting the amplitude of mIPSCs in both bipolar and multipolar neurons, indicating the direct action on the respective receptors. In bipolar neurons, ACh plus 100 mM ethanol greatly increased the frequency of mIPSCs beyond the levels achieved by ACh alone, while no such increases were observed in multipolar neurons. CONCLUSIONS: It is concluded that ethanol stimulation of nAChRs modulates the activity of both glutamate and GABA receptors in rat cortical bipolar neurons.     

Anti-Inflammatory Effects of Pravastatin on Helicobacter Pylori-Induced Gastritis in Mice

Pravastatin, an HMG-CoA reductase inhibitor, exerts anti-inflammatory effects via several mechanisms including induction of endothelial nitric oxide synthase (eNOS). We investigated the effect of pravastatin on Helicobacter pylori-induced gastritis in mice. Mice with or without H. pylori infection received intraperitoneal pravastatin daily for 1 week. Expression of eNOS mRNA and tumor necrosis factor-α mRNA and myeloperoxidase activity in gastric tissue was determined. Myeloperoxidase activity was reduced in a dose-dependent manner by pravastatin, with activity inhibited by 53.5 and 73.7% at doses of 0.3 and 1 mg/kg, respectively. At a dose of 1 mg/kg, pravastatin reduced the level of tumor necrosis factor-α mRNA by 52.7%, while it did not affect eNOS expression. Pravastatin had no effects on these inflammatory parameters in uninfected mice. Pravastatin did not affect the viability of H. pylori. In conclusion, pravastatin exerts an anti-inflammatory effect on H. pylori-induced gastritis in mice without affecting eNOS expression.     

Present state of diabetes management in the elderly, Japan

A recent dramatic increase in elderly patients with diabetes mellitus has made the proper management of the disease in this population more important. Here, we discuss the present status of diabetes management in the elderly in Japan. As a characteristic feature of elderly persons, body weight reduction is difficult, because of the profound adaptive reduction in resting energy expenditure under calorie restriction in the elderly. However, hyperglycemia increases the risk for diabetic complications, except proliferative retinopathy, similarly in elderly and non-elderly. Of note, there is marked clinical heterogeneity in this generation in the following aspects: duration, complication status (past aspect), insulin secretion, insulin sensitivity, familial support and physical exercise/activity (present aspect), as well as the expected lifespan (future aspect). This heterogeneity among the elderly should render diabetes treatment diverse, and in fact, one of the largest surveys in Japan demonstrated significant diversity in diabetes management in the elderly. In Japan, thus, the present management of diabetes in the elderly is considerably diverse, reflecting the clinical heterogeneity among elderly patients with diabetes. Further clinical evidence is awaited for the establishment of proper and safe management of diabetes in the elderly.     

Genetics of type 1 diabetes in Asian and Caucasian populations

Among candidate genes for type 1 diabetes, HLA, INS, CTLA4, PTPN22 and SUMO4 have been shown to be associated with the disease in Caucasian populations. To clarify the similarities and differences in the contribution of these genes to type 1 diabetes between Asian and Caucasian populations, association of these genes with type 1 diabetes was studied in a large number of samples in Japanese and Korean populations. Class II HLA was strongly associated with type 1 diabetes in both Asian and Caucasian populations, but haplotypes associated with type 1 diabetes were markedly different due to difference in the presence and absence of haplotypes in each population. INS was consistently associated with type 1 diabetes in both Japanese and Caucasian populations, but frequency of disease-associated haplotype was markedly high in Japanese general population. CTLA4 was associated with type 1 diabetes only in a subset of patients with type 1 diabetes complicated with AITD in Japanese. A variant (R620W) of PTPN22 was associated with type 1 diabetes and other autoimmune diseases in Caucasians, but the variant was absent in Asians. SUMO4 was associated with type 1 diabetes in Asians, but not in Caucasian, suggesting a genetic heterogeneity among diverse ethnic groups. Trans-racial study with a large number of samples in both Asian and Caucasian populations will contribute to genetic dissection of type 1 diabetes and identification of causative variants.     

Phase II study of 2-week TS-1 administration followed by 1-week rest for gastric cancer

BACKGROUND/AIMS: TS-1 monotherapy with 4-week administration followed by 2-week rest is used as the community standard treatment for metastatic gastric cancer in Japan. However, according to a postmarketing survey, the percentage of patients who received three or more courses was only 44.6%; for the reasons of discontinuation due to exacerbation of symptoms or adverse reactions during the first or second course. Therefore, we conducted the phase II study of 2-weeks administration with TS-1 followed by a 1-week rest against metastatic gastric cancer, aiming for mitigation of adverse reactions without reduction of antitumor effect. METHODOLOGY: Thirty-five patients were enrolled between 2001 and 2003 at nine institutes in Japan. One cycle of TS-1 treatment whose dosage was 80 mg/m2/day consisted of administration for 2 weeks followed by a 1-week rest. The primary endpoint was overall response rate and the secondary endpoints were safety and feasibility. RESULTS: There were 6 PRs, 13 NCs, 11 PDs, and 5 patients were not evaluable (NE), yielding a response rate of 17%. The median survival time of all patients was 290 days. Severe adverse Grade 3 or 4 reactions were observed in 8 (23%) patients. The rate of patients who received six or more courses was 43%. The cumulative rate of the relative total administration days was 93%. CONCLUSIONS: We concluded that the schedule of TS-1 administration for 2 weeks followed by a 1-week rest might not be superior to the conventional schedule (4 weeks on and 2 weeks off) with regard to the antitumor effect, adverse reactions and prolonged medication, although it was acceptable from the point of view of survival.     

The validity of sentinel lymph node biopsy using dye technique alone in patients with gastric cancer

BACKGROUND/AIMS: We investigated whether sentinel lymph node biopsy using dye technique alone is useful or not in decision-making for less invasive surgery in patients with gastric cancer. METHODOLOGY: The subjects were 43 patients who had undergone laparotomy for gastric cancer and consented to undergo sentinel lymph node biopsy using patent blue dye. The patients enrolled were 26 males and 17 females, with a mean age of 62.5 years. The tumor sites were upper third of the stomach in 14, middle third in 16, and lower third in 13 patients. The depth of invasion was mucosa in eight, submucosa in 19, muscularis propria in five, subserosa in five, and serosa in six patients. Total gastrectomy was performed in 12, subtotal gastrectomy in 28, and proximal gastrectomy in three patients. RESULTS: The mean number of sentinel lymph node biopsies per surgery was 3.5 +/- 4.1. We were able to perform blue node biopsy in 40 out of 43 patients, but could not find any blue nodes in three patients. Among the 40 patients in whom blue nodes were identified, 29 patients with no metastasis in blue nodes had no evidence of lymph node metastasis (NO). The depth of invasion was not deeper than subserosa in all these patients. Metastasis was observed in one out of the three patients in whom no blue nodes were found. CONCLUSIONS: When the depth of invasion was not deeper than the subserosa and blue nodes were identified, no metastases in either non-blue nodes or blue nodes could be found in the absence of metastatic blue nodes. Therefore, if the depth of invasion is not deeper than the subserosa in gastric cancer, metastatic search in blue nodes seems sufficient and less invasive surgery can be performed safely. Even when the invasion depth is not deeper than the submucosa, the tumor could be metastatic to Group 2 lymph nodes in patients in whom blue node biopsy revealed metastases. When metastasis is found in lymph nodes by intraoperative frozen section diagnosis, less invasive surgery for gastric cancer is not indicated.