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Mathur A.D.,Microbiology | Devesh S.,Microbiology
Journal of Association of Physicians of India | Year: 2017

Introduction: There are conflicting reports in literature about correlation of CSF viral RNA levels with neurological manifestations in HIV positive patients. Many studies in animals and human subjects have shown that CSF HIV-1 RNA can be useful as a specific marker of HIV induced neuropathology. To the contrary there are studies which show that neurological disease states can occur in absence of significant increase of CSF HIV RNA. Materials and Methods: This was a prospective study conducted at Base hospital Delhi Cantt, New Delhi, a tertiary care hospitals for HIV patients. The study period was from 16 May 2006 to16 Jun 2011. The current study included forty (40) patients (Twenty HIV positive patients with neurological manifestations and twenty HIV positive patients clinically without any neurological manifestation). All potential study subjects and controls were explained the nature of this study and enrolled thereafter with written consent. Results: In our study all the cases (HIV/AIDS patients with Neuro AIDS) and controls (HIV/AIDS patients without Neuro manifestations) were males only. 45% of the cases and 60% of controls were in the age group of 25 to 35 yrs and 35 % of cases and 40% of controls were in age group of 36 to 45 yrs. Among cases (HIV patients with neurological manifestations), The neurological manifestations in our 20 patients included; dementia-5, cryptococcal meningitis-4, Tubercular meningitis-4, CVA-3, Headache-3, (without CSF abnormality), 1 case each of pyogenic meningitis, Candida meningitis, Tremors and Herpes Zoster. Among the 20 cases fourteen patients had abnormal CSF (70%) whereas only one patient among the controls showed CSF abnormality (5%). Out of 20 cases, radio-imaging (CT Scan/ MRI) of brain was done in 18 cases. Twelve cases (66.66) had some abnormality on CT/ MRI. Various abnormalities seen were as under Calcified granuloma-1, Infarcts-5, Hydrocephalus-2, TBM (meningeal enhancement)-2, Candidiasis (Focal hypodensities in subcortical white matter of cerebral hemispheres)-1, Cryptococcoma-1, Cerebral atrophy-1, Focal enhancing lesions-2. In our study, mean CSF viral load in cases was 5236.3 copies/ml and in controls 502.4 copies/ml. Viral load in CSF among cases was significantly higher than viral load in CSF among controls. Conclusion: CSF HIV-1 RNA viral load estimation can be a useful tool for clinicians in confirming neurological involvement in HIV infected patients. Hence, in HIV positive patients suspected to have neurological involvement, CSF viral load studies should be done. Serial estimations of CSF HIV-1 RNA levels can be of prognostic significance. Estimation of CSF HIV-1 RNA level before and serially during administration of HAART can be useful to judge the efficacy of HAART. © 2017, Journal of Association of Physicians of India. All rights reserved.

Lehmann B.A.,Maastricht University | Ruiter R.A.C.,Maastricht University | Wicker S.,Goethe University Frankfurt | Van Dam D.,Microbiology | Kok G.,Maastricht University
BMC Public Health | Year: 2014

Background: Health Authorities recommend influenza vaccination of healthcare personnel (HCP) to decrease the transmission of influenza to vulnerable patients. Recent studies have almost exclusively used quantitative questionnaires in order to identify determinants of vaccination behaviour. Interviews enable HCP to express freely why they think they are (not) willing to get vaccinated against influenza. Methods. By means of semi-structured one-on-one interviews with 123 Belgian, Dutch and German HCP, reasons for and against vaccination, experiences with influenza vaccination, intention to get vaccinated and possible barriers, as well as willingness to advice influenza vaccination to patients were investigated. Data were processed with QSR NVivo 8.0 and analysed using a combination of a deductive and a general inductive approach. Results: Across countries, self-protection, patient protection, and protection of family members were reported as most important reasons to get vaccinated against influenza. Reasons to not get vaccinated against influenza were fear of side effects caused by the vaccine, a low risk-perception, the disbelief in the effectiveness of influenza vaccination, organizational barriers, misconceptions, and undefined negative emotions. Conclusions: The social cognitive variables underlying the decision of HCP to get vaccinated against influenza (or not) seem to be similar in Belgium, Germany, and the Netherlands, even though some differences surfaced. A quantitative investigation of those social cognitive variables is needed in order to determine the importance of the social cognitive variables in explaining the intention to get vaccinated and the importance of the similarities and differences between countries that have been found in this study. © 2014 Lehmann et al.; licensee BioMed Central Ltd.

