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Elnaz Abbasi,1 Mahdieh Mondanizadeh,2 Alex van Belkum,3 Ehsanollah Ghaznavi-Rad4,5 1Department of Microbiology & Immunology, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran; 2Department of Biotechnology and Molecular Medicine, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran; 3Data Analytics Department, BioMérieux, La Balme les Grottes, France; 4Molecular and Medicine Research Center, Faculty of Medicine Arak University of Medical Sciences, Arak, Iran; 5Department of Medical Laboratory Sciences, Arak School of Paramedicine, Arak University of Medical Sciences, Arak, IranCorrespondence: Ehsanollah Ghaznavi-Rad Tel/Fax +98-8634173526Email ghaznaviehs@yahoo.comBackground: Diarrheagenic Escherichia coli (DEC) is a significant cause of gastroenteritis and a major public health problem. This study investigates the prevalence and the antibiotic resistance patterns of DEC that were isolated from infectious diarrhea samples of pediatric patients from central Iran.Patients and Methods: Pediatric diarrhea samples were collected from 230 pediatric patients visiting the hospital. E. coli pathotypes were diagnosed by using conventional culture methods and PCR. Antibiotic resistance profiles, the frequency of multi-drug resistance (MDR), and the phenotypic and genotypic characteristics of extended spectrum-β-lactamase (ESBL), AmpC and integron-associated genes were analyzed.Results: Of the 230 samples of infectious diarrhea, 91 (39.5%) produced E. coli isolates. Of these, 32 cases (35.1%) were identified as DEC by culture and PCR. The frequency of the E. coli pathotypes obtained was as follows: EAEC 11/32 (34.3%), EPEC 9/32 (28.1%), ETEC 6/32 (18.7%), EIEC 3/32 (9.3%), and EHEC 3/32 (9.3%). The antibiotic resistance rates were greater for nalidixic acid (30/32; 93.7%), ampicillin (29/32; 90.6%), and tetracycline (25/32; 78.1%) than for any of the other tested antibiotics. High levels of MDR (25/32; 78.1%) and the presence of ESBL (18/32; 56.2%) and AmpC (9/32; 28.1%) were observed in the DEC isolates. The isolates showed a higher frequency of the ESBL genes [blaTEM (18/18; 100%), blaCTX-M15 (17/18; 94.4%)], and AmpC [blaCIT (4/9; 44.4%) and blaDHA (4/9; 44.4%)] than of the other ESBL and AmpC genes.Conclusion: Compared to the previous study, DEC appeared to be the second-most abundant agent of diarrhea in pediatric patients after Campylobacter jejuni, with frequent MDR and ESBL presence.Keywords: diarrheagenic Escherichia coli pathotypes, pediatrics, diarrhea, antibiotic resistance, MDR, Iran

Elnaz Abbasi,1,2 Hamid Abtahi,3 Alex van Belkum,4 Ehsanollah Ghaznavi-Rad1,31Department of Microbiology & Immunology, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran; 2Department of Microbiology & Immunology, Khomein University of Medical Sciences, Khomein, Iran; 3Molecular and Medicine Research Center, Faculty of Medicine, Arak University of Medical Sciences, Arak, Iran; 4Data Analytics Department, BioMérieux, La Balme les Grottes, FranceBackground: Shigella spp. are primary pathogens of diarrhea in children worldwide. Emergence of resistance to fluoroquinolones and third-generation cephalosporins is crucial in the management of pediatric shigellosis. We determined the prevalence and the antibiotic resistance patterns of Shigella species isolated from pediatric patients in central Iran.Materials and methods: Pediatric diarrhea samples (n=230) were cultured on MacConkey and XLD agar media and in GN broth. Genus-specific PCR for ipaH was also used for detection directly from fecal specimens. Antibiotic resistance and the frequency of ESBL and AmpC genes were determined.Results: Out of the 230 samples, 19 (8.2%) cases of Shigella spp. were identified using culture. Twenty-six samples were positive by PCR (11.3%), S. flexneri (4/19; 21%) and S. sonnei (15/19; 78.9%) being the most detected. The highest antibiotic resistance rates were found for cotrimoxazole (19/19; 100%), ampicillin (16/19; 84.2%), cefixime (13/19; 68.4%) and ceftriaxone (12/19; 63.1%). Ten cases showed phenotypic ESBL presence and all these strains were positive for blaTEM, blaCTX-M-1, and blaCTX-M-15. Three strains were AmpC positive, all of which harbored blaCMY-2 and two contained blaCIT. Of the 19 Shigella isolates 5 (26.3%), 2 (10.5%), and 1 (5.2%) were phenotypically resistant to nalidixic acid, ciprofloxacin, and norfloxacin, respectively. Class 1 integron was found in 18 (94.7%) isolates whereas class 2 integron was found in 19 (100%) strains.Conclusion: We found a considerable presence of Shigella species with elevated antibiotic resistance levels. In particular, the resistance to third-generation cephalosporins (ESBL) and ciprofloxacin must be taken seriously.Keywords: Shigella, dysentery, antibiotic resistance, MDR, integrons, Iran

OBJECTIVE: From the beginning of the novel coronavirus infection (COVID-19) pandemic in the world, much efforts have been accomplished to explain a precise clinical feature for the disease and to find the best therapeutic approach for the patients. Although coagulation abnormalities have found in novel coronavirus infection (COVID-19) patients, still little is known about the association between the disease and changes in coagulation parameters. Our purpose is to evaluate the differences between the coagulation parameters between COVID-19 patients and healthy counterparts. PATIENTS AND METHODS: 63 patients with confirmed COVID-19 infection were admitted to the present study. We evaluated coagulation value in these patients and in 40 healthy individuals. RESULTS: We found that although there was no significant difference between PT and PTT values in patients and healthy counterparts, the fibrinogen values in patients were higher than the control group (p < 0.05). Moreover, the values of fibrin/fibrinogen degradation products (FDP) and D-dimer in all COVID-19 cases were considerably higher than those in control people (p < 0.05). Of note, FDP and D-dimer in patients with regular COVID-19 infection were lower than patients with severe forms. CONCLUSIONS: It seems that the conduction of routine blood coagulation test could be a beneficial supplementary approach for early diagnosis of COVID-19. In addition, our study shed more light on the therapeutic value of anti- coagulant-based treatment for COVID-19 patients, especially for those with severe type of the disease. [ABSTRACT FROM AUTHOR]

