Accessory gene regulator types of Staphylococcus aureus isolated in Gorgan, North of Iran

Background: Staphylococcus aureus is a gram-positive bacterium that has remained a persistent pathogen, causing infections such as endocarditis, meningitis, and toxic shock syndrome in humans. The accessory gene regulator (agr) system of Staphylococcus aureus is responsible for controlling the expression of many genes that code for virulence factors. In this study, we assessed the S.aureus agr Group, based on their source of isolation, in Gorgan, North of Iran. Materials and Methods: DNA of 194 S. aureus isolates was extracted by lysozyme-phenol chloroform method, which included 85 clinical samples, 58 samples which were isolated from noses of health care workers and 51 cases which were obtained from food products in Gorgan, northern Iran. PCR-based assays were used to evaluate agr locus nucleotide polymorphism for the identification of agr specificity Group. Distributions of each agr Group were determined and comparison between different sources was assessed by X2. A p-value of <0.05 was considered as significant. Results: The majority of isolates belonged to agr Group I (43.3%), followed by agr Group III (28.87%), agr Group II (22.68%), and agr Group IV (5.15%). In our study, a majority of S. aureus isolates were recovered from health care workers and food product specimens were of agr Group I and isolates which were recovered from patients were of agr Group III. These differences were statistically significant (P=0.005). There was no statistical difference between the source of isolation of clinical samples of S.aureus and agr type. Conclusion: Agr Group I was predominant among health care workers and food product specimens in Gorgan, North of Iran, but in strains which were isolated from patients, agr Group III was predominant. Investigating the possible role of agr Group III in Staphylococcus aureus infection in future studies is recommended

Airborne Fungi Spores in Different Wards of Hospitals Affiliated to Kerman University of Medical Sciences

Abstract: Introduction: The infections resulting from opportunist invasive fungi, such as Aspergillus, are an increasingly developing problem in hospitalized patients especially those suffering from immunosuppressive deficiencies. Method: In this study, air sampling from selected wards of three hospitals affiliated to Kerman University of Medical Sciences was carried out 2 times/day over 7 months (May – October, 2003) by using 500 plates containing saborodextrose-agar. Results: According to the results, 89% of the plates proved to be positive for fungal growth. Of this pool of fungi plates, 1034 fungal colonies containing 16 different fungi were sorted out which in regard to the frequency were Penicillium, Rhizopus, Aspergillus flavus, yeast, and Alternaria respectively. Higher rate of fungi colonies was observed in Bahonar hospital comparing to the two others. The most contaminated settings were ICU wards in Bahonar and Shafa hospitals, respectively. Comparison of the common wards of Bahonar and Afzalipour hospitals, revealed that the emergency room in Bahonar hospital had the highest rate of contamination. Finally, among the isolated Aspergillus and fungi, Aspergillus flavus and yeast species showed the highest frequencies. Conclusion: Considering the results of the present study, control of fungal contamination in hospital wards, especially those in which patients with immunosuppressive deficiency are hospitalized is highly necessary. Keywords: Airborne fungal spores, Hospita

Staphylococcus aureus typing by digestion of protein A coding gene using Bsp143I

Background: Protein A is the virulence factors of Staphylococcus aureus rolling in its pathogenesis, and its gene is used for typing. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with different enzymes has been used for this action. Objectives: In this study, we used Bsp143I enzyme for digestion of the gene, coding protein A (spa gene) in S. aureus. The bacteria were isolated from patients and healthy carriers in Gorgan, north of Iran. Patients and Methods: DNAs of 128 S. aureus subjects (53 from healthy carriers and 75 from patients) were extracted and amplified using specific primers of the spa gene. The product was digested by Bsp143I enzyme and its pattern was assessed by gel electrophoresis. Results: There were seven spa types among the tested S. aureus samples, among which six types differed in the repeated X region of the spa gene, but the seventh type had a deletion on one of BSP143I restriction sites. The frequency of spa types among isolated S. aureus samples as well as healthy carriers was six and five, respectively. S. aureus isolated from wounds showed the most diverse spa types (five) among clinical samples. Types 1, 2 and 4 were observed in all clinical samples, while only one case of type 3 was identified among patients, whereas this type constituted over 32% of the isolates among carriers. We found seven and four spa types among methicillin-resistant S. aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) isolates, respectively. Conclusions: Our results showed that typing the spa gene using PCR-RFLP using Bsp143I was an acceptable method for typing S. aureus. Furthermore, this survey showed that the types in healthy carriers and MSSA were more variable than patient and MRSA isolates, respectively. We used the Bsp143I enzyme, which was not used in any previous studies on the spa gene. The results of this study suggested that we can use PCR-RFLP of spa gene by Bsp143I for molecular typing and sequencing of S. aureus, instead of relatively expensive methods. This method is relatively rapid and inexpensive, and can be accomplished in centers with conventional molecular facilities. © 2014, Ahvaz Jundishapur University of Medical Sciences

Prevalence of Panton-valentine gene in Staphylococcus aureus isolated from clinical samples and healthy carriers in Gorgan city, north of Iran

