Upregulation of pmrA, pmrB, pmrC, phoQ, phoP, and arnT genes contributing to resistance to colistin in superbug Klebsiella pneumoniae isolates from human clinical samples in Tehran, Iran.

Background: Antibiotic resistance in Klebsiella pneumoniae isolates, particularly resistance to colistin, has become a growing concern. This study seeks to investigate the upregulation of specific genes (pmrA, pmrB, pmrC, phoQ, phoP, and arnT) that contribute to colistin resistance in K. pneumoniae isolates collected from human clinical samples in Tehran, Iran. Methods: Thirty eight K. pneumoniae isolates were obtained and subjected to antibiotic susceptibility testing, as well as evaluation for phenotypic AmpC and ESBL production according to CLSI guidelines. The investigation of antibiotic resistance genes was conducted using polymerase chain reaction (PCR), whereas the quantification of colistin resistance related genes expressions was performed via Real-Time PCR. Results: The highest and lowest antibiotics resistance were observed for cefotaxime 33 (86.8%) and minocycline 8 (21.1%), respectively. Twenty-four (63.2%) and 31 (81.6%) isolates carried AmpC and ESBLs, respectively. Also, antibiotic resistance genes containing blaNDM, blaIMP, blaVIM, blaSHV, blaTEM, blaCTXM, qnrA, qnrB, qnrS, and aac(6')-Ib were detected in K. pneumoniae isolates. Only 5 (13.1%) isolates were resistant to colistin and the MIC range of these isolates was between 4 and 64 µg ml-1. Upregulation of the pmrA, pmrB, pmrC, phoQ, phoP, and arnT genes was observed in colistin-resistant isolates. The colistin-resistant isolates were found to possess a simultaneous presence of ESBLs, AmpC, fluoroquinolone, aminoglycoside, and carbapenem resistant genes. Conclusions: This study reveals escalating antibiotic resistance in K. pneumoniae, with notable coexistence of various resistance traits, emphasizing the need for vigilant surveillance and innovative interventions.

Unraveling the complex relationship: Multiple sclerosis, urinary tract infections, and infertility.

BACKGROUND: Multiple sclerosis (MS) is an immune system disorder that affects the central nervous system (CNS) and progressively damages nerve fibers and protective myelin. People with MS often experience a wide range of complications, including lower urinary tract dysfunction, urinary tract infections (UTIs) and sexual dysfunction. MS is common in young people and can lead to sexual dysfunction (SD) and infertility, which becomes more pronounced as the disease progresses. RESULTS: Over the past two decades, significant advances have been made in the management of MS, which may slow the progression of the disease and alter its course. However, UTI and SD remain significant challenges for these patients. Awareness of the underlying complications of MS, such as UTIs and infertility, is crucial for prevention, early detection and appropriate treatment, as there is a causal relationship between UTIs and the use of corticosteroids during an attack. CONCLUSION: This article provides an overview of potential microbial pathogens that contribute to the development of MS, as well as an assessment of people with MS who report UTIs and infertility.

Prevalence of colistin resistance in clinical isolates of Acinetobacter baumannii: a systematic review and meta-analysis.

INTRODUCTION: The development of colistin resistance in Acinetobacter baumannii during treatment has been identified in certain patients, often leading to prolonged or recurrent infections. As colistin, is the last line of therapy for A. baumannii infections that are resistant to almost all other antibiotics, colistin-resistant A. baumannii strains currently represent a significant public health threat, particularly in healthcare settings where there is significant selective pressure. AIM: The aim of this study was to comprehensively determine the prevalence of colistin resistance in A. baumannii from clinical samples. Regional differences in these rates were also investigated using subgroup analyses. METHOD: The comprehensive search was conducted using "Acinetobacter baumannii", "Colistin resistant" and all relevant keywords. A systematic literature search was performed after searching in PubMed, Embase, Web of Science, and Scopus databases up to April 25, 2023. Statistical analysis was performed using Stata software version 17 and sources of heterogeneity were evaluated using I2. The potential for publication bias was explored using Egger's tests. A total of 30,307 articles were retrieved. After a thorough evaluation, 734 studies were finally eligible for inclusion in the present systematic review and meta-analysis. RESULT: According to the results, the prevalence of resistance to colistin among A. baumannii isolates was 4% (95% CI 3-5%), which has increased significantly from 2% before 2011 to 5% after 2012. South America had the highest resistance rate to this antibiotic. The broth microdilution method had the highest level of resistance, while the agar dilution showed the lowest level. CONCLUSIONS: This meta-analysis found a low prevalence of colistin resistance among A. baumannii isolates responsible for infections worldwide from 2000 to 2023. However, there is a high prevalence of colistin-resistant isolates in certain countries. This implies an urgent public health threat, as colistin is one of the last antibiotics available for the treatment of infections caused by XDR strains of A. baumannii.

