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Phenotypic methods for detection of various β-lactamases in Gram-negative clinical isolates: Need of the hour 革兰氏阴性临床分离株中各种β-内酰胺酶的表型检测方法:时势需要
Pub Date : 2012-10-01 DOI: 10.4103/2229-5186.103098
N. Nagdeo, N. Kaore, V. R. Thombare
Background: Many clinical laboratories have problems detecting various β-lactamases. Confusion exists about the importance of these resistance mechanisms, optimal test methods, and appropriate reporting conventions. It is more imperative to use various phenotypic methods for detection of various β-lactamases in routine microbiology laboratory on day-to-day basis to prevent antimicrobial resistance by evidence-based judicious use of antimicrobials. Aims: In view of the multidrug-resistant organisms being reported world over, we planned a cross-sectional prospective analytical study to determine resistance mechanism by various β-lactamases in Gram-negative clinical isolates using various phenotypic methods. Materials and Methods: All nonrepeat, nonenteric clinical isolates of Gram-negative bacilli, resistant to at least two third-generation cephalosporins, were first screened by Novel disc placement method, and isolates showing multiple mechanisms of resistance and reduced zone of inhibition for imipenem were further confirmed for AmpC and metallo β-lactamases. Statistical Analysis: All the data was managed and analyzed in Microsoft Excel. Results: Out of 807 isolates tested, as many as 795 (98.51%) revealed the presence of extended-spectrum β-lactamases (ESBLs). Only 10 isolates of Escherichia coli and 2 of Klebsiella pneumoniae did not show production of ESBL. A total of 450 (55.76%) isolates produced single enzyme,while 345 (42.75%) strains revealed multiple enzyme production simultaneously. Only ESBL production was seen in 315 (39.03%) strains, only AmpC in 75 (9.29%) and only MBL in 60 (7.44%) strains, while ESBL and AmpC together were seen in 219 (27.14%) and AmpC plus MBL in 92 (11.40%) strains. However, ESBL plus MBL were never observed together. All three enzymes were simultaneously detected in 34 (4.21%) strains. Conclusion: This innovative method of disc placement makes it easy, affordable, and reliable method for routine use by basic microbiology laboratories for detection of various β-lactamases, pending confirmation for AmpC and metallo β-lactamase by three-dimensional test and double disc potentiation test, respectively.
背景:许多临床实验室在检测各种β-内酰胺酶方面存在问题。对于这些抗性机制、最佳测试方法和适当的报告惯例的重要性存在混淆。日常常规微生物实验室应采用多种表型方法检测各种β-内酰胺酶,以循证用药预防耐药。目的:鉴于世界范围内多药耐药菌的报道,我们计划采用横断面前瞻性分析研究,利用各种表型方法确定革兰氏阴性临床分离株中各种β-内酰胺酶的耐药机制。材料与方法:首先采用Novel盘片法筛选对至少两种第三代头孢菌素耐药的革兰氏阴性杆菌临床非重复、非肠源性分离株,进一步证实其对AmpC和金属β-内酰胺酶具有多种耐药机制和抑制区缩小。统计分析:所有数据在Microsoft Excel中进行管理和分析。结果:807株分离菌中,795株(98.51%)检出广谱β-内酰胺酶(ESBLs)。只有10株大肠埃希菌和2株肺炎克雷伯菌没有产生ESBL。450株(55.76%)产生单酶,345株(42.75%)同时产生多种酶。315株(39.03%)、75株(9.29%)和60株(7.44%)仅产ESBL, 219株(27.14%)和92株(11.40%)分别产AmpC和MBL。然而,ESBL和MBL从未同时观察到。同时检出3种酶的菌株34株(4.21%)。结论:该创新的圆盘放置方法可作为基础微生物实验室常规检测各种β-内酰胺酶的简便、经济、可靠的方法,有待于AmpC和金属β-内酰胺酶分别通过三维试验和双圆盘强化试验进行确认。
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引用次数: 23
A validated stability indicating high-performance liquid chromatographic method for simultaneous estimation of cefuroxime sodium and sulbactam sodium in injection dosage form 建立了同时测定注射剂型头孢呋辛钠和舒巴坦钠含量的高效液相色谱法
Pub Date : 2012-10-01 DOI: 10.4103/2229-5186.103096
