Cell culture isolation can miss the laboratory diagnosis of HSV ocular infection

AIMWe compared polymerase chain reaction (PCR) to cell culture isolation for the laboratory diagnosis of ocular herpes simplex virus (HSV) disease.

Evaluation of the Antibacterial and Antifungal Capacity of Nanoemulsions Loaded with Synthetic Chalcone Derivatives Di-Benzyl Cinnamaldehyde and Benzyl 4-Aminochalcone

With the increase in antimicrobial resistance,it has become necessary to explore alternative approaches for combating and preventing diseases.DB-cinnamaldehyde(CNM)and Benzyl4-amino(B4AM)are bioactive compounds derived from chalcones but with restricted solubility in aqueous media.Nanoemulsions can enhance the solubility of compounds and can be a promising alternative in the development of novel antimicrobials,with reduced side effects and prolonged release.The objective of this study was to evaluate the stability of oil-in-water nanoemulsions loaded with two distinct types of chalcones at two different dosages,to propose a stable formulation with antimicrobial properties.Results showed that nanoemulsions presented high encapsulation efficiency,low polydispersity index(PDI)and particle size below 200 nm,indicating that emulsification was a suitable method for nanoemulsion preparation.Nanoemulsions with higher dosages exhibited significant antimicrobial effects when compared to free chalcones and positive controls.Notably,B4AM nanoemulsions at higher dosages showed expressive activity against Salmonella minnesota,with a 420%greater inhibitory response compared to the free form and showing equivalence to the positive control.CNM nanoemulsions showed excellent inhibitory activity at the highest dosage,equivalent to the positive control against S.minnesota and Staphylococcus aureus.The greater number of conjugated bonds in CNM increased the antimicrobial activity in comparison with B4AM,and the formation of nanometric domains enhanced the bioavailability,being a promising alternative for antimicrobial applications.

psk1 virulence gene-induced pulmonary and systemic tuberculosis in a young woman with normal immune function:A case report

BACKGROUND Tuberculosis is a chronic infectious disease and an important public health pro-blem.Despite progress in controlling tuberculosis,the incidence of tuberculosis in China is still very high,with 895000 new cases annually.This case report des-cribes the investigation of a case of severe disseminated tuberculosis in a young adult with normal immune function,conducted to ascertain why a Mycobacterium tuberculosis(M.tuberculosis)strain caused such severe disease.CASE SUMMARY A previously healthy 28-year-old woman presented to our hospital with a 1-mo-nth history of fever and fatigue.She was diagnosed with severe disseminated pulmonary tuberculosis,spinal tuberculosis with paravertebral abscesses,and tuberculous meningitis.M.tuberculosis was isolated from bronchoal-veolar lavage fluid.She was treated with standard antituberculous therapy and underwent debridement,bone graft,and internal fixation surgery for spinal tuberculosis.She responded to therapy and regained her ability to walk following the surgery.We analysed the whole-genome sequence of the strain and designated it BLM-A21.Additional M.tuberculosis genomes were selected from the Virulence Factor Database(http://www.mgc.ac.cn/cgi-bin/VFs/genus.cgi?Genus=Mycobacterium)for comparison.An evolutionary tree of the BLM-A21 strain was built using PhyML maximum likelihood software.Further gene analysis revealed that,except for the pks1 gene,BLM-A21 had similar virulence genes to the CDC 1551 and H37Rv strains,which have lower dissemination.CONCLUSION We speculate that the pks1 virulence gene in BLM-A21 may be the key virulence gene responsible for the wide-spread dissemination of M.tuberculosis infection in this previously healthy adult with normal immune function.

