An in vitro comparative study on the reactivation of nerve agent-inhibited guinea pig and human acetylcholinesterases by oximes

The reactivation of nerve agent-inhibited acetylcholinesterase (AChE) by oxime is the most important step in the treatment of nerve agent poisoning. Since the evaluation of nerve agent antidotes cannot be conducted in humans, results from animal experiments are extrapolated to humans. Guinea pig is one of the animal models that is frequently used for conducting nerve agent antidote evaluations. Several investigations have demonstrated that the efficacy of an oxime primarily depends on its ability to reactivate nerve agent-inhibited AChE. If the in vitro oxime reactivation of nerve agent-inhibited animal AChE is similar to that of human AChE, it is likely that the results of an in vivo animal study will reliably extrapolate to humans. Therefore, the goal of this study was to compare the reactivation of guinea pig and human AChEs inhibited by six different G and V type nerve agents. Reactivation kinetic studies with five mono- and bis-pyridinium oximes showed that oxime reactivation of nerve agent-inhibited human AChE in most cases was faster than guinea pig AChE. The most significant enhancement was observed in the reactivation of human AChE inhibited by nerve agents containing bulky side chains GF, GD, and VR, by H-series oximes HLo-7, HI-6, and ICD-585. In these cases, species-related differences observed between the two AChEs, based on the second-order reactivation rate constants, were 90- to over 400-fold. On the other hand, less than 3-fold differences were observed in the rates of aging of nerve agent-inhibited guinea pig and human AChEs. These results suggest that the remarkable species-related differences observed in the reactivation of nerve agent-inhibited guinea pig and human AChEs were not due to differences in the rates of aging. These results also suggest that guinea pig may not be an appropriate animal model for the in vivo evaluation of oxime therapy.     

Efficacy of the entomopathogenic nematode, Steinernema feltiae, against sweetpotato whitefly Bemisia tabaci (Homoptera: Aleyrodidae) under laboratory and glasshouse conditions

The potential of using the entomopathogenic nematode Steinernema feltiae to control the sweetpotato whitefly Bemisia tabaci (Gennadius) has been established in previous laboratory studies. However, laboratory studies can overestimate the level of control achieved by biocontrol agents in the glasshouse. Glasshouse trials are therefore required to confirm laboratory results before full-scale commercial development is considered. Under both controlled laboratory and glasshouse conditions high mortality of second instar B. tabaci (>90% and >80%, respectively) was recorded after application of S. feltiae. The efficacy of the biocontrol agent at various application rates was also investigated, where halving the rate of S. feltiae application caused no significant reduction in B. tabaci mortality on tomato foliage. Steinernema feltiae has shown much potential for incorporation into integrated pest management strategies for the control of B. tabaci.     

Panicle Water Potential, a Physiological Trait to Identify Drought Tolerance in Rice

Two upland rice varieties （IRAT109, IAPAR9） and one lowland rice variety （Zhenshan 97B） were planted in summer and treated with both normal （full water） and drought stress in the reproductive stage. Panicle water potential （PWP） and leaf water potential （LWP） were measured every 1.0-1.5 h over 24 h on sunny days. Both PWP and LWP of upland varieties started to decrease later, maintained a higher level and recovered more quickly than that of the lowland variety. The results show that PWP can be used as an indicator of plant water status based on the parallel daily changes, and the high correlation between PWP and LWP. Similar correlations were also observed between PWP, LWP and eight traits related to plant growth and grain yield formation. PWP seemed to be more effective for distinguishing the upland rice varieties with different drought-tolerant ability. Differences in PWP and LWP between upland and lowland rice varieties were also observed at noon even under normal water conditions, implying the incorporation of the drought-tolerant mechanism to improve the photosynthesis and yield of traditional paddy rice.     

Cloning, purification, and characterization of thermostable hypoxanthine-guanine phosphoribosyltransferase from Thermoanaerobacter tengcongensis

Hypoxanthine-guanine phosphoribosyltransferase (HGPRT, EC 2.4.2.8) from a newly characterized thermophile Thermoanaerobacter tengcongensis was expressed in Escherichia coli and purified. Analytical gel filtration suggested that the enzyme exist as a homotetramer in solution. The optimal pH for the forward reaction was found to be 8.0 and the optimal temperature 70 degrees C. The steady-state kinetic characteristics suggest that hypoxanthine is the most effective substrate. This enzyme showed a half-life of 75min at 50 degrees C and no apparent loss of activity after 3 months at 4 degrees C.     

