High frequency of carbapenemase in extensively drug-resistant Acinetobacter baumannii isolates in central Iran.

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-ß-lactamase (MBL) by phenotypic diagnosis. The most abundant genes were blaPER (32/87; 36.7%), blaTEM (29/87; 33.3%), blaVEB (26/87; 29.8%) for ESBL and Ambler class D ß -lactamases included blaOXA-23 (69/84; 82.1%), blaOXA-24 (46/84; 54.7%), MBLs included blaVIM (51/84; 60.7%), and blaIMP (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.

In vivo efficiency of the produced recombinant lysostaphin antimicrobial peptide in treatment of methicillin-resistant Staphylococcus aureus (MRSA) skin infection in a mouse model.

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-ß-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.

High frequency of NDM-1 and OXA-48 carbapenemase genes among Klebsiella pneumoniae isolates in central Iran.

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 ß-lactamase (ESBL), AMPC ß-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 blaTEM (77/77; 100%), blaCTX-M1 (76/77; 98.7%), blaSHV (76/77; 98.7%), blaCTX-M15 (73/77; 94.8%) for ESBL; blaCIT 28 (48.3%) and blaCMY-2 26 (44.8%) for AMPC ß-lactamase; and blaOXA-48 46 (90.1%) and blaNDM 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.

The significant role of Carbapenems-resistant Acinetobacter Baumannii in mortality rate of patients with COVID-19.

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.

Antecedentes: Las infecciones causadas por Acinetobacter baumannii, especialmente las cepas resistentes a carbapenem (CR), plantean desafíos importantes en pacientes con infección por COVID-19. Por lo tanto, en el presente estudio investigamos los patrones de coinfección y resistencia a los antimicrobianos, así como el papel de A. baumannii en el desenlace de los pacientes con COVID-19.Materiales y métodos: entre febrero de 2019 y enero de 2021, se detectaron 141 pacientes con infecciones por A. baumannii de siete hospitales diferentes (A a G) en Arak, Irán, y se evaluó el patrón de susceptibilidad antibacteriana de estos aislamientos mediante difusión en disco y E- métodos de prueba. Cuarenta y siete de estos pacientes estaban coinfectados con COVID-19. Durante el estudio, se recopilaron datos sobre datos demográficos, curso clínico y el papel de las infecciones por A. baumannii en la tasa de mortalidad de los pacientes con COVID-19.Resultados: Los hospitales A y B reportaron mayor número de pacientes con 53 (38%) y 47 (44%), respectivamente. Además, la mayoría de los casos (105 casos, 75%) se informaron desde la UCI quirúrgica y general. Los ventiladores mecánicos se detectaron como factores predisponentes en 95 (67%) pacientes y la infección se detectó en el 20% de los pacientes al décimo día después de la intubación. Todos los aislamientos de A. baumannii fueron resistentes a diferentes clases de antibióticos, como los carbapenémicos. El 33,6% (47 pacientes) también dieron positivo a COVID-19, y el 68% (32 pacientes) fallecieron a causa de la infección. El análisis estadístico mostró un papel significativo de la coinfección por A. baumannii en la tasa de mortalidad de los pacientes con COVID-19 (valor de p 0,05).Conclusión: la coinfección con A. baumannii es uno de los desafíos más importantes en pacientes positivos para COVID-19. Nuestros resultados mostraron que todas las bacterias aisladas eran CR y aumentaron significativamente las tasas de mortalidad en pacientes con COVID-19.

The LL-37 Antimicrobial Peptide as a Treatment for Systematic Infection of Acinetobacter baumannii in a Mouse Model.

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.

The importance of using routine laboratory tests in the diagnosis and prognosis of patients with coronavirus disease 2019: Shedding light on clinical laboratory data in COVID-19.

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.

Common Etiological Agents in Adult Patients with Gastroenteritis from Central Iran.

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-ß-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 blaTEM (100% in both species), blaCTX-M15 (91.6% and 100%, respectively), AmpC blaCIT (80% and 100%, respectively), and blaDHA (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.

PLGA-Based Nanoplatforms in Drug Delivery for Inhibition and Destruction of Microbial Biofilm.

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.

SARS-CoV-2 Viral Shedding and Associated Factors among COVID-19 Inpatients and Outpatients.

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.

Ib-AMP4 antimicrobial peptide as a treatment for skin and systematic infection of methicillin-resistant Staphylococcus aureus (MRSA).

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.

Isolation of a lytic bacteriophage against extensively drug-resistant Acinetobacter baumannii infections and its dramatic effect in rat model of burn infection.

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.

The role of microRNAs in diseases and related signaling pathways.

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.

Frequency of Cardiovascular Complications and Its Association with Prognosis of COVID-19 Patients.

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.

Comparison of three interventional approaches to prevent ventilator-associated pneumonia in intensive care units (ICUs): A clinical trial study.

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.