Background: Multi-drug resistance due to Acinetobacter baumannii strains has become a significant challenge. Efflux pump plays a vital role in the development of resistance in this bacteria. The aim of this study was to evaluate the frequency of the AdeABC efflux pump genes and its role in resistance to imipenem in clinical isolates of A.baumannii. Materials and methods: A total of 56 isolates of A.baumannii were collected from different clinical specimens of Valiasr hospital in the Arak -Iran and all isolates were identified by standard biochemical tests. The Antimicrobial susceptibility patterns were determined by disk diffusion method and minimum inhibitory concentrations of imipenemin via E-test strips with and without CCCP efflux pump inhibitor were determined according to CLSI guidelines. The PCR test was used to detect the AdeABC efflux pump genes in isolates. Results: All A.bumannii isolates were resistant to cefotaxim, ceftazidim, cefepim, cefoxitin, azteronam, piperacillin-tazobactam and ciprofloxacin, as well as all isolates were resistant to imipenem according to the results of the E-test method. Imipenem MIC with efflux pump inhibitor not reduced in all isolates and showed no differences in imipenem activity. The adeA, adeB and adeC genes were found in 100%, 100% and 96.5% of isolates, respectively. Conclusion: AdeABC efflux system contributes to resistance to other antibiotics and resistance to imipenem has not been involved with this efflux system in A.baumanni isolates in current study and other mechanism such as carbapenemase enzymes play vital role to imipenem resistance in A. baumanni isolates. [ABSTRACT FROM AUTHOR]

The role of opioid receptor and voltage dependent calcium channels on the kindling induced by the convulsant pentylenetetrazole (PTZ) were investigated in the rats. Experiment involved 24 rats which were divided into four groups. Kindling was established with PTZ in subconvulsive dose (37.5 mg/kg i.p.) every 48 h and effects were observed within 20 min using five-point scoring system. All animals were kindled to three consecutive-stage five seizures and their stability was tested. Saline, verapamil (calcium channel blocker), naloxone (opioid antagonist) or both of them were then administrated 20 min before PTZ application. Convulsant parameters were significantly (P<0.05) reduced by verapamil. Naloxone had no significant effect on the seizure expression of fully kindled animals, whereas simultaneous application of naloxone and verapamil had profound inhibitory effect on all seizure parameters. The results of the present study suggest that naloxane increased the inhibitory effect of verapamil on the seizure induced by PTZ kindling [ABSTRACT FROM AUTHOR]

Background: The aim of present study was evaluation and comparison of widely used molecular techniques including pulsed-field gel electrophoresis (PFGE), multi locus sequence typing (MLST), Staphylococcus aureus protein A typing (spa), Staphylococcal Cassette Chromosome typing (SCCmec), and SCCmec typing with recently introduced method using direct repeat unit (dru) sequencing in the hyper-variable region of the SCCmec cassette for clustering and discrimination of MRSA isolates. Material and Methods: Out of 389 already molecularly established methicillin-resistant Staphylococcus aureus (MRSA) isolates 61 representative samples were subjected to PFGE and dru typing methods according to guideline. Results: The results showed that isolates were typed by PFGE typing, MLST typing, spa typing, SCCmec was typing and dru typing to 7,6,6,12,19 different types respectively. Evaluation of the discriminatory power for each method identified dru typing and PFGE as the most discriminatory methods. Conclusion: Although the discriminatory ability of dru typing, especially with closely related MRSA ST239 strains underscore its utility as a feasible and cheap method in epidemiological investigation of MRSA, we suggest the use of the conjugation of dru typing and PFGE typing for epidemiological surveillance studies, since this combination provides more discriminatory and typeability which is useful for the control of nosocomial infection. [ABSTRACT FROM AUTHOR]

Background: Increasing prevalence of Methicillin-resistant Staphylococcus aureus (MRSA) in different communities is clearly visible .Because of this, treatment of patients with infections caused by those bacteria has fallen into critical troubles .Current study, therefore, is aimed to compare phenotypic (disk diffusion) and genotypic (PCR) methods for fast diagnosis of methicillin-resistant strains, isolated from patients of Arak Central Hospital Materials and Methods: In a cross sectional study within one year of period, a total of 100 samples were taken and tested from the patients of Arak hospital (located in the central part of Iran). Isolates' sensitivity to Cefoxitin Disk and Oxacillin was confirmed through disk diffusion. Using PCR, the isolates were tested for the presence of mecA gene. Results were compared from the points of sensitivity and specificity by application of chi square test in SPSS software.. Results: Seventy five (75%) out of the total 100 samples (through oxacillin disk diffusion method, already isolated from patients were resistant to oxacillin. Meanwhile, 83(83%) of cefoxitin disk diffusion method samples' were resistant to cefoxitin. Three resistant samples to cefoxitin were negative for mecA gene and 80 (80%) samples were positive for mecA gene using PCR. Sensitivity were respectively 93.75%, 100%, and specificity were 100% and 100%, 85%, 100 Conclusion: Findings indicate that oxacillin disk diffusion method is a simple phenotypic method, however, it has lower sensitivity compared to cefoxitin disk diffusion and polymerase chain reaction (PCR) methods. Therefore, it is not recommended for detection of Methicillin-resistant Staphylococcus aureus (MRSA). Existence of strains resistant to cefoxitin without mecA gene, shows the outset of another type of resistance or mutation in Methicillin-resistant Staphylococcus aureus (MRSA) . [ABSTRACT FROM AUTHOR]

