Fukui-shi, Japan
Fukui-shi, Japan

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Kitagawa Y.,Shiga University of Medical Science | Zhou M.,Shiga University of Medical Science | Zhou M.,Nagahama Institute of Bio-Science and Technology | Yamaguchi M.,Shiga University of Medical Science | And 5 more authors.
Microbes and Infection | Year: 2010

M2-2 protein of human metapneumovirus (HMPV) is encoded by one of two overlapping open reading frames within M2 mRNA. The precise function of HMPV M2-2 protein remains unknown. We here examined effect of M2-2 protein on HMPV transcription and replication using a minigenome construct and monitoring luciferase reporter gene expression. The minigenome assays demonstrated that M2-2 protein inhibited both transcription and RNA replication. The inhibitory function of M2-2 protein was completely abrogated by removal of eight or four amino acids from its N- or C-terminus, respectively, demonstrating importance of both short terminal sequences for maintaining its functional structure. Immunoprecipitation experiments revealed interaction of M2-2 protein with L protein, which might be involved in inhibition of HMPV transcription and replication. Prior accumulation of intracellular M2-2 protein severely restrained HMPV from replicating. Thus inherent viral control of the M2-2 gene expression in infected cells seems to be essential for efficient HMPV replication. © 2009 Elsevier Masson SAS. All rights reserved.


Castro-Silva A.N.,University of Sao Paulo | Castro-Silva A.N.,Hospital Beneficencia Portuguesa Sao Paulo | Freire A.O.,Federal University of São Paulo | Grinbaum R.S.,Hospital Beneficencia Portuguesa Sao Paulo | And 7 more authors.
Transplant Infectious Disease | Year: 2011

Mycobacterium haemophilum is a slow-growing nontuberculous mycobacterium that can cause disease in both immunocompetent and immunocompromised patients. The most common clinical presentations of infection are the appearance of suppurative and ulcerated skin nodules. For the diagnosis, samples collected from suspected cases must be processed under the appropriate conditions, because M. haemophilum requires lower incubation temperatures and iron supplementation in order to grow in culture. In this case report, we describe the occurrence of skin lesions in a kidney transplant recipient, caused by M. haemophilum, associated with acupuncture treatment. The diagnosis was established by direct smear and culture of material aspirated from cutaneous lesions. Species identification was achieved by characterization of the growth requirements and by partial sequencing of the hsp65 gene. The patient was successfully treated with clarithromycin and ciprofloxacin for 12 months. Considering that the number of patients receiving acupuncture treatment is widely increasing, the implications of this potential complication should be recognized, particularly in immunosuppressed patients. © 2010 John Wiley & Sons A/S.


Roda A.,University of Bologna | Roda A.,Italian National Institute of Biosystems and Biostructures | Mirasoli M.,University of Bologna | Dolci L.S.,University of Bologna | And 5 more authors.
Analytical Chemistry | Year: 2011

A simple and versatile analytical device designed to perform, even simultaneously, different types of bioassays has been developed and optimized. A transparent microfluidics-based reaction chip, where analytes were quantitatively detected by means of biospecific reactions and chemiluminescence detection, was placed in contact with a thermoelectrically cooled CCD sensor through a fiber optic taper. Such a lensless contact imaging configuration combined adequate spatial resolution and high light collection efficiency within a small size portable device. The miniaturization of the reaction chamber ensured short analysis times (in the minutes range), while the use of chemiluminescence detection provided wide signal dynamic range and high detectability, down to attomole levels of protein and femtomole levels of nucleic acid analytes. A model hybrid panel test was realized by combining an enzyme assay for alkaline phosphatase activity, a nucleic acid hybridization assay for Parvovirus B19 DNA, and an immunoassay for horseradish peroxidase as a model antigen. The successful simultaneous quantification of the three targets demonstrated that a range of analytes, from enzymes to antigens, antibodies, and nucleic acids, can be measured in a single run, thus enabling the realization of a complete, personalized diagnostic panel test for early diagnosis of a given disease and patient follow-up. © 2011 American Chemical Society.