Keim S.A.,Research Institute at Nationwide Childrens Hospital | Hogan J.S.,Microbiology | Mcnamara K.A.,Research Institute at Nationwide Childrens Hospital | Gudimetla V.,Microbial Infection and Immunity | And 4 more authors.
Pediatrics | Year: 2013

OBJECTIVE: To quantify microbial contamination of human milk purchased via the Internet as an indicator of disease risk to recipient infants. METHODS: Cross-sectional sample of human milk purchased via a popular US milk-sharing Web site (2012). Individuals advertising milk were contacted to arrange purchase, and milk was shipped to a rented mailbox in Ohio. The Internet milk samples (n = 101) were compared with unpasteurized samples of milk donated to a milk bank (n = 20). RESULTS: Most (74%) Internet milk samples were colonized with Gramnegative bacteria or had .104 colony-forming units/mL total aerobic count. They exhibited higher mean total aerobic, total Gram-negative, coliform, and Staphylococcus sp counts than milk bank samples. Growth of most species was positively associated with days in transit (total aerobic count [log10 colony-forming units/mL] β = 0.71 [95% confidence interval: 0.38-1.05]), and negatively associated with number of months since the milk was expressed (β = 20.36 [95% confidence interval: 20.55 to 20.16]), per simple linear regression. No samples were HIV type 1 RNA-positive; 21% of Internet samples were cytomegalovirus DNA-positive. CONCLUSIONS: Human milk purchased via the Internet exhibited high overall bacterial growth and frequent contamination with pathogenic bacteria, reflecting poor collection, storage, or shipping practices. Infants consuming this milk are at risk for negative outcomes, particularly if born preterm or are medically compromised. Increased use of lactation support services may begin to address the milk supply gap for women who want to feed their child human milk but cannot meet his or her needs. Pediatrics 2013;132:e1227-e1235. Copyright © 2013 by the American Academy of Pediatrics.

Lafaurie M.,Infectious Diseases Intervention Unit U2i | Porcher R.,University Paris Diderot | Donay J.-L.,Microbiology | Touratier S.,Pharmacy | Molina J.-M.,University Paris Diderot
Journal of Antimicrobial Chemotherapy | Year: 2012

Objectives: High rates of methicillin-resistant Staphylococcus aureus (MRSA) and fluoroquinolone-resistant Pseudomonas aeruginosa may be related, in part, to the overuse of fluoroquinolones. The objective was to analyse and correlate long-term surveillance data on MRSA and fluoroquinolone-resistant P. aeruginosa rates and antibiotic consumption after implementation of an institution-wide programme to reduce fluoroquinolone use. Methods: An interrupted time series/quasi-experimental study of monthly fluoroquinolone use and MRSA and fluoroquinolone-resistant P. aeruginosa isolation rates was carried out in a tertiary hospital during three periods: pre-intervention (January 2000-August 2005), intervention (September 2005-March 2006), and post-intervention (March 2006-March 2010). The effect of the intervention on the consumption of fluoroquinolones and bacterial resistance was assessed using segmented regression analyses. Results: Mean monthly fluoroquinolone consumption dropped by 29.1 defined daily doses per 1000 patient-days (DDD/1000 PD) (95% CI 13.1-45.9; P = 0.0005) from a mean of 148.2 to 119.1 DDD/1000 PD during the intervention period. A sustained and significant decrease in fluoroquinolone consumption of -0.95 DDD/1000 PD/month was also observed during the post-intervention period (P = 0.0002). During the post-intervention period the rate of fluoroquinolone-resistant P. aeruginosa continuously decreased, from a mean of 42% to 26%, with a constant relative change rate of -13%/year (95% CI -19 to -5, P = 0.001). A decrease in the MRSA rate was observed during the intervention period, from a mean resistance rate of 27% to 21% (P < 0.0001). Conclusions: We showed the sustained impact of a fluoroquinolone control programme on the reduction of fluoroquinolone use with a significant decrease in fluoroquinolone-resistant P. aeruginosa and MRSA rates over 4 years. © The Author 2012. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved.