Background: Increasing frequency of Staphylococcus aureus infections and changes in antimicrobial resistance pattern have led to renewed interest in the use of lincosamide- streptogramin B (MLSB) antibiotics for treatment of infections. Since no study has focused on the molecular epidemiology of community -acquired staphylococcus aureus isolates in Iran, the aim of this study was to determine the molecular typing and prevalence of the macrolides-lincosamides-streptogramins B (MLSB) resistance in community associated s.aureus isolated from healthy students at Arak university of Medical sciences. Material and Methods: 568 healthy students from Arak university of Medical sciences were subjected to this study. All samples were subjected to S. aureus-specific isolation procedures. D test was performed to determine various phenotypes as well as spa typing done for molecular typing of these strains. Results: Of 568 the 84 community acquired Staphylococcus aureus, six (7%) were Methiclicin resistant Staphylococcus aureus (CA-MRSA) and 78(93%) were Methicillin sensitive Staphylococcus aureus (CAMSSA) of the 84 s.aureus strains, eight (9.5%) showed constitutive resistance with spa type t660, t701, t304, t5598, t012, t3204, t084 and t1944. Two strains (2.5%) demonstrated inducible resistance with spa type t9024, t077, two strains (2.5%) were D test negative with spa type t084 and t1149. 72(85.5%) strains. Illustrated susceptible Phenotype. Among CA-MRSA isolates, two strains had constitutive resistance and four remaining CA-MRSA had susceptible phenotype Conclusion: The result of this study indicates that in community associated s.aureus strains, constitutive MLSB resistance rate is higher than the rate of inducible resistance. Presence of inducible resistance to clindamycin in CA-MRSA strains, warrants that D test should be performed to detect this type of resistance. All isolates with inducible and constitutive resistance and D zone negative strains had different molecular typings. [ABSTRACT FROM AUTHOR]

The prevalence of methicillin resistant Staphylococcus aureus (MRSA) and methicillin sensitive Staphylococcus aureus (MSSA) carriage among poultry and poultry farmers in Malaysia is largely unknown. In the current investigation, chickens and chicken farmers from 30 chicken farms were screened for MRSA and S. aureus carriage. The genetic characteristics of the isolates were determined through multi locus sequence typing (MLST), Staphylococcus protein A (spa) typing and virulent gene profiling. The outcome of the study showed lack of MRSA and extremely low S. aureus prevalence (n=7 of 503, 1.4%) among chicken flocks and the poultry farmers in Malaysia. Staphylococcus aureus isolates belonged to 4 sequence types (ST): ST97 (spa type t359), ST1179 (t359), ST 692 (t2247) and ST188 (t189). It can be concluded that MRSA/MSSA prevalence is very low among chicken and chicken farmers, human and chicken cross transmission of S. aureus does not seem to be a threat in Malaysia. [ABSTRACT FROM AUTHOR]

Background: Escherichia coli (E. coli) is an indicator of potential human fecal contamination. Polymerase Chain Reaction (PCR) as an ideal detection method for detecting E. coli has some advantages like rapidity, high sensitivity and accuracy, easy performance, ability to run in high numbers and inexpensiveness. On the other hand, the disadvantage of PCR is possibility of its false positive results. In this study, reverse transcriptase polymerase chain reaction (RT-PCR) method was used to overcome the problem, and the results were compared to most probable number (MPN). Materials and Methods: 16srRNA forward (SF) and 16srRNA reverse (SR) primers were designed using E. coli 16srRNA sequence. After preparing different diluted samples of E. coli in distilled water, the bacteria were separated by FHLP and HAWP filters and its 16srRNA was propagated using mentioned primers. To confirm the sensitivity of the RTPCR method compared to MPN one, samples obtained from 15 water sources in Arak city were examined. Results: The number of bacteria in dilutions were confirmed with culture. RT-PCR data showed that FHLP compared to HAWP filters have a higher capability in separating of bacteria in different dilutions. Also there was a higher sensitivity of RT-PCR compared to RT- PCR and MPN. Conclusion: RT-PCR can detect the bacteria in lower dilutions of bacterial suspension. Hydrophobic filters (e.g. FHLP) compared to hydrophilic filters (e.g. HAWP) have higher capability in separating bacteria. To detect all coliform bacteria RT-PCR amplifications achieved by cells concentrated with hydrophobic filters are recommended. [ABSTRACT FROM AUTHOR]

Methicillin-resistant Staphylococcus aureus (MRSA) from Malaysia were shown to possess staphylococcal cassette chromosome mec (SCC mec)-III and IIIA. Spa sequencing and multi-locus sequence typing (MLST) documented t037 and ST 239 (CC8) for 83.3% of the isolates. This confirms observations in several other Far Eastern countries and corroborates the epidemicity of this clone. [ABSTRACT FROM AUTHOR]

