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Background: Non- Tuberculous Mycobacteria are environmental opportunistic pathogens that can be found in various terrestrial and aquatic habitats. There are an epidemiological links between species isolated in tap water and those isolated from patients. hsp65 gene has more variability in its sequences, compared to the some more conserved genes in NTM, for identification of mycobacteria to species level. In this study, the prevalence of NTM in Isfahan City water samples was determined using culture, biochemical tests and PCR-RFLP analyses of hsp65 gene.Methods: Eighty-five water samples were collected and cultured. The mycobacterial isolates were identified by conventional biochemical tests. A 441 bp fragment of hsp65 genes was amplified and digested by two restriction enzymes, BstEII and HaeII. Digested products were analyzed using polyacrilamid gel electrophoresis (PAGE).Results: 25.9% of the water samples contained different species of NTM. Dominant isolates were M. fortuitum (26.7%), M. chelonae like organism (13.3%) and M. mucogenicum (13.3%). Nineteen isolates of Mycobacteria were differentiated using hsp65 genes PCR-RFLP. Three isolates could not be identified at the species level because their RFLP patterns were different from other known PCR-RFLP profiles. There were different hsp65 gene PCR-RFLP profiles produced by digestion with BstEII and HaeIII. Conclusion: This study showed that PCR-RFLP of hsp65 gene in mycobacteria is more reliable method for identification of NTM at the specie level than conventional phenotypic methods (P

Background&Aims: Vancomycin Resistant Enterococci (VRE) are among the most common nosocomial pathogens worldwide. The intestinal tract provides a major source for transmission of these bacteria. Probiotics are living microorganisms that moderate use of them has inhibitory effect on intestinal colonization by enteric pathogens. We examined the effect of Lactobacillus rhamnosus GG (LGG) on inhibition of VRE colonization in mouse model. Methods: Twenty four mice (Balb/C) were controlled for a week and then were infected to VRE by daily receiving of 1ml oral vancomycin (250µg/ml) and 1ml VRE suspension in MHB ( 5 10 CFU / ml 8 × ) for one week. Mice were randomly divided into two groups of treatment and control, and the effect of LGG probiotic was compared in the tow groups. VRE, total Enterococci, and enteric gram-negative bacilli counts in feces were determined before and after colonization by VRE. Results: At first, all mice were colonized by non –Vancomycin Resistant Enterococci (mean 5×105CFU/g for 7 days), and Vancomycin resistance was not detectable. Following gastric inoculation of VRE and receiving oral vancomycin, VRE was colonized in gastrointestinal tract of all mice (mean 1.6×106CFU/g for 7 days). Oral administration of LGG suppressed growth of all Enterococci, including the vancomycin-resistant strain in treatment group feces (P

Background and Objectives: Cystic fibrosis (CF) is an autosomal recessive genetic disease. Infections in these patients gene.are mostly caused by three bacteria: Staphylococcus aureus, Haemophilus influenza and particularly Pseudomonas aeruginosa. Carbapenems including antibiotics are used to combat infections with Pseudomonas aeruginosa. In recent years, carbapenems resistant strains of P. aeruginosa isolated from clinical specimens are being reported. Decrease in drug penetration and production of metalobeta lactamase (MBLS) have been proposed as mechanisms of resistance. Materials and Methods: In this descriptive study, the population under investigation was 27 patients suffering from CF in Alzahra hospital of Isfahan. Clinical specimens were taken by deep swabbing from throat and data from every patient was recorded in a questionnaire. The specimens were cultured and isolated organisms were identified as P. aeruginosa using standard tests. Kirby-Bauer disk diffusion method was used to determine the bacterial drug resistance pattern. Strains of P. aeruginosa were checked for production of MBLS using disk impregnated with IPM-EDTA and PCR targeting of bla. Results: Among the 27 patients, 7 (26%) had P. aeruginosa infection. In total, 11 P. gene. aeruginosa isolates were taken. All isolates were susceptible to imipenem, ticarcillin, ciprofloxacin and piperacillin. The lowest scale of susceptibility belonged VIM to eftazidime (72.2%) followed by tobramycin (45.4%). None of the strains were positive for the bla Conclusion: Isolates of P. aeruginosa from CF patients in Isfahan were susceptible to antibiotics during the study period.

Background: Non- Tuberculous Mycobacteria are environmental opportunistic pathogens that can be found in various terrestrial and aquatic habitats. There are an epidemiological links between species isolated in tap water and those isolated from patients. hsp65 gene has more variability in its sequences, compared to the some more conserved genes in NTM, for identification of mycobacteria to species level. In this study, the prevalence of NTM in Isfahan City water samples was determined using culture, biochemical tests and PCR-RFLP analyses of hsp65 gene. Methods: Eighty-five water samples were collected and cultured. The mycobacterial isolates were identified by conventional biochemical tests. A 441 bp fragment of hsp65 genes was amplified and digested by two restriction enzymes, BstEII and HaeII. Digested products were analyzed using polyacrilamid gel electrophoresis (PAGE). Results: 25.9% of the water samples contained different species of NTM. Dominant isolates were M. fortuitum (26.7%), M. chelonae like organism (13.3%) and M. mucogenicum (13.3%). Nineteen isolates of Mycobacteria were differentiated using hsp65 genes PCR-RFLP. Three isolates could not be identified at the species level because their RFLP patterns were different from other known PCR-RFLP profiles. There were different hsp65 gene PCR-RFLP profiles produced by digestion with BstEII and HaeIII. Conclusion: This study showed that PCR-RFLP of hsp65 gene in mycobacteria is more reliable method for identification of NTM at the specie level than conventional phenotypic methods (P

Purpose of Review: This review evaluates the therapeutic potential of Ziziphus jujuba and its main components in managing complications of metabolic syndrome, including diabetes, dyslipidemia, obesity, and hypertension., Recent Findings: The reviewed studies provide evidence supporting the use of Z. jujuba and its main components (lupeol and betulinic acid) as natural treatments for complications of metabolic syndrome. These substances enhance glucose uptake through the activation of signaling pathways such as phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt), reduce hepatic glucose synthesis, and increase glucose uptake by adipocytes and skeletal muscle cells. They also improve insulin sensitivity by modulating AMP-activated protein kinase (AMPK) activity and regulating insulin signaling proteins and glucose transporters. In the field of dyslipidemia, they inhibit triglyceride synthesis, lipid accumulation, and adipogenic enzymes, while influencing key signaling pathways involved in adipogenesis. Z. jujuba and its constituents demonstrate anti-adipogenic effects, inhibiting lipid accumulation and modulating adipogenic enzymes and transcription factors. They also exhibit positive effects on endothelial function and vascular health by enhancing endothelial nitric oxide synthase (eNOS) expression, NO production, and antioxidant enzyme activity. Z. jujuba, lupeol, and betulinic acid hold promise as natural treatments for complications of metabolic syndrome. They improve glucose metabolism, insulin sensitivity, and lipid profiles while exerting anti-adipogenic effects and enhancing endothelial function. However, further research is needed to elucidate the mechanisms and confirm their efficacy in clinical trials. These natural compounds offer potential as alternative therapies for metabolic disorders and contribute to the growing body of evidence supporting the use of natural medicines in their management., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The Caucasus and surrounding areas, with their rich metal resources, became a crucible of the Bronze Age 1 and the birthplace of the earliest steppe pastoralist societies 2 . Yet, despite this region having a large influence on the subsequent development of Europe and Asia, questions remain regarding its hunter-gatherer past and its formation of expansionist mobile steppe societies 3-5 . Here we present new genome-wide data for 131 individuals from 38 archaeological sites spanning 6,000 years. We find a strong genetic differentiation between populations north and south of the Caucasus mountains during the Mesolithic, with Eastern hunter-gatherer ancestry 4,6 in the north, and a distinct Caucasus hunter-gatherer ancestry 7 with increasing East Anatolian farmer admixture in the south. During the subsequent Eneolithic period, we observe the formation of the characteristic West Eurasian steppe ancestry and heightened interaction between the mountain and steppe regions, facilitated by technological developments of the Maykop cultural complex 8 . By contrast, the peak of pastoralist activities and territorial expansions during the Early and Middle Bronze Age is characterized by long-term genetic stability. The Late Bronze Age marks another period of gene flow from multiple distinct sources that coincides with a decline of steppe cultures, followed by a transformation and absorption of the steppe ancestry into highland populations., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

