Chrysin loaded polycaprolactone-chitosan electrospun nanofibers as potential antimicrobial wound dressing.

In this study, various concentrations of chrysin (chry) were loaded into polycaprolactone-chitosan (PCL-CTS) nanofibers to develop a potential wound dressing materials using electrospinning method. The structural composition and the morphology of the produced PCL-CTS5, PCL-CTS10 and PCL-CTS15 were analyzed by FE-SEM and FTIR, respectively. By increasing the amount of chry, the average diameter of the nanofibres was also increased to 191 ± 65 nm, 203 ± 72 nm, and 313 ± 69 nm for PCL-CTS5, PCL-CTS10, and PCL-CTS15, respectively. Moreover, the physicochemical characteristics and biological properties of synthesized nanofibers such as tensile testing, in-vitro drug release, porosity, decomposition rate, water absorption rate, water vapor permeability rate, cell viability, antioxidant and antibacterial activity were evaluated. By using Korsmeyer-Peppas and Higuchi kinetic models, the chry release mechanism in all nanofibers was studied in PBS solution, which suggested a Fick's diffusion. In-vitro antioxidant experiments by DPPH assay indicated 24, 43, 61 and 78 % free radical scavenging activity for PCL-CTS, PCL-CTS5, PCL-CTS10 and PCL-CTS15. In-vitro antibacterial examination showed that chry-loaded nanofibers had high antibacterial activity in which were comparable with the standard reagents. In-vitro cytotoxicity results obtained by MTT assay indicated a desired cytocompatibility towards fibroblast cells.

Epidemiological characterization of clinical isolates of meticillin resistant Staphylococcus aureus through multilocus sequence typing and staphylococcal cassette chromosome mec typing in Northwest Iran.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA), is considered a potential and aggressive nosocomial pathogen. It accounts for 50% of S. aureus isolates in tertiary hospitals in Iran, however, there is no sufficient evolutionary and epidemiological investigation about this medically important bacterium. We aimed to study the lineage and evolution of MRSA in Northwest Iran during 2021-2022 based on the obtained phenotypic and genotypic characteristics. MATERIALS AND METHODS: Seventy-two non-duplicate MRSA isolates were collected from 3 referral hospitals in Tabriz, Ardebil, and Urmia cities. The antimicrobial susceptibility patterns were determined by disk diffusion test and micro broth dilution methods. Thereafter 4 virulence genes (eta, etb, pvl, tst) and 5 types of staphylococcal cassette chromosome mec (SCCmec) were detected by PCR. In the final step, representative isolates were selected to be studied by Multilocus sequence typing (MLST). RESULTS: The highest resistance was observed to erythromycin and clindamycin at a rate of 76.4%, followed by ciprofloxacin (61.1%), gentamicin (54.2%), rifampin (38.9%), and co-trimoxazole (27.8%). All isolates were susceptible to vancomycin. The virulence genes of etb, pvl, tst, and eta were detected in 50%, 29.2%, 21.8%, and 13.9% of isolates, respectively. SCCmec types III and I were the most prevalent types, followed by types IV, II, and V. MLST analysis revealed 6 sequence types: ST6854, ST5282, ST127, ST7804, ST1607, and ST7784. Two MLST-based clonal complexes (CC8, and CC97) were identified as well. CONCLUSION: The ST numbers were non-repetitive. CC8 as a pandemic clone and an individual lineage and clinically significant clade was reported as the most prevalent clonal complex. It is essential periodic evaluations of antibiotic susceptibility patterns and study the evolutionary characteristics of medical-challenging microorganisms in particular MRSA to effectively treat and restrict the outbreaks.

Fabrication and characterization of tretinoin-loaded nanofiber for topical skin delivery.