F. M. Patel, J. Dave, P. J. Vyas, C. Patel
Background: A fixed dose combination of cefuroxime sodium (β lactam antibiotic) and sulbactam sodium (β Lactamase inhibitor) is used in ratio of 2:1 as powder for injection for the treatment of resistant lower respiratory tract and other infections. Aims: A simple, precise, and accurate ion-pair reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for determination of cefuroxime Na(CEF) and sulbactam Na(SUL) in injection. Materials and Methods: Isocratic RP-HPLC separation was achieved on an ACE C 18 column (150×4.6 mm id, 5 μm particle size) using the mobile phase 0.002 M tetrabutylammonium hydroxide sulfate (TBAH) in 10 mm potassium di-hydrogen phosphate buffer-acetonitrile (86:14 v/v, pH 3.7) at a flow rate of 1.0 ml/min. Results and Conclusion: The retention time of sulbactam Na and cefuroxime Na were 3.2 min and 10.2 min, respectively. The ion-pairing reagent improved the retention of highly polar sulbactam Na on reverse-phase column. The detection was performed at 210 nm. The method was validated for linearity, precision, accuracy, robustness, solution stability, and specificity. The method was validated for linearity, precision, accuracy, robustness, solution stability, and specificity. The method was linear in the concentration range of 10-100 μg/ml for cefuroxime Na and 5-50 μg/ml for sulbactam Na, with a correlation coefficient of 0.9999 and 0.9998 for the respective drugs. The intraday precision was 0.13-0.21% and 0.48-0.65%, and the interday precision was 0.32-0.81% and 0.60-0.83% for cefuroxime Na and sulbactam Na, respectively. The accuracy (recovery) was found to be in the range of 98.76-100.61% and 98.99-100.30% for cefuroxime Na and sulbactam Na, respectively. The drugs were found to degrade under hydrolytic and oxidative conditions. The drugs could be effectively separated from different degradation products, and hence the method can be used for stability analysis.
背景:采用头孢呋辛钠(β内酰胺类抗生素)和舒巴坦钠(β内酰胺酶抑制剂)的固定剂量组合,以2:1的比例作为粉剂用于注射治疗耐药下呼吸道感染和其他感染。目的:建立一种简便、精确、准确的离子对反相高效液相色谱(RP-HPLC)测定注射用头孢呋辛钠(CEF)和舒巴坦钠(SUL)的方法。材料与方法:采用ACE C - 18色谱柱(150×4.6 mm id, 5 μm粒径),流动相为0.002 M四丁基氢氧化铵硫酸铵(TBAH),溶液为10 mm磷酸二氢钾缓冲液-乙腈(86:14 v/v, pH 3.7),流速为1.0 ml/min。结果与结论:舒巴坦钠和头孢呋辛钠的滞留时间分别为3.2 min和10.2 min。离子配对试剂提高了高极性舒巴坦钠在反相柱上的保留率。在210 nm处进行检测。方法具有线性、精密度、准确度、鲁棒性、溶液稳定性和特异性。方法具有线性、精密度、准确度、鲁棒性、溶液稳定性和特异性。头孢呋辛钠和舒巴坦钠在10 ~ 100 μg/ml和5 ~ 50 μg/ml浓度范围内呈线性关系,相关系数分别为0.9999和0.9998。头孢呋辛钠和舒巴坦钠日内精密度分别为0.13 ~ 0.21%和0.48 ~ 0.65%,日内精密度分别为0.32 ~ 0.81%和0.60 ~ 0.83%。头孢呋辛钠和舒巴坦钠的准确度(回收率)分别为98.76 ~ 100.61%和98.99 ~ 100.30%。发现这些药物在水解和氧化条件下会降解。该方法能有效地将药物从不同的降解产物中分离出来,可用于稳定性分析。
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引用次数: 2
An account of phytochemicals from Plumbago zeylanica (Family: Plumbaginaceae): A natural gift to human being # 白桦花属植物化学物质记述:天赐给人类的礼物#
Pub Date : 2012-07-01 DOI: 10.4103/2229-5186.99564
N. Kishore, B. B. Mishra, V. Tiwari, V. Tripathi
The semi-climbing sub-shrub Plumbago zeylanica (family: Plumbaginaceae) is a widely accepted ethnomedicine around the world including India, Pakistan, Bangladesh, Sri Lanka, and Australia. The plant is credited with potential therapeutic properties including antiatherogenic, cardiotonic, hepatoprotective, and neuroprotective properties. The present review highlights the various medicinal and pharmacological aspects along with recent updates on phytochemical contents of the plant.