Re M.C.,Italian National Institute of Biosystems and Biostructures | Vitone F.,Microbiology Section | Biagetti C.,University of Bologna | Schiavone P.,Microbiology Section | And 4 more authors.
Clinical Microbiology and Infection | Year: 2010

As proviral human immunodeficiency virus type 1 (HIV-1) DNA can replenish and revive viral infection upon activation, its detection might offer significant therapeutic information, complementing the input provided by plasma RNA determination in the follow-up of infected individuals. A selected group of acutely infected subjects was studied to verify both total and 2-long terminal repeat (2-LTR) DNA proviral load during the acute phase of infection and thereafter. Patients were divided in two sex- and age-matched groups: 19 naive individuals who did not receive antiretroviral therapy during the observation period and 20 subjects treated according to current guidelines. Total and 2-LTR HIV-1 DNA proviral load, in addition to RNA viral load and CD4 cell count, were determined in peripheral blood mononuclear cells (PBMC) at baseline, 6 and 12 months after the first sampling. Total and 2-LTR HIV-1 DNA proviral load exhibited no significant variation at any time in the naive patients (total HIV-1 DNA ranging from 896 ± 731 to 715 ± 673 copies/105 PBMC and 2-LTR HIV-1 DNA ranging from 94 ± 105 to 65 ± 44 copies/105 PBMC), whereas a significant reduction in both total HIV-1 DNA (ranging from 997 ± 676 to 262 ± 174 copies/105 PBMC) and 2-LTR HIV-1 DNA proviral load (ranging from 116 ± 55 to 26 ± 35 copies/105 PBMC) was detected in highly active antiretroviral therapy (HAART) patients, together with a CD4+ T cell count increase and RNA load decrease. HAART negatively affects both the labile HIV burden and the integrated proviral DNA, at least in the initial period of successful treatment, suggesting that quantification of HIV-1 DNA proviral load may be an important parameter in monitoring HIV infection. © 2009 The Authors. Journal Compilation.


Kumar V.,Microbiology Section | Singh Solanki A.,Plant Wealth Sector | Sharma S.,Motilal Nehru National Institute of Technology
Thai Journal of Agricultural Science | Year: 2011

The yield and nutrient uptake of Plantago ovata were studied using dual inoculation with AM fungi (Glomus fasciculatum) and Azotobacter chroococcum. The experiment was conducted for two consecutive years, with two levels of chemical fertilizers (F1: N-P-K: 24-24-32 kg ha -1) and (F2: N-P-K: 30-30-40) kg ha -1). Inoculation with the AM fungi and Azotobacter chroococcum generated encouraging results; seed yield (1120.7 kg ha -1), husk yield (297.1 kg ha -1), plant height (40.4 cm), spike plant-1 (33.6), and 1000 seeds weight (1.793 g) were highest with treatment T12 (F2 fertilizer level + both bioinoculants) followed by treatment T9 (F1 fertilizer level + both bioinoculants), T10 (F2 + A. chroococcum), T11 (F2 + AM fungi), as compared to T1 (control), T2-T4 (bioinoculants) and T5-T6 (F1 and F2 fertilizer levels). The nitrogen uptake was more in Azotobacter treated plants, while higher P and K uptake was attained in AM fungi inoculated plants. The survival count of inoculated bacteria was highest on 60 days after inoculation and declined thereafter. Economic analysis revealed the net profit from seed and husk was highest in T9 using dual inoculation of microbes with 80% of recommended dose of fertilizer followed by T12 and T10.


Centonze A.R.,Microbiology Section | Tonolli E.,Microbiology Section | Fontana R.,Microbiology Section
Clinical and Vaccine Immunology | Year: 2013

The performances of seven Immulite 2000 (Siemens Healthcare Diagnostics) TORCH (Toxoplasma gondii, other microorganisms, rubella virus, cytomegalovirus, and herpes simplex virus) assays were evaluated in comparison with the performances of the ETI-MAX 3000 (DiaSorin) TORCH assays. The two systems demonstrated good agreement, and given their sensitivity, specificity, and positive predictive value, they can be used with confidence for TORCH prenatal screening. Copyright © 2013, American Society for Microbiology. All Rights Reserved.


Varalakshmi S.,National Dairy Research Institute | Kirubaharan J.J.,Microbiology Section
Research Journal of Biotechnology | Year: 2014

The effect of Cyclophosphamide (CY) on the immune status of birds was studied by administering the chickens with live Newcastle disease vaccine. Normal and immunosuppressed chickens were vaccinated with the Thermostable live Newcastle disease vaccine by ocular-nasal route at 10th day of age. The immune suppressed chickens had significantly low serum antibody titers and histopathological studies showed that there is depletion of bursal tissue in the CY treated chickens compared to CY untreated chickens. There is no significant difference in the cell mediated immunity between ablated and non ablated group. This study concludes that there is suppression of only humoral immunity by administering the cyclophosphamide and there is no effect on cell mediated immunity. CY will be used to study the role of cell mediated immunity and mucosal immunity in the absence of humoral immunity.


