Comparative Genomic Hybridization Detects the Chromosomal Gains and Losses in Early Tongue Carcinomas

Comparative genomic hybridization (以下,CGHと略す)法は,腫瘍細胞における全ゲノムの相対的なコピー数の増減を網羅的に検出する手法である。本研究の目的はCGH法を用いて,早期舌癌(T1-2NOMO)の染色体異常と予後の関連性を解析することである。対象は1998年4月1日から2001年3月31日に,広島大学医学部放射線科において舌癌の組織内照射前に原発巣から組織が採取できた早期舌癌26例(I期6例,II期20例)とした。男女比は15:11,平均年齢61.7才,初診からの観察期間の中央値は25.5ヶ月であった。実験方法は腫瘍細胞DNAと正常細胞DNAをそれぞれ異なる蛍光色素で標識し,スライドガラス上で正常分裂中期(染色体)細胞とハイブリダイゼーションを行った。その後,CCDカメラ付の顕微鏡にて画像を取り込み,染色体上のそれぞれの蛍光色素の量を測定し,コピー数の増減を解析した。結果は,lq(62%),5q(62%),16p(65%),19p(77%)にコピー数の増加を認め,3p(31%),21q(35%)にコピー数の減少を認めた。I期とII期の病期別の比較では,I期よりもII期で1qのコピー数の顕著な増加が認められた(17% vs. 75%,p<0.05)。一方,患者の観察期間中に認められた8例の後発頸部リンパ節転移の内7例は病期II期であった。特に,II期での後発頸部リンパ節転移症例はリンパ節転移のない症例に比べ,3qのコピー数増加が高頻度に認められた(86% vs. 31%,p<0.05)。この結果,CGH法を用いた早期舌癌の染色体異常の解析は,早期舌癌の予後因子としての診断に有用であることが示された。特にII期の舌癌にみられる3qの増幅は後発頸部リンパ節転移と相間していることから,患者治療法の選択に際して重要な判断材料である。The aim of this study was to detect the genomic deletions and amplifications in early tongue carcinomas. Genetic imbalances were assessed in twenty-six primary early tongue carcinomas using comparative genomic hybridization (CGH). Early tongue carcinomas were consisted of 6 patients of stage I, and 20 patients of stage II. The median follow-up time for all patients was 25.5 months (range 9 to 42 months). Copy number increases were most frequently observed on chromosomes 19p (77%), 16p (65%), Iq (62%) and 5q (62%). Copy number decreases were occurred mostly frequently at 21p(35%) and 3p(31%). Gains of Iq was significantly higher in stage II patients than in stage I patients (17% vs. 75%, p<0.05). In comparison to eight patients with lymph node metastasis (stage I; 1 patient, stage II; 7 patients), it was noted that the gains of 3q was frequently observed in tumors metastatic lymph nodes in stage II patients (86 0n patient with metastasis vs. 31 0n without metastasis, p<0.05). These results showed that CGH detected chromosomal imbalances in early tongue carcinomas and gain of 3q may help to improve the therapeutic result defining lymph node matastasis of stage II patients of tongue carcinomas

A sacrificial millipede altruistically protects its swarm using a drone blood enzyme, mandelonitrile oxidase

Soldiers of some eusocial insects exhibit an altruistic self-destructive defense behavior in emergency situations when attacked by large enemies. The swarm-forming invasive millipede, Chamberlinius hualienensis, which is not classified as eusocial animal, exudes irritant chemicals such as benzoyl cyanide as a defensive secretion. Although it has been thought that this defensive chemical was converted from mandelonitrile, identification of the biocatalyst has remained unidentified for 40 years. Here, we identify the novel blood enzyme, mandelonitrile oxidase (ChuaMOX), which stoichiometrically catalyzes oxygen consumption and synthesis of benzoyl cyanide and hydrogen peroxide from mandelonitrile. Interestingly the enzymatic activity is suppressed at a blood pH of 7, and the enzyme is segregated by membranes of defensive sacs from mandelonitrile which has a pH of 4.6, the optimum pH for ChuaMOX activity. In addition, strong body muscle contractions are necessary for de novo synthesis of benzoyl cyanide. We propose that, to protect its swarm, the sacrificial millipede also applies a self-destructive defense strategy—the endogenous rupturing of the defensive sacs to mix ChuaMOX and mandelonitrile at an optimum pH. Further study of defensive systems in primitive arthropods will pave the way to elucidate the evolution of altruistic defenses in the animal kingdom

Ablation of the N-type calcium channel ameliorates diabetic nephropathy with improved glycemic control and reduced blood pressure