半攀缘亚灌木白桦花(plumumbago zeylanica)(科:白桦花科)是世界上广泛接受的民族药,包括印度,巴基斯坦,孟加拉国,斯里兰卡和澳大利亚。该植物被认为具有潜在的治疗特性,包括抗动脉粥样硬化、强心剂、肝保护和神经保护特性。目前的综述重点介绍了该植物的各种药用和药理学方面以及植物化学成分的最新进展。
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引用次数: 28
In vitro antimicrobial status of methanolic extract of Citrus sinensis Linn. fruit peel 柑桔醇提物体外抑菌性能研究。果皮
Pub Date : 2012-07-01 DOI: 10.4103/2229-5186.99573
A. Dhiman, A. Nanda, Sayeed Ahmad, B. Narasimhan
Aim : The present investigation evaluated the antimicrobial potential of methanolic extract of Citrus sinensis Linn. (Rutaceae) fruit peel. There is a basis for the traditional use of this plant for local health remedies. Materials and Methods: The antimicrobial activity of methanolic extract of C. sinensis fruit peel was tested against three bacterial and two fungal strains. Turbidimetric or tube dilution method and paper disc diffusion method were followed. Results are expressed as mean ± standard deviation. Results: The C. sinensis fruit peel methanolic extract exhibited antibacterial activity against Escherichia coli with minimum inhibitory concentration of 0.78 μg/ ml and minimum bactericidal concentration of 6.25 μg/ml, and appreciable antifungal activity with minimum inhibitory concentration of 12.5 μg/ml. The phytochemistry of C. sinensis fruit peel methanolic extract revealed the presence of carbohydrates (reducing sugars, hexose sugars, non-reducing polysaccharides, gums, and mucilages), flavonoid glycosides, coumarin glycosides, volatile oils, organic acids, fats and fixed oils. Conclusion: Most of the organic chemical constituents reported are aromatic phenolic compounds, which are known for their wide spectra of antimicrobial activity. Therefore, the bacteriostatic and fungistatic action of the tested extract may be attributed to the presence of polyphenolic compounds. In short, C. sinensis fruit peel methanolic extract is a potential source of natural antimicrobials.