Alcohol solvent effect on the self-assembly behaviors of lignin oligomers

The interactions between lignin oligomers and solvents determine the behaviors of lignin oligomers self-assembling into uniform lignin nanoparticles(LNPs).Herein,several alcohol solvents,which readily interact with the lignin oligomers,were adopted to study their effects during solvent shifting process for LNPs’production.The lignin oligomers with widely distributed molecular weight and abundant guaiacyl units were extracted from wood waste(mainly consists of pine wood),exerting outstanding self-assembly capability.Uniform and spherical LNPs were generated in H_(2)O-n-propanol cosolvent,whereas irregular LNPs were obtained in H_(2)O-methanol cosolvent.The unsatisfactory self-assembly performance of the lignin oligomers in H_(2)O-methanol cosolvent could be attributed to two aspects.On one hand,for the initial dissolution state,the distinguishing Hansen solubility parameter and polarity between methanol solvent and lignin oligomers resulted in the poor dispersion of the lignin oligomers.On the other hand,strong hydrogen bonds between methanol solvent and lignin oligomers during solvent shifting process,hindered the interactions among the lignin oligomers for self-assembly.

用δ^(13)C方法研究玉米秸秆分解期间土壤有机质数量动态变化

通过室内培养实验 ,应用δ1 3C方法研究了玉米秸秆分解期间 ,土壤中胡敏酸 (HA)和富里酸 (FA)数量的动态变化。结果表明 :培养期间 ,新加入的玉米秸秆以及原土有机C都减少 ,但后者分解速度较慢。培养初期 ,FA的形成速度大于HA ;随培养时间延长 ,FA转化为HA或相互转化。原土有机质中 ,HA、FA也发生了相互转化 ,但与新形成的HA、FA相比转化速度较慢。用δ1 3 C方法研究短期培养 (几个月～几年 )条件下新加入有机质在土壤中的分解动力学是可行的。

药物流出泵基因在白假丝酵母菌生物膜耐药性产生机制中的作用

目的利用药物流出泵基因缺陷株,初步阐明药物流出泵基因CDR1、CDR2、MDR1在生物膜耐药性产生和表现型耐药菌株产生中的作用。方法以3种咪唑类抗真菌药物以及CDR1药物流出泵抑制剂作用于药物流出泵基因缺陷株形成的生物膜,采用XTT法和菌落计数的方法,分析药物流出泵基因在生物膜耐药性产生中的作用。结果药物流出泵基因对缺陷株形成的24h生物膜SMIC80影响不大,生物膜表现型耐药株的产生与药物流出泵基因有关,抗真菌药物联合CDR1基因抑制剂在一定程度上能够帮助杀灭白假丝酵母菌生物膜耐药株。结论药物流出泵基因对24h生物膜表现出来的耐药性影响不大,而对生物膜产生的耐药株有重要作用。

日本凋毛藻(Griffithsiajaponica)硫氧还蛋白基因及其编码的蛋白特征

从日本凋毛藻的表达序列标记 EST库中筛选到 2 条全长的编码硫氧还蛋白的 cDNA序列,分别命名为GjTRX1和GjTRX2。将这2条基因编码的蛋白序列与其它藻类(包括酵母)进行比较,结果表明不同物种及同一物种间TRX的相似性都较差,除GjTRX2编码的蛋白序列外,所有蛋白在活性位点序列 WCGPC处都完全保守。推测GjTRX1属于细胞质内分布的硫氧还蛋白。GjTRX1序列推导的蛋白二级结构单元与以往报道的传统上保守的TRX的二级结构单元类型一致,但是顺序有所不同。

Helicobacter pylori in human health and disease:Mechanisms for local gastric and systemic effects

Helicobacter pylori(H. pylori) is present in roughly 50% of the human population worldwide and infection levels reach over 70% in developing countries. The infection has classically been associated with different gastro-intestinal diseases, but also with extra gastric diseases. Despite such associations, the bacterium frequently persists in the human host without inducing disease, and it has been suggested that H. pylori may also play a beneficial role in health. To understand how H. pylori can produce such diverse effects in the human host, several studies have focused on understanding the local and systemic effects triggered by this bacterium. One of the main mechanisms by which H. pylori is thought to damage the host is by inducing local and systemic inflammation. However, more recently, studies are beginning to focus on the effects of H. pylori and its metabolism on the gastric and intestinal microbiome. The objective of this review is to discuss how H. pylori has co-evolved with humans, how H. pylori presence is associated with positive and negative effects in human health and how inflammation and/or changes in the microbiome are associated with the observed outcomes.