Efflux pumps and biofilm play significant roles in bacterial antibiotic resistance. This study investigates the potential of chlorogenic acid (CGA) and carnosol (CL), as phenolic and diterpene compounds, respectively, for their inhibitory effects on efflux pumps. Among the 12 multidrug-resistant (MDR) strains of Staphylococcus aureus and Pseudomonas aeruginosa isolated from nosocomial skin infections, eight strains were identified as extensively drug resistant (XDR) using the disc diffusion method. The presence of efflux pumps in MDR strains of S. aureus and P. aeruginosa was screened using carbonyl cyanide-m-chlorophenylhydrazone. Between the 12 MDR strains of S. aureus and P. aeruginosa , 80% (4 out of 5) of the S. aureus strains and 85.7% (6 out of 7) of the P. aeruginosa strains exhibited active efflux pumps associated with gentamicin resistance. The checkerboard assay results, in combination with gentamicin, demonstrated that CGA exhibited a reduction in the minimum inhibitory concentration (MIC) for XDR S. aureus strain. Similarly, CL showed a synergistic effect and reduced the MIC for both XDR strains of S. aureus and P. aeruginosa . Flow cytometry was used to examine efflux pump activity at sub-MIC concentrations of 1/8, 1/4, and 1/2 MIC in comparison to the control. In XDR S. aureus , CGA demonstrated 39%, 70%, and 19% inhibition, while CL exhibited 74%, 73.5%, and 62% suppression. In XDR P. aeruginosa , CL exhibited inhibition rates of 25%, 10%, and 15%. The inhibition of biofilm formation was assessed using the microtiter plate method, resulting in successful inhibition of biofilm formation. Finally, the MTT assay was conducted, and it confirmed minimal cytotoxicity. Given the significant reduction in efflux pump activity and biofilm formation observed with CGA and CL in this study, these compounds can be considered as potential inhibitors of efflux pumps and biofilm formation, offering potential strategies to overcome antimicrobial resistance., Importance: In summary, CGA and CL demonstrated promising potentiating antimicrobial effects against XDR strains of Staphylococcus aureus and Pseudomonas aeruginosa , suggesting their probably potential as candidates for addressing nosocomial pathogens. They exhibited significant suppression of efflux pump activity, indicating a possible successful inhibition of this mechanism. Moreover, all substances effectively inhibited biofilm formation, while showing minimal cytotoxicity. However, further advancement to clinical trials is needed to evaluate the feasibility of utilizing CGA and CL for reversing bacterial XDR efflux and determining their efficacy against biofilms. These trials will provide valuable insights into the practical applications of these compounds in combating drug-resistant infections., Competing Interests: The authors declare no conflict of interest.

Introduction: Tuberculosis (TB) remains a serious public health problem worldwide. Drug-resistant TB is considered a major and growing global threat. Despite the great variety of described mutations in Mycobacterium tuberculosis (MTB) resistance genes, the mechanisms of drug resistance are still controversial. Recently, a report on the role of efflux pump genes in drug resistance added to this complexity. Therefore, a thorough understanding of efflux pump genes in drug-resistant TB clinical isolates is needed., Methodology: We performed molecular analysis of the efflux pump gene (Rv1258c) in 33 drug-resistant and 20 drug-sensitive clinical MTB isolates by sequencing the amplicons' targets in both the forward and reverse directions., Results: A novel mutation of the Rv1258c gene was identified at G442A (Ala148Thr) in rifampicin mono-resistant clinical strain, as compared to the H37Rv reference strain. In addition, a cytosine nucleotide insertion was found between the positions 580 and 581 (denominated Tap580) in two drug-sensitive strains at identical gene positions., Conclusions: These results indicated the possibility of mutation in the efflux pump genes and the important role of Tap efflux pump genes in drug-resistant MTB isolates. However, further research is required to determine the direct association of these mutations with resistant MTB., Competing Interests: No Conflict of Interest is declared, (Copyright (c) 2024 ShimaSadat Farzaneh, Fatemeh Norouzi, Hossein Fazeli, Mahshid Salehi, Marzieh Safari, Bahram Nasr Esfahani.)

Background: Acrylamide (ACR) can induce neurotoxicity through different pathways, including oxidative stress and apoptosis. Azithromycin is well-known for its antioxidant and anti-apoptotic properties., Objective: To evaluate the potential neuroprotective effect of azithromycin in an in vivo model of ACR-induced neurotoxicity, by investigating its impact on oxidative stress and apoptosis pathways., Methods: Male rats were divided into eleven groups at random (n = 6). 1:control (vehicle), 2:ACR (50 mg/kg, 11 days, I.P.), 3-7:ACR+ azithromycin (3.1, 6.25, 12.5, 25, 50 mg/kg, 11 days, I.P.), 8-9:ACR+ azithromycin (3.1, 6.25 mg/kg, from day 3-11), 10: ACR+ vitamin E (200 mg/kg, every other day, I.P.), 11. Azithromycin (50 mg/kg). Following the treatment period, a gait score examination was performed, and malondialdehyde (MDA), glutathione (GSH), Bcl-2-associated X protein (Bax)/B-cell lymphoma 2 (Bcl-2) ratio and caspase-3 levels in the cerebral cortex were measured., Results: Gait abnormality, a drop in GSH, and an increase in lipid peroxidation, Bax/Bcl-2 ratio, and caspase-3 levels were all significantly triggered by ACR in the cerebral cortex versus the control group. Azithromycin 3.1 and 6.25 mg/kg with ACR and azithromycin 6.25 mg/kg with ACR from day 3-11 ameliorated movement disorders caused by ACR. Azithromycin in all doses and both protocols along with ACR decreased the MDA level. Azithromycin (3.1, 6.25 mg/kg) along with ACR in both protocols increased the level of GSH, reduced the Bax/Bcl-2 ratio and caspase-3 amounts in the brain tissue versus the ACR group., Conclusions: Administration of azithromycin had both preventive and therapeutic effects on ACR-induced neurotoxicity through its antioxidant and antiapoptotic properties., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

There is increasing evidence showing that microbial dysbiosis impacts the health and cancer risk of the host. An association between adherent-invasive Escherichia coli (AIEC) and colorectal cancer (CRC) has been revealed. Cyclomodulins (CMs) have been receiving increasing attention for carcinogenic changes. In this study, the incidence and features of intracellular AIEC and cyclomodulin-encoding genes were investigated and the phylogenetic grouping and genetic relatedness were evaluated. E. coli strains were isolated from the colorectal biopsies. Adhesion and invasion assays and intramacrophage cell survival test were performed to separate the AIEC isolates. Virulence genotyping for the genes htrA , dsbA , chuA , and lpfA and the cyclomodulin toxins was also conducted. In addition, phylogenetic grouping of the isolates was determined. Subsequently, repetitive element sequence-based PCR (rep-PCR) fingerprinting was performed. A total of 24 AIEC pathovars were isolated from 150 patients. The prevalence rates of htr , dsbA , and lpfA were 70.83% and that of chuA was 91.66%. The frequencies of the cyclomodulin toxins were as follows: cnf1 , 29.2%; cnf2 , 25%; colibactin, 29.2%; and cdt , 4.2%; cif was not found. Among the AIEC isolates, 4.2%, 4.2%, 54.2%, 29.2%, and 8.3% with phylotypes A or C, B1, B2, D, and E were identified, respectively. Left-sided colon carcinoma and adenocarcinoma T≥1 stage (CRC2) were colonized by B2 phylogroup AIEC-producing CMs more often than the samples from the other groups. Close genetic relatedness was observed in AIEC isolates with rep-PCR., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Kamali Dolatabadi, Fazeli, Emami, Karbasizade, Maghool, Fahim and Rahimi.)

Tuberculosis (TB) is a global threat due to the emergence and spread of drug-resistant Mycobacterium tuberculosis (MTB). Isoniazid (INH) is the main antibiotic used for prevention and treatment of TB. Evidence shows that accumulated mutations can produce INH resistant (INHR) strains, resulting in the progression of multidrug-resistant (MDR) TB. Since point mutations in katG gene, inhA gene, and oxyR-ahpC region correlated with the INH resistance, in this study, we aimed to identify mutations in these three genes in INHR and MDR clinical isolates of MTB by Sanger DNA sequencing analysis. Thirty-three out of 438 isolates were resistant, including 66.7% INHR and 30.3% MDR isolates. In the katG gene, 68.2% INHR isolates had non-synonymous point mutations, mainly R463L (63.6%), and non-synonymous point mutation KatG L587P was seen in one of the MDR isolate. A novel silent substitution L649L was identified in the inhA gene of the MDR isolates. The oxyR-ahpC intergenic region g-88a common mutations (63.6%) in INHR and two distinct novel mutations were found at positions -76 and -77 of the oxyR-ahpC intergenic region. The coexistence of katG non-codon 315 with oxyR-ahpC intergenic region mutations was highly frequent in INHR 59.1% and MDR isolates 70%. Since mutations of all three genes 95.5% lead to the detection of INHR, they might be useful for molecular detection. Our results indicated the continuous evolution and region-specific prevalence of INH resistance. Overall, identification of new mutations in INH resistance can improve the available strategies for diagnosis and control of TB.