Aim. Staphylococcus aureus (S. aureus) is a nosocomial and community acquired pathogen. S. aureus is a pathogen that causes several types of disease from skin infections to systemic diseases that is because of having several virulence factors such as enzymes, toxins, superantigens and Panton-Valentine leukocidin (pvl). pvl is a bi-component leukotoxin that destroy PMNs and monocytes and causes furunculosis, abscesses and necrotizing soft tissue infections in patients without any risk factors for such infections. The goal of this study was determine the prevalence of pvl gene in S. aureus isolated from patients and healthy carriers in Gorgan city, north of Iran. Methods. One hundred seventy isolates of S. aureus, 95 from patients and 75 healthy carriers, were collected during one year. After identification and purification, DNA extraction was done by phenol-chloroform method. Amplification of pvl gene was done by specific primer and polymerase chain reaction method. Results. Among the 170 isolates of S. aureus, 20 contained pvl gene. The frequency of isolates contained pvl gene in MRSA and MSSA isolates were 21.6, 19.3, which was not statistically significant. The frequency of these genes was not related to age, sex and source of isolation from patients. Conclusion. The frequency of pvl gene in this region were much higher than expected. © Copyright 2016 Edizioni Minerva Medica

The Frequency of MRSA carriers in health care workers in Gorgan, North of Iran

Methicillin resistant Staphylococcus aureus (MRSA) is one of the most important pathogen in hospitals. Healthcare personnel are the main source of nosocomial infections and identification and control of MRSA carriers can reduce incidence of infections. The aim of this study was to determine the frequency of methicillin resistant Staphylococcus aureus (MRSA) and their antibiotic susceptibility profile among healthcare workers in Gorgan located in northern Iran. Three hundred and thirty three of healthcare workers were participated in this cross-sectional study in 2010. Samples were taken with sterile cotton swabs from both anterior nares. Swabs were plated onto Mannitol salt agar. S. aureus were identified by Gram stain, Catalase, Coagulase and DNase tests. MIC (micro dilution broth) method was used to determine resistance of strains to methicillin. Antimicrobial susceptibility pattern to other antibiotics was performed by diffusion method. Frequency of S. aureus and MRSA carriers among healthcare workers was 24% (80.33) and 3% (10.33) respectively. MIC of isolates was varied between 0.5 and 65.31 (39%) of cases were showed MIC of intermediate that ranged between 4 and 8. Penicillin and Imipenem resistance were seen in 97.5% and 1.4% of isolated S. aureus strains, respectively. Frequency of S. aureus carriers in healthcare workers in our area was median in compare with other region in Iran but the MRSA carriage in healthy staff was lower than most part of Iran. It would be considering to monitor healthy carrier staff because of high rate intermediate MIC in this group to prevent conversion to MRSA

The Effect of Melatonin on Behavioral, Molecular, and Histopathological Changes in Cuprizone Model of Demyelination

Multiple sclerosis (MS) is an autoimmune, demyelinating disease of the central nervous system. The protective effects of melatonin (MLT) on various neurodegenerative diseases, including MS, have been suggested. In the present study, we examined the effect of MLT on demyelination, apoptosis, inflammation, and behavioral dysfunctions in the cuprizone toxic model of demyelination. C57BL/6J mice were fed a chaw containing 0.2 % cuprizone for 5 weeks and received two doses of MLT (50 and 100 mg/kg) intraperitoneally for the last 7 days of cuprizone diet. Administration of MLT improved motor behavior deficits induced by cuprizone diet. MLT dose-dependently decreased the mean number of apoptotic cells via decreasing caspase-3 and Bax as well as increasing Bcl-2 levels. In addition, MLT significantly enhanced nuclear factor-κB activation and decreased heme oxygenase-1 level. However, MLT had no effect on interleukin-6 and myelin protein production. Our data revealed that MLT improved neurological deficits and enhanced cell survival but was not able to initiate myelin production in the cuprizone model of demyelination. These findings may be important for the design of potential MLT therapy in demyelinating disorders, such as MS. © 2015, Springer Science+Business Media New York

Effects of intermittent fasting on experimental autoimune encephalomyelitis in C57BL/6 mice

Several religions recommend periods of fasting. One of the most frequently asked questions of MS patients before the holy month of Ramadan is weather fasting might have an unfavorable effect on their disease course. This debate became more challenging after the publication of experimental studies suggesting that calorie restriction prior to disease induction attenuates disease severity. We conducted this study to assess early and late effects of fasting on the animal model of MS, known as autoimmune encephalomyelitis. EAE was induced in the C57BL/6 mice, using Myelin Oligodendrocyte Glycopeptide (MOG) 35-55 and they fasted every other day either after the appearance of the first clinical sign or 30 days after disease induction for ten days. Thereafter, the mice were sacrificed for further histological and immunological evaluations. Intermittent fasting after the establishment of EAE did not have any unfavorable effect on the course of disease. Moreover, fasting at the early phase of disease alleviated EAE severity by ameliorating spinal cord demyelination. Fasting suppressed the secretion of IFN-γ, TNF-α and raised IL-10 production in splenocytes. Fasting was also associated with a lower percent of cytotoxicity. Intermittent fasting not only had no unfavorable effect on EAE but also reduced EAE severity if started at early phase of disease. © Summer 2016, Iran J Allergy Asthma Immunol. All rights reserved