Biochemical characters and antibiotic susceptibility of Staphylococcus aureus isolates

Objective:To observe the biochemical characters and antibiotic susceptibility of isolated Staphylococcia aureus(S.auerus) strains against some conventional and traditional antibiotics. Methods:Thirty post operative pathogenic isolated S.aureus strains were used in this study. Bacterial culture was done in Mueller-Hinton broth at 37 ℃.Characters of these strains were determined by traditional biochemical tests such as hydrolysis test of gelatin,urea,galactose, starch and protein,and fermentation of lactose and sucrose.Antibiotic susceptibility were carried out by minimum inhibilory concentration test,minium bactericidal concentration test,disc agar diffusion test and brain heart infusion oxacillin screening agar.Results:Prom this study,it was observed that 100%S.aureus isolates showed positive results in gelatin,urea and galactose hydrolysis test.50%isolates were positive in starch hydrolysis test,35%in protein hydrolysis test. 100%isolates in lactose fermenting test,but no isolate was positive in sucrose fermenting test. Antibiotic susceptibility testing suggested that 20%of isolates were resistant to kanamycin and 46.67%were resistant to oxacillin.Conclusions:These findings show that all these isolates have gelatin,urea,galactose hydrolysis and lactose fermenting activity.20%of these isolates were resistant to kanamvcin and 46.67%were resistant to oxacillin.

Using the polymerase chain reaction coupled with denaturing gradient gel electrophoresis to investigate the association between bacterial translocation and systemic inflammatory response syndrome in predicted acute severe pancreatitis

AIM: To investigate the use of PCR and DGGE to investigate the association between bacterial translocation and systemic inflammatory response syndrome in predicted severe AP.METHODS: Patients with biochemical and clinical evidence of acute pancreatitis and an APACHE Ⅱ score ≥8 were enrolled. PCR and DGGE were employed to detect bacterial translocation in blood samples collected on d1,3, and 8 after the admission. Standard microbial blood cultures were taken when there was clinical evidence of sepsis or when felt to be clinically indicated by the supervising team.RESULTS: Six patients were included. Of all the patients investigated, only one developed septic complications;the others had uneventful illness. Bacteria were detected using PCR in 4 of the 17 collected blood samples. The patient with sepsis was PCR-positive in two samples (taken on d 1 and 3), despite three negative blood cultures. Using DGGE and specific primers, the bacteria in all blood specimens which tested positive for the presence of bacterial DNA were identified as E coli.CONCLUSION: Our study confirmed thatunlike traditional microbiological techniques, PCR can detect the presence of bacteria in the blood of patients with severe AP. Therefore, this latter method in conjunction with DGGE is potentially an extremely useful tool in predicting septic morbidity and evaluating patients with the disease. Further research using increased numbers of patients, in particular those patients with necrosis and sepsis, is required to assess the reliability of PCR and DGGE in the rapid diagnosis of infection in AP.

Nitric oxide mediated Staphylococcus aureus pathogenesis and protective role of nanoconjugated vancomycin

Objective:To investigate the relation between fruit seeds,plants residuals and appendicitis. Methods:Among cases that underwent appendectomy,the appendicitis cases having fruit seeds and undigested plant residuals in their etiology were examined retrospectively.Also, histopathological features,age,sex,and parameters of morbidity and mortality were used. Results:Fruit seed was found in one case(0.05%) with presence of pus in appendix lumen, undigested plant residuals in 7 cases(0.35%).It was determined that there were appendix inflammation in 2 of the plant residuals cases,while there were obstruction and lymphoid hyperplasia in the appendix lumen of 5 cases.No mortality was observed.Conclusions:The ratio of acute appendicitis caused by plants is minimal among all appendectomised patients, but avoidence of eating undigested fruit seeds and chewing plants well may help to prevent appendicitis.