Inflammatory bowel diseases (IBD), including Crohn's disease (CD) and ulcerative colitis (UC), are known as chronic gastrointestinal inflammatory disorders. The present systematic review and meta analysis was conducted to estimate the prevalence of adherent-invasive Escherichia coli (AIEC) isolates and their phylogenetic grouping among IBD patients compared with the controls. A systematic literature search was conducted among published papers by international authors until April 30, 2020 in Web of Science, Scopus, EMBASE, and PubMed databases. The pooled prevalence of AIEC isolates and their phylogenetic grouping among IBD patients as well as in controls was estimated using fixed or random effects models. Furthermore, for estimating the association of colonization by AIEC with IBD, odds ratio along with 95% confidence interval was reported. A total of 205 articles retrieved by the initial search of databases, 13 case-control studies met the eligibility criteria for inclusion in the meta analysis. There were 465 IBD cases (348 CD and 117 UC) and 307 controls. The pooled prevalence of AIEC isolates were 28% (95% CI: 18-39%), 29% (95% CI: 20-40%), 13% (95% CI: 1-30%), and 9% (95% CI: 3-19%), respectively among IBD, CD, UC, and control group, respectively. Our results revealed that the most frequent AIEC phylogroup in the IBD, CD, and control groups was B 2 . Fixed-effects meta analysis showed that colonization of AIEC is significantly associated with IBD (OR: 2.93; 95% CI: 1.90-4.52; P < 0.001) and CD (OR: 3.07; 95% CI: 1.99-4.74; P < 0.001), but not with UC (OR: 2.29; 95% CI: 0.81-6.51; P = 0.11). In summary, this meta analysis revealed that colonization by AIEC is more frequent in IBD and is associated with IBD (CD and UC). Our results suggested that the affects of IBD in patients colonized with the AIEC pathovar is not random, it is in fact a specific disease-related pathovar., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Kamali Dolatabadi, Feizi, Halaji, Fazeli and Adibi.)

Background: Documented streptococcal resistance to erythromycin has recently been raised. The aim of this study is to identify the molecular mechanism of erythromycin resistance among group B Streptococcus (GBS) strains and to correlate with the clinical origin of strains., Materials and Methods: A total number of 134 colonizing ( n = 36), invasive ( n = 36), noninvasive ( n = 46), and asymptomatic ( n = 16) GBS isolates were characterized by the detection of dltS gene, capsular serotyping, antibiotic susceptibility profiles using disc diffusion method, and screening of the ermB , ermTR , and mefA resistance genes., Results: The distribution of capsular serotypes was as follow: serotype III (24.6%), Ia (21.6%), V (17.9%), Ib (14.9%), II (8.9%), IV (8.9%), VI (1.5%), and VII (1.5%). From 134 GBS isolates, 51 (38%) isolates were resistant to erythromycin. The constitutive macrolide lincosamide streptogrmin B (MLSB) was the most common resistance phenotype (62.7%), followed by inducible MLSB (27.4%) and M phenotype (9.8%). Erythromycin resistance rate was higher among asymptomatic GBS strains (13/16, 81.2%). Serotype III was the most prevalent type among resistant isolates (41.1%). The ermB gene highly distributed among resistant strains (64.7%), followed by ermTR (21.5%) and mefA (9.8%). The ermB gene was related to constitutive MLSB phenotype (84.3%, P < 0.05) and serotypes III (61.9%), Ib (87.5%), and V (83.3%). All M phenotype strains harbored mefA gene and were in association with serotype Ia (90%)., Conclusion: The current study suggests that ribosomal modification with erm genes is the main mechanism of erythromycin resistance. Because of relatively high prevalence of erythromycin resistance, double disc test highly recommended for GBS disease treatment and intrapartum prophylaxis among penicillin intolerant patients in our region., Competing Interests: There are no conflicts of interest., (Copyright: © 2021 Advanced Biomedical Research.)

Background: The information on antibiotic resistance and molecular features of Group B Streptococcus (GBS) are essential for epidemiological purposes as well as vaccine development. Therefore, we aimed to assess the antimicrobial resistance profiles and molecular characteristics of GBS isolates in Isfahan, Iran. A total number of 72 colonizing and invasive GBS were collected from pregnant and non-pregnant women. The GBS isolates were analyzed for resistance profiles, capsular genotyping, and detection of PI-1, PI-2a, PI-2b, hvgA, ermB, ermTR, lnuB and, mefA genes. Besides, erythromycin-resistant strains were subjected to multilocus sequence typing (MLST)., Results: The prevalence of colonizing and invasive GBS were 11 and 0.05%, respectively. The frequency of capsular serotypes was as follows: III (26.3%), Ia (20.83%), Ib and V (each 15.2%), IV (9.7%), II (8.3%), VII (2.7%), and VI (1.3%). Overall frequencies of PIs were as follows: PI-1, 37.5%, PI-1 + PI-2a, 30.5%, PI-1 + PI-2b, 29.1% and PI-2b, 2.7%. Two maternal colonizing GBS (2.6%) were hvgA positive and were belonged to ST-17/CPS-III/PI-1 + PI-2b lineage. Among 30(41.6%) erythromycin resistant GBS, 21 isolates (70%) harbored ermB gene, followed by ermTR (23.3%) and mefA (10%). One clindamycin-resistant isolate harbored the lnuB gene. MLST analysis revealed the following five clonal complexes (CCs) and nine STs: (CC-19/ST-335, ST-19, and ST-197), (CC-12/ST-43, ST-12), (CC-23/ST-163, ST-23), (CC-17/ST-17) and (CC-4/ST-16)., Conclusion: The study shows an alarmingly high prevalence of erythromycin-resistant GBS in Iran. In addition, we report dissemination of ST-335/CPS-III clone associated with tetracycline and erythromycin resistance in our region. The distribution of capsular and pilus genotypes varies between invasive and colonizing GBS that could be helpful for vaccine development.

Purpose: Leishmaniasis, a widespread parasitic disease, is a public health concern that is endemic in more than 90 countries. Owing to the drug resistance and also undesirable complications, designing new therapeutic methods are essential. C-reactive protein (CRP) is an acute phase protein of plasma with several immune modulatory functions. This study aimed to evaluate the effect of human recombinant CRP (hrCRP) on treating cutaneous leishmaniasis in mice models., Methods: hrCRP was expressed in E. coli Rosetta-gami and extracted from the SDS-PAGE gel. Male BALB/c mice were inoculated subcutaneously at the base of their tails by 1 × 10 5 stationary-phase of Leishmania major promastigotes (MHRO/IR/75/ER) suspended in sterile phosphate buffered saline (PBS). Nodules and subsequently, ulcers developed 14 days post-injection. 1.5 µg of the purified protein was administered on lesions of pre-infected mice by Leishmania major in the intervention group for five consecutive days., Results: The mean area of the lesions was decreased by about seven folds in the intervention group as compared to the control group after two weeks of the treatment (p = 0.024). The results were verified by the real-time polymerase chain reaction so that the parasite burden was determined 2 7 times in the control group as compared to the intervention group (p = 0.02). Two weeks after treatment, the conversion of the lesions to scars in the intervention group was observed., Conclusion: The results indicate a potential therapeutic role for hrCRP in improving cutaneous leishmaniasis due to Leishmania major in mice models. The healing was in a stage-dependent manner.

Background and Objectives: Bacterial antibiotic resistance is one of the most important threats for public health around the world. Carbapenemase-producing Gram-negative bacteria have resistance to most antibiotics including carbapenems complicating the treatment of infections. The aim of this study was to determine the antimicrobial susceptibility pattern of carbapenemase-producing nosocomial Gram-negative pathogens at a referral teaching hospital to reveal the best options for treatment of related infections., Materials and Methods: Gram-negative bacteria, isolated from hospitalized patients with nosocomial infections, underwent meropenem susceptibility test by disk diffusion method. Meropenem-resistant strains were evaluated for the presence of carbapenemase using Modified Hodge test (MHT). Finally, the antibiotic susceptibility test was performed to determine the sensitivity of each carbapenemase-positive strain against various antimicrobial agents according to the guidelines of Clinical and Laboratory Standards Institute (CLSI)., Results: Over the study period, 155 carbapenemase-positive isolates were detected. Pneumonia was the most frequent related nosocomial infection (67.1%) followed by UTI (23.2%). Acinetobacter baumannii (53.5%) and Klebsiella pneumoniae (40%) were the most frequently isolated pathogens. The pathogens had high rate of resistance to all antibiotics. Colistin had the most in vitro effect against all pathogens. Also, K. pneumoniae had a co-trimoxazole sensitivity rate equal to colistin (30.6%)., Conclusion: Carbapenemase-positive Gram-negative bacteria causing nosocomial infections are common in our hospital and have high rate of resistance to most antibiotics. Improvement in the pattern of antibiotic use and infection control measures are necessary to overcome this resistance., (Copyright © 2021 The Authors. Published by Tehran University of Medical Sciences.)