BACKGROUND: Tretinoin or all-trans retinoic acid is used in the treatment of acne vulgaris and photo-aging. This work aims to develop tretinoin-loaded nanofibers as a potential anti-acne patch and to investigate its physicochemical characteristics. METHOD: Nanofibers were produced via electrospinning method and surface topography was evaluated by Field Emission Scanning Electron Microscopy (FESEM). The functional groups of polymer and the drug molecule and the possible interactions were studied by Fourier Transform Infrared Spectroscopy (FTIR). Drug release studies were carried out by total immersion method at 25 °C and 32 °C. Tretinoin stability was evaluated at room temperature and fridge for 45 days. The possibility of synergistic antibacterial activity of tretinoin and erythromycin combination was investigated on Staphylococcus aureus (ATCC® 25923™) and (ATCC® 29213™) by Kirby Bauer disc diffusion method. RESULTS: Uniform fibers without drug crystals were fabricated via electrospinning. Drug-loaded nanofibers show inherent stability under various storage conditions. Electrospun nanofibers showed a prolonged release of tretinoin. The stability of formulations in FT was higher than RT. Disc diffusion tests did not show any synergism in the antibacterial activity of erythromycin when used in combination with tretinoin. CONCLUSION: It can be anticipated that the easy fabrication, low costs and dosing frequency of the construct reported here provide a platform that can be adapted for on-demand delivery of tretinoin.

Comparison of biofilm production and virulence gene distribution among community- and hospital-acquired Staphylococcus aureus isolates from northwestern Iran.

INTRODUCTION: The emergence of antimicrobial-resistant isolates among Staphylococcus aureus and their genetic variations has become a major concern worldwide. The present study aims at comparing the biofilm formation and the genes encoding adhesion molecules in methicillin-susceptible, community- and hospital-acquired methicillin-resistant, vancomycin-intermediate and vancomycin-resistant S. aureus isolates. METHODOLOGY: The current study was conducted on 60 S.aureus isolates, collected at Urmia University of Medical Sciences, Iran, between the years 2014 and 2015. The modified Congo-red agar and Microtiter plate methods were used to determine biofilm production. PCR was used to detect the genes which were associated with a protein family of staphylococcal microbial surface components recognizing adhesive matrix molecules. The data were analyzed using SPSS (IBM SPSS Statistics, version 16). RESULTS: Of 60 isolates, 57 (95%) were biofilm producers. Unlike the bbp gene, which was only detected in 3 (5%) isolates, the eno and icaD genes were identified as the most prevalent as they were detected in 53 (88.3%) and 50 (85%) of 60 isolates, respectively. The dominant virulotype comprised eight genes (icaA, icaD, clfA, clfB, fnbA, cna, eno, ebpS) in eight isolates, six of which were community-acquired-MRSAs. CONCLUSION: A high percentage of the S. aureus isolates could produce a biofilm which is more common among methicillin-susceptible isolates. The high frequency of eno and icaD genes suggests that these genes may synergistically function in the onset and progression of bacterial colonization and biofilm formation. Meanwhile, this ability may help the bacteria resist the exposure of antibacterial agents and cause severe infections.

Comparative analysis of Enterococcus faecalis and Enterococcus faecium strains isolated from clinical samples and traditional cheese types in the Northwest of Iran: antimicrobial susceptibility and virulence traits.

This research was conducted using 50 samples of popular traditional cheeses and 160 enterococcal clinical isolates. Phenotypic and genotypic methods used for identification of enterococci. Then, the incidences of antibacterial resistance and virulence traits were investigated. In total, 165 E. faecalis and 43 E. faecium obtained from traditional cheeses and different clinical isolates were analyzed in the study. Antibiotic susceptibility testing revealed 175(84.1%) isolates with multi-drug resistance (MDR) patterns, which was more common among clinical sources. The predominant virulence profile, including gelE, asa1 and cpd was detected within 47 (22.6%) of the MDR isolates. Our results showed that traditional cheeses and clinical E. faecalis isolates have distinct patterns of virulence traits. The identified enterococci with antibiotic resistance and associated virulence factors, could provide a potential risk to the public health.

One piece biopsy for both rapid urease test and cultivation of Helicobacter pylori.

BACKGROUND & AIMS: Increasing antibiotic resistance among Helicobacter pylori isolates and the unsuccessful attempts at eradication can impose many costs to both healthcare systems and patients. The present study intended to find a way from which H. pylori could be isolated from biopsies with less invasive procedures undertaken on infected patients. METHODS: A biopsy specimen from each patient with gastric disorders was put into urea-containing agar. After two hours, the specimens were removed from agar and placed into tubes containing 1 ml 20% glucose solution. Then, the specimens were inoculated onto the Columbia agar and incubated under microaerophilic conditions. The grown colonies were identified as H.pylori based on the microbiology tests and PCR. RESULTS: Overall, 449 biopsy specimens were collected from the patients. Of all biopsies, 219 (48.8%) revealed positive results in the rapid urease test. Using the aforementioned method, 158 (35.2%) culture positive biopsy specimens were obtained. CONCLUSION: The researchers attempted to use one biopsy specimen for both rapid urease and culture tests. This method causes fewer injuries of gastric tissue and allows antimicrobial susceptibility testing and characterization in detail of the isolated organism.

