Re-examination of inhibitor resistance conferred by Qo-site mutations in cytochrome b using yeast as a model system

Cytochrome b from yeast (Saccharomyces cerevisiae Meyer ex Hansen) provides a convenient model system for the study of Qo-site inhibitor (QoI) resistance mutations from a variety of organisms. QoI resistance mutations from fungal plant pathogens (G143A and F129L), malaria agent Plasmodium sp (Y279C/S), and Pneumocystis carinii (L275F), an opportunistic pathogenic fungus of man, were introduced into yeast cytochrome b and their effect on the binding of a variety of natural (myxothiazol and stigmatellin) and synthetic (atovaquone, azoxystrobin and pyraclostrobin) inhibitors to the bc1 complex monitored. L275S (from a myxothiazol-resistant yeast) was also re-examined. Stigmatellin binding was relatively unaffected by the introduction of these mutations. Significant increases in resistance were observed for the strobilurin-class inhibitors myxothiazol, azoxystrobin and pyraclostrobin, with the largest increase in resistance conferred by G143A. In contrast, atovaquone binding was most effected by Y279C/S and L275S. Notably, F129L, G143A and L275S had a minor effect on bc1 activity, and so are unlikely to confer significant fitness penalties in vivo. These data are discussed in the light of the atomic structures for myxothiazol- and azoxystrobin-inhibited bovine bc1 which have recently become available. We propose that QoI resistance due to G143A arises from steric hindrance between the inhibitor and cytochrome b, whereas the mechanism of resistance for the other mutations is due to an increase in binding energy between the protein and inhibitor molecule. Site-directed mutagenesis was also used to model selected regions of the mammalian Qo site in yeast cytochrome b in order to further understand the differential efficacy of these QoI in the mammalian and pathogen bc1 complexes.     

Mechanisms of resistance to acetyl-coenzyme A carboxylase inhibitors: a review

Resistance to acetyl-coenzyme A carboxylase (ACCase) inhibitors has developed in at least 10 grass weed species in recent years. In most instances, resistance is conferred by an ACCase alteration in the resistant biotypes that reduces sensitivity to aryloxyphenoxypropionate (AOPP) and cyclohexanedione (CHD) herbicides. Analysis of ACCase from many of these resistant weed biotypes suggests the presence of different mutations, each conferring a different pattern and level of resistance to various AOPP and CHD herbicides. In all cases analyzed to date, resistance is controlled by a single dominant or semi-dominant nuclear gene. In several weed biotypes, resistance is conferred by enhanced herbicide detoxification, primarily through elevated expression or activity of cytochrome P450 monooxygenase(s). This mechanism can confer cross-resistance to herbicides from other chemical classes with different modes of action. Finally, multiple herbicide resistance, i.e. the acquisition of several different resistance mechanisms, has been reported in some weed biotypes. ©1997 SCI     

Evaluation of efflux pump inhibitory activity of some plant extracts and using them as adjuvants to potentiate the inhibitory activity of some antibiotics against Staphylococcus aureus

Background:Antibiotic-resistant pathogens became a real global threat to human and animal health. This needs to concentrate the efforts to minimize and control these organisms. Efflux pumps are considered one of the important strategies used by bacteria to exclude harmful materials from the cell. Inhibition of these pumps can be an active strategy against multidrug resistance pathogens. There are two sources of efflux pump inhibitors that can be used, chemical and natural inhibitors. The chemical origin efflux pump inhibitors have many toxic side effects while the natural origin is characterized by a wide margin of safety for the host cell.Aim:In this study, the ability of some plant extracts like (propolis show rosemary, clove, capsaicin, and cumin) to potentiate the inhibitory activity of some antibiotics such as (ciprofloxacin, erythromycin, gentamycin, tetracycline, and ampicillin) against Staphylococcus aureus pathogen were tested.Methods:Efflux pump inhibitory activity of the selected plant extracts was tested using an ethidium bromide (EtBr) accumulation assay.Results:The results have shown that Propolis has a significant synergistic effect in combination with ciprofloxacin, erythromycin, and gentamycin. While it has no effect with tetracycline or ampicillin. Also, no synergic effect was noticed in a combination of the minimum inhibitory concentration for the selected plant extracts (rosemary, clove, capsaicin, and cumin) with any of the tested antibiotics. Interestingly, according to the results of the EtBr accumulation assay, Propolis has potent inhibitory activity against the S. aureus (MRS usa300) pump system.Conclusion:This study suggests that Propolis might act as a resistance breaker that is able to restore the activity of ciprofloxacin, erythromycin, and gentamycin against S. aureus strains, in case of the efflux-mediated antimicrobial resistance mechanisms.     

Study on bone marrow angiogenic mediators and inhibitors in aplastic anemia

AIM: Through detecting bone marrow angiogenic mediators and inhibitors in aplastic anemia (AA) patients,the value of angionesis in AA pathogenesis was elucidated.METHODS: The patients were divided into severe AA group (SAA,8 patients),non severe AA group (NSAA,10 patients),and normal control group (7 persons),5 patients were observed before treating (group beginning) and getting improvement (group improving).The angiogenic mediators vascular endothelial growth factor (VEGF) and bFGF were detected by ELISA,angiogenic inhibitors IFN-γ and TSP were detected by ELISA and flow cytometry,respectively.RESULTS: The levels of VEGF were lower in SAA group and NSAA group than those in control group significantly (P<0.05),the levels of IFN-γ and TSP were higher than those in control group (P<0.05),especially in SAA group (P<0.01).Compared with group beginning,the level of VEGF was higher in group improving (P<0.05),the levels of IFN-γ,TSP were lower (P<0.05),there was no obviously difference between group beginning and group improving except IFN-γ.CONCLUSION: The dropping of angiogenic mediators and the rising of angiogenic inhibitors may be one reason of reducing the number of microvessel,which result in deficiency in supporting hemapoietic stem cells by bone marrow microenvironment.     