Chronic wounds have become a growing concern as they can have a profound impact on individuals, potentially resulting in mortality. It is crucial to prevent and manage bacterial infections, particularly drug-resistant ones. Antimicrobial peptides, such as LL-37, can firmly eliminate pathogens. Additionally, the process of angiogenesis, facilitated by growth factors like VEGF, is essential for tissue repair and wound healing. To enhance the stability and bioavailability of therapeutic agents, targeted delivery strategies utilizing Chitosan-based carriers have been employed. Electrospun biopolymers in advanced wound dressings have revolutionized wound care by providing a more effective and efficient solution for promoting tissue regeneration and speeding up the healing process. The present investigation utilized Chitosan nanoparticles to encapsulate the recombinant LL37 peptide and VEGF. An in-depth investigation was carried out to analyze the biophysical and morphological traits of the LL37-CSNPs and VEGF-CSNPs. The first support layer consisted of PCL electrospun nanofiber, followed by the electrospinning of PVA/CsLL37, PVA/CsVEGF, and PVA/CsLL37/CsVEGF onto the PCL layer. An in vitro examination assessed the fabricated nanofibers' morphological, mechanical, and biological characteristics. The antimicrobial effects were tested on methicillin-resistant Staphylococcus aureus (MRSA). The in vivo experiments assessed the antibacterial and wound-healing capabilities of the nanofibers. The findings validated the continuous release of LL37 and VEGF. The composite material PCL/PVA/CsLL37/CsVEGF demonstrated potent bactericidal and antioxidant characteristics. The cytotoxic assay demonstrated the biocompatibility of the fabricated nano mats and their potential to accelerate fibroblast cell proliferation. The efficacy of PVA/CsLL37/CsVEGF in promoting wound healing was confirmed through an in vivo wound healing assay. Furthermore, the histological analysis provided evidence of faster epidermal formation and improved antibacterial activity in wounds covered with PVA/CsLL37/CsVEGF. Adding LL37 and VEGF to the composite material improves the immune response and promotes blood vessel formation, accelerating wound healing and decreasing inflammation., 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 © 2024 Elsevier B.V. All rights reserved.)

Background: The goal was to assess the antimicrobial efficacy of two commonly used biocides, chlorhexidine, and benzalkonium chloride, against MDR isolates of Pseudomonas aeruginosa, Acinetobacter baumannii, and Escherichia coli ST131, as well as the prevalence of resistance genes., Methods: MIC of chlorhexidine and benzalkonium chloride and their effects on both the planktonic phase and biofilm were determined. Finally, the presence of genes responsible for resistance to quaternary ammonium compounds was investigated by PCR., Results: No significant relationship was observed between the presence of resistance genes and different concentrations of quaternary ammonium compounds (benzalkonium chloride). There was no association between biofilm formation and the presence of resistance genes., Conclusions: Chlorhexidine digluconate and benzalkonium chloride at appropriate concentrations could prevent biofilm formation.

Objectives: We assessed the frequency of occurrence for infections caused by wild-type A. baumannii, multidrug-resistant (MDR) or XDR A. baumannii, and CRAB. We detected different antibiotic resistance genes in the genomes of infectious A. baumannii strains from central Iran., Methods: This study investigated 546 clinical patient samples for the presence of A. baumannii by using conventional culture methods and PCR. Antibiotic resistance profiles, and the phenotypic and genotypic characteristics of various antibiotic genes were analyzed., Results: Out of 546 samples, 87 (15.9%) A. baumannii isolates were obtained using culture and all culture positive samples were also positive by PCR. The most effective antibiotics were polymyxin B (n = 84 strains) (96.6% susceptibility), colistin (n = 81) (93.1%), and ampicillin/sulbactam (n = 18) (20.7%). All clinical A. baumannii isolates were ESBL-positive. The number of CRAB was 84 (96.5%). All CRAB isolates were both MDR and XDR. Of all CRAB isolates, 78 out of 84 (92.4%) produced metallo-β-lactamase (MBL) by phenotypic diagnosis. The most abundant genes were bla PER (32/87; 36.7%), bla TEM (29/87; 33.3%), bla VEB (26/87; 29.8%) for ESBL and Ambler class D β -lactamases included bla OXA-23 (69/84; 82.1%), bla OXA-24 (46/84; 54.7%), MBLs included bla VIM (51/84; 60.7%), and bla IMP (28/84; 33.3%) for carbapenemase., Conclusion: High frequencies of XDR A. baumannii and CRAB (96.5%) were detected in central Iran. Quick and accurate diagnosis, appropriate isolation of patients colonized or infected by CRAB isolates, application of accurate and effective infection control policies and programs, and appropriate preventive measures are deemed helpful in preventing the further spread of these resistant and clinically highly relevant strains., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: The emergence and distribution of multidrug-resistant (MDR) and carbapenem-resistant Klebsiella pneumoniae (CRKP) has become a global health threat. Therefore, this study aimed to investigate the frequency and antibiotic resistance patterns of MDR, extensively drug-resistant (XDR), and CRKP, as well as the antibiotic resistance genes of Klebsiella pneumoniae (K. pneumoniae) isolates from patients' infectious samples from central Iran., Methods: This study examined 546 clinical samples of patients to identify K. pneumoniae. The isolates were investigated for their antibiotic resistance profile, extended-spectrum β-lactamase (ESBL), AMPC β-lactamase, carbapenemase resistance, sulfonamide, tetracycline, plasmid-mediated quinolone resistance (PMQR) along with their resistance genes, integrase, and quaternary ammonium compounds (qac) by polymerase chain reaction (PCR)., Results: Out of 546 clinical samples, 121 (22.1%) cases of K. pneumoniae were identified using culture and PCR methods. The highest antibiotic resistance rates were found for ampicillin (119/121; 98.3%), cotrimoxazole (78/121; 64.4%), and cefixime, cefotaxime, ceftriaxone, and ceftazidime as a group (77/121; 63.6%). Tigecycline, colistin, and fosfomycin were the most effective antimicrobial agents with 98.4%, 96.7%, and 95.9% susceptibility, respectively. The amount of CRKP was 51 (42.1%). All CRKP isolates were MDR. The most abundant genes were bla TEM (77/77; 100%), bla CTX-M1 (76/77; 98.7%), bla SHV (76/77; 98.7%), bla CTX-M15 (73/77; 94.8%) for ESBL; bla CIT 28 (48.3%) and bla CMY-2 26 (44.8%) for AMPC β-lactamase; and bla OXA-48 46 (90.1%) and bla NDM 36 (70.5%) for carbapenemase. Among the PMQR determinants, qnrB (25/52; 48%), qnrS (19/52; 36.5%), and qnrA (11/52; 21.1%) were positive from the isolates. TetA and tetB were recognized in 25 (44.6%) and 17 (30.3%) isolates, respectively. Class 1 and 2 integrons were recognized in 97 (80.1%) and 53 (43.8%) isolates, respectively., Conclusions: Due to the high prevalence of MDR and CRKP in central Iran, tracking and immediate intervention are necessary for control and inhibition of K. pneumoniae resistant isolates. Tigecycline, colistin, and fosfomycin are the best treatment options for treatment of patients with CRKP in this geographical area., (© 2023. The Author(s).)