Molecular Epidemiology of Vancomycin-Resistant Enterococcus faecalis and Enterococcus faecium Isolated from Clinical Specimens in the Northwest of Iran.

This study was conducted to investigate the phenotypic and genotypic characteristics of vancomycin-resistant Enterococcus faecalis and Enterococcus faecium. Antibiotic resistance and virulence genes in the aforementioned resistant isolates were studied using the epsilometer (E)-test and polymerase chain reaction (PCR). These isolates were subjected to typing by pulsed-field gel electrophoresis (PFGE). Thirty vancomycin-resistant enterococci (VRE; 18.75%) were isolated from a total of 160 various clinical specimens cultured for any bacterial growth. Of these, 11 (36.7%) isolates were identified as E. faecalis and 19 (63.3%) as E. faecium. Minimum inhibitory concentrations (MICs) of vancomycin, teicoplanin, and three alternative therapeutic options (linezolid, daptomycin, and quinupristin/dalfopristin) were determined using the E-test. Multiplex PCR was done for confirming species, identification of the resistant genotypes, and the detection of the virulence genes. Finally, the clonal relationship of all VRE strains was studied by PFGE. All VRE strains showed vancomycin MIC ≥256 µg/mL, and 27 (90%) isolates carried the vanA gene, whereas none of the isolates carried vanB. The most common resistance antibiotic pattern observed was toward rifampicin (n = 30 [100%]). Among all virulence genes studied, gelE (n = 28 [93.33%]) was found as the most prevalent virulent gene. VRE isolates exhibited 90%, 46.67%, 100%, and 66.67% resistance to teicoplanin, linezolid, quinupristin/dalfopristin, and daptomycin, respectively. Molecular typing demonstrated 16 PFGE types of VRE isolates (A-P). Although vanA was carried by most of the isolates, PFGE displayed small clonal dissemination among VR E. faecium and VR E. faecalis species.

Emergence of vancomycin-intermediate and -resistant Staphylococcus aureus among methicillin-resistant S. aureus isolated from clinical specimens in the northwest of Iran.

OBJECTIVES: The aim of this study was to evaluate the frequency as well as the phenotypic and molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant S. aureus (VRSA) isolates from clinical specimens at three university teaching hospitals in Urmia, Northwest Iran, from 2012-2015. METHODS: Following identification of the isolates, antibiotic susceptibility testing was performed. The presence of the mecA, vanA and pvl genes was evaluated, and staphylococcal cassette chromosome mec (SCCmec) typing was performed. RESULTS: A total of 177 S. aureus isolates were collected from various clinical specimens. Antibiotic susceptibility testing revealed high resistance rates to penicillin (98.9%), followed by erythromycin (61.6%). A total of 95 isolates (53.7%) were confirmed as MRSA. Among the initially screened vancomycin-intermediate S. aureus (VISA) isolates, one isolate with a minimum inhibitory concentration (MIC) of 6µg/mL harboured the vanA gene. Eleven MRSA isolates (11.6%) were also VRSA. A majority (23/95; 24.2%) of MRSA were classified as SCCmec type III. Only 6 MRSA isolates (6.3%) harboured the pvl gene. CONCLUSIONS: This study highlights the presence of MRSA along with VISA and VRSA in our setting. To our knowledge, this is the first report showing that a strain can be defined as VISA phenotypically and as VRSA by molecular analysis. Such a finding raises major concerns with regard to control measures and reliable laboratory tests for screening of resistant strains.

Occurrence of Enterococcus faecalis and Enterococcus faecium in Various Clinical Infections: Detection of Their Drug Resistance and Virulence Determinants.