This study investigated carbapenem resistance among Acinetobacter baumannii isolated from respiratory specimens. Epidemiological relationship of the isolates was also evaluated. In this study, 81 respiratory specimens of A. baumannii from AL Zahra Hospital were confirmed by phenotypic and genotypic methods. Antimicrobial susceptibility was performed by disc diffusion method. Carbapenem resistance genes were identified by PCR. The isolates were typed by RAPD-PCR and multilocus sequence typing (MLST) methods. All isolates were resistant to imipenem and 80 isolates to meropenem. Frequency of oxacillinase genes was as follows: blaOXA-23 gene was positive in 74 (91.3%), blaOXA-24 gene in 50 (61.7%) and blaOXA-58 was not found in any isolates. On the other hand 22 (27.2%) isolates contained blaIMP-1, 3 (3.7%) isolates contained blaIMP-2 gene, 5 (6.2%) isolates contained blaVIM-1, 4 (5%) isolates had blaVIM-2 and none of the isolates had blaSIM-1 gene. RAPD-PCR typing identified 16 different patterns, with one pattern being the most frequent one in 26 isolates. In MLST 6 different sequence types were identified, the most predominant being ST2 belonging to clonal complex 2. The results of this study showed high resistance to carbapenems as well as high abundance of oxacillinase genes.

Background: The increasing incidence of Group B Streptococcus (GBS) infection among nonpregnant adults has become of growing clinical and public health concern. The current study investigated the distribution of important virulence determinants and antibiotic susceptibility of GBS isolates causing community acquired (CA) and hospital acquired (HA) infections among nonpregnant adults., Materials and Methods: A total of 62 GBS, including 31 CA GBS and 31 HA GBS, were collected from a teaching hospital in Isfahan, Iran. Capsular polysaccharide genotypes (CPS), PI 1, PI 2a, PI 2b, and hypervirulent GBS adhesin (hvgA) virulence genes and antibiotic resistance profiling were determined., Results: There were 19 (30.6%) cases of underlying disease that diabetes mellitus (20.9%) was most common. The rate of multidrug resistant GBS strains was accounted for 29%. Distribution of macrolide resistant phenotypes was as follows: constitutive macrolides, lincosamides, and streptogramin B (MLSB) (15 isolates); inducible resistance to MLSB; and L phenotype (each 5 isolates) and M phenotype (1 isolate). V and Ia serotypes were the most predominant capsular type in HA GBS and CA GBS isolates, respectively. The most frequent pilus types were PI 1, PI 1+PI 2a, PI 1+PI 2b, and PI 2a. PI 1 and PI 1+PI 2a had significantly different distributions between CA and HA GBS isolates. Three CA GBS isolates (9.6%) were positive for hvgA gene that belonged to clonal complex 17/sequence type 17/CPS III/PI 1+PI 2b lineage., Conclusion: There was a significant difference in the distribution of PIs among CA GBS and HA GBS isolates in our region., Competing Interests: There are no conflicts of interest., (Copyright: © 2020 Advanced Biomedical Research.)

Introduction: This study was conducted to identify the hypermucoviscosity, iron acquisition, and capsule serotypes of K. pneumoniae strains isolated from urinary tract infections among community-acquired patients (CA) and assess the frequency of plasmid-mediated quinolone resistance (PMQR) and extended-spectrum beta-lactamases (ESBL) genes between classic and hypervirulent strains., Materials and Methods: A total of 105 K. pneumoniae were isolated from CA-UTI. Demographic data related to the underlying diseases and clinical manifestations were further collected. Antibiotic resistance pattern and molecular characterization were compared among ESBL-positive, ESBL-negative, hypervirulent, and classic isolates., Results: The results revealed that 52.4% of the isolates were confirmed as ESBL producers and 11 (10.5%) were considered as hypervirulent K. pneumoniae (hvKp). Ciprofloxacin and nalidixic acid were the most inactive antibiotics with resistance rates of 68.6% and 64.8%, respectively. Molecular characterization revealed that 7.6% of all the isolates carried k1 and 66.6% carried K2 genes. The most frequent ESBL gene was bla SHV 63.8%, followed by bla TEM 59.0%, and bla CTX-M 58.1%. ESBL genes were significantly more in hvKp than in cKp. Moreover, 61 (84.7%), 47 (65.2%), and 16 (22.2%) of isolates harbored qnrB, qnrS , and qnrA . ESBL genes were detected in all hvKps, and bla SHV was observed in 90.9% of hvKp ( P value= 0.048, 95%)., Discussion: This study reported the high frequency of antimicrobial and multidrug resistance among hvKp isolates. Coexistence of PMQR and ESBL genes in hvkp indicates the necessity to enhance the clinical knowledge and management of hvKp infections., Competing Interests: The authors report no conflicts of interest in this work., (© 2020 Taraghian et al.)

Aim: The purpose of this study was to analyze the sequence of azurin gene in relation to its expression in Pseudomanas aeruginosa strains isolated from different clinical specimens of burn patients. Moreover, in silico sequence analysis of azurin gene using globally reported sequences was intended., Materials and Methods: Fifty-nine multidrug-resistant P. aeruginosa isolates were selected from different clinical specimens of patients suffering from burn wound infections in two university hospitals and subjected to antibacterial susceptibility testing. The frequency and genetic diversity of the azurin gene was determined by polymerase chain reaction (PCR) and Sanger sequencing. The azurin gene sequences were compared with the sequence data from other countries. The expression level of azurin gene in P. aeruginosa isolates with different azurin sequences from different clinical specimens was evaluated by real-time PCR., Results and Conclusion: About 98%-100% of the isolates were resistant to gentamicin, tobramycin, cefoxitin, ciprofloxacin, amikacin, and imipenem, while 100% and 23.9% of the isolates were susceptible to colistin and ceftazidime, respectively. Only eight point mutations were detected with amino acid substitutions in only two positions (81 and 102). In global analysis, 93% of strains showed missense mutation at positions 81 (alanine to threonine). The majority (81%) of Iranian strains were allocated to two major clusters distinct from the rest of world, which may suggest that strains from Iran have made a distinct genetic stockpile through point mutations which has established them separate from the other counties. However, 19% were distributed in different clusters together with the strains from different countries of North and South America, Europe, South and East Asia. The expression level of the azurin gene was statistically higher in the isolates collected from the blood of burns patients with systemic infection compared to the isolates collected from other specimens (wound, catheter and tissue), which shows a positive correlation between azurin gene expression and increased pathogenicity and capability for dissemination. This study may open new insight about azurin genetic variation and significance in P. aeruginosa pathogenesis., Competing Interests: The authors report no conflicts of interest in this work., (© 2020 Mohammadi Barzelighi et al.)

Introduction: Some countries have implemented reuse of laparoscopic instruments for cost-effective purposes. An accurate cleaning as the first step of reprocessing would lead to the effective sterilization. The purpose was to evaluate the effect of cleaning guidelines implementation on microbial load of laparoscopic instruments which were used in laparoscopic cholecystectomy surgery., Methods: This experimental study was done in an educational hospital, in 2017 and included a total of 128 laparoscopic instruments randomly selected from cholecystectomy surgeries and divided into two cleaning groups. The instruments were checked out in terms of number (colony-forming units [CFU]/mL) and type of microorganisms in two groups of routine cleaning and according to guideline cleaning. This guideline was indigenous and taken from successful instruction in this context that was presented by the Association for the Advancement of Medical Instrumentation (AAMI). The appropriate statistical analysis was conducted by SPSS version 19., Results: The average microbial load was 2.4 × 10 6 CFU/100 mL after clinical use. It was reduced to 7.2 × 10 5 CFU/100 mL in the control group and 3.4 × 10 4 CFU/100 mL in the intervention group, after the cleaning process. The most common microorganisms that were isolated immediately after clinical use were Escherichia coli 81.2%, Pseudomonas 68.8%, Klebsiella 57.8%, and spp., and so on., Conclusion: The AAMI cleaning method is recommended to be utilized by operating room nurses for laparoscopic instruments., (© 2019 Japan Society for Endoscopic Surgery, Asia Endosurgery Task Force and John Wiley & Sons Australia, Ltd.)

This study proposed to investigate the effect of azurin on the major stages of pathogenesis (adhesion and invasion) of intestinal bacterial pathogens (Salmonella spp. and Escherichia coli) and epithelial pathogens (Staphylococcus aureus and Pseudomonas aeruginosa) on the human colorectal adenocarcinoma (Caco-2) cell line. Azurin protein was produced by cloning the azurin gene into pET21a and heterologous expression in E. coli BL21. The protein was purified using affinity chromatography and confirmed by Western blotting. The purified protein was evaluated by three experiments of adhesion and invasion assays, including exclusion, competition, and replacement. Azurin was observed to significantly inhibit the attachment and invasion of S. aureus, Salmonella spp., and E. coli, while no such inhibitory effects were observed on P. aeruginosa. In fact, the protein increased the adhesion of P. aeruginosa to the cell. In conclusion, our study proposes that azurin is a potential prophylactic or preventive helper candidate to inhibit the attachment and invasion of pathogenic bacteria to host cells and reduce the progression of the infection process. Our study also reveals the involvement of azurin in bacteria-host cell interactions, providing novel and important insights toward the elucidation of its biological function in this field. Thus, this study provides new opportunities to use azurin as an adjunct therapy against critical stages of infection by a wide range of pathogenic bacteria.