Background and Objectives: Staphylococcus simulans secretes an antimicrobial compound called lysostaphin, which has bactericidal properties. It destroys staphylococci through the hydrolysis of peptidoglycan in the cell wall. Therefore, this unique property indicates the high ability of lysostaphin in the treatment of staphylococcal infections and is considered as an anti-staphylococcal agent., Materials and Methods: Escherichia coli BL21 (DE3) competent cells were transformed with pET32a-lysostaphin clone and induced by isopropyl-β-D-thio-galactoside (IPTG). The recombinant protein was purified by affinity chromatography. Recombinant lysostaphin -A-based ointment was used for external wound healing in animal model. In vivo activity of ointment was evaluated by clinical evidences and cytological microscopic assessment., Results: Our results showed the recombinant protein was produced exactly. The results of checkerboard tests showed MIC, MBC and antibacterial activity test an acute reduction of cell viability during the use of lysostaphin, and SEM results approved the intense wrecking effects of lysostaphin in combination on bacterial cells. Macroscopic findings and microscopic data showed that the recombinant lysostaphin ointment was effective on excisional wound healing., Conclusion: Our findings proved that the recombinant lysostaphin ointment was effective on wound healing due to Staphylococcus aureus infection., (Copyright © 2023 The Authors. Published by Tehran University of Medical Sciences.)

Background: Infections caused by Acinetobacter baumannii , especially carbapenem-resistant (CR) strains, pose important challenges in patients with COVID-19 infection. Therefore, in the present study, we investigated co-infection and antimicrobial resistance patterns, as well as the role of A. baumannii in the outcome of patients with COVID-19. Materials and methods: Between February 2019 and January 2021, 141 patients with A. baumannii infections were detected from seven different hospitals (A to G) in Arak, Iran, and the antibacterial susceptibility pattern of these isolates was evaluated using disk diffusion and E-test methods. Forty-seven of these patients were co-infected with COVID-19. During the study, the data about the clinical course, demographic data, and the role of A. baumannii infections in the mortality rate of COVID-19 patients were collected. Results: Hospitals A and B reported the most patients, with 53 (38%) and 47 (33%), respectively. Additionally, most cases (105 cases, 75%) were reported from surgical and general ICUs. Mechanical ventilators were detected as predisposing factors in 95 (67%) patients, and infection was detected in 20% of patients on the 10th day after intubation. All of the A. baumannii isolates were resistant to different classes of antibiotics, such as carbapenems. Notably, 33% (47 patients) were also positive for COVID-19, and 68% (32 patients) died due to the infection. Statistical analysis showed a significant role of A. baumannii co-infection in the mortality rate of COVID-19 patients (p-value 0.05). Conclusion: co-infection with A. baumannii is one of the most important challenges in COVID-19 patients. Our results showed that all isolated bacteria were CR and significantly increased mortality rates in COVID-19 patients., (© 2022 Elsevier España, S.L.U. All rights reserved.)

Background: The antimicrobial peptides (AMPs) played a critical role in the innate immunity of the host and are considered natural sources illustrating a broad-spectrum antimicrobial activity with high specificity and low cytotoxicity. AMPs generally possess a net positive charge and have amphipathic structures. Thus, AMPs can bind and interact with negatively charged bacterial cell membranes, leading to destructive defects in biomembranes and ending in cell death. LL37 is the only human cathelicidin-derived antimicrobial peptide that shows a broad spectrum of antimicrobial activity., Materials and Methods: To determine the antibacterial efficiency of LL37 in a mouse model of systemic A. baumannii infection, LL37 corresponding gene was expressed in E. coli, purification and refolding situations were optimized. The antimicrobial performance of produced LL-37 against A. baumannii was evaluated in vitro via MIC and Time Kill assays, and its destructive effects on the bacterial cell were confirmed by SEM image., Results: The recombinant LL37 showed strong antibacterial function against A. baumannii at 1.5 μg/mL concentration. Time kill assay showed a sharp reduction of cell viability during the first period of exposure, and complete cell death was recorded after 40 min exposure., Conclusion: Furthermore, in vivo results represented a significant ability of LL37 in the treatment of systematic infected mouse models, and all infected mice receiving LL37 protein survived without no trace of bacteria in their blood samples., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Background: Novel coronavirus (COVID-19) pandemic has become a global concern and requires early detection, isolation, and treatment. Our purpose is to find some beneficial information by analyzing the COVID-19 laboratory data to provide guidance for clinical practice., Material and Methods: In this study, 174 patients with confirmed COVID-19 infection were admitted. We evaluated the hematological and biochemical parameters in these patients and in 80 healthy individuals., Results: We found that there was significant difference between WBC, LYM, RBC, HB, and HCT parameters of patients and healthy counterparts (p < .001), though there was no remarkable change between NEU, MONO, PLT, and other characteristics of RBC values of patients and the control group (p ≥ .09). Among the evaluated biochemical parameters, the values of CK-MB and LDH in the patient group were significantly different from the control group (p < .01), while other biochemical indicators were in the normal range., Conclusion: Several hematological and biochemistry parameters, in particular WBC, LYM, RBC, HB, HCT, CK-MB, and LDH, could be beneficial supplementary approach for COVID-19 infection evaluation to confirm risk stratification and effective management., (© 2022 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC.)