Potential antibacterial activity and healing effect of topical administration of bone marrow and adipose mesenchymal stem cells encapsulated in collagen-fibrin hydrogel scaffold on full-thickness burn wound infection caused by Pseudomonas aeruginosa.

Burns injuries are prone to hospital-acquired infections, and Pseudomonas aeruginosa is one of the most common causes of mortality and morbidity in patients with burn injuries. Thus, this study aimed to analyze the effects of topical treatment with bone marrow (BM-MSC) and adipose mesenchymal stem cells (AD-MSC) encapsulated in collagen and fibrin scaffolds in a Balb/c model of burn wound infection. Extraction of stem cells from adipose and bone marrow tissue of rats was performed and cells were characterized using standard methods. Then, collagen, fibrin and collagen-fibrin scaffolds were constructed and the extracted cells were encapsulated in all three scaffolds. Then, 3rd degree burn was induced in mice and 1.5 × 108 (CFU/ml) of P. aeruginosa was introduced to the burn wound. Subsequently, after 24 h of inducing wound infection, encapsulated MSCs were introduced as dressings to burn wound infection and microbial load as well as rate of wound infection healing was measured. The results of this study showed that the use of BM-MSC and AD-MSC encapsulated in collagen-fibrin scaffold reduced the bacteria load down to 54 and 21 CFU/gr, respectively (P < 0.05). Moreover, BM-MSC and AD-MSC encapsulated in collagen-fibrin showed 80% and 75% wound healing, respectively (P < 0.05). Also, we found no significant between cell origin and healing. Encapsulation of MSCs into collagen-fibrin scaffolds could be effective not only against P. aeruginosa infection, but also healing and regeneration of burn wound.

Evaluation of biosynthesized silver nanoparticles effects on expression levels of virulence and biofilm-related genes of multidrug-resistant Klebsiella pneumoniae isolates.

The emergence of multidrug-resistant (MDR) strains of Klebsiella pneumoniae is associated with high morbidity and mortality due to limited treatment options. This study attempts to biologically synthesize silver nanoparticles (AgNPs) and investigate their effect on expression levels of virulence and biofilm-related genes in clinically isolated K. pneumoniae. In this study, biofilm formation ability, antibiotic resistance pattern, extended-spectrum ß-lactamases (ESBLs), and carbapenemases production were investigated for 200 clinical isolates of K. pneumoniae using phenotypic methods. Polymerase chain reaction (PCR) was used to detect virulence and biofilm-related genes, ESBL-encoding genes, and carbapenem resistance genes. AgNPs were synthesized using the bio-reduction method. The antibacterial effects of AgNPs were investigated by microdilution broth. In addition, the cytotoxic effect of AgNPs on L929 fibroblast cell lines was determined. The effects of AgNPs on K. pneumoniae virulence and biofilm-related genes (fimH, rmpA, and mrkA) were determined using quantitative real-time PCR. Thirty percent of the isolates produced a strong biofilm. The highest and lowest levels of resistance were observed against amoxicillin/clavulanic acid (95.4%) and tigecycline (96%), respectively. About 31% of isolates were considered positive for carbapenemases, and 75% of the isolates produced an ESBLs enzyme. Different frequencies of mentioned genes were observed. The synthesized AgNPs had a spherical morphology and varied in size. AgNPs inhibited the growth of MDR K. pneumoniae at 128 µg/ml. In addition, AgNPs downregulated the expression of fimH, rmpA, and mrkA genes by 10, 7, and 14-fold, respectively (p < 0.05), also exerted no cytotoxic effect on L929 fibroblast cell lines. It was revealed that AgNPs lead to a decrease in expression levels of virulence and biofilm-related genes; therefore, it was concluded that AgNPs had an excellent antibacterial effect on MDR K. pneumoniae.

Zinc oxide nanoparticles impact the expression of the genes involved in toxin-antitoxin systems in multidrug-resistant Acinetobacter baumannii.