Vaginal infections caused by bacteria, Candida and Trichomonas vaginalis, affect millions of women annually worldwide. Symptoms and signs have limited value in differential diagnosis of three causes of vaginitis. Current laboratory methods for differential diagnosis are either expensive or time consuming. Therefore, in this work, development of a method based on gold nanoparticles has been investigated for rapid diagnosis of vaginal infections. Specific antibodies against three main causes of vaginal infections were raised in rabbits. The antibodies were then purified and conjugated to gold nanoparticles and used in an agglutination test for detection of vaginal infections. Finally, sensitivity and specificity of this test for diagnosis of vaginal infections were estimated using culture method as gold standard. Purification of antibodies from sera was confirmed by electrophoresis. Construction of nanoparticles was proved by TEM and FT-IR methods. Conjugation of antibodies to gold nanoparticles was confirmed using XPS method. Sensitivity and specificity of gold nanoparticles for diagnosis of Candida species were 100%, for Gardnerella were 100% and 93%, and for T. vaginalis was 53.3% and 100%, respectively. Gold nanoparticle-based method is a simple, rapid, accurate, and cost-effective test for differential laboratory diagnosis of vaginal infections.

Aim: It has been reported that spiroaminopyrimidine derivatives inhibited the growth and proliferation of various cancer cell lines. In the present study, we evaluated cytotoxic and apoptosis-inducing effects of 2,4-diamino-1,3-diazaspiro[5.5]-9-tert-butyl-2, 4-diene-5-carbonitril (9-tBAP) on NB4 acute promyelocytic leukemia (APL) cells., Materials and Methods: The cells were treated with 10-100 µM of 9-tBAP. Cytotoxic activity of the compound was measured using the MTT assay. Apoptosis was investigated by Hoechst 33258 staining as well as by Annexin V/PI double staining., Results: The compound under study was found to be highly active cell growth inhibitor with IC 50 of 30.0 ± 3.5 µM inducing apoptosis in NB4 cells. Cell cycle analysis by flow cytometry showed a time-dependent increase in sub-G 1 cell population. Real-time polymerase chain reaction analysis revealed that the treatment with the compound down-regulated the BIRC5 expression in a time-dependent manner., Conclusion: 9-tBAP displayed potent anti-leukemic activity in vitro thus warranting further investigation.

Objectives: Group B Streptococcus (GBS) is an important opportunistic bacteria that causes a wide range of infections including neonatal sepsis, meningitis, pneumonia, soft tissue and urinary tract infections (UTI). The aim of this study was to evaluate the antimicrobial susceptibility patterns, surface proteins and capsular types of GBS isolates., Results: 100 of UTI isolates were confirmed as GBS. Antimicrobial susceptibility pattern showed that 95% of GBS isolates were resistant to tetracycline, followed by erythromycin (52%), clindamycin (47%), levofloxacin (9%) and penicillin, cefepime, cefotaxime, and ceftriaxone each with (8%), and vancomycin 1%. Common capsular types were III, Ib, V, II, Ia and IV respectively and the distribution of surface protein genes was as follows: rib (40%), alpha-c (22%), alp2/3 (18%) and epsilon (15%), and alp4 gene was not detected in the isolates. Our findings showed the relationship between capsular types with Alpha-like proteins, as well as reduced sensitivity to antibiotics, so the performance of antibiotic surveillance programs is recommended.

Fractures in caprocks overlying CO 2 storage reservoirs can adversely affect the sealing capacity of the rocks. Interactions between acidified fluid and minerals with different reactivities along a fracture pathway can affect the chemically induced changes in hydrodynamic properties of fractures. To study porosity and permeability evolution of small-scale (millimeter scale) fractures, a three-dimensional pore-scale reactive transport model based on the lattice Boltzmann method has been developed. The model simulates the evolution of two different fractured carbonate-rich caprock samples subjected to a flow of CO 2 -rich brine. The results show that the existence of nonreactive minerals along the flow path can restrict the increase in permeability and the cubic law used to relate porosity and permeability in monomineral fractured systems is therefore not valid in multimineral systems. Moreover, the injection of CO 2 -acidified brine at high rates resulted in a more permeable fractured media in comparison to the case with lower injection rates. The overall rate of calcite dissolution along the fracture decreased over time, confirming similar observations from previous continuum scale models. The presented 3D pore-scale model can be used to provide inputs for continuum scale models, such as improved porosity-permeability relationships for heterogeneous rocks, and also to investigate other reactive transport processes in the context of CO 2 leakage in fractured seals.

Background and Objectives: The aim of this study was to evaluate the antibacterial and antibiofilm activity of recombinant Azurin from Pseudomonas aeruginosa against different bacterial species., Materials and Methods: The azurin gene was cloned in the pET21a vector. The pET21a- azurin construct was transformed into Escherichia coli BL21. The recombinant Azurin was expressed and purified using affinity chromatography and confirmed by Western blotting. The cytotoxicity of rAzurin was assessed on peripheral blood mononuclear cells. Antibacterial and antibiofilm activity of rAzurin with different concentrations were determined by micro-broth dilution and crystal violet methods, respectively. The effect of rAzurin on bacterial species was statistically analyzed by t-test and spearman correlation., Results: The identity of purified protein was confirmed by blotting and distinguished as a 14 kDa band on 15% SDS-PAGE. The IC50 of rAzurin on Peripheral Blood Mononuclear Cell (PBMC) was determined as 377.91±0.5 μg/mL in 24 h. Vibrio cholerae and Campilobacter jejuni displayed the most sensitivity to rAzurin (27.5 and 55 μg/mL, respectively) and the highest resistance (220 μg/mL) was displayed by P. aeruginosa and E. coli. The MIC for other species was 110 μg/mL. The Minimum Biofilm Inhibition Concentration (MBIC) was determined as 220 μg/mL for Salmonella enterica and V. cholerae , 300 μg/mL for Shigella sonnei, Shigella flexneri and P. aeruginosa and 440 μg/mL for the other species. The antimicrobial effect of rAzurin on bacterial species were significant (p value<0.05) and correlation coefficient was negative., Conclusion: The rAzurin appears to be an appropriate choice and a new strategy for prevention of bacterial infection. It inhibits bacterial growth and biofilm formation and candidates as antimicrobial peptides.

Background: The aim of this study was to investigate the effect of the incise drape (ID) on surgical wound bacterial contamination during lumbar spine surgical procedures in treatment group (with ID) and control group (without ID)., Materials and Methods: The present study was conducted on 88 patients who were a candidate for lumbar spine surgery. The patients were randomly assigned to one of the two groups, treatment and control. The ID was only used in the treatment group. The surgical wound sampling for bacterial culture was done in two steps, immediately after surgical incision (IASI) and immediately prior to the surgical wound closure (IPSWC). The samples were then sent to the laboratory., Results: The mean total bacterial count of the surgical wound in the stage IASI was not significantly different between treatment and control groups (0.09 vs. 0.02, P = 0.31). However, this means in the stage IPSWC in treatment group was significantly more than the control group (18.6 vs. 0.41, P = 0.04). The frequency distribution of Staphylococcus aureus (25% vs. 3%, P = 0.02) and Staphylococcus epidermidis (36.4% vs. 9.1%, P = 0.002) was significantly higher in the treatment group compared with control group in the stage IPSWC., Conclusion: The results suggest that the use of ID is unable to reduce surgical wound bacterial contamination in clean lumbar spine surgery. Therefore, based on the results obtained in our study, the application of ID is not recommended as an essential action for the prevention of surgical wound contamination., Competing Interests: There are no conflicts of interest.

Background: Hospital-associated infections, recently renamed Healthcare-associated infections, are among the most common life-threatening complications of hospitalized patients, especially the immunocompromised patients. Regarding the significant role of Enterobacteriaceae in nosocomial infections and also the increasing trends of carbapenem-resistant strains, the present study aimed to evaluate the antibiotic resistance pattern and the occurrence of metallo-beta-lactamases (MBLs) in Enterobacteriaceae strains from Iranian cancer patients., Methods: This hospital-based cross-sectional study was conducted in teaching hospitals of two cities in the central parts of Iran during the 6 months period from December 2015 to May 2016. The Enterobacteriaceae isolates were obtained from different clinical specimens and were identified using standard microbiological methods. Antimicrobial susceptibility pattern for the bacterial isolates was determined using the disk diffusion method. The presence of antibiotic resistance genes was determined by PCR method., Results: The distribution of Enterobacteriaceae isolates were 74 (71.8%) E. coli, 23 (22.3%) Klebsiella spp., 3 (2.9%) Proteus spp., 2 (1.9%) Salmonella spp., and 1 (1%) Shigella spp. The results of antibiotic susceptibility revealed that all of the isolates were multiple-drug resistant (MDR) and 60% of them were (excluded Salmonella and Shigella) carbapenem-resistant. Of all the carbapenem-resistant isolates, 31.7% were MBL-positive. Meanwhile, fosfomycin and minocycline were the most effective antibiotics against MBL-positive bacteria. Moreover, none of the investigated carbapenemases genes were found in MBL-positive isolates., Conclusions: This study highlights the importance of MBLs producing Enterobacteriaceae in causing nosocomial infections in cancer patients. However, carbapenem resistance was not associated with the presence of MBL genes such as IMP, VIM, and SPM. Vatus haeque crent Catilium ausatem nendactui scerem clere forum dicaur hili consceri plin ternul ut audam que factus, que ad ponis. Go vicaet L. Legilici pos.