Aims: This study represents the first analysis from Iran for both the frequency of the most common causes of infectious diarrhoea and their antibiotic resistance patterns in adult patients. Methods: Adult stool specimens ( n  = 211) were analyzed. Stool specimens were analyzed using standard microbiological, polymerase chain reaction, and reverse transcription polymerase chain reaction tests to identify bacterial, parasitic, and viral enteropathogens. Antibiotic resistance profiles were determined. Results: Enteropathogens were identified in 46.4% (98/211) of the surveyed samples. This included 33.1% (70/211) bacterial infections, including 9.9% (21/211) diarrheagenic Escherichia coli (DEC) and 8.5% (18/211) Shigella spp. We detected 7.1% (15/211) parasitic infections (mostly Giardia lamblia ) and 6.1% (13/211) viral infections (mostly adenovirus). The DEC and Shigella spp. isolates included many multi-drug resistant (MDR) isolates (95.2% and 77.7%, respectively), and extended spectrum-β-lactamase (ESBL) genes were often present (57.1% and 61.1%, respectively). The most commonly identified ESBL genes in the DEC and Shigella spp. isolates were bla TEM (100% in both species), bla CTX-M15 (91.6% and 100%, respectively), AmpC bla CIT (80% and 100%, respectively), and bla DHA (80% and 100%, respectively). Conclusions: Bacterial infection was the primary cause of infectious diarrhea, affecting one-third of the adults. The frequency of DEC and Shigella spp. was higher than for other enteropathogens. The high prevalence of MDR, the elevated incidence of ESBL genes among Shigella spp. and DEC isolates, and the presence of quinolone resistance in the Salmonella spp. isolates represent a significant challenge for gastroenteritis diagnosis and treatment in this region.

Objectives: Acinetobacter Baumannii is an opportunistic nosocomial pathogen belonging to the Moraxellaceae family. The emergence of multidrug resistant strains of this pathogen caused many problems for hospitals and patients. The aim of the current study was to isolate, identify, and morphologically, physiologically, and in vivo analyze a new lytic bacteriophage targeting extensively drug-resistant (XDR) A. baumannii., Materials and Methods: Different wastewater samples were tested for isolation of lytic bacteriophage against 19 A. baumannii isolates obtained from patients hospitalized in a hospital in Arak, Iran, from January 2019 to March 2019. The phenotypic and genotypic characteristics of A. baumannii strains (resistance genes including: adeA, adeB, adeC, adeR, adeS, ISAba1, blaOXA-23, blaOXA-24) were analyzed. The isolated phage characteristics including adsorption time, pH and thermal stability, host range, one-step growth rate, electron microscopy examination, and therapeutic efficacy of the phage were also investigated. Therapeutic efficacy of the phage was evaluated in a rat model with burn infection of XDR A. baumannii. The lesion image was taken on different days after burning and infection induction and was compared with phage untreated lesions., Results: The results showed unique characteristics of the isolated phage (vB-AbauM-Arak1) including high specificity for Acinetobacter baumannii, stability at a relatively wide range of temperatures and pH values, short adsorption time, short latent period, and large burst size. In relation to the therapeutic efficacy of the phage, the lesion area decreased in phage-treated groups over 14 days than in those untreated, significantly (p < 0.05)., Conclusion: Our findings demonstrated that isolated lytic phage was able to eliminate burn infections caused by XDR A. baumannii in a rat model. So, it may be recommended as alternative options toward to developing a treatment for extensively drug resistant Acinetobacter infections., (© 2022 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC.)

MicroRNAs (miRNAs) are epigenetic regulators of the gene expression and act through posttranslational modification. They bind to 3'-UTR of target mRNAs to inhibit translation or increase the degradation mRNA in many tissues. Any alteration in the level of miRNA expression in many human diseases indicates their involvement in the pathogenesis of many diseases. On the other hand, the regulation of the signaling pathways is necessary for the maintenance of natural and physiological characteristics of any cell. It is worth mentioning that dysfunction of the signaling pathways manifests itself as a disorder or disease. The significant evidence report that miRNAs regulate the several signaling pathways in many diseases. Base on previous studies, miRNAs can be used for therapeutic or diagnostic purposes. According to the important role of miRNAs on the cell signaling pathways, this article reviews miRNAs involvement in incidence of diseases by changing signaling pathways., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

The biofilm community of microorganisms has been identified as the dominant mode of microbial growth in nature and a common characteristic of different microorganisms such as Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis. The biofilm structure helps in the protection from environmental threats including host immune system and antimicrobial agents. Thus, the biofilm community has led to a higher prevalence of multidrug-resistant (MDR) strains in recent years. In this regard, the use of a new class of antibiotics, natural compounds, and anti-biofilm enzymes has been considered for the destruction of the microbial biofilm. However, different drawbacks such as low penetration, high susceptibility to degradation, instability, and poor solubility in aqueous solutions limit the use of anti-biofilm agents (ABAs) in a clinical setting. As such, recent studies have been using poly lactic- co -glycolic acid (PLGA)-based nanoplatforms (PLGA NPFs) for delivery of ABAs that have reported promising results. These particles, due to proper drug loading and release kinetics, could suppress microbial attachment, colonization, and biofilm formation for a long time. Additionally, PLGA NPFs, because of the high drug-loading efficiencies, hydrophilic surface, negative charge, and electrostatic interaction, lead to effective penetration of antibiotics to the deeper layer of the biofilm, thereby eliminating the microbial biofilm. Thus, PLGA NPFs could be considered as a potential candidate for coating catheters and other medical material surfaces for inhibition and destruction of the microbial biofilm. However, the exact interaction of PLGA NPFs and the microbial biofilm should be evaluated in animal studies. Additionally, a future goal will be to develop PLGA formulations as systems that can be used for the treatment of the MDR microbial biofilm, since the exact interactions of PLGA NPFs and these biofilm structures are not elucidated. In the present review article, we have discussed various aspects of PLGA usage for inhibition and destruction of the microbial biofilm along with different methods and procedures that have been used for improving PLGA NPF efficacy against the microbial biofilm., 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 Shariati, Chegini, Ghaznavi-Rad, Zare and Hosseini.)