目的:评价柑桔醇提物的抑菌潜力。(芸香科)果皮。有一个基础的传统使用这种植物的地方保健疗法。材料与方法:对枳实果皮甲醇提取物进行了抑菌试验,测定其对3株细菌和2株真菌的抑菌活性。采用比浊法、管稀释法和纸片扩散法。结果以均数±标准差表示。结果:枸杞果皮甲醇提取物对大肠杆菌具有较强的抑菌活性,最低抑菌浓度为0.78 μg/ml,最低杀菌浓度为6.25 μg/ml;最低抑菌浓度为12.5 μg/ml,抑菌活性明显。果皮甲醇提取物的植物化学成分分析表明,果皮甲醇提取物中含有碳水化合物(还原糖、己糖、非还原性多糖、树胶和粘液)、类黄酮苷、香豆素苷、挥发油、有机酸、脂肪和固定油。结论:报道的有机化学成分多为芳香酚类化合物,具有广泛的抗菌活性。因此,所测提取物的抑菌和抑菌作用可能归因于多酚化合物的存在。总之,柑橘果皮甲醇提取物是天然抗菌剂的潜在来源。
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引用次数: 31
Trace element structure of the most widespread plants of genus Pulmonaria 最广泛分布的肺属植物的微量元素结构
Pub Date : 2012-07-01 DOI: 10.4103/2229-5186.99587
D. Kruglov
Aim: The aim of this work was a comparative research of trace element structure of various organs of three Pulmonaria species. Materials and Methods: The aerial parts of the most widespread plants of genus Pulmonaria such as Pulmonaria officinalis L., Pulmonaria obscura Dumort. and Pulmonaria mollis Wulf. ex Hornem., which were collected in ending of flowering and were used as the research objects. The amount of trace elements (B, K, P, V, Ca, Co, Cu, Fe, Mg, Mn, Mo, Na, Si, Zn, Ag, Al, Ba, Br, Cr, I, Ni, Se, Sr, and Ti) was determined by means of mass spectroscopy with inductively coupled plasma. Results: The data clustering has shown that floral shoots and rosellate leaves possess essentially various trace element status. At the same time, the trace elements' status of organs of researched plants poorly depends on a taxonomic position of the plant. Thereupon, it is obvious that pharmacological activity is defined by organs of plants from which medicines were made, but not by a species of the used plant. Conclusions: The significant distinction in pharmacological activity of preparations depends on the trace elements' status of used medicinal vegetative raw materials.
目的:对三种肺属植物各器官的微量元素结构进行比较研究。材料与方法:选取分布最广的肺属植物的地上部分,如铁皮肺、暗肺等。和肺孢菌。Hornem交货。,作为研究对象,采集于开花末期。采用电感耦合等离子体质谱法测定了样品中微量元素(B、K、P、V、Ca、Co、Cu、Fe、Mg、Mn、Mo、Na、Si、Zn、Ag、Al、Ba、Br、Cr、I、Ni、Se、Sr和Ti)的含量。结果:数据聚类表明,花芽和莲座叶具有基本不同的微量元素状态。同时,所研究植物器官中微量元素的地位与植物的分类位置关系不大。因此,很明显,药理学活性是由制造药物的植物器官决定的,而不是由所用植物的种类决定的。结论:药用植物原料中微量元素的状态决定了制剂药理活性的显著差异。
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引用次数: 1
Cytotoxicity study of plant Aloe vera (Linn) 植物芦荟的细胞毒性研究
Pub Date : 2012-07-01 DOI: 10.4103/2229-5186.99595
A. Chandu, Santhosh Kumar, C. Bhattacharjee, S. Debnath
Background: The objective of this study has been to evaluate the in-vitro antitumor activity of Aloe vera extract of in cultured B16F10 melanoma cell line by measuring cell viability using "Trypan blue exclusion assay" method. Aim: To find out such kind of anticancer drug which is a cheap, safe, less toxic, and more potent drug compared to chemotherapy drug. Materials and Methods: In-vitro antitumor activity cell culture1, drug treatment (standard and test extract) and Trypan blue exclusion assay growth and viability test 1 were used. Treatment of Aloe vera extract against B16F10 melanoma cell line, in all concentration range, showed decrease in percent cell viability, as compared to that of negative when examined by "Trypan blue exclusion assay". Results: In overall variation of test samples, Aloe vera extract showed its best activity in the concentration of 300 μg/ml, which was approximately equal to the activity of standard drug doxorubicin. Evaluation of in-vitro antitumor activity revealed that Aloe vera extract exhibits good cytotoxic activity. The best cytotoxic activity by Aloe vera was shown at 200 μg/ml concentration. Conclusion: The study of cytoprotection against normal cells by micronucleus assay has shown that the herbal extracts have less toxic effects to the normal blood lymphocytes, as compared to that of standard anticancer drug.