Genetic analysis of IREB2, FAM13A and XRCC5 variants in Chinese Han patients with chronic obstructive pulmonary disease

Recently, variants (rs2568494, rs2869967 and rs3821104) in the IREB2, FAM13A and XRCC5 genes were found to be associated with chronic obstructive pulmonary disease (COPD) in non-Asian populations by genome-wide association study (GWAS) analysis. To evaluate whether variants in these genes are related to COPD in Chinese Han population, we investigated COPD patients of Chinese Han ethnicity from Mainland China. Significant differences in genotypic distributions (χ² = 6.319, p = 0.042 for rs2869967; χ² = 6.062, p = 0.048 for rs3821104) and allele distributions (χ² = 4.014, p = 0.045 for rs2869967; χ² = 5.607, p = 0.018 for rs3821104) were observed between patients and control subjects for variants rs2869967 and rs3821104, whereas no statistically significant associations for genotypic and allelic distribution between IREB2 rs2568494 and COPD phenotype (p > 0.05) were identified. Our results support that FAM13A rs2869967 and XRCC5 rs3821104 are associated with COPD in Chinese Han population.     

重组人S100A6促进人乳腺癌细胞MCF-7的增殖和迁移侵袭并抑制其凋亡

该研究旨在探讨重组人S100A6蛋白对乳腺癌细胞株MCF-7的增殖、凋亡、迁移及侵袭能力的影响。利用原核表达制备重组人S100A6蛋白(GST-hS100A6),SDS-PAGE显示其大小为36 kDa,Western blot显示其可以被S100A6抗体特异识别,BCA法测定1 L菌液共收获约16.7 mg蛋白;将其作用于人乳腺癌细胞MCF-7,MTT显示细胞培养48 h时,浓度为100μg/mL和300μg/mL的GST-hS100A6组的D_(492)值较GST组增加29.1%和84.6%(P<0.05),提示S100A6促进MCF-7细胞增殖;平板克隆形成实验显示GST-hS100A6组的克隆形成率较GST组高38.7%(P<0.05),提示S100A6促进MCF-7的克隆形成;Hoechst染色显示GST-hS100A6组在24 h时细胞凋亡率较GST组减少67.8%(P<0.05),48 h时细胞凋亡率较GST组减少58.4%(P<0.05),提示S100A6抑制MCF-7细胞凋亡;划痕实验显示在24 h时GST-hS100A6组的划痕愈合率为GST组的2.2倍(P<0.05),提示S100A6促...     

Role of phosphodiesterase in cyclic AMP signaling in cultured rat granulosa cells

Inactivation of the cyclic nucleotide signal in granulosa cells depends on a complex array of cyclic nucleotide phosphodiesterases (PDE). In order to examine the role of PDE in cyclic AMP (cAMP) signaling in granulosa cells, the present study examined the expression of PDE4D proteins and regulation of cAMP-PDE activities in cultured rat granulosa cells. The results of immunoblot analyses showed that two predominant PDE4D subtypes of approximately 80 and 70 kDa appeared when immature rat granulosa cells were treated with FSH. However, these two new subtypes presumed to be PDE4D proteins were not influenced by treatments of DETA/NO, cGMP and PKB inhibitor, LY294002. Immature rat granulosa cells treated with medium alone displayed low cAMP-PDE activity throughout 48 h of culture while those treated with FSH (2 ng.mL-1) showed a marked increase in cAMP-PDE activity between 6 and 12 h of culture, followed by a decline. The findings from the present study indicate that the increased cAMP-PDE activity by FSH is mainly related to the changes of PDE4D protein levels. However, the inhibitory effects of NO on cAMP accumulation in rat granulosa cells are not via the increased cAMP-PDE activity.     

Protein structure preference, tRNA copy number, and mRNA stem/loop content

From statistical analyses of protein sequences for humans and Escherichia coli we found that the messenger RNA segment of m-codons (for m=2 to 6) with average high tRNA copy number (TCN) (larger than approximately 10.5 for humans or approximately 1.95 for E. coli) preferably code for the alpha helix and that with low TCN (smaller than approximately 7.5 for humans or approximately 1.7 for E. coli) preferably code for coil. Between them there is an intermediate region without correlation to structure preference. For the beta strand the preference/ avoidance tendency is not obvious. All strong preference-modes of TCN for protein secondary structures have been deduced. The mutual interaction between two factors--protein secondary structural type and codon TCN--is tested by F distribution. A phenomenological model on the relation between structure preference and translational efficiency or accuracy is proposed. It is pointed out that the structure preference of codons is related to the distribution of mRNA stem/loop content in three TCN regions.