Background: Changes in the motor control of the spine were found in patients with chronic low back pain (CLBP). Sudden loading of the spine is supposed to be the cause of about 12% of lower back injuries. However, some aspects of this problem, such as alterations in the sensory-motor control of the spine, remain questionable., Objective: To investigate the effects of familiarization with loading, weight and size of loading on neuromuscular responses during sudden upper limb loading in CLBP patients., Methods: In this quasi-experimental study surface electromyography of the erector spinae (ES) and transverses abdominis/internal oblique (TrA/IO) and external oblique (EOA) muscles were recorded in 7 men and 13 women with CLBP and 20 asymptomatic subjects (10 men and 10 women) aged 18-45 years from the general community familiarization. Moreover, investigating control of the posture measurements of the center of pressure (COP) and vertical ground reaction force (GRF) or Fz were recorded using a force plate. Data were analyzed using paired t-test and independent t-test with the significance level of 0.05., Results: Data analyses were performed using SPSS version 18. Some electromyography and force plate variables were significantly different for different conditions in each group and between the asymptomatic and low back pain groups (p⩽ 0.05)., Conclusion: Several motor control changes were observed in the CLBP patients. These patients showed decreased trunk muscle activity as well as too early and too delayed responses compared to asymptomatic subjects.

Competing Interests: There are no conflicts of interest.

A Novel poly [2-hydroxyethyl methacrylate-Lactide-dimethylaminoethyl methacrylate quaternary ammonium alkyl halide] [P(HEMA-LA-MADQUAT)] copolymer was synthesized through combination of ring opening polymerization (ROP) and 'free' radical initiated polymerization methods. This newly developed copolymer was fully characterized by FT-IR, 1 HNMR and 13 CNMR spectroscopy. Micellization of the copolymer was performed by dialysis membrane method and obtained micelles were characterized by FESEM, dynamic light scattering (DLS), zeta potential (ξ), and critical micelle concentration (CMC) measurements. This copolymer was developed with the aim of co-delivering two different anticancer drugs: methotrexate (MTX) and chrysin. In vitro cytotoxicity effect of MTX@Chrysin-loaded P(HEMA-LA-MADQUAT) was also studied through assessing the survival rate of breast cancer cell line (MCF-7) and DAPI staining assays. Cationic micelle (and surface charge of + 7.6) with spherical morphology and an average diameter of 55 nm and CMC of 0.023 gL -1 was successfully obtained. Micelles showed the drug loaded capacity around 87.6 and 86.5% for MTX and Chrysin, respectively. The cytotoxicity assay of a drug-free nanocarrier on MCF-7 cell lines indicated that this developed micelles were suitable nanocarriers for anticancer drugs. Furthermore, the MTX@Chrysin-loaded micelle had more efficient anticancer performance than free dual anticancer drugs (MTX @ chrysin), confirmed by MTT assay and DAPI stainingmethods. Therefore, we envision that this recently developed novel micelle can enhance the efficacy of chemotherapeutic agents, MTX and Chrysin, combination chemotherapy and has the potential to be used as an anticancer drug delivery system for in vivo studies. Therefore, this recently developed novel micelle can enhance the efficacy of chemotherapeutic agents, MTX and Chrysin, combination chemotherapy and has the potential to be used as an anticancer drug delivery system for in vivo studies.

Introduction: The emergence of Metallo-beta-lactamase (MBL)-producing Acinetobacter baumannii has become a global concern in nosocomial infections. The aim of this study is to determine the prevalence of MBL producing genes among clinical isolates of A. baumannii from hospitalized patients., Methods: This study was performed from October 2015 to October 2016 at three teaching hospitals located in Isfahan, Iran. Totally, 100 A-baumannii isolates were collected from clinical specimens and identified as A-baumannii using standard microbiological methods. Antimicrobial susceptibility test was determined by disc diffusion method according to the CLSI. Furthermore, the determination of bla IMP-1 , bla IMP-2 , bla VIM-1 , bla VIM-2 and bla SIM-1 was detected by PCR., Results: Totally, Sixty-eight percent (68%) of isolates of A. baumannii were recovered from tracheal aspirate. According to the antibiotic susceptibility pattern, the highest level of resistance was against ciprofloxacin (99%), while among tested antibiotics amikacin (10%) was found to be the most effective. 21%, 4%, 7% and 6% isolates carried bla IMP-1 , bla IMP-2 , bla VIM-1 and bla VIM-2 genes, respectively. Also, bla SIM-1 was not detected in any of the isolates., Conclusion: The results of this study showed high rate of the MBL producing A-baumannii isolates in our region and displayed that MBLs producing A-baumannii strains are emerging threats to ICUs.

Fracture networks inside the caprock for CO 2 storage reservoirs may serve as leakage pathways. Fluid flow through fractured caprocks and bypass conduits, however, can be restrained or diminished by mineral precipitations. This study investigates precipitation of salt crystals in an artificial fracture network as a function of pressure-temperature conditions and CO 2 phase states. The impact of CO 2 flow rate on salt precipitation was also studied. The primary research objective was to examine whether salt precipitation can block potential CO 2 leakage pathways. In this study, we developed a novel microfluidic high-pressure high-temperature vessel to house geomaterial micromodels. A fracture network was laser-scribed on the organic-rich shales of the Draupne Formation, the primary caprock for the Smeaheia CO 2 storage in Norway. Experimental observations demonstrated that CO 2 phase states influence the magnitude, distribution, and precipitation patterns of salt accumulations. The CO 2 phase states also affect the relationship between injection rate and extent of precipitated salts due to differences in solubility of water in CO 2 and density of different CO 2 phases. Injection of gaseous CO 2 resulted in higher salt precipitation compared to liquid and supercritical CO 2 . It is shown that micrometer-sized halite crystals have the potential to partially or entirely clog fracture apertures.

Background: Nosocomial infections are one of the most leading causes of morbidity and mortality in patients with cancer. The emergence of multiple-drug-resistant (MDR) strains of Gram-negative bacteria causing nosocomial infection has become a serious concern in cancer patients. Therefore, the present study aimed to determine the spectrum and antibiotic resistance pattern of Gram-negative bacteria related nosocomial infections among Iranian cancer patients., Materials and Methods: This descriptive cross-sectional study was conducted during the 6 months from December 2015 to May 2016 in two tertiary care centers located in Isfahan and Arak Province. Gram-negative bacteria obtained from different clinical specimens from hospitalized patients with cancer and were identified using standard microbiological methods. Antibiotic susceptibility pattern was determined by the disk diffusion method according to the Clinical and Laboratory Standards Institute (CLSI) recommendation., Results: Of totally 259 culture positive cases, Escherichia coli showed the highest isolation rate (60.6%) followed by Klebsiella pneumoniae (26.6%) and Proteus spp (11.2%). The rate of MDR isolates were 91.5% (237/259). Overall, the most frequent source of bacterial isolation was urinary tract infection (65.6%) followed by skin and soft-tissue infection (23.6%). The antibiotic susceptibility results showed meropenem (MEN) and ceftazidime as the most effective antibiotics for E. coli , K. pneumoniae , and Proteus spp. isolates. Moreover, MEN was the most effective antibiotic against MDR isolates., Conclusion: The study findings showed a significant distribution of MDR Gram-negative bacteria which may increase the burden of healthcare-associated infections in cancer patients. Although, carbapenem can be considered as effective agents toward MDR strains for empirical antibiotic therapy in our region., Competing Interests: There are no conflicts of interest.

Background: Pseudomonas aeruginosa is a biofilm-forming bacterium which can result in serious health problems, particularly in burn patients. Biofilm has been assumed to protect the bacteria from environmental fluctuations such as antimicrobial agent. Mucoid strains generate extensive levels of the alginate exopolysaccharide, which is an important factor of its biofilm., Materials and Methods: Totally, 100 isolates of P. aeruginosa has been gathered from wound infections of burn patients. Polymerase chain reaction of exoA gene has been carried out to confirm the bacteriologic identification of isolates. The biofilm-forming capacity has been specified by capsule staining and microtiter plate test as qualitative and quantitative determination, respectively. Antimicrobial susceptibility of the isolates has been specified by disk diffusion method., Results: All the isolates carried the exoA gene. The antibiotic resistance was imipenem (90%); levofloxacin (93%); aztreonam (87%); piperacillin-tazobactam (85%); tobramycin (92%); polymyxin b (PB) (2%); and ceftazidime (CAZ) (32%). Totally, multidrug-resistant (MDR) and extended drug-resistant (XDR) isolates were 19% and 75%, respectively. Fortunately, pan drug-resistant (PDR) strain has not been observed. The assessment of biofilm formation has shown that 7% of the isolates were nonbiofilm (N), weak (W) 67%, moderate (M) 22%, and strong (S) 4%., Conclusions: As a result, the findings of this survey indicated that PB and CAZ were the most effective antibiotics against P. aeruginosa , which of course indicate a serious problem about the emergence of the PDR strains. There was no relationship between the patterns of biofilm production and antibiotic susceptibility, but high frequency of MDR/XDR and biofilm producer strains has been detected., Competing Interests: There are no conflicts of interest.