RamaRao G.,Canadian Department of National Defence | Afley P.,Microbiology | Acharya J.,Canadian Department of National Defence | Bhattacharya B.K.,Canadian Department of National Defence
BMC Neuroscience | Year: 2014

Background: Recent alleged attacks with nerve agent sarin on civilians in Syria indicate their potential threat to both civilian and military population. Acute nerve agent exposure can cause rapid death or leads to multiple and long term neurological effects. The biochemical changes that occur following nerve agent exposure needs to be elucidated to understand the mechanisms behind their long term neurological effects and to design better therapeutic drugs to block their multiple neurotoxic effects. In the present study, we intend to study the efficacy of antidotes comprising of HI-6 (1-[[[4-(aminocarbonyl)-pyridinio]-methoxy]-methyl]-2-[(hydroxyimino) methyl] pyridinium dichloride), atropine and midazolam on soman induced neurodegeneration and the expression of c-Fos, Calpain, and Bax levels in discrete rat brain areas.Results: Therapeutic regime consisting of HI-6 (50 mg/kg, i.m), atropine (10 mg/kg, i.m) and midazolam (5 mg/kg, i.m) protected animals against soman (2 × LD50, s.c) lethality completely at 2 h and 80% at 24 h. HI-6 treatment reactivated soman inhibited plasma and RBC cholinesterase up to 40%. Fluoro-Jade B (FJ-B) staining of neurodegenerative neurons showed that soman induced significant necrotic neuronal cell death, which was reduced by this antidotal treatment. Soman increased the expression of neuronal proteins including c-Fos, Bax and Calpain levels in the hippocampus, cerebral cortex and cerebellum regions of the brain. This therapeutic regime also reduced the soman induced Bax, Calpain expression levels to near control levels in the different brain regions studied, except a mild induction of c-Fos expression in the hippocampus.Conclusion: Rats that received antidotal treatment after soman exposure were protected from mortality and showed reduction in the soman induced expression of c-Fos, Bax and Calpain and necrosis. Results highlight the need for timely administration of better antidotes than standard therapy in order to prevent the molecular and biochemical changes and subsequent long term neurological effects induced by nerve agents. © 2014 RamaRao et al.; licensee BioMed Central Ltd.

Adav S.S.,National Taiwan University | Lin J.C.-T.,National Taiwan University | Yang Z.,Tsinghua University | Whiteley C.G.,Microbiology | And 3 more authors.
Biotechnology Advances | Year: 2010

This review addresses the introduction of fluorescent molecular tags into exo-enzymes and extra polymeric substances of bioaggregates and the use of confocal laser scanning microscopy (CLSM) to map their role, purpose and quantitative description of the biological processes they undertake. Multiple color staining coupled with CLSM and fluorescent in situ hybridisation (FISH) and flow cytometry have identified the individual polymeric substances, whether they are proteins, lipids, polysaccharides, nucleic acids or antibodies, as well as the microorganisms in the bioaggregate. Procedures are presented for simultaneous multicolor staining with seven different fluorochromes - SYTOX Blue for nucleic acids; Nile red for lipids; Calcofluor white [CW] for β-polysaccharides; concanavalin A [Con A] for α-poly-saccharides; fluorescein-isothiocyanate [FITC] for proteins; SYTO 63 for live microbial cells and Calcium Green for monitoring calcium levels in the microbial cells. For the distribution of certain microbial strains, metabolic enzymes and extrapolymeric substances to be quantitatively described the generated colored images are converted into digital forms under specific predefined criteria. Procedures and computer software programs (Amira; MATLAB) are presented in order to quantitatively establish grid patterns from the CLSM images. The image is digitized using a threshholding algorithm followed by a reconstruction of the image as a volumetric grid for finite element simulation. The original color image is first converted to a grey followed by resizing, detection and modification of bilevel images and finally a total reversal of the image colors. The grid file is then used by specific computer software (Gambit, Fluent) for further numerical studies incorporating chemical reactions, transport processes and computational fluid dynamics including intra-bioaggregate fluid flow, and heat and mass transfer within the bioaggregate matrix. © 2009 Elsevier Inc. All rights reserved.

Shaw C.K.,Pediatrics | Shaw P.,Microbiology
International Journal of Pharma and Bio Sciences | Year: 2012

Catheter related bloodstream infections are a major problem in most tertiary care hospitals. Among the various organisms associated with nosocomial infections, coagulase negative staphylococci are responsible for majority of the catheter related infections. They are usually resistant to standard antibiotics necessitating prolonged hospital stay and amplifying the cost of treatment manifold which usually becomes a vicious cycle difficult to break and ultimately contributing to increased morbidity and mortality. Coagulase negative staphylococci are skin commensals but the strains producing biofilms manage to evade the host immune system. Biofilms consist of a microbially derived sessile community characterized by cells that are irreversibly attached to a substratum or each other, embedded in a matrix of extracellular polymeric substances, exhibiting an altered phenotype with respect to growth rate and gene transcription. This unnatural yet favourable ecological niche protects the organisms from host immune responses and antimicrobials. In the following account we present the characteristics of biofilms and the latter's relationship with catheter related blood stream infections particularly by coagulase negative staphylococcus and vice versa. Together, biofilms and coagulase negative staphylococci dominate the saga of catheter related sepsis and strict asepsis protocols related to catheter placement and maintenance and rational antibiotic policy are the only hope as other approaches to inhibition of biofilm formation are still experimental.