Role of antibiotics for treatment of inflammatory boweldisease

Inflammatory bowel disease is thought to be caused by an aberrant immune response to gut bacteria in a genetically susceptible host. The gut microbiota plays an important role in the pathogenesis and complications of the two main inflammatory bowel diseases: Crohn's disease(CD) and ulcerative colitis. Alterations in gut microbiota, and specifically reduced intestinal microbial diversity, have been found to be associated with chronic gut inflammation in these disorders. Specific bacterial pathogens, such as virulent Escherichia coli strains, Bacteroides spp, and Mycobacterium avium subspecies paratuberculosis, have been linked to the pathogenesis of inflammatory bowel disease. Antibiotics may influence the course of these diseases by decreasing concentrations of bacteria in the gut lumen and altering the composition of intestinal microbiota. Different antibiotics, including ciprofloxacin, metronidazole, the combination of both, rifaximin, and anti-tuberculous regimens have been evaluated in clinical trials for the treatment of inflammatory bowel disease. For the treatment of active luminal CD, antibiotics may have a modest effect in decreasing disease activity and achieving remission, and are more effective in patients with disease involving the colon. Rifamixin, a non absorbable rifamycin has shown promising results. Treatment of suppurative complications of CD such as abscesses and fistulas, includes drainage and antibiotic therapy, most often ciprofloxacin, metronidazole, or a combination of both. Antibiotics might also play a role in maintenance of remission and prevention of post operative recurrence of CD. Data is more sparse for ulcerative colitis, and mostly consists of small trials evaluating ciprofloxacin, metronidazole and rifaximin. Most trials did not show a benefit for the treatment of active ulcerative colitis with antibiotics, though 2 meta-analyses concluded that antibiotic therapy is associated with a modest improvement in clinical symptoms. Antibiotics show a clinical benefit when used for the treatment of pouchitis. The downsides of antibiotic treatment, especially with recurrent or prolonged courses such as used in inflammatory bowel disease, are significant side effects that often cause intolerance to treatment, Clostridium dificile infection, and increasing antibiotic resistance. More studies are needed to define the exact role of antibiotics in inflammatory bowel diseases.

Prevalence of hepatitis C infection among intravenous drug users in Shanghai

AIM:To characterize the prevalence of hepatitis C virus(HCV)infection among Chinese intravenous drug users(IDUs).METHODS:A total of 432 adult IDUs(95 women and337 men)in Shanghai were included in the study.The third-generation Elecsys Anti-HCV assay(Roche Diagnostics GmbH,Sandhofer Strasse 116,D-68305,Mannheim,Germany)was used to screen for antibodies against HCV.The RIBA strip,a supplemental antiHCV test with high specificity,was performed on all of the samples that tested positive during the initial screening.All of the anti-HCV positive samples were analyzed with a Cobas TaqMan 48 Analyzer(Roche Diagnostics)for direct detection of HCV RNA.All of the HCV RNA-positive samples were sequenced for genotype determination.RESULTS:The preliminary screening identified 262(60.6%)subjects who were seropositive for HCV.Of the 62 females and 200 males seropositive subjects,16(16.7%)and 65(19.3%),respectively,were confirmed by RIBA,yielding an overall HCV seropositive rate of18.8%.Four female(6.5%)and 14 male(7.0%)subjects tested positive for HCV RNA,indicating an active infection rate of 4.2%for the entire study population.The 18 HCV RNA-positive serum samples were genotyped.Seven individuals were genotype 1b,and four were genotype 1a.One individual each was infected with genotypes 2a,2b and 3a.Four subjects were coinfected with multiple strains:two with genotypes 1a and 2a,and two with genotypes 1b and 2a.The active infection rate among HCV-seropositive individuals was22.2%,which was significantly lower than most estimates.CONCLUSION:The prevalence of HCV is relatively low among IDUs in Shanghai,with a spontaneous recovery rate much higher than previous estimates.

