Targeted delivery of a short antimicrobial peptide (CM11) against Helicobacter pylori gastric infection using concanavalin A-coated chitosan nanoparticles.

Helicobacter pylori is the cause of most cases of stomach ulcers and also causes some digestive cancers. The emergence and spread of antibiotic-resistant strains of H. pylori is one of the most important challenges in the treatment of its infections. The present study aims to develop a concanavalin A (ConA) coated chitosan (CS) nanocarrier-based drug delivery for the targeted release of peptides to the site of H. pylori infection. Accordingly, chitosan was used as an encapsulating agent for CM11 peptide delivery by applying ionotropic gelation method. Con-A was used for coating CS nanoparticles to target H. pylori. The CS NPs and ConA-CS NPs were characterized by FTIR, dynamic light scattering (DLS), and scanning electron microscopy (SEM). The MIC of CM11-loaded ConA-CS NPs against H. pylori SS1 strain was analyzed in vitro. In order to evaluate the treatment efficiency in vivo, a gastric infection model of H. pylori SS1 strain was established in mice and histopathological studies and IL-1ß cytokine assay were performed. Based on the results, the size frequency for CS NPs and ConA-CS NPs was about 200 and 350 nm, respectively. The prepared CM11-loaded ConA-CS NPs exhibited antibacterial activity against H. pylori SS1 strain with a concentration of 32 µg/ml. The highest healing process was observed in synthesized CM11-loaded ConA-CS NPs treatments and a significant decrease in IL-1ß was observed. Our findings highlight the potential of chitosan nanoparticles as a drug delivery vehicle in the treatment of gastric infection model of H. pylori SS1 strain.

The effects of functional foods mixture on inflammatory cytokines and biochemical findings in hospitalized patients with COVID-19: a randomized double-blind controlled trial.

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic has been shown to affect nutritional recommendations. Some functional foods have been demonstrated to be useful in the treatment of people with COVID-19. However, little is known about the impact of combining functional foods on disease control. This study aimed to investigate the effects of functional foods mixture on serum levels of inflammatory cytokines and biochemical findings in patients with COVID-19. METHODS: A randomized double-blind controlled trial was conducted in Baqiyatallah Al-Azam hospital in Tehran, Iran. Sixty patients were randomly assigned to receive either a soup containing functional foods (n = 30) or a usual soup (control group) (n = 30). Participants' sociodemographic information was gathered using a general questionnaire. Blood levels of inflammatory markers and biochemical findings were assessed using standard protocols. RESULTS: The results showed that soup containing functional foods was more effective in controlling serum levels of D-dimer, blood urea nitrogen, and creatinine than the control group (P < 0.05). Also, more significant improvement was found in the intervention group vs control group in terms of interleukin (IL)-1ß, IL-6, IL-17, IL-10, and tumor necrose factor-α (P < 0.05). In contrast, the control intervention more efficiently controlled potassium levels and reduced quantitative C-reactive protein than the intervention group (P < 0.05). CONCLUSIONS: This study indicates a soup containing functional foods could alleviate biomarkers of inflammation in patients with COVID-19. However, its effectiveness on biochemical findings remained inconclusive which warranted further research. TRIAL REGISTRATION: IRCT, IRCT20180201038585N11. Registered 23 August 2021, https://www.irct.ir/trial/57338.

Molecular Dynamics Simulation of Antimicrobial Peptide CM15 in Staphylococcus Aureus and Escherichia coli Model Bilayer Lipid.

Background: In animals and plants, antimicrobial peptides (AMPs) are crucial components of defense mechanisms, as they play a crucial role in innate immunity, which protects hosts from pathogenic bacteria. The CM15 has attracted considerable interest as a novel antibiotic against gram-negative and positive pathogens. Objective: The aim of this study was to investigate the permeation potential of the CM15 with membrane bilayers of Staphylococcus aureus and Escherichia coli. Material and Methods: The bilayer membranes of Escherichia coli and Staphylococcus aureus were modelled with the resemblance in lipid composition to its biological sample. This study followed Protein-Membrane Interaction (PMI) through successive applications of molecular dynamics simulation by GROMACS and CHARMM36 force field for two sets of 120-ns simulations. Results: Significant results were obtained from analyzing the trajectory of the unsuccessful insertion of CM15 during simulation. Our data suggested that Lysine residues in CM15 and Cardiolipins in membrane leaflets play a crucial role in stability and interaction terms. Conclusion: The obtained results strengthen the insertion possibility through the toroidal model, which should consider for further studies on AMPs interaction.

Evaluation of an antibacterial peptide-loaded amniotic membrane/silk fibroin electrospun nanofiber in wound healing.

Antimicrobial peptides (AMPs) are among the compounds that have significant potential to deal with infectious skin wounds. Using wound dressings or skin scaffolds containing AMPs can be an effective way to overcome infections caused by antibiotic-resistant strains. In this study, we developed an amniotic membrane-based skin scaffold using