The aim of this study was to characterize virulence determinants and antibiotic resistance profiles in enterococci obtained from various clinical sources in the northwest of Iran. A total of 160 enterococcal clinical isolates from various wards of University Teaching Hospitals were collected and specified by biochemical test, from September 2014 to July 2015. Identification of enterococci was confirmed by multiplex PCR in the genus and species level. Antibiotic resistance properties and virulence determinants were examined by phenotypic and molecular methods. Of 160 enterococcal isolates, 125 (78.12%) and 35 (21.88%) isolates were identified as Enterococcus faecalis and Enterococcus faecium, respectively. The most common antibiotic nonsusceptible pattern observed was resistance toward rifampicin [n = 122 (76.25%)] followed by erythromycin [n = 117 (73.12%)]. Among all isolates, gelE [n = 140 (87.5%)], cpd [n = 137 (85.6%)], and asa1 [n = 118 (73.8%)] were the most prevalent virulence genes studied. Thirty isolates (11 E. faecalis, 19 E. faecium) were found to be resistant to vancomycin, with minimum inhibitory concentration of ≥256 µg/ml. Twenty-seven isolates carried the vanA gene, whereas none of the isolates carried vanB. E. faecalis had a considerable ability to show virulence genes and drug resistance. Emergence of antibiotic-resistant enterococci and the high prevalence of virulence traits in our study could be regarded as an alarming situation.

Methicillin-resistant Staphylococcus aureus nasal carriage between healthy students of medical and nonmedical universities.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is a challenge for public health, and community-acquired (CA) infections seem to be increasing among people in different areas. METHODS: A total of 700 healthy student volunteers residing in dormitories of universities in Urmia, Iran, were enrolled in this study. After identification of the isolates, antibiotic susceptibility, presence of mecA and pvl genes, and staphylococcal cassette chromosome mec (SCCmec) typing were evaluated. RESULTS: Nasal screening identified 137 (19.6%) carriers of S aureus, and 18 (13.14%) were MRSA isolates. The antimicrobial susceptibility patterns of isolates revealed high resistance to penicillin (93.4%). All isolates were sensitive to vancomycin. The SCCmec typing showed that most MRSA strains belonged to SCCmec type IV (n = 14; 77.8%). Only 1 (5.56%) MRSA isolates carried the pvl gene. CONCLUSIONS: Our findings revealed the relatively high frequency of S aureus nasal carriers and the advent of multidrug resistance among these isolates. Most MRSA isolates were SCCmec type IV; the transfer of such MRSA strains from carriers to other individuals in crowded living conditions such as dormitories can act as a risk factor for outbreak of CA MRSA and is a serious threat for the study groups.

Immunology and vaccines and nanovaccines for Helicobacter pylori infection.

Helicobacter pylori infection is very common worldwide and is an important cause of gastritis, peptic ulcer disease, gastric mucosa-associated lymphoid tissue lymphoma, and gastric adenocarcinoma. Since the eradication requires treatment with multidrug regimens, prevention of primary infection by a suitable vaccine is attractive. Developing vaccines on the spot when and where an infection is breaking out might be possible, thanks to engineered nanoparticles. In this review, the nature of the host immune response to H. pylori infection is considered. We explain recent candidate vaccines and prophylactic or therapeutic immunization strategies for use against H. pylori. We also describe identification of different types of immune responses that may be related to protection against H. pylori infection. Thus, it seems that there is still a strong need to clarify the main protective immune response against H. pylori.

Virulence and antimicrobial resistance in enterococci isolated from urinary tract infections.

PURPOSE: Urinary tract infection (UTI) is the most common nosocomial infection among hospitalized patients. Meanwhile, most frequent infections involving enterococci affect the urinary tract. The aims of this study were to investigate the susceptibility pattern of isolated enterococci from UTI and the prevalence of virulence genes. METHODS: The study used enterococci isolated from urinary tract infections obtained from 3 university teaching hospitals in Northwest Iran. The antimicrobial susceptibility of the strains was determined using the disc diffusion method. Multiplex PCR was performed for the detection of genus- species specific targets, and potential virulence genes. RESULTS: Of 188 enterococcal isolates, 138 (73.4%) and 50 (26.6%) were Enterococcus faecalis and E. faecium, respectively. Antibiotic susceptibility testing showed high resistance to amikacin (86.2%), rifampicin (86.2%) and erythromycin (73.9%), irrespective of species. In total, 68.1% were positive for gelE, and 57.4%, 53.2%, 56.4%, and 52.1% of isolates were positive for cpd, asa1, ace, and esp, respectively. CONCLUSION: The study revealed that most of UTI isolates were multidrug resistance against the antibiotics tested and antibiotic resistance was more common among E. faecium isolates than E. faecalis. A significant correlation was found between UTI and the presence of gelE among E. faecalis strains (p < 0.001).

