MICROORGANISMS VARIANTS FOR HEALTHCARE-ASSOCIATED INFECTIONS IN A SELECTED TERTIARY CARE HOSPITAL

Objective: Microorganisms are minute and can be only in microscope and these are not visible to naked eyes. Various types of microbe include bacteria, virus, fungi, and protozoa. These microorganisms are subclassified and these are disease causing leading to mortality and morbidity. Healthcare-associated infections (HAIs) arise from different variants of microbes and knowing the category of microbes for treating the diseases with specific antibiotics is important for better patient outcome. Methods: Using secondary data, all the patients who had HAI for 3 years were taken into consideration by considering the different variants of microorganisms. Results: Retrospective data collected for the period of 3 years the inpatients who got admitted for more than 48 h of duration, the data collected included the parameters for various microorganisms such as Bacilli, cocci, Klebsiella, Acinetobacter, and Aures, other micro-organisms such as Escherichia coli, Citrobacter, and Pseudomonas microorganisms. Bacilli group of microorganisms was more common for urinary tract infection, blood stream infection, and ventilator-associated pneumonia. Aures was more common among surgical site infection infections. Conclusions: Most of the patients who had an HAI had two or more different kind of microorganisms which are responsible for spreading infection. There is a need to control microbial flora in the hospital set up as the rate of HAI increases with microbial flora

IdeS: A Bacterial Proteolytic Enzyme with Therapeutic Potential

Background: IdeS, a proteinase from Streptococcus pyogenes, cleaves immunoglobulin (Ig)G antibodies with a unique degree of specificity. Pathogenic IgG antibodies constitute an important clinical problem contributing to the pathogenesis of a number of autoimmune conditions and acute transplant rejection. To be able to effectively remove such antibodies is therefore an important clinical challenge. Methodology/Principal Findings: IdeS was found to specifically and efficiently cleave IgG in human blood in vitro (20 mg of IdeS caused a complete degradation of IgG in one ml of human whole blood in 15 minutes) and to clear IgG from the blood stream of rabbits in vivo (no IgG was detected six hours following an intravenous injection of 5 mg of IdeS) without any side effects. In a mouse model of immune thrombocytopenic purpura (ITP), polyclonal IgG antibodies against platelet surface antigens were used to induce a lethal disease. These profoundly thrombocytopenic animals were treated and cured by a single injection of IdeS. Conclusions/Significance: Novel information is provided concerning the IgG-cleaving activity of IdeS in vitro and in vivo. The highly specific and rapid elimination of IgG in vivo, the dramatic effect in a mouse model of ITP, and the lack of sid

A Recombinant Vaccine Effectively Induces C5a-Specific Neutralizing Antibodies and Prevents Arthritis

OBJECTIVES: To develop and validate a recombinant vaccine to attenuate inflammation in arthritis by sustained neutralization of the anaphylatoxin C5a. METHODS: We constructed and expressed fusion protein of C5a and maltose binding protein. Efficacy of specific C5a neutralization was tested using the fusion protein as vaccine in three different arthritis mouse models: collagen induced arthritis (CIA), chronic relapsing CIA and collagen antibody induced arthritis (CAIA). Levels of anti-C5a antibodies and anti-collagen type II were measured by ELISA. C5a neutralization assay was done using a rat basophilic leukemia cell-line transfected with the human C5aR. Complement activity was determined using a hemolytic assay and joint morphology was assessed by histology. RESULTS: Vaccination of mice with MBP-C5a led to significant reduction of arthritis incidence and severity but not anti-collagen antibody synthesis. Histology of the MBP-C5a and control (MBP or PBS) vaccinated mice paws confirmed the vaccination effect. Sera from the vaccinated mice developed C5a-specific neutralizing antibodies, however C5 activation and formation of the membrane attack complex by C5b were not significantly altered. CONCLUSIONS: Exploitation of host immune response to generate sustained C5a neutralizing antibodies without significantly compromising C5/C5b activity is a useful strategy for developing an effective vaccine for antibody mediated and C5a dependent inflammatory diseases. Further developing of such a therapeutic vaccine would be more optimal and cost effective to attenuate inflammation without affecting host immunity

B Cell Depletion Reduces the Number of Autoreactive T Helper Cells and Prevents Glucose-6-Phosphate Isomerase-Induced Arthritis

The therapeutic benefit of B cell depletion in patients with rheumatoid arthritis has provided proof of concept that B cells are relevant for the pathogenesis of arthritis. It remains unknown which B cell effector functions contribute to the induction or chronification of arthritis. We studied the clinical and immunological effects of B cell depletion in glucose-6-phosphate isomerase-induced arthritis. We targeted CD22 to deplete B cells. Mice were depleted of B cells before or after immunization with glucose-6-phosphate isomerase (G6PI). The clinical and histological effects were studied. G6PI-specific antibody responses were measured by ELISA. G6PI-specific T helper (Th) cell responses were assayed by polychromatic flow cytometry. B cell depletion prior to G6PI-immunization prevented arthritis. B cell depletion after immunization ameliorated arthritis, whereas B cell depletion in arthritic mice was ineffective. Transfer of antibodies from arthritic mice into B cell depleted recipients did not reconstitute arthritis. B cell depleted mice harbored much fewer G6PI-specific Th cells than control animals. B cell depletion prevents but does not cure G6PI-induced arthritis. Arthritis prevention upon B cell depletion is associated with a drastic reduction in the number of G6PI-specific effector Th cells