Pharmacological blockade of the N-and L-type calcium channel lessens renal injury in kidney disease patients. The significance of specific blockade of α1 subunit of N-type calcium channel, Cav2.2, in diabetic nephropathy, however, remains to be clarified. To examine functional roles, we mated Cav2.2-/- mice with db/db (diabetic) mice on the C57BLKS background. Cav2.2 was localized in glomeruli including podocytes and in distal tubular cells. Diabetic Cav2.2-/- mice significantly reduced urinary albumin excretion, glomerular hyperfiltration, blood glucose levels, histological deterioration and systolic blood pressure (SBP) with decreased urinary catecholamine compared to diabetic Cav2.2+/+ mice. Interestingly, diabetic heterozygous Cav2.2+/- mice also decreased albuminuria, although they exhibited comparable systolic blood pressure, sympathetic nerve activity and creatinine clearance to diabetic Cav2.2+/+ mice. Consistently, diabetic mice with cilnidipine, an N-/L-type calcium channel blocker, showed a reduction in albuminuria and improvement of glomerular changes compared to diabetic mice with nitrendipine. In cultured podocytes, depolarization-dependent calcium responses were decreased by Ω-conotoxin, a Cav2.2-specific inhibitor. Furthermore, reduction of nephrin by transforming growth factor-β (TGF-β) in podocytes was abolished with Ω-conotoxin, cilnidipine or mitogen-activated protein kinase kinase inhibitor. In conclusion, Cav2.2 inhibition exerts renoprotective effects against the progression of diabetic nephropathy, partly by protecting podocytes.</p

Rapid and sensitive XAFS using a tunable X-ray undulator at BL10XU of SPring-8

The design and performance of the high-brilliance XAFS facility at BL10XU of SPring-8, aimed at rapid and sensitive measurement of X-ray absorption fine structure (XAFS), is reported. Both undulator gap and double-crystal monochromator have been successfully controlled covering a wide energy range (5-30 keV). A versatile goniometer system, consisting of two independent high-precision goniometers. is capable of polarized XAFS in fluorescence mode and surface-sensitive experiments using a grazing-incidence geometry. By sharing major components, i.e. a monolithic Ge 100-pixel array detector and a closed-cycle He cryostat, both polarized XAFS and X-ray standing wave (XSW) experiments can be performed at low temperature (15-300 K). The performance of the spectrometer has been evaluated by recording XAFS spectra in transmission mode

Preventative Effect of a Flavonoid, Enzymatically Modified Isoquercitrin on Ocular Symptoms of Japanese Cedar Pollinosis

ABSTRACTBackgroundFlavonoids are nutrients that exert anti-allergic effects. We investigated the preventative effect of enzymatically modified isoquercitrin (EMIQ), a flavonoid, to relieve the symptoms of Japanese cedar pollinosis.MethodsIn a parallel-group, double-blind placebo-controlled study design, 24 subjects with Japanese cedar pollinosis took 100 mg EMIQ or a placebo for 8 weeks, starting 4 weeks prior to the onset of pollen release. Subjective symptoms, ADL scores and the usage of drugs were recorded daily, and the QOL score was obtained every 4 weeks. Blood sampling was performed before and after the study to measure serum levels of IgE and flavonoids.ResultsDuring the entire study period, ocular symptom + medication score for the EMIQ group was significantly lower (p < 0.05) than that of the placebo group. When limited to the period, ocular symptom scores (p < 0.05, weeks 5–6), and ocular congestion scores (p < 0.05, weeks 5–6) for the EMIQ group was significantly lower than that for the placebo group while other scores for the EMIQ group, such as ocular itching scores (p = 0.09, weeks 4–5), lacrimation scores (p = 0.07, weeks 5–6), and ocular congestion scores (p = 0.06, weeks 45), all tended to be lower. However no significant differences were found in nasal symptoms between the two groups. Serum concentrations of IgE were not significantly downregulated but the serum concentrations of quercetin and its derivatives were elevated significantly by the intake of EMIQ.ConclusionsIntake of the quercetin glycoside EMIQ proved to be effective for the relief of ocular symptoms caused by Japanese cedar pollinosis

Increase of Total Nephron Albumin Filtration and Reabsorption in Diabetic Nephropathy

There is a hot debate concerning actual amount of albumin filtered through glomeruli and reabsorbed at proximal tubules in normal kidneys and diabetic conditions. To overcome current technical problems, we generated a drug-inducible megalin knockout mouse line, megalin(lox/lox);Ndrg1-CreER[T2] (or iMegKO), whose protein reabsorption can be shut off anytime by tamoxifen (Tam). After Tam administration, renal megalin protein expression was reduced by 92% compared to wild-type C57BL/6J mice, and renal reabsorption of intravenously-injected retinol binding protein was almost completely abrogated. Urinary albumin excretion increased to 175 μg/day (0.460 mg/mg-creatinine), suggesting that this was the amount of total nephron albumin filtration. Glomerular sieving coefficient of albumin was 1.7 x 10[-5]. By comparing streptozotocin-induced, Tam-treated, diabetic STZ;iMegKO mice with non-STZ;iMegKO mice, we estimated that daily albumin filtration was increased by 1.9-fold, reabsorption was increased by 1.8-fold, and reabsorption efficiency was reduced to 86% by development of diabetes (versus 96% in control). Such abnormalities were well normalized after insulin treatment. Another type 1 diabetic model of Akita;iMegKO mice showed equivalent results. This study reveals actual values and changes of albumin filtration and reabsorption in early diabetic nephropathy, bringing new insights into our understanding of renal albumin dynamics in hyperfiltration status of diabetic nephropath