Background: According to the contagious ability of the new virus, SARS-CoV-2, characterization of viral shedding duration in the period of infection is highly valuable in terms of providing quarantine guidelines and isolation policies. Therefore, we aimed at viral shedding determination in 58 COVID-19 confirmed Iranian subjects in different stages., Methods: 58 COVID-19 confirmed Iranian subjects including 21 outpatients and 37 inpatients were investigated. The analytical data and clinical properties were documented in the standard questionnaire. The RT-PCR tests were done two and three weeks after the symptoms initiation., Results: Viral eradication occurred in 44.8% two weeks after illness initiation whereas in 71% who achieved a negative PCR test in the third week. Moreover, prolonged viral shedding was observed in hospitalized cases in comparison to outpatients. Almost 30% of patients continued viral shedding three weeks after disease initiation., Conclusion: A longer duration of viral shedding in hospitalized cases rather than outpatients was observed in this study. The results similar to other investigations call into question if the current policies are enough to prevent the viral spread or not. This study should be done on a larger sample to provide an appropriate time in isolation policy., Competing Interests: All authors declare that there are no conflicts of interest., (Copyright © 2022 Masoomeh Sofian et al.)

Objectives: Antimicrobial peptide compounds (AMPs) play important roles in the immune system. They also exhibit significant anti-tumor and antibacterial properties. Most AMPs are cationic and are able to bind bacterial cell membranes through electrostatic affinity. Ib-AMP4 is a plant-derived AMP that exerts rapid bactericidal functions. In the present study, the antibacterial efficiency of the produced recombinant Ib-AMP4 in elimination of Methicillin-resistant Staphylococcus aureus ( MRSA ) bacterial infection, was investigated under in vitro and in vivo situations., Materials and Methods: The synthesized Escherichia coli codon-optimized gene sequences of the Ib-AMP4 were expressed in E. coli BL21 (DE3) pLysS . The recombinant Ib-AMP4 was purified and refolding conditions were optimized. The antibacterial efficiency of the refolded peptide against MRSA was tested under in vivo and in vitro situations for treatment of skin and systematic infection of MRSA in a mouse model., Results: Antibacterial assays confirmed the antibacterial function of Ib-AMP4 against MRSA . SEM results proved the destructive effects of applying Ib-AMP4 on MRSA biomembrane. Time-kill curve and growth kinetic assay illustrated rapid antibacterial activity of the produced Ib-AMP4. Moreover, Ib-AMP4 showed significant infection treatment ability in a mouse model and all infected mice receiving Ib-AMP4 protein survived and there was no trace of bacteria in their blood samples., Conclusion: The results confirmed the rapid antibacterial potential of the produced recombinant Ib-AMP4 to be used for efficient treatment of MRSA infection., Competing Interests: The authors declare that no conflict of interest exists.

Coronavirus disease 2019 (COVID-19) may lead to acute respiratory disease; cardiovascular, gastrointestinal, and coagulation complications; and even death. One of the major complications is cardiovascular disorders, including arrhythmias, myocarditis, pericarditis, and acute coronary artery disease. The aim of this study was to evaluate the frequency of cardiovascular complications and to determine its association with the prognosis of COVID-19 patients. In a prospective analytic study, 137 hospitalized COVID-19 patients were enrolled. During hospitalization, an electrocardiogram (ECG) was performed every other day, and laboratory tests such as cardiac troponin I (cTnI) and creatine kinase-MB (CK-MB) were done 0, 6, and 12 hours after admission. These tests were repeated for patients with chest pain or ECG changes. Patients were categorized into three groups (improved, complicated, and expired patients) and assessed for the rate and type of arrhythmias, cardiac complications, lab tests, and outcomes of treatments. There was no significant relationship among the three groups related to primary arrhythmia and arrhythmias during treatment. The most common arrhythmia during hospitalization and after treatment was ST-T fragment changes. There was a significant age difference between the three groups ( P = 0.001). There was a significant difference among the three groups for some underlying diseases, including diabetes mellitus ( P = 0.003) and hyperlipidemia ( P = 0.004). In our study, different types of arrhythmias had no association with patients' outcomes but age over 60 years, diabetes mellitus, and hyperlipidemia played an important role in the prognosis of COVID-19 cases., Competing Interests: The authors declare no competing interests., (Copyright © 2021 Hossein Mazaherpour et al.)