The aim of this study was to investigate the effect of zinc oxide nanoparticles (ZnO-NPs) on the expression of genes involved in toxin-antitoxin (TA) systems in multidrug-resistant (MDR) Acinetobacter baumannii. Seventy clinical isolates of A. baumannii were collected from variuos clinical samples. Antimicrobial susceptibility test was determined by disk diffusion. Type II TA system-related genes including GNAT, XRE-like, hipA, hipB, hicA, hicB were screened using polymerase chain reaction (PCR). ZnO-NPs prepared and characterized by field emission scanning electron microscopy and X-ray diffraction. MIC of ZnO-NPs of A. baumannii isolates was performed using the microdilution method. The expression of type II TA systems-related genes were assessed with and without exposure to ZnO-NPs using real-time PCR. The highest rate of resistance and sensitivity was observed against cefepime (77.14%), and ampicillin/sulbactam (42.85%), respectively. All A. baumannii isolates were considered as MDR. In this study, three TA loci were identified for A. baumannii including GNAT/XRE-like, HicA/HicB, and HipA/HipB and their prevalence was 100%, 42%, and 27.1%, respectively. There was no significant relationship between the prevalence of these systems and the origin of A. baumannii. Our data showed significant correlations between the presence of HicA/HicB system and resistance to ceftazidime, meropenem, imipenem, and cefepime (p < 0.05), and the presence of HipA/HipB system and resistance to ceftazidime, meropenem, imipenem, and cefepime (p < 0.05). In presence of ZnO-NPs, the expression of all studied genes decreased. GNAT and hicB showed the highest and lowest expression changes by 2.4 folds (p < 0.001) and 1.3 folds (p < 0.05), respectively. This study demonstrates the promising potential of nanoparticles to impact the expression of the genes involved in TA Systems. So, the application of ZnO-NPs may be helpful to design target-based strategies towards MDRs pathogens for empowered clinical applications by microbiologists and nanotechnologists.

The expression of type II TA system genes following persister cell formation in Pseudomonas aeruginosa isolates in the exponential and stationary phases.

Failure of infection therapy in the presence of antibiotics has become a major problem which has been mostly attributed to the ability of bacterial persister cell formation. Bacteria use various mechanisms to form persister cells in different phases, among which is the toxin-antitoxin (TA) systems. This study aimed at investigating the expression of type II TA system genes under the stress of ciprofloxacin and colistin antibiotics in the exponential and stationary phases. To determine the effects of ciprofloxacin and colistin on persister cell formation in the exponential and stationary phases of Pseudomonas aeruginosa strains, colony counting was performed at different time intervals in the presence of fivefold MIC of ciprofloxacin and colistin. In addition, the expression of relBE, Xre-COG5654, vapBC, and Xre-GNAT genes in P. aeruginosa isolates was assessed 3.5 h after antibiotic treatment in the exponential and stationary phases using qRT-PCR. Our results indicated the presence of persister phenotype of P. aeruginosa strains in the presence of fivefold MIC of ciprofloxacin and colistin compared to the control after 3.5 h of incubation in the exponential and stationary phases. Also, the number of persister cells in the stationary phase was higher than that of the exponential phase. According to the results of qRT-PCR, ciprofloxacin and colistin may induce persister cells by increasing the expression of type II TA systems in stationary and exponential phases. Ciprofloxacin and colistin may increase the formation of persister cells by affecting the expression of type II TA systems.

Molecular analysis of dominant paranasal sinus bacteria in patients with and without chronic rhinosinusitis.

Recent studies have established the possible role of microbiota in developing various diseases. In this regard, attention has shifted to the evaluation of microbiota changes in the paranasal sinuses and its relationship to chronic rhinosinusitis (CRS), especially CRS with nasal polyposis (CRSwNP). This study aimed to examine the bacterial communities of the sphenoidal sinus in Iranian patients with and without CRS. The investigation included 36 subjects, including 18 patients with CRSwNP who underwent Functional Endoscopic Sinus Surgery (FESS) and 18 non-CRS patients who underwent Endoscopic Endonasal Approach (EEA) for pituitary adenoma. The surgeries were performed under general anesthesia, and the sphenoidal sinus was sampled using sterile rayon-tipped swabs coated with a sheet. TaqMan quantitative real-time polymerase chain reaction (qPCR) method (the 16S rDNA gene from bacteria) was used for detection of bacterial communities in different samples. Staphylococcus haemolyticus and Pseudomonas aeruginosa were significantly more prevalent in CRS patients than non-CRS patients (P value ≤ 0.05). However, no significant difference in the frequency of Corynebacterium spp. and Staphylococcus aureus was observed between the two groups, and no Streptococcus pneumoniae or Haemophilus influenza species were isolated from any of the samples. The current study's findings indicated a significant difference in the frequency of certain bacterial species in patients with CRS vs. non-CRS patients. By establishing a link between microbial burden and CRS, it is possible to develop effective treatments or even prevent disorders in this body area.