四种SBIs类杀菌剂对不同发育阶段小麦赤霉病菌的毒力及其作用方式

为明确不同发育阶段小麦赤霉病菌对甾醇生物合成抑制剂类(SBIs)杀菌剂的敏感性差异及杀菌剂的作用方式,对4种SBIs类杀菌剂进行了室内毒力测定及田间药效试验。室内毒力测定结果表明:不同发育阶段小麦赤霉病菌对4种SBIs类药剂的敏感性存在较大差异,其敏感性由高到低依次为菌丝生长阶段和产孢阶段芽管伸长阶段孢子萌发阶段;其中,戊唑醇对菌丝生长、产孢、芽管伸长和孢子萌发阶段的EC50值分别为0.686、0.191、3.532和3.825μg/m L,咪鲜胺分别为0.063、0.305、0.827和36.581μg/m L,苯醚甲环唑分别为0.873、3.659、5.687和79.465μg/m L,烯唑醇分别为1.961、33.658、41.881和54.986μg/m L。显微观察4种SBIs类杀菌剂对小麦赤霉病菌菌丝生长和孢子萌发形态的影响,发现杀菌剂处理后菌丝末端分枝增多,新生菌丝伸长受阻,菌丝体局部膨大,有时可见菌丝及芽管的消解。田间试验结果表明:用量为有效成分100 g/hm2的戊唑醇、200 g/hm2的咪鲜胺和200 g/hm2的苯醚甲环唑对小麦赤霉病的保护效果为77.15%~79.26%,治疗效果为70.64%~72.77%,于发病前后使用均可有效防治该病害。     

植物源血管紧张素转换酶抑制剂的研究进展

血管紧张素转换酶抑制剂(Angiotensin converting enzyme inhibitors,ACEI)通过肾素-血管紧张素系统(RAS)和激肽释放酶-肽酶系统(KKS),抑制血管紧张素II的生成,减少缓激肽的降解,从而达到降血压的目的;植物来源的天然化合物具有来源广泛、结构多样、特异性高、毒性低等特点,因而从植物中筛选安全有效的ACEI成为研究热点。在此综述了血管紧张素转换酶(ACE)的活性测定方法以及植物来源的不同结构类型的ACEI研究进展。     

EFFECT OF NEEM-OIL COATED PRILLED UREA WITH VARYING THICKNESS OF NEEM-OIL COATING AND NITROGEN RATES ON PRODUCTIVITY AND NITROGEN-USE EFFICIENCY OF LOWLAND IRRIGATED RICE UNDER INDO-GANGETIC PLAINS

The nitrogen (N) fertilizer-use efficiency (20–50%) is low in rice fields in India. The neem-oil coated urea can increase N-use efficiency in lowland rice, but the desirable thickness of neem-oil coating onto urea is not known yet. Therefore, field experiments were conducted during kharif (rainy) season years 2004 and 2005 at the Research Farm of Indian Agricultural Research Institute, New Delhi to know the suitable thickness of neem-oil coating on prilled urea (PU) for increased N-use efficiency and yield. The treatments comprised of twelve combinations of four N sources (PU coated with neem-oil thickness of 0, 500, 1000 and 2000 mg kg^?1 PU) and three N levels (50, 100, and 150 kg N ha^?1) plus a no-N control. Prilled urea (PU) refers to the common urea available commercially in prills, which is different from urea super granules. Application of urea coated with neem-oil thickness of 1000 mg kg^?1 PU resulted in significantly higher growth, yield parameters, grain yield, N uptake, and efficiency of aromatic rice (Oryza sativa L.) over uncoated PU. Nitrogen application at 122 kg ha^?1 was optimum for increased yield of rice. Nitrogen-use efficiency decreased significantly and substantially with each successive increase in levels of N from 50 to 150 kg ha^?1.     

信号转导抑制剂对囊素刺激杂交瘤细胞抗体分泌及其mRNA表达的影响

以杂交瘤细胞为模型 ,在培养液中加入不同的信号转导抑制剂和囊素 ,用间接ELISA法测定抗体分泌水平 ,用定量RT PCR法检测抗体mRNA的表达水平。结果表明 ,加入Gα 蛋白选择性抑制剂NF 0 2 3、蛋白激酶C抑制剂GF10 92 0 3X、磷脂酰肌醇 3激酶抑制剂heparin、Ca2 /calmodulin依赖蛋白激酶Ⅱ抑制剂KN 6 2、磷脂酶C抑制剂U 7312 2和蛋白激酶G抑制剂后 ,囊素对杂交瘤细胞抗体分泌速度的影响明显 (P <0 0 1)。而加入蛋白激酶A抑制剂 4 Cyano 3 M thylisoquinoline、酪氨酸蛋白激酶抑制剂 genistein和丝裂原活化的蛋白激酶抑制剂PD 980 5 9后 ,囊素对杂交瘤细胞抗体分泌速度的影响不明显 (P >0 0 5 ) ,这 3种抑制剂对杂交瘤细胞抗体分泌速度的抑制率分别为 74 5 5 % ,91 13%和80 6 9%。加入 4 Cyano 3 M thylisoquinoline、genistein和PD 980 5 9后 ,囊素处理组与对照组γ1mRNA的表达水平差异不显著 (P >0 0 5 ) ,说明这 3种抑制剂阻断了囊素刺激的杂交瘤细胞抗体mRNA的表达。以上结果表明 ,蛋白激酶A、酪氨酸蛋白激酶和丝裂原活化的蛋白激酶参与了囊素在杂交瘤细胞中的信号转导。