The status of antimicrobial resistance of Helicobacter pylori in Eastern Azerbaijan, Iran: comparative study according to demographics.

Helicobacter pylori-associated infections are extremely common in Iran, but few data about antibiotic sensitivity of H. pylori are available for this region. The purpose of this study was to investigate the prevalence of resistance in isolates against commonly used antibiotics in Eastern Azerbaijan, Iran, and the dependence of prevalence on the sex and age of patients. H. pylori isolates were collected by culture from gastric biopsies. Antibiotic susceptibility of isolates was determined by use of the disk agar diffusion test, and the minimum inhibitory concentration of clarithromycin was established by use of the Etest. A total 395 of biopsy specimens were studied; 112 samples of H. pylori were isolated (28.3 %), 55 (49 %) from males and 57 (51 %) from females. The prevalence of resistance to clarithromycin, metronidazole, erythromycin, amoxicillin, ciprofloxacin, rifampin, nitrofurantoin, and tetracycline were 16 (14.3 %), 86 (76.8 %), 29 (26.0 %), 32 (28.6 %), 37 (33.0 %), 32 (28.6 %), 13 (11.6 %), and 21 (18.7 %), respectively. Antimicrobial resistance was not statistically significantly associated with sex or age. Furthermore, the prevalence of resistance to metronidazole was high and that to clarithromycin was reasonable, consistent with reported low success in H. pylori treatment in this area. Therefore, continuous surveillance of antibiotic resistance of H. pylori is essential.

Survey of Virulence Determinants among Vancomycin Resistant Enterococcus faecalis and Enterococcus faecium Isolated from Clinical Specimens of Hospitalized Patients of North west of Iran.

Recent data indicates an increasing rate of vancomycin resistance in clinical enterococcal isolates worldwide. The nosocomial enterococci are likely to harbor virulence elements that increase their ability to colonize hospitalized patients. The aim of this study was to characterize virulence determinants in vancomycin-resistant enterococci (VRE) obtained from various clinical sources.During the years 2008 to 2010, a total of 48 VRE isolates were obtained from three University teaching hospitals in Northwest, Iran. Initially, phenotypic speciation was done and minimum inhibitory concentrations (MICs) of vancomycin were determined by agar dilution method and E-test. Then, species identification and resistance genotypes along with detection of virulence genes (asa1, esp, gelE, ace and cpd) of the isolates were performed by multiplex PCR.Thirty eight isolates were identified as vancomycin-resistant Enterococcus faecium (VREfm) and ten as E. faecalis (VREfs). Irrespective of the species, vanA gene (89.58%) was dominant and three phenotypically vancomycin susceptible E. faecium isolates carried the vanB gene. Among virulence genes investigated, the esp was found in 27(71%) VREfm strains, but did not in any VREfs. Other virulence determinants were highly detected in VREfs strains. Our data indicate a high prevalence of E. faecium harboring vancomycin resistance with vanA genotype and the two VRE species displayed different virulence genes.

Prevalence of bacteriocin activities and bacteriocin-encoding genes in enterococcal clinical isolates in Iran.

The aim of the project was to isolate and characterize bacteriocin-producing enterococci, as well as determine the prevalence of enterocin structural genes in 187 enterococcal clinical isolates from the northwest of Iran. The isolates were screened for antibacterial activity against 15 different indicator strains. The proteinaceous nature of the antimicrobial substances was confirmed by sensitivity to proteinase K; their stability to heat treatment was tested at 60 °C and 100 °C for 20 and 10 min, respectively. The PCR method was applied to detect previously identified enterocin genes. Our results showed that 38 (20.3%) of the enterococcal isolates were considered to be potential bacteriocinogenic strains. Furthermore, genes encoding diverse bacteriocin are highly distributed among clinical enterococci, and the strains with multi-bacteriocin genes displayed high antimicrobial activity. Enterocin A, enterolysin A, and enterocin L50A/B were the most abundant structural genes detected in bacteriocinogenic strains. This work is the first survey on the prevalence of bacteriocin genes among clinical enterococci in Iran that has isolated a strain with high antimicrobial activity and sensitivity to clinically relevant antibiotics.