Acinetobacter baumannii is one of the most important bacterial species with the ability to produce OXA-type carbapenemases. We aimed to evaluate the prevalence of OXA-type carbapenemases among clinical isolates of A. baumannii in three major hospitals of Isfahan. In this cross-sectional descriptive study, 153 non-repeated strains of A. baumannii were isolated from various clinical samples of hospitalized patients in Al-Zahra, Imam Mousa Kazem, and Shariati hospitals from October 2015 to October 2016. Antimicrobial susceptibility testing for imipenem, meropenem, ertapenem, cefepime, ceftazidime, ceftriaxone, piperacillin-tazobactam, gentamicin, amikacin, ciprofloxacin, and tetracycline was performed using the disk diffusion method. In order to identify bla-oxa genes, a multiplex polymerase chain reaction was used. The resistance rates in A. baumannii isolates to beta-lactam antibiotics including imipenem, ertapenem, meropenem, cefepime, ceftazidime, ceftriaxone, and piperacillin/tazobactam were 100%, 100%, 99.3%, 97.4%, 96.7%, 97.4%, and 98.6%, respectively. PCR assay showed the presence of bla-oxa genes in all isolates. The bla-oxa-51 gene was recognized in all (100%) isolates, 90.8% and 62.1% of isolates possessed the bla-oxa-23 and bla-oxa-24 genes, respectively, while the bla-oxa-58 gene was not detected in any of the isolates. Also, 56.2% of isolates had both the bla-oxa-23 and bla-oxa-24 genes simultaneously. We found that the prevalence of OXA-type carbapenemases among carbapenem-resistant A. baumannii isolates is high in Isfahan, with OXA-23 being the major carbapenemase mechanism responsible for the resistance phenotype.

Overuse and misuse of Carbapenems among Klebsiella pneumoniae isolates have caused Carbapenem-Resistant Klebsiella Pneumonia (CRKP) during recent years. Colistin is one of the last available options, and there are increasing concerns about the dosage and resistance to this agent in long-term monotherapies. This study was designed to identification of carbapenemase producing isolates of K. pneumoniae via phenotypic and genotypic methods as well as evaluation of colistin-meropenem combination therapy potential. This study was carried out in Isfahan, of Iran on 100 samples from Alzahra and Khorshid hospitals in 2017. The Modified Hodge Test (MHT) was used to investigate the carbapenemase presence. The minimum inhibitory concentration (MIC) and the Fractional Inhibitory Concentration (FIC) were determined using broth macrodilution and checkerboard assays (respectively) for both meropenem and colistin. The bla-KPC gene was studied by polymerase chain reaction (PCR).The highest and the lowest rate of resistance were observed for piperacillin (84%) and ertapenem (50%) respectively. 68 isolates by MHT were CRKP, but None of them were positive for bla-KPC gene. 21 isolates from CRKP cases were high resistant to used antimicrobial agents in the study that both MIC and FIC results showed significant synergy for this antibiotics in checkerboard test (p-value &lt; 0.05). 21 resistant isolates from CRKP cases showed statistically significant synergy potential for meropenem and colistin. The meropenem-colistin combination therapy can be applied as a suitable antibiotic synergy but it requires further investigation in clinical assay. Regarding to our findings, Probably other mechanisms of resistance to Carbapenems ,except bla-kpc genes are involved.

Antibiotic resistance is an important problem in antibiotic treatment of infections, particularly in hospitals. Tomatidine is a plant secondary metabolite with antimicrobial and antifungal effects. This study examined the possible synergistic effect tomatidine with several antibiotics against standard and clinical strains of Staphylococcus aureus , Enterococcus faecalis , Pseudomonas aeruginosa and Escherichia coli . After determining the minimum inhibitory concentrations (MICs) of antibiotics and tomatidine against the bacterial isolates using broth microdilution method, the synergistic effect between tomatidine and antibiotics was evaluated by checkerboard method and calculation of FIC indices. Tomatidine alone did not show any antimicrobial effect. However, it had synergistic effect with gentamicin and cefepime against standard and clinical isolates of S. aureus and P. aeruginosa , respectively. It also had synergistic effect with ampicillin and ciprofloxacin only against standard strains of E. faecalis and P. aeruginosa , respectively. In conclusion, tomatidine could be considered as a potential antibiotic potentiator for gentamicin, cefepime and ciprofloxacin, and ampicillin against Staphylococcus aureus , Pseudomonas aeruginosa , and Enterococcus faecalis infections, respectively. However, the toxicological and pharmacological properties of tomatidine for use as a therapeutic agent remain to be determined.

Background: Nontuberculous mycobacteria (NTM) are a group of opportunistic pathogens and these are widely dispersed in water and soil resources. Identification of mycobacteria isolates by conventional methods including biochemical tests, growth rates, colony pigmentation, and presence of acid-fast bacilli is widely used, but these methods are time-consuming, labor-intensive, and may sometimes remain inconclusive., Materials and Methods: The DNA was extracted from NTM cultures using CTAB, Chelex, Chelex + Nonidet P-40, FTA ® Elute card, and boiling The quantity and quality of the DNA extracted via these methods were determined using UV-photometer at 260 and 280 nm, and polymerase chain reaction (PCR) amplification of the heat-shock protein 65 gene with serially diluted DNA samples., Results: The CTAB method showed more positive results at 1:10-1:100,000 at which the DNA amount was substantial. With the Chelex method of DNA extraction, PCR amplification was detected at 1:10 and 1:1000 dilutions., Conclusions: According to the electrophoresis results, the CTAB and Chelex DNA extraction methods were more successful in comparison with the others as regard producing suitable concentrations of DNA with the minimum use of PCR inhibitor., Competing Interests: There are no conflicts of interest.

Background: Coliform bacteria are used as indicator organisms for detecting fecal pollution in water. Traditional methods including microbial culture tests in lactose-containing media and enzyme-based tests for the detection of β-galactosidase; however, these methods are time-consuming and less specific. The aim of this study was to evaluate polymerase chain reaction (PCR) for detecting coliform., Materials and Methods: Totally, 100 of water samples from Isfahan drinking water source were collected. Coliform bacteria and Escherichia coli were detected in drinking water using LacZ and LamB genes in PCR method performed in comparison with biochemical tests for all samples., Results: Using phenotyping, 80 coliform isolates were found. The results of the biochemical tests illustrated 78.7% coliform bacteria and 21.2% E. coli . PCR results for LacZ and LamB genes were 67.5% and 17.5%, respectively., Conclusion: The PCR method was shown to be an effective, sensitive, and rapid method for detecting coliform and E. coli in drinking water from the Isfahan drinking water sources., Competing Interests: There are no conflicts of interest.

Increasing multiple drug resistant (MDR) strains of Acinetobacter baumannii has aggravated curiosity in development of alternative therapy. Bacteriophages are often considered as alternative agents for controlling A. baumannii infections. In the present study two lytic phages for MDR A. baumannii were isolated and their efficacy and host ranges were evaluated. The phages were isolated from hospital wastewater. Electron microscopy revealed that IsfAB78 might belong to Myoviridae and IsfAB39 to Podoviridae . Initial characterization of phages showed that they have narrow host range and failed to infect relative and non-relative bacteria. Both phages decreased the A. baumannii turbidity significantly, indicating that these isolated phages may be considered as candidates for phage therapy.

Background: Pseudomonas aeruginosa is an opportunistic and Gram-negative pathogen that is used as the most important factor in burn wound infections, and due to the rapid acquisition of multidrug resistance (MDR), it causes high mortality rates in these sectors. Thus, diagnosis and assessment of antibiotic resistance patterns are very important in these patients. The aim of this study was to evaluate antibiotic resistance pattern and determining P. aeruginosa MDR., Materials and Methods: In this study, phenotypic, biochemical, and polymerase chain reaction tests were used to identify P. aeruginosa from 120 wound burn samples that 96 samples were detected to P. aeruginosa species. In the next step, according to the Clinical and Laboratory Standard Institute standard guidelines, antibiogram test was performed by disk diffusion method for amikacin, ciprofloxacin, norfloxacin, gentamicin, cefepime, aztreonam, meropenem, colistin, ceftazidime, and piperacillin-tazobactam antibiotics. Antibiotic data were analyzed by WHONET software; finally, the rate of antibiotic resistance and MDR strains was determined., Results: The highest antibiotic resistance belonged to amikacin (94.8%) and norfloxacin (90.6%); in contrast, colistin (8.3%) had the lowest and the MDR strains were MDR (95.8%) and extensively drug resistance (XDR) (87.5%)., Conclusion: In this study, there was MDR with an alarming rate including MDR (95.8%), XDR (87.5%), and pan-drug resistance (0%). As a result, given antibiotics to patients should be controlled by the antibiogram results to avoid increasing MDR strains., Competing Interests: There are no conflicts of interest.