Effect of oral genistein administration in early and late phases of allergic encephalomyelitis

Objective(s): Experimental allergic encephalomyelitis (EAE) is an autoimmune disease validated as animal model of multiple sclerosis (MS). Administration of genistein, a phytoestrogenic component of soy, to mice at the onset of EAE is known to attenuate the clinical signs of the disease. The potential effects of genistein on established EAE is less studied. In the current study, we aimed to compare the effects of genistein administration on EAE severity in early and late phases of the disease. Materials and Methods: The C57BL/6 mice were induced with EAE, using MOG 35-55 and gavaged with genistein (300 mg/kg) either after the appearance of the first clinical sign or 30 days post disease induction for ten days. 24 hr after the last gavage, mice were sacrificed. Brains and spleens were removed for assessing lymphocyte proliferation, cell cytotoxicity, and cytokine profile. Spinal cords were dissected to assess the amount of demyelination using Luxol fast blue/cresyl violet staining. Results: Administering mice with genistein, after the establishment of EAE, did not reverse the clinical signs of disease. However, treating with genistein at the onset of disease alleviated the clinical signs by reducing neuronal demyelination. Genistein suppressed the production of IFN-γ and enhanced IL-10 secretion in splenocyte and brain. Genistein also reduced IL-12 and TNF-α secretion in splenocytes, suppressed the proliferation of T-cells, and reduced the cell cytotoxicity. Conclusion: Genistein oral therapy might only reduce EAE severity if started in early phases of the disease

Comparison of ferment sugars, produce hemolysis and measuring growth in methicillin-resistant and methicillin-sensitive Staphylococcus aureus isolates from inpatients and healthcare workers in Gorgan Hospitals, North of Iran

The mec A gene in Staphylococcus aureus leads to production of new penicillin-binding protein called PBP2a.This change may follow some changes in other phenotypes. The aim of this study was the comparison of Ferment Sugars, Produce Hemolysis and Measuring Growth in MRSA and MSSA isolates. 188 Staphylococcus aureus isolates separated from inpatients and healthcare workers (healthy carriers)were studied.Bacterialcultures in blood agar environment at 37°C during 24h and at 4°C during other 24h were applied for studying hemolysis. Sugar fermentation carried out in phenol red Broth medium, containing glucose, galactose, arabinose, fructose, xylose, ramnose, mannose, sucrose, trehalose, raffinose or maltose. For determining bacterial growth,bacterial concentration of 103was taken each hour during 12 cultured in MHAand colonies were counted after 24h.The mean amount of hemolysis diameter in MRSA isolates was rather more than that of MSSA isolates. The difference between MRSA and MSSA isolates were significant as to fermenting ramnose, trehalose, galactose and xylose. The mean rate of growth in MRSAwere significantly different from that of MSSAisolates (p&lt;0.05).Resistance to methicillin in Staphylococcus aureus isolates accompanies the increase of ability to ferment sugars. This phenomenon may be one of reasons for increased pathogenicity of MRSA isolates; So results shows the logarithmic phase is longer in MRSA isolates, This may implicate that PBP2a production in methicillin-resistant isolates follows slowing down nutrients entrance into the bacterium that in turn may causes slow growth

Antitumor effect of therapeutic HPV DNA vaccines with chitosan-based nanodelivery systems

Cervical cancer is the second-most-common cause of malignancies in women worldwide, and the oncogenic activity of the human papilloma virus types (HPV) E7 protein has a crucial role in anogenital tumors. In this study, we have designed a therapeutic vaccine based on chitosan nanodelivery systems to deliver HPV-16 E7 DNA vaccine, considered as a tumor specific antigen for immunotherapy of HPV-associated cervical cancer. We have developed a Nano-chitosan (NCS) as a carrier system for intramuscular administration using a recombinant DNA vaccine expressing HPV-16 E7 (NCS-DNA E7 vaccine). NCS were characterized in vitro for their gene transfection ability. Results: The transfection of CS-pEGFP NPs was efficient in CHO cells and the expression of green fluorescent proteins was well observed. In addition, NCS-DNA E7 vaccine induced the strongest E7-specific CD8+ T cell and interferon γ responses in C57BL/6 mice. Mice vaccinated with NCS-DNA E7 vaccine were able to generate potent protective and therapeutic antitumor effects against challenge with E7-expressing tumor cell line, TC-1. Conclusions: The strong therapeutic effect induced by the Chitosan-based nanodelivery suggest that nanoparticles may be an efficient carrier to improve the immunogenicity of DNA vaccination upon intramuscular administration and the platform could be further exploited as a potential cancer vaccine candidate in humans. © 2014 Tahamtan et al