Electrodeposition and characterisation of CdS thin films using thiourea precursor for application in solar cells

CdS thin films have been successfully electrodeposited on glass/FTO substrates using acidic and aqueous solution of CdCl2.xH2O and thiourea (SC(NH2)2). The electrodeposition of CdS thin films were carried out potentiostatically using a 2-electrode system. The prepared films were characterised using X-ray diffraction (XRD), Raman spectroscopy, Scanning electron microscopy (SEM), Atomic force microscopy (AFM), Photoelectrochemical (PEC) cell measurements, Electrical resistivity measurements and UV-Vis spectrophotometry to study their structural, compositional, morphological, electrical and optical properties, respectively. The structural studies show that the as-deposited and annealed CdS layers are polycrystalline with hexagonal crystal structure and preferentially oriented along (200) planes. The optical studies indicate that the ED-CdS layers have direct bandgaps in the range (2.53-2.58) eV for the as-deposited and (2.42-2.48) eV after annealing at 400oC for 20 minutes in air. The morphological studies show the good coverage of the FTO surface by the CdS grains. The average grain sizes for the as-deposited and annealed layers were in the range (60-225) nm. These grains or clusters are made out of smaller nano crystallites with the sizes in the range ~(11-33) nm. The electrical resistivity shows reduction as thickness increases. The resistivity values for the as-deposited and annealed layers were in the range (0.82-4.92)×105 Ωcm. The optimum growth voltage for the CdS thin films was found to be at the cathodic potential of 797 mV with respect to the graphite anode. No visible precipitations of elemental S or CdS particles were observed in the deposition electrolyte showing a stable bath using thiourea during the growth

Premolis semirufa (Walker, 1856) Envenomation, Disease Affecting Rubber Tappers of the Amazon: Searching for Caterpillar-Bristles Toxic Components

Pararama, the popular name of the larval form of the moth Premolis semirufa inhabits rubber plantations in the Amazon region and the accidental contact of the skin with the caterpillar's bristles or cocoons results in immediate and intense heat, pain, edema, and itching. In many cases a chronic inflammatory reaction with immobilization of the joints occurs. The current study has evaluated the biological and immunochemical characteristics of the Pararama caterpillar bristles extract. Electrophoretic analysis showed the presence of several components, including a very intense 82 kDa band. This latter component was endowed with intense gelatinolytic activity, as observed in zymography assays. Further analysis revealed that the extract also contained hyaluronidase activity but is devoid of phospholipase A2 activity. In vivo assays, using mice, showed that the extract was not lethal, but caused significant edema and induced intense infiltration of inflammatory cells to the envenomation site. The extract also induced high specific antibody titers, but no autoantibodies were detected. The data obtained, so far, demonstrate the existence of a mixture of different enzymes in the bristles of Premolis semirufa caterpillar, which can act together in the generation and development of the clinical manifestations of the Pararama envenomation

Human IgG/FcγR Interactions Are Modulated by Streptococcal IgG Glycan Hydrolysis

BACKGROUND: The human pathogen Streptococcus pyogenes produces an endoglycosidase, EndoS that hydrolyzes the chitobiose core of the asparagine-linked glycan on the heavy chain of human IgG. IgG-binding to Fc gamma receptors (FcgammaR) on leukocytes triggers effector functions including phagocytosis, oxidative burst and the release of inflammatory mediators. The interactions between FcgammaR and the Fc domain of IgG depend on the IgG glycosylation state. METHODOLOGY/PRINCIPAL FINDINGS: Here we show for the first time that EndoS hydrolyzes the heavy chain glycan of all four human IgG subclasses (IgG1-4), in purified form and in a plasma environment. An inactive form of EndoS, obtained by site-directed mutagenesis, binds IgG with high affinity, in contrast to wild type EndoS that only transiently interacts with IgG, as shown by Slot-blotting and surface plasmon resonance technology. Furthermore, EndoS hydrolysis of the IgG glycan influences the binding of IgG to immobilized soluble FcgammaR and to an erythroleukemic cell line, K562, expressing FcgammaRIIa. Incubation of whole blood with EndoS results in a dramatic decrease of IgG binding to activated monocytes as analyzed by flow cytometry. Moreover, the IgG bound to K562 cells dissociates when cells are treated with EndoS. Likewise, IgG bound to immobilized FcgammaRIIa and subsequently treated with EndoS, dissociates from the receptor as analyzed by surface plasmon resonance and Western blot. CONCLUSIONS/SIGNIFICANCE: We provide novel information about bacterial enzymatic modulation of the IgG/FcgammaR interaction that emphasizes the importance of glycosylation for antibody effector functions. Moreover, EndoS could be used as a biochemical tool for specific IgG N-glycan hydrolysis and IgG purification/detection, or as a potential immunosuppressing agent for treatment of antibody-mediated pathological processes