In this study, we aimed to identify the genetic lineages of Mycobacterium tuberculosis isolates in Isfahan via the mycobacterial interspersed repetitive-unit-variable number tandem repeat typing method based on 15 loci. Forty-nine M. tuberculosis isolates were collected between 2013 and 2015 from Tuberculosis patients in Mollahadi Sabzevari Tuberculosis Center in Isfahan. All isolates were typed by 15-locus MIRU-VNTR typing. The highest percentage of isolates, 44.89 % (22/49), belonged to the Euro-American lineage, while the frequencies of the East-African-Indian, East-Asian, and Indo-Oceanic lineages were 28.57 % (14/49), 24.4 % (12/49), and 2.04 % (1/49), respectively. Among the 22 isolates of the Euro-American lineage, those belonging to the NEW-1 sub-lineage were most prevalent (24.4 %). Approximately, the same proportion of isolates belonging to the Delhi/CAS, Beijing, and NEW-1 sub-lineages were identified in Iranian and Afghan immigrant patients. The Delhi/CAS and Beijing sub-lineage isolates were prevalent among patients who had been previously treated for TB. Results showed that all of the 49 MIRU-VNTR patterns were unique and the clustering rate of the 15-locus MIRU-VNTR was 0.0 (minimum recent transmission). The results of this study show that the lineages of M. tuberculosis isolates in Isfahan are similar to those reported in the Eastern Mediterranean region (indicative of the epidemiological relationship between the countries in the region). The low clustering rate in our results reveals that transmission of tuberculosis in Isfahan is, in most cases, a reactivation of previous tuberculosis infection and the role of recently transmitted disease is minor.

Background: During the past several years, nontuberculous mycobacteria (NTM) have been reported as some of the most important agents of infection in immunocompromised patients., Objectives: The aim of this study was to evaluate the ciprofloxacin susceptibility of clinical and environmental NTM species isolated from Isfahan province, Iran, using the agar dilution method, and to perform an analysis of gyrA gene-related ciprofloxacin resistance., Materials and Methods: A total of 41 clinical and environmental isolates of NTM were identified by conventional and multiplex PCR techniques. The isolates were separated out of water, blood, abscess, and bronchial samples. The susceptibility of the isolates to 1 µg/mL, 2 µg/mL and 4 µg/mL of ciprofloxacin concentrations was determined by the agar dilution method according to CLSI guidelines. A 120-bp area of the gyrA gene was amplified, and PCR-SSCP templates were defined using polyacrylamide gel electrophoresis. The 120-bp of gyrA amplicons with different PCR-SSCP patterns were sequenced., Results: The frequency of the identified isolates was as follows: Mycobacterium fortuitum, 27 cases; M. gordonae, 10 cases; M. smegmatis, one case; M. conceptionense, one case; and M. abscessus, two cases. All isolates except for M. abscessus were sensitive to all three concentrations of ciprofloxacin. The PCR-SSCP pattern of the gyrA gene of resistant M. abscessus isolates showed four different bands. The gyrA sequencing of resistant M. abscessus isolates showed 12 alterations in nucleotides compared to the M. abscessus ATCC 19977 resistant strain; however, the amino acid sequences were similar., Conclusions: This study demonstrated the specificity and sensitivity of the PCR-SSCP method for finding mutations in the gyrA gene. Due to the sensitivity of most isolates to ciprofloxacin, this antibiotic should be considered an appropriate drug for the treatment of related diseases.

Objective: To study on antibiotic susceptibility and identify coagulase-negative Staphylococcus (CoNS) species based on tuf gene sequencing from keratitis followed by using soft contact lenses in Isfahan, Iran, 2013., Methods: This study examined 77 keratitis cases. The samples were cultured and the isolation of CoNS was done by phenotypic tests, and in vitro sensitivity testing was done by Kirby-Bauer disk diffusion susceptibility method., Results: Thirty-eight of isolates were conveniently identified as CoNS. In this study, 27 (71.1%), 21 (55.3%), and 16 (42.1%) were resistant to penicillin, erythromycin, and tetracycline, respectively. One hundred percent of isolates were sensitive to gentamicin, and 36 (94.7%) and 33 (86.8%) of isolates were sensitive to chloramphenicol and ciprofloxacin, respectively. Also, resistances to cefoxitin were 7 (18.4%). Analysis of tuf gene proved to be discriminative and sensitive in which all the isolates were identified with 99.0% similarity to reference strains, and Staphylococcus epidermidis had the highest prevalence among other species., Conclusions: Results of this study showed that CoNS are the most common agents causing contact lens-associated microbial keratitis, and the tuf gene sequencing analysis is a reliable method for distinguishing CoNS species. Also gentamycin, chloramphenicol, and ciprofloxacin are more effective than the other antibacterial agents against these types of bacteria.

Background: Helicobacter pylori infection is highly prevalent in the developing countries. It causes gastritis, peptic ulcer disease, and gastrocarcinoma. Treatment with drugs and antibiotics is problematic due to the following reasons: cost, resistance to antibiotics, prolonged treatment and using multiple drugs. Catalase is highly conserved among the Helicobacter species and is important to the survival of the organism. It is expressed in high amounts and is exposed to the surface of this bacterium; therefore it represents a suitable candidate vaccine antigen., Objectives: A suitable approach in H. pylori vaccinology is the administration of epitope based vaccines. Therefore the responses of T-cells (IFN-γ and IL-4 production) against the catalase of H. pylori were determined. Then the quality of the immune responses against intact catalase and three epitopes of catalase were compared., Materials and Methods: In this study, a composition of three epitopes of the H. pylori catalase was selected based on Propred software. The effect of catalase epitopes on T-cells were assayed and immune responses identified., Results: The results of IFN-γ, IL-4 production against antigens, epitopes, and recombinant catalase by T-cells were compared for better understanding of epitope efficiency., Conclusions: The current research demonstrated that epitope sequence stimulates cellular immune responses effectively. In addition, increased safety and potency as well as a reduction in time and cost were advantages of this method. Authors are going to use this sequence as a suitable vaccine candidate for further research on animal models and humans in future.

Background: Pseudomonas aeruginosa might be converted to coccoid bacteria under antibiotic stress. Bacterial conversion would increase resistance to antibiotics due to changes in cell wall crosslink or decreased metabolic activity. Morphology of P. aeruginosa under stress conditions (presence of antibiotics) can be changed to elongated bacilli, U shape and finally coccoid bacteria. Results of several researches showed that coccoid bacteria are one of the most important aspects of drug resistance. It would be the major reason for treatment failure., Objectives: The aim of this study was to determine in vitro morphological and bactericidal effects of amikacin, meropenem and imipenem on P. aeruginosa isolated from clinical specimens., Materials and Methods: Eight P. aeruginosa isolates obtained from clinical samples of burned patients and standard strain ATCC 27853 were used in this study. Isolates were identified by biochemical tests and confirmed by PCR method using ITS specific primer. Minimum inhibitory concentrations (MICs) of three antibiotics were determined by E-test method. Bacteria were exposed to antibiotics at different concentrations. Bacterial morphology in different days was examined by specific microscope and viability of isolates was examined by flow cytometry., Results: All used antibiotics at sub MIC concentration had capability to induce coccoid bacteria. The highest rate of induced coccoid bacteria was 98.2% after 8 days, with contribution of imipenem and meropenem at 2 μg/mL concentration. Amikacin at 4 μg/mL concentration induced lower rate of coccoid bacteria (55.05%). Amikacin had a strong bactericidal effect on coccoid bacteria at 8 μg/mL concentration. Imipenem and meropenem showed very weak bactericidal effect on coccoid bacteria., Conclusions: Induction of coccoid form of P. aeruginosa may be one of the important reasons for antibiotic treatment failure; therefore, prescribed dose of antibiotics should be carefully managed to prevent increasing antibiotic resistance and coccoid bacteria induction.

Background: Pseudomonas aeruginosa is a well-known opportunistic pathogen, which affects hospitalized patients in different wards due to its natural resistance to drugs., Objectives: The purpose of the current study was to determine the antibiotic susceptibility profiles and genetic relatedness in P. aeruginosa isolated from patients admitted to a referral hospital in Isfahan, Iran., Materials and Methods: Out of 150 analyzed samples, 54 P. aeruginosa isolates were recovered and were subjected to antibiotic resistance patterns and genetic diversity determination by Kirby-Bauer's disk diffusion method and RAPD-PCR, respectively., Results: The highest percentage of resistance was observed against ceftazidime and imipenem with 30 (55.6%) isolates; meanwhile all isolates were sensitive to polymyxin B. Twenty-eight (51.8%) isolates revealed resistance to all applied antibiotics. RAPD-PCR (Random Amplified Polymorphic DNA- Polymerase Chain Reaction) results showed 54 unique genotypes, which were divided into 39 clusters., Conclusions: Although different source of P. aeruginosa may involve in patient colonization, genetically related strains were isolated from different wards and or the same ward of the hospital. Our results pointed to the restriction of currently used antibiotics in studied hospital. We hope that our results cast light on the control and transmission of the infection in the investigated hospital.