Recombinant production of Trx-Ib-AMP4 and Trx-E50-52 antimicrobial peptides and antimicrobial synergistic assessment on the treatment of methicillin-resistant Staphylococcus aureus under in vitro and in vivo situations.

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_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 log10 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.

Association between colorectal cancer and Fusobacterium nucleatum and Bacteroides fragilis bacteria in Iranian patients: a preliminary study.

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.

Investigation into antibacterial and wound healing properties of platelets lysate against Acinetobacter baumannii and Klebsiella pneumoniae burn wound infections.

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.

Wound healing performance of PCL/chitosan based electrospun nanofiber electrosprayed with curcumin loaded chitosan nanoparticles.

In this study, the electrospun poly(ε-caprolactone) (PCL)/Chitosan (CS)/curcumin (CUR) nanofiber was fabricated successfully with curcumin loaded chitosan nano-encapsulated particles (CURCSNPs). The morphology of the produced CURCSNPs, PCL, PCL/CS, PCL/CS/CUR, and PCL/CS/CUR electrosprayed with CURCSNPs were analyzed by scanning electron microscopy (SEM). The physicochemical properties and biological characteristics of fabricated nanofibers such as antibacterial, antioxidant, cell viability, and in vivo wound healing efficiency and histological assay were tested. The electrospraying of CURCSNPs on surface PCL/CS/CUR nanofiber resulted in the enhanced antibacterial, antioxidant, cell proliferation efficiencies and higher swelling and water vapor transition rates. In vivo examination and Histological analysis showed PCL/CS/CUR electrosprayed with CURCSNPs led to significant improvement of complete well-organized wound healing process in MRSA infected wounds. These results suggest that the application of PCL/CS/CUR electrosprayed with CURCSNPs as a wound dressing significantly facilitates wound healing with notable antibacterial, antioxidant, and cell proliferation properties.

Quinolone resistant Salmonella species isolated from pediatric patients with diarrhea in central Iran.

BACKGROUND: This study aimed to investigate the frequency and the antibiotic resistance patterns of Salmonella species that were isolated from infectious diarrhea samples taken from pediatric patients in central Iran. METHODS: The study analyzed 230 stool specimens that were cultured on XLD, MacConkey agar and GN broth. Polymerase chain reaction (PCR) assay was used to identify the Salmonella genus. The antibiotic resistance profiles and the frequency of quinolone and integron genes were obtained. RESULTS: Out of 230 samples of infectious diarrhea, 21 (9.1%) cases of Salmonella spp. were identified using culture methods. Another 28 (12.1%) samples had positive PCR results, with S. serovar Paratyphi B and C (9/21; 42.8%) and S. Typhi (3/21; 14.3%) being the most recognized. The highest antibiotic resistance rates were found for nalidixic acid (15/21; 71.4%), tetracycline (9/21; 42.8%). However, six (28.5%) of isolates were found resistant to cotrimoxazole, ampicillin and chloramphenicol. Among the plasmid-mediated quinolone resistance (PMQR) determinants, qnrS, qnrA, and qnrB were positive in (9/15; 60%), (6/15; 40%) and (3/15; 20%) of the isolates, respectively. Class 1 and 2 integrons were identified in 15 (71.4%) and 3 (14.3%) isolates, respectively. CONCLUSION: High rates of quinolone resistant and low frequency of MDR Salmonella spp. isolates were identified in central Iran, similar to findings in other parts of Asia. To prevent the spread of these resistant strains, the antimicrobial resistance of Salmonella spp. isolates should be under constant surveillance, and empiric antibiotic therapy should be adapted appropriately.

Prevalence and molecular epidemiology of ceftaroline non-susceptible methicillin-resistant Staphylococcus aureus isolates, first clinical report from Iran.

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major pathogens in Iran with a high prevalence and a high level of antibiotic resistance. Ceftaroline is a fifth generation cephalosporin binding and inhibiting penicillin binding protein (PBP2a). METHODS: In the present study, 228 clinical MRSA isolates were collected from four cities of Iran and their susceptibility to ceftaroline was evaluated by E-test and the disk diffusion method. RESULTS: Our results showed a high susceptibility rate (97.3%) to ceftaroline in MRSA strains from Iran. Six isolates were found to be ceftaroline non-susceptible (CPT-NS) with Minimum inhibitory concentration (MIC) ≥2 µg/mL. All CPT-NS isolates were isolated from blood and tracheal aspirate and belonged to SCCmec type III as well as agr type I and were all susceptible to vancomycin. Out of six isolates, three, two and one belonged to spa type t030, t4864, and t969, respectively. Vancomycin, quinupristin/dalfopristin, linezolid, chloramphenicol, and tigecycline were the most active agents against CPT-NS isolates. CONCLUSION: Due to the broad-spectrum activity and low toxicity of ceftaroline as well as the increased rate of vancomycin resistance among MRSA strains in recent years, ceftaroline can be considered as a novel approach to treat MRSA-induced infections.