Association between virulence factors and biofilm formation in Enterococcus faecalis isolated from semen of infertile men.

PROBLEM: Enterococcus faecalis is a common microbial semen contaminant. Although virulence factors and biofilm formation have often been analyzed in Enterococcus spp., there is little information about these features in isolates obtained from the genitourinary tract. This study was intended to characterize and determine the relationship between biofilm-forming ability and the presence of E. faecalis virulence factors isolated from human semen. METHOD OF STUDY: A total of 32 patients diagnosed with primary infertility and 28 healthy men were included in the study. Semen analyses were performed according to the WHO guidelines. PCR reactions were applied for the detection of ace, esp, efeA, gelE, asa1, and cylA genes. Microtiter plate assay, via measurement of OD560, was used to measure the biofilm-forming ability of the isolates. RESULTS: Sixty E. faecalis isolates from semen of infertile and fertile men were characterized by phenotypic and genotypic methods. The prevalence of ace, esp, efeA, gelE, asa1 and cylA were reported to be 81.3%/100.0%, 81.3%/89.3%, 81.3%/85.7%, 71.9%/53.6%, 8.8%/75.0%, and 62.5%/67.9% in infertile/fertile groups; respectively. Strong, weak, and non-biofilm reactions were reported to be 50.0%/21.4%, 40.6%/64.3%, and 9.4%/14.3% in infertile and fertile groups; respectively. CONCLUSIONS: There was a significant relationship between fertility and weak biofilm reaction and also between biofilm formation and possession of the esp gene (P < .05). It could be speculated that colonization with E. faecalis with a strong ability for biofilm formation could become a potential threat to men's fertility.

Bacteriophage as a Novel Therapeutic Weapon for Killing Colistin-Resistant Multi-Drug-Resistant and Extensively Drug-Resistant Gram-Negative Bacteria.

Colistin-resistant multidrug-resistant (MDR), extensively drug-resistant (XDR), and pan-drug-resistant (PDR) bacteria are highly lethal and many researchers have tried hard to combat these microorganisms around the world. Infections caused by these bacteria are resistant to the last resort of antibiotic therapy and have posed a major challenge in clinical and public health. Since the production of new antibiotics is very expensive and also very slow compared to the increasing rate of antibiotic resistance, researchers are suggesting the use of natural substances with high antibacterial potential. Bacteriophages are one of the most effective therapeutic measures that are known to exist for use for incurable and highly resistant infections. Phages are highly taken into consideration due to the lack of side effects, potential spread to various body organs, distinct modes of action from antibiotics, and proliferation at the site of infection. Although the effects of phages on MDR and XDR bacteria have been demonstrated in various studies, only a few have investigated the effect of phage therapy on colistin-resistant isolates. Therefore, in this review, we discuss the problems caused by colistin-resistant MDR and XDR bacteria in the clinics, explain the different mechanisms associated with colistin resistance, introduce bacteriophage therapy as a powerful remedy, and finally present new studies that have used bacteriophages against colistin-resistant isolates.

Pan drug-resistant Acinetobacter baumannii causing nosocomial infections among burnt children.

