Largazole and its derivatives selectively inhibit ubiquitin activating enzyme (e1)

Protein ubiquitination plays an important role in the regulation of almost every aspect of eukaryotic cellular function; therefore, its destabilization is often observed in most human diseases and cancers. Consequently, developing inhibitors of the ubiquitination system for the treatment of cancer has been a recent area of interest. Currently, only a few classes of compounds have been discovered to inhibit the ubiquitin-activating enzyme (E1) and only one class is relatively selective in E1 inhibition in cells. We now report that Largazole and its ester and ketone analogs selectively inhibit ubiquitin conjugation to p27(Kip1) and TRF1 in vitro. The inhibitory activity of these small molecules on ubiquitin conjugation has been traced to their inhibition of the ubiquitin E1 enzyme. To further dissect the mechanism of E1 inhibition, we analyzed the effects of these inhibitors on each of the two steps of E1 activation. We show that Largazole and its derivatives specifically inhibit the adenylation step of the E1 reaction while having no effect on thioester bond formation between ubiquitin and E1. E1 inhibition appears to be specific to human E1 as Largazole ketone fails to inhibit the activation of Uba1p, a homolog of E1 in Schizosaccharomyces pombe. Moreover, Largazole analogs do not significantly inhibit SUMO E1. Thus, Largazole and select analogs are a novel class of ubiquitin E1 inhibitors and valuable tools for studying ubiquitination in vitro. This class of compounds could be further developed and potentially be a useful tool in cells.     

Cardiomyocyte Antihypertrophic Effect of Adipose Tissue Conditioned Medium from Rats and Its Abrogation by Obesity is Mediated by the Leptin to Adiponectin Ratio

White adipocytes are known to function as endocrine organs by secreting a plethora of bioactive adipokines which can regulate cardiac function including the development of hypertrophy. We determined whether adipose tissue conditioned medium (ATCM) generated from the epididymal regions of normal rats can affect the hypertrophic response of cultured rat ventricular myocytes to endothelin-1 (ET-1) administration. Myocytes were treated with ET-1 (10 nM) for 24 hours in the absence or presence of increasing ATCM concentrations. ATCM supressed the hypertrophic response to ET-1 in a concentration-dependent manner, an effect enhanced by the leptin receptor antagonist and attenuated by an antibody against the adiponectin AdipoR1 receptor. Antihypertrophic effects were also observed with ATCM generated from perirenal-derived adipose tissue. However, this effect was absent in ATCM from adipose tissue harvested from corpulent JCR:LA-cp rats. Detailed analyses of adipokine content in ATCM from normal and corpulent rats revealed no differences in the majority of products assayed, although a significant increase in leptin concentrations concomitant with decreased adiponectin levels was observed, resulting in a 11 fold increase in the leptin to adiponectin ratio in ATCM from JCR:LA-cp. The antihypertrophic effect of ATCM was associated with increased phosphorylation of AMP-activated protein kinase (AMPK), an effect abrogated by the AdipoR1 antibody. Moreover, the antihypertrophic effect of ATCM was mimicked by an AMPK activator. There was no effect of ET-1 on mitogen-activated protein kinase (MAPK) activities 24 hour after its addition either in the presence or absence of ATCM. Our study suggests that adipose tissue from healthy subjects exerts antihypertrophic effects via an adiponectin–dependent pathway which is impaired in obesity, most likely due to adipocyte remodelling resulting in enhanced leptin and reduced adiponectin levels.     

Phase II trial of hu14.18-IL2 for patients with metastatic melanoma

Phase I testing of the hu14.18-IL2 immunocytokine in melanoma patients showed immune activation, reversible toxicities, and a maximal tolerated dose of 7.5?mg/m²/day. In this phase II study, 14 patients with measurable metastatic melanoma were scheduled to receive hu14.18-IL2 at 6?mg/m²/day as 4-h intravenous infusions on Days 1, 2, and 3 of each 28?day cycle. Patients with stable disease (SD) or regression following cycle 2 could receive two additional treatment cycles. The primary objective was to evaluate antitumor activity and response duration. Secondary objectives evaluated adverse events and immunologic activation. All patients received two cycles of treatment. One patient had a partial response (PR) [1 PR of 14 patients?=?response rate of 7.1?%; confidence interval, 0.2?C33.9?%], and 4 patients had SD and received cycles 3 and 4. The PR and SD responses lasted 3?C4?months. All toxicities were reversible and those resulting in dose reduction included grade 3 hypotension (2 patients) and grade 2 renal insufficiency with oliguria (1 patient). Patients had a peripheral blood lymphocytosis on Day 8 and increased C-reactive protein. While one PR in 14 patients met protocol criteria to proceed to stage 2 and enter 16 additional patients, we suspended stage 2 due to limited availability of hu14.18-IL2 at that time and the brief duration of PR and SD. We conclude that subsequent testing of hu14.18-IL2 should involve melanoma patients with minimal residual disease based on compelling preclinical data and the confirmed immune activation with some antitumor activity in this study.     

江苏沿江生态防护林树种评价体系的建立与树种的初步筛选

采用层次分析法(AHP),以生长适应性、生态防护能力、景观价值和经济价值为Ⅰ级指标,以耐水湿性、抗污染性、生长速率、树冠特征、枝条特征和根系特征以及叶、花、果、树干和树形的观赏价值及材用、药用、食用和工业价值为Ⅱ级指标,确定各评价指标的权重值和赋值,建立了江苏沿江生态防护林树种综合评价体系.将江苏沿江生态防护林划分为水源涵养和水土保持林、景观防护林、污染隔离林和农林复合防护林4种类型,以耐水湿性、抗污染性、抗风性、水土保持能力、景观价值、经济价值和生长速率为评价指标,并设置各指标的权重值,建立了江苏沿江生态防护林树种分类评价体系.利用综合评价体系对223个树种进行评价,分别筛选出适用于江苏沿江生态防护林建设的综合得分前20位的落叶乔木、常绿乔木和灌木种类,其中,落叶乔木包括中山杉(Taxodium distichum 'Zhongshansha' )、池杉(T. ascendens Brongn. )、落羽杉[T. distichum (L. ) Rich. ]、墨西哥落羽杉(T. mucronatum Tenore)和水杉(Metasequoia glyptostroboides Hu et Cheng)等,常绿乔木包括樟树[Cinnamomum camphora (L. ) Presl]、北美红杉[Sequoia sempervirens (Lamb. ) Lindl. ]、浙江樟(C. chekiangensis Nakai)和青冈[Cyclobalanopsis glauca (Thunb. ) Oerst. ]等,灌木种类包括杞柳(Salix suchowensis Cheng)、木芙蓉(Hibiscus mutabilis L. )、夹竹桃(Nerium oleander L. )和栀子(Gardenia jasminoides Ellis)等.分类评价结果显示,在223个树种中有许多树种都可在4种类型的防护林中通用,这些通用树种包括中山杉、水杉、落羽杉、池杉、墨西哥落羽杉、樟树、杞柳、木芙蓉和栀子等;另外,从生活型角度,针对不同的防护林类型筛选出了一些适宜的树种.研究结果显示,该评价体系在江苏沿江生态防护林建设中具有一定的实用价值.     

红腹锦鸡和白腹锦鸡卵壳的超微结构

本文报道了锦鸡属——白腹锦鸡和红腹锦鸡卵壳的气孔、外壳膜、锥体层、木栅层的超微结构。并对两者的卵壳进行了比较。