Dalfino L.,University of Bari | Puntillo F.,University of Bari | Ondok M.J.M.,University of Bari | Mosca A.,Microbiology Section | And 5 more authors.
Clinical Infectious Diseases | Year: 2015

Background. Critically ill patients with severe sepsis or septic shock may need relatively high colistin daily doses for efficacy against multidrug-resistant and extensively drug-resistant gram-negative rods. However, acute kidney injury (AKI) may represent a major dose-limiting adverse effect of colistin. We sought to determine AKI occurrence and to identify factors influencing AKI risk in severely ill patients receiving colistin according to a recently proposed dosing strategy. Methods. A prospective, observational, cohort study involving patients with severe sepsis or septic shock who received colistin was performed. AKI was defined according to Acute Kidney Injury Network criteria. Colistin administration was driven by a modified pharmacokinetics-pharmacodynamics (PK/PD)-based dosing approach. Results. Of 70 patients who received colistin at a median daily dose of 9 million IU (MIU; interquartile range, 5.87-11.1 MIU), 31 (44%) developed AKI. In univariate analysis, age, Acute Physiology and Chronic Health Evaluation (APACHE) II score, Sequential Organ Failure Assessment (SOFA), score and baseline renal impairment were significantly associated with AKI. Moreover, patients with AKI were less frequently treated with adjuvant ascorbic acid (P =. 003). In multivariate analysis, independent predictors of AKI were baseline renal impairment (adjusted hazard ratio, 4.15; 95% confidence interval, 1.9-9.2; P <. 001) and age (1.03; 1.0-1.05; P =. 028), whereas a strong independent renal-protective role emerged for ascorbic acid (0.27;. 12-.57; P <. 001). Conclusions. In severely ill patients receiving colistin according to a PK/PD-driven dosing approach, baseline renal impairment and older age strongly predict AKI occurrence, but concomitant administration of ascorbic acid markedly reduces AKI risk, allowing safer use of colistin. © 2015 The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved.


Vecchiarelli A.,Microbiology Section | d'Enfert C.,Institute Pasteur Paris
Virulence | Year: 2012

Bioluminescence imaging allows the visualization of the temporal and spatial progression of biological phenomena, in particular infection, by non-invasive methods in vivo. This nature-borrowed technology has been successfully used to monitor bacterial infections but recent studies have also succeeded in tracking fungal infections such as those caused by the two major opportunistic fungal pathogens Candida albicans and Aspergillus fumigatus. The findings of Donat and collaborators published in this issue now show that by combining the sensitivity of the Gaussia princeps luciferase with a surface display expression system it is possible to perform longitudinal infection studies on cutaneous forms of aspergillosis with a small number of animals. Besides providing new and valuable information in the field of aspergillosis, the findings of Donat et al. offer a new perspective on the general applicability of bioluminescence methodologies for eukaryotic pathogens where the bacterial lux operon cannot be exploited. © 2012 Landes Bioscience.


PubMed | Anesthesia and Intensive Care Unit, Microbiology Section and University of Bari
Type: Journal Article | Journal: Clinical infectious diseases : an official publication of the Infectious Diseases Society of America | Year: 2015

Critically ill patients with severe sepsis or septic shock may need relatively high colistin daily doses for efficacy against multidrug-resistant and extensively drug-resistant gram-negative rods. However, acute kidney injury (AKI) may represent a major dose-limiting adverse effect of colistin. We sought to determine AKI occurrence and to identify factors influencing AKI risk in severely ill patients receiving colistin according to a recently proposed dosing strategy.A prospective, observational, cohort study involving patients with severe sepsis or septic shock who received colistin was performed. AKI was defined according to Acute Kidney Injury Network criteria. Colistin administration was driven by a modified pharmacokinetics-pharmacodynamics (PK/PD)-based dosing approach.Of 70 patients who received colistin at a median daily dose of 9 million IU (MIU; interquartile range, 5.87-11.1 MIU), 31 (44%) developed AKI. In univariate analysis, age, Acute Physiology and Chronic Health Evaluation (APACHE) II score, Sequential Organ Failure Assessment (SOFA), score and baseline renal impairment were significantly associated with AKI. Moreover, patients with AKI were less frequently treated with adjuvant ascorbic acid (P = .003). In multivariate analysis, independent predictors of AKI were baseline renal impairment (adjusted hazard ratio, 4.15; 95% confidence interval, 1.9-9.2; P < .001) and age (1.03; 1.0-1.05; P = .028), whereas a strong independent renal-protective role emerged for ascorbic acid (0.27; .12-.57; P < .001).In severely ill patients receiving colistin according to a PK/PD-driven dosing approach, baseline renal impairment and older age strongly predict AKI occurrence, but concomitant administration of ascorbic acid markedly reduces AKI risk, allowing safer use of colistin.

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