背景:本研究的目的是通过“台盼蓝排斥法”测定体外培养的B16F10黑色素瘤细胞株的细胞活力,评价芦荟提取物的体外抗肿瘤活性。目的:寻找一种比化疗药物更便宜、安全、毒性更小、更有效的抗癌药物。材料和方法:体外抗肿瘤活性细胞培养1、药物处理(标准品和试验提取物)、台盼蓝排除法生长和活力试验1。在所有浓度范围内,芦荟提取物对B16F10黑色素瘤细胞系的细胞存活率都比阴性的细胞存活率低。结果:在所测样品的总体变化中,芦荟提取物在300 μg/ml的浓度下活性最佳,与标准药物阿霉素的活性大致相当。体外抗肿瘤活性评价表明,芦荟提取物具有良好的细胞毒活性。芦荟在200 μg/ml浓度下的细胞毒活性最佳。结论:微核实验对正常细胞的细胞保护作用的研究表明,与标准抗癌药物相比,中药提取物对正常血液淋巴细胞的毒性作用较小。
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引用次数: 16
Antihyperlipidemic and antioxidant activities of Bruguiera cylindrinca (L) 白茅的降血脂及抗氧化活性研究
Pub Date : 2012-07-01 DOI: 10.4103/2229-5186.99596
V. Lakshmi, Ravi Sonkar, A. Khanna
Background: Atheroclerosis is the major cause of morbidity and mortality world wide. The drugs used these days have some or other side effects. Aim: Therefore it is an urgent need to develop a safe and cheaper drugs for the society. We have planned this study to investigate lipid lowering and antioxidant activities of Bruguiera cylindrinca in triton and cholesterol fed rats. Materials and Methods: Different extract and fractions were collected from B. cylindrica. Two major compounds were isolated from there and have been tested for their lipid lowering activity along with increase in high density lipoprotein,cholesterol, compared with the cholesterol fed animals. Results: On chronic feeding of active fraction and pure compounds (at 100 mg / kg b.w.) for 30 days. The pure compounds isolated from the active fraction were found showing lipid lowering activity similar to the fraction. Conclusion: Structre modification of the two pure compounds isolated from the active fraction is required for further enhacement of biological activity.
背景:动脉粥样硬化是世界范围内发病率和死亡率的主要原因。现在使用的药物都有这样或那样的副作用。目的:因此,开发一种安全、廉价的药物是社会迫切需要的。本实验旨在探讨白茅对高脂和高胆固醇大鼠的降脂和抗氧化活性。材料与方法:从白茅中提取不同部位的提取物。从那里分离出两种主要化合物,并对它们的降脂活性进行了测试,同时与喂食胆固醇的动物相比,它们的高密度脂蛋白和胆固醇也有所增加。结果:活性部分和纯化合物(剂量为100 mg / kg b.w)长期饲喂30 d。从活性部位分离得到的纯化合物具有与该部位相似的降脂活性。结论:从活性部位分离得到的两种纯化化合物需要进行结构修饰以进一步提高生物活性。
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引用次数: 7
Steroidal saponin from Chlorophytum nimonii (Grah) with lipid-lowering and antioxidant activity 从吊兰中提取具有降脂和抗氧化活性的甾体皂苷
Pub Date : 2012-07-01 DOI: 10.4103/2229-5186.99592
V. Lakshmi, A. Mahdi, S. K. Agarwal, A. Khanna
Aim: Since drugs used these days to lower the lipids are all synthetic drugs, they have some or the other side effects, therefore in search of cheaper lipid-lowering drugs with no side effects, we have conducted a study on Chlorophytum nimonii for its lipid-lowering and antioxidant properties. Materials and Methods: Chloragin and Gemfibrozil both caused a significant decrease in the serum level of lipids in triton-induced hyperlipidemic rats, and this model has been successfully used for the evaluation of lipid-lowering activity of chloragin in the rats. Results and Discussion: The lipid-lowering action of steroidal saponin and chloragin of the C. nimonii has been studied in triton model (in cholesterol-fed hyperlipidemic rats) in vivo and antioxidant activity in vitro model. Serum lipids were found to be lowered by the steroidal saponin (100 mg/kg body weight) in triton WR-1339-induced hyperlipidemia. Chronic feeding of this compound (50 mg/kg) in animals simultaneously fed with high-fat diet for 30 days caused lowering in the lipid and lipoproteins levels of low-density lipoproteins in experimental animals. Conclusion: Chloragin activates lipolytic enzymes in plasma and liver. Chloragin is mediated through inhibition of hepatic lipids, increased fecal bile acid excretion, and enhanced plasma lecithin cholesterol acyl transferase activities. Chloragin from the C. nimonii showed potent antioxidant activity as well.