Background: Nosocomial infection caused by Acinetobacter baumannii has emerged as a world-wide serious problem in the emergence of multidrug-resistant (MDR). Infections caused by antibiotic-resistant strains of A. baumannii cannot be completely eliminated among the infected patients. This study aimed to monitor antibiotic resistance among A. baumannii strains isolated from burnt children. Methods: After performing biochemical identification tests on 115 isolates, 62 were detected as A. baumannii . Minimum inhibitory concentration (MIC) was used to test susceptibility to colistin, and disk agar diffusion was used for the susceptibility of the isolates to the antibiotics Ciprofloxacin, Amikacin, Gentamicin, Cefepime, Meropenem, Imipenem, Ceftazidime, Levofloxacin and Piperacillin/Tazobactam. Bacterial species were isolated and identified as multidrug-resistant (MDR), extensively drug-resistant (XDR) and pan drug-resistant (PDR), based on the susceptibility patterns to elected antibiotics, deputing different classes of antimicrobial. Results: The antibiotic susceptibility pattern out of a total of 62 bacterial strains used in this study. Thirty-six (58%) strains were categorized as MDR, 17 (27.5%) as XDR, and nine (14.5%) as PDR. Conclusion: To reduce the threat of antimicrobial resistance, MDR, XDR and PDR A. baumannii strains must be evaluated by all clinical microbiology laboratories.

The effects of probiotics on reducing the colorectal cancer surgery complications: A periodic review during 2007-2017.

BACKGROUND & AIMS: To assess the effects of pro-/synbiotic treatment on patients with colorectal cancer (CRC), a systematic review was conducted on randomized controlled trials. METHODS: International databanks (ISI Web of Science, PubMed, Scopus, and Google Scholar) were searched from January 2007 to December 2017 using the following keywords: 'colorectal cancer' and 'probiotics'. The search was restricted to original articles published in English. Reference lists of all related studies were also reviewed to find other relevant publications. The statistical analysis was performed using SPSS software version 18.0 (IBM, NY, USA). Also, p < .05 was regarded as statistically significant. RESULTS: A total of 21 clinical trials were retrieved, involving 1831 patients subjected to elective colorectal surgery. The studies included in this review have investigated the effects of probiotics on different aspects of colorectal cancer treatment (p < .05). According to the present study results, probiotics could significantly decrease inflammatory factors, chemotherapy side effects, severe diarrhea, postoperative infectious complications, and duration of antibiotic therapy; shift fecal microbiota in favor of Actinobacteria; and change the tumor tissue microbiota (p < .05). CONCLUSION: Based on the present review, the preoperative use of pro-/synbiotics as prophylaxis for patients with CRC could improve clinical outcomes. More detailed data about the types of probiotic species and the optimal consumption dose of pro-/synbiotics should be taken in to account in future meta-analysis reviews.

Diversity of Class 1 Integrons, and Disruption of carO and dacD by Insertion Sequences Among Acinetobacter baumannii Isolates in Tehran, Iran.

Acinetobacter baumannii is an important nosocomial pathogen which causes a wide range of infections. In this study, we addressed the role of class 1 integron, ISAba1 and ISAba125 associated with antimicrobial resistance in 72 clinical isolates of A. baumannii collected from clinical settings in Tehran, Iran. Moreover, to study the clonal relatedness of strains, repetitive extragenic palindromic-PCR (rep-PCR) assay was carried out. PCR revealed that blaOXA-51-like, blaOXA-23-like, blaOXA-24/40-like, blaOXA-58-like, blaNDM, integrase gene (intI1), ISAba1, and ISAba125 were present in 86.11% (62/72), 84.72% (61/72), 30.55% (22/72), 0% (0/72), 0% (0/72), 58.33% (42/72), 97.22% (70/72), and 65.27% (47/72) of the strains, respectively. Sequencing of 39 internal variable regions of class 1 integrons showed seven gene cassette arrays, including aadA4-catB8-aadA1 (2.77%), aadA1-aadA4 (1.38%), aacC4-aadA1 (2.77%), aacC4 (22.22%), aadA1 (13.88%), aadA4 (5.55%), and catB8 (5.55%). We detected ISAba1 in the upstream of blaOXA-23-like, blaOXA-51-like, and blaADC in 54.16% (39/72), 9.72% (7/72), and 56.94% (41/72) of the strains, respectively. Whereas, there was a low frequency of disruptions in carO and dacD genes: 5.55% (4/72) and 4.16% (3/72). Rep-PCR analysis revealed that the isolates were genetically diverse. However, Cl-12 and Cl-15 were the largest clusters and they were recovered from various hospitals. Our analysis showed the high rates of class 1 integrons as a repertoire of aminoglycoside-modifying enzymes. It seems that linkages of ISAba1-blaOXA-23-like and ISAba1-blaOXA-69, and disruptions in carO or dacD can develop resistance to carbapenems among clinical isolates of A. baumannii.