Purpose: The production of alternative novel antimicrobial agents is considered an efficient way to cope with multidrug resistance among pathogenic bacteria. E50-52 and Ib-AMP4 antimicrobial peptides (AMPs) have illustrated great proven antibacterial effects. The aim of this study was recombinant production of these AMPs and investigation of their synergistic effects on methicillin-resistant Staphylococcus aureus (MRSA)., Method: At first, the codon optimized sequences of the Ib-AMP4 (UniProt: 024006 (PRO&#95;0000020721), and E50-52 (UniProtKB: P85148) were individually ligated into the pET-32α vector and transformed into E. coli. After the optimization of production and purification steps, the MIC (Minimum inhibitory concentration), time kill and growth kinetic tests of recombinant proteins were determined against MRSA. Finally, the in vivo wound healing efficiency was tested., Results and Conclusion: The recorded MIC of recombinant Trx-Ib-AMP4, Trx-E50-52 against MRSA bacterium were 0.375 and 0.0875 mg/mL respectively. The combination application of the produced AMPs by the checkerboard method confirmed their synergic activity. The results of the time-kill showed sharply decrease of the number of viable cells with over five time reductions in log 10  CFU/mL by the combination of Trx-E50-52 and Trx-IbAMP4 at 2 × MIC within 240 min. The growth kinetic results confirmed the combination of Trx-E50-52 and Trx-IbAMP4 had much greater success in the reduction of over 50 % of MRSA suspensions' turbidity within the first hour. Wound healing assay and histological analysis of infected mice treated with Trx-Ib-AMP4 or Trx-E50-52 compared with those treated with a combination of Trx-Ib-AMP4 and Trx-E50-52 showed significant synergic effects., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background: Ventilator-associated pneumonia (VAP) is an infectious pulmonary disease that develops after 48 hours of ventilation. To date, several methods have been proposed to reduce VAP occurrence, such as the VAP prevention bundle, which involves raising the head of the bed, reducing sedation, avoiding deep vein thrombosis, and preventing peptic ulcer in the gastrointestinal system. The purpose of this study was to evaluate the role of personnel in hand washing, case airway suctioning, and systematic monitoring in the prevention of VAP., Methods: In the current clinical trial, 129 patients hospitalized and intubated at Vali-e-Asr Hospital ICU in Arak, Iran, were included in the study and randomized to one of the three VAP prevention methods: group A, VAP prevention bundle measures; group B, group A measures plus washing of patients' mouth with 0.12% chlorhexidine and suction of secretion every six hours; and finally group C, group B measures plus 72-hour suction package. Demographic information, VAP diagnosis, and outcome of each patient were recorded in the special checklist., Results: The age of the patients ranged from 18 years to 93 years with a mean of 54.6 ± 21.8 years. There was no significant difference in age, sex, Clinical Pulmonary Infection Score (CPIS), and Glasgow Coma Scale (GCS) between the three groups. However, there is a significant relationship between chest X-ray (CXR) index and pneumonia in the three groups ( p  < 0.05). The prevalence of pneumonia is generally seen to be higher in patients who were local, diffuse, or patchy than those who had no infiltration ( p  < 0.05)., Conclusion: This study showed that the application of VAP prevention bundle measures, mouthwash with chlorhexidine, personnel hand washing, airway suctioning, and systematic monitoring is an efficient approach to the prevention of VAP in ICUs., (© 2021 Zarinfar, Ghaznavi-Rad, Mahmoodiyeh, Reyhani, licensee HBKU Press.)

Background and Aim: Recent studies have proposed that commensal bacteria might be involved in the development and progression of gastrointestinal disorders such as colorectal cancer (CRC). Therefore, in this study, the relative abundance of Fusobacterium nucleatum, Bacteroides fragilis, Streptococcus bovis/gallolyticus, and Enteropathogenic Escherichia coli (EPEC) in CRC tissues, and their association with clinicopathologic characteristics of CRC was investigated in Iranian patients. Moreover, the role of these bacteria in the CRC-associated mutations including PIK3CA, KRAS, and BRAF was studied., Method: To these ends, the noted bacteria were quantified in paired tumors and normal tissue specimens of 30 CRC patients, by TaqMan quantitative Real-Time Polymerase Chain Reaction (qPCR). Next, possible correlations between clinicopathologic factors and mutations in PIK3CA, KRAS, and BRAF genes were analyzed., Results: In studied samples, B. fragilis was the most abundant bacteria that was detected in 66 and 60% of paired tumor and normal samples, respectively. Furthermore, 15% of the B. fragilis-positive patients were infected with Enterotoxigenic B. fragilis (ETBF) in both adenocarcinoma and matched adjacent normal samples. F. nucleatum was also identified in 23% of tumors and 13% of adjacent normal tissue samples. Moreover, the relative abundance of these bacteria determined by 2 -ΔCT was significantly higher in CRC samples than in adjacent normal mucosa (p < 0.05). On the other hand, our findings indicated that S. gallolyticus and EPEC, compared to adjacent normal mucosa, were not prevalent in CRC tissues. Finally, our results revealed a correlation between F. nucleatum-positive patients and the KRAS mutation (p = 0.02), while analyses did not show any association between bacteria and mutation in PIK3CA and BRAF genes., Conclusion: The present study is the first report on the analysis of different bacteria in CRC tissue samples of Iranian patients. Our findings revealed that F. nucleatum and B. fragilis might be linked to CRC. However, any link between gut microbiome dysbiosis and CRC remains unknown.

Background and Aim: Treatment of burn wound infections has become a global challenge due to the spread of multidrug-resistant bacteria; therefore, the development of new treatment options for the mentioned infections is essential. Platelets have drawn much attention for this purpose because they are a safe and cost-effective source of different antimicrobial peptides and growth factors. The present study evaluated antibacterial effects and wound healing properties of Platelet-derived Biomaterial (PdB) against Acinetobacter baumannii and Klebsiella pneumoniae burn wound infections., Methods: PdB was prepared through the freezing and thawing process and then, in vitro antibacterial effect was determined by disk diffusion and broth microdilution methods. Afterward, burn wound was inflicted on 56 rats, infected with both bacteria, and topical administration was performed to evaluate antibacterial effects and wound healing properties of PdB., Results: In vitro results showed that PdB inhibited the growth of A. baumannii in the highest dose (0.5), while we did not detect any inhibitory effects against K. pneumoniae. By contrast, PdB significantly inhibited the growth of bacteria in treated animal wounds compared to the control groups (P value < 0.05). Macroscopic assessments pointed to the significant enhancement of wound closure in the treated animals. In addition, histopathological examination demonstrated that treatment of rats with PdB led to a considerable increase in re-epithelialization and attenuated the formation of granulation tissue (P value < 0.05)., Conclusion: The use of topical PdB is an attractive strategy for treating A. baumannii and K. pneumoniae burn wound infections because it inhibits bacterial growth and promotes wound healing properties.