目的:由于目前使用的降脂药物都是合成药物,它们都有这样或那样的副作用,因此为了寻找更便宜的无副作用的降脂药物,我们对吊兰的降脂和抗氧化特性进行了研究。材料与方法:氯霉素和吉非罗齐均能显著降低triton诱导的高脂血症大鼠的血清脂质水平,该模型已成功用于评价氯霉素对大鼠的降脂活性。结果与讨论:在triton模型(高胆固醇喂养的高脂血症大鼠)中,研究了山参甾体皂苷和氯代金的体内降脂作用和体外抗氧化活性。在triton wr -1339诱导的高脂血症中,类固醇皂苷(100 mg/kg体重)可降低血脂。与高脂饲料同时长期饲喂该化合物(50 mg/kg) 30 d,可降低实验动物低密度脂蛋白的脂质和脂蛋白水平。结论:氯霉素可激活血浆和肝脏的脂溶酶。氯agin通过抑制肝脏脂质、增加粪胆汁酸排泄和增强血浆卵磷脂-胆固醇酰基转移酶活性来介导。同时,绿僵菌的氯agin也具有较强的抗氧化活性。
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引用次数: 11
A novel protein tyrosine phosphatase 1B inhibitor from Tinospora sinensis 中华Tinospora sinensis蛋白酪氨酸磷酸酶1B抑制剂
Pub Date : 2012-07-01 DOI: 10.4103/2229-5186.99569
Prasoon Gupta, Upasana Sharma, P. Gupta, R. Maurya
Bioassay-directed fractionation led to the identification of a new compound, 4-hydroxy-heptadec-6-enoic acid ethyl ester (1) together with three known compounds (2-4) from Tinospora sinensis. The structure of 1 was determined by analysis of spectroscopic data. The isolated compounds were evaluated for their protein tyrosine phosphatase 1B (PTP1B) inhibitory activity. Compounds 1 and 2 displayed significant inhibitory activity with IC 50 values 61.1 and 74.2 μM, respectively
生物测定定向分离鉴定了一个新的化合物,4-羟基十七烷基-6-烯酸乙酯(1)和三个已知的化合物(2-4)从Tinospora sinensis。通过光谱数据分析确定了1的结构。对分离得到的化合物进行了蛋白酪氨酸磷酸酶1B (PTP1B)抑制活性评价。化合物1和2的ic50分别为61.1 μM和74.2 μM,具有显著的抑制活性
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引用次数: 5
Special issue in honor of Dr. Norman R. Farnsworth 纪念诺曼·法恩斯沃斯博士的特刊
Pub Date : 2012-07-01 DOI: 10.4103/2229-5186.99555
Prasoon Gupta
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引用次数: 0
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Chronicles of Young Scientists
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