Peter H.,University of Edinburgh | Berggrav K.,University of Edinburgh | Thomas P.,University of Edinburgh | Pfeifer Y.,Robert Koch Institute | And 3 more authors.
Journal of Clinical Microbiology | Year: 2012

Klebsiella pneumoniae carbapenemases (KPCs) are considered a serious threat to antibiotic therapy, as they confer resistance to carbapenems, which are used to treat extended-spectrum beta-lactamase (ESBL)-producing bacteria. Here, we describe the development and evaluation of a DNA microarray for the detection and genotyping of KPC genes (blaKPC) within a 5-h period. To test the whole assay procedure (DNA extraction plus a DNA microarray assay) directly from clinical specimens, we compared two commercial DNA extraction kits (the QIAprep Spin miniprep kit [Qiagen] and the urine bacterial DNA isolation kit [Norgen]) for the direct DNA extraction from urine samples (dilution series spiked in human urine). Reliable single nucleotide polymorphism (SNP) typing was demonstrated using 1×105 CFU/ml urine for Escherichia coli (Qiagen and Norgen) and 80 CFU/ml urine, on average, for K. pneumoniae (Norgen). This study presents, for the first time, the combination of a new KPC microarray with commercial sample preparation for detecting and genotyping microbial pathogens directly from clinical specimens; this paves the way toward tests providing epidemiological and diagnostic data, enabling better antimicrobial stewardship. Copyright © 2012, American Society for Microbiology. All Rights Reserved.

Chilton C.H.,University of Leeds | Freeman J.,Microbiology | Crowther G.S.,University of Leeds | Todhunter S.L.,University of Leeds | And 2 more authors.
Journal of Antimicrobial Chemotherapy | Year: 2012

Objectives: Co-amoxiclav is widely prescribed in hospitals. Although reports have suggested it may be linked to onset of Clostridium difficile infection (CDI), data on the risk of CDI associated with specific antibiotics is difficult to obtain, due to confounding clinical factors. We have examined the propensity of co-amoxiclav to induce CDI using a human gut model. Methods: We used a triple-stage chemostat human gut model to study the effects of co-amoxiclav on indigenous gut microorganisms and C. difficile PCR ribotype 027. C. difficile viable counts and spores were evaluated, and cytotoxin titres were assayed. Co-amoxiclav concentrations were measured using a large plate bioassay. Results: Co-amoxiclav induced rapid C. difficile germination and high toxin production in the gut model, from 5 days after commencement of instillation. Cell proliferation and toxin production were prolonged and continued throughout the duration of the experiment. Only very low levels of co-amoxiclav antimicrobial activity could be detected within the gut model, despite having a marked effect on gut flora microorganisms. Conclusions: Co-amoxiclav induced CDI within the gut model, supporting clinical observations linking co-amoxiclav treatment with CDI onset. This reinforces the value of the gut model as a clinically relevant means of studying CDI. Caution should be exercised in the prescription of co-amoxiclav to patients in high CDI risk settings. © The Author 2012. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved.

Marquart T.J.,Saint Louis University | Wu J.,Microbiology | Lusis A.J.,Microbiology | Baldan A.,Saint Louis University
Arteriosclerosis, Thrombosis, and Vascular Biology | Year: 2013

Objective-To determine the efficacy of long-term anti-miR-33 therapy on the progression of atherosclerosis in high-fat, high-cholesterol-fed Ldlr mice. Methods and Results-Ldlr mice received saline, or control or anti-miR-33 oligonucleotides once a week for 14 weeks. The treatment was effective, as measured by reduced levels of hepatic miR-33 and increased hepatic expression of miR-33 targets. Analysis of plasma samples revealed an initial elevation in high-density lipoprotein cholesterol after 2 weeks of treatment that was not sustained by the end of the experiment. Additionally, we found a significant increase in circulating triglycerides in anti-miR-33-treated mice, compared with controls. Finally, examination of atheromata revealed no significant changes in the size or composition of lesions between the 3 groups. Conclusion-Prolonged silencing of miR-33 fails to maintain elevated plasma high-density lipoprotein cholesterol and does not prevent the progression of atherosclerosis in Ldlr mice. © 2013 American Heart Association, Inc.

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