In-vitro antimicrobial activity of marine actinobacteria against multidrug resistance Staphylococcus aureus

Objective:To investigate the antibacterial aclivily of marine actinobacteria against multidrug resistance Staphylococcus aureus(MDRSA).Methods:Fifty one actinobacterial strains were isolated from salt pans soil,costal area in Kothapattanam,Ongole,Andhra Pradesh.Primary screening was done using cross-streak method against MDRSA.The bioaclive compounds are extracted from efficient actinobacteria using solvent extraction.The antimicrobial activity of crude and solvent extracts was perfomied using Kirby-Bauer method.MIC for ethyl acetate extract was determined by modified agar well diffusion method.The potent actinobacteria are identified using Nonomura key,Shirling and Gottlieb 1966 with Bergey's manual of determinative bacteriology.Results:Among the fifty one isolates screened for antibacterial activity,SRB25were found efficient against MDRSA.The ethyl acetate extracts showed high inhibition against test organism.MIC test was performed with the ethyl acetate extract against MDRSA and found to be 1 000μg/mL.The isolaled actinobacteria are identified as Streptomyces sp with the help of Nonomura key.Conclusions:The current investigation reveals that the marine actinobacteria from salt pan environment can be able to produce new drug molecules against drug resistant microorganisms.

Bacteria as indicators for finding oil and gas reservoirs:A case study of the Bikaner-Nagaur Basin,Rajasthan,India

Geo-microbial prospecting for hydrocarbons is an exploration method based on the seepage of light gaseous hydrocarbons from oil/gas reservoirs to the surface and their utilization by hydrocarbon oxidizing bacteria. These bacteria utilize hydrocarbon gases as their sole source of food and are found to be enriched in the near surface soils/sediments above the oil and gas reservoirs. The detection of anomalous populations of n-pentane and n-hexane oxidizing bacteria in the surface soils can help to evaluate the prospects for hydrocarbon exploration. A geo-microbial survey has been carried out in the Bikaner Nagaur basin to investigate the prospects for hydrocarbon exploration. In the present study, bacterial counts for n-pentane utilizing bacteria range between 2.0× 10^2 and 1.26× 10^6 cfu/gm and n-hexane utilizing bacteria range between 2.0× 10^2 and 1.21 × 10^6 cfu/gm. The bacterial concentration distribution maps show four distinct anomalies in the study area. The possibility of discovering oil or gas reservoirs using the microbiological method is emphasized by the tact that the hydrocarbon-oxidizing bacteria range between 103 and 106 cfu/gm in soil/sediment receiving hydrocarbon micro-seepages. In the present study area of the Bikaner Nagaur basin, n-pentane and n-hexane utilizing bacteria are found between 105 and 106 cfu/gm of soil sample, which is significant and thereby substantiates the seepage of lighter hydrocarbon accumulations from oil and gas reservoirs. Geo-microbial prospecting studies suggest that hydrocarbon micro-seepage of subsurface origin is present in the study area and indicate that the area has positive prospects for petroleum exploration.

Antioxidative effect of folate-modified chitosan nanoparticles

Objective:To evaluate the potency of carboxymethyl chitosan-2,2' ethylenedioxy bisethylamine-folate(CMC-EDBE-FA) on tissue injury,antioxidant status and glutathione system in tissue mitochondria and serum against nicotine-induced oxidative stress in mice.Methods: CMC-EDBE-FA was prepared on basis of carboxymethyl chitosan tagged with folic acid by covalently linkage through 2,2' ethylenedioxy bis-ethylamine.Animals were divided into four groups,i.e.,control,nicotine(1 mg/kg bw/day),CMC-EDBE-FA(1 mg/kg bw/day) and nicotine(1 mg/kg bw/day) and CMC-EDBE-FA(1 mg/kg bw/day) for 7 days.Levels of lipid peroxidation, oxidized glutathione level,antioxidant enzyme status and DNA damage were observed and compared.Results:The significantly increase of lipid peroxidation,oxidized glutathione levels and DNA damage was observed in nicotine treated group as compared with control group;those were significantly reduced in CMC-EDBE-FA supplemented group.Moreover,significantly reduced antioxidant status in nicotine treated group was effectively ameliorated by the supplementation of CMC-EDBE-FA.Only CMC-EDBE-FA treated groups showed no significant change as compared with control group;rather than it repairs the tissue damage of nicotine treated group.Conclusions:These findings suggest that CMC-EDBE-FA is non-toxic and ameliorates nicotine-induced toxicity.

Detection of class 1 integron in Acinetobacter baumannii isolates collected from nine hospitals in Turkey

Objective:To investigate the antibiotic resistance genes inserted into class I and class 2integrons in Acinetobacler baumannii[A.baumannii)isolates obtained from nine different cities in Turkey.Methods:A collection of 281 A.baumannii clinical isolates were collected from nine diferenl state hospitals in Turkey and were confirmed as A.baumannU by conventional biochemical,API testing and bla_(-OXA-51)specific PCR.The isolates were examined by PCR for existence of class I and2 integron gene cassettes.Results:They were characterized by antimicrobial susceptibility testing and the highest resistance rates were determined for piperacillin(90.03%),ciprofloxacin(87.54%),cefepime and trimethoprim/sulfamethoxazole(81.13%).The lowest resistance rates was for cefotaxime(3.55%).class 1 integrons were detected in 6.4%(18/281)of A.baumannii strains and no class 2 integron was detected.The gene casselles of class 1 inlegrons AacCI-AAC(3)l-aadAI,AacCI-aadA1,AAC(3)-I,AAC(3)-I-AAC(3)-I-aadA1,TEM-1.AAC(3)-I-aadA1-AAC(3)-I-AAC(3)-I,AAC(3)-I-AAC(3)-I-AAC(3)-I-aadA1,AAC(3)-I-aadA1,AAC(3)-I-AAC(3)-I,AAC(3)-I-aadA1-AAC(3)-I-aadA1,AAC(3)-I-AAC(3)-I-aadA1-AAC(3)-I-aadA1 were detected in eighteen strains.The aac genes family were most frequently found integrated into the class 1 inlegrons and it was followed by aadA genes and TEM-1 genes.Conclusions:This is an extensive study on the distribulion of class 1 integron among A.baumannii in Turkey.In addition to these,two new alleles were observed.Their percentage rates of similarity to other cassettes are 95%aadA1(TK A18)and 89%,aadA 1(ANKA3).

Protective effect of Polyalthia longifolia var.pendula leaves on ethanol and ethanol/HCl induced ulcer in rats and its antimicrobial potency

Objective:To explore antiulcer and antimicrobial properties of methanolic extract of Polyalthia longifolia var.pendula.Methods:Gastroprotective potential of Polyalthia longifolia was studied on ethanol and ethanol/HCl induced ulcers at 2 different doses(270 and 540 mg/kg/body weight). Antimicrobial efficacy of Polyalthia longifolia(25 mg/mL) was also studied against six gram positive,seven gram negative bacteria and five fungi by agar well diffusion method.Minimum inhibitory concentration was determined by agar well diffusion method in two fold serial dilution, in the range of 97-25 000μg/mL.Results:The reduction of ulcer index in Polyalthia longifolia treated animals was found to be statistically significant with respect to control animals.The Polyalthia longifolia exhibited ulcer protection activity in dose dependent manner and was also better than the standard.In antimicrobial activity,gram positive bacteria were more susceptible to Polyalthia longifolia than gram negative bacteria and fungal strains.Conclusions:Results obtained confirm the antiulcer and antimicrobial potential of the Polyalthia longifolia.

Hepatoprotective effect of Woodfordia fruticosa Kurz flowers on diclofenac sodium induced liver toxicity in rats

Objective:To evaluate the protective effect of Woodfordia fruticosa Kurz flowers against experimentally induced liver toxicity in rats.Methods:Two different doses of methanol extract of Woodfordia fruticosa(WFM) were evaluated for the hepatoprotective activity against diclofenac sodium induced hepatotoxicity in rats.Various biochemical parameters like alanine aminotransferase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),total protein(TP),albumin(ALB),blood urea nitrogen(BUN) from serum;total protein(TP),glutathione (GSH) levels,catalase(CAT) and glutathione peroxidase(GPx) activities from liver were studied; histopathologic changes of liver were also evaluated.Results:WFM effectively reduced the elevated levels of serum ALT,AST,ALP and BUN,enhanced the reduced TP,ALB and hepatic GSH,CAT,GPx activity.The histopathological analysis suggested that WFM decreased the degree of liver fibrosis induced by diclofenac.Conclusions:This study demonstrates the hepatoprotective activity of WFM and thus scientifically support the use of this plant in traditional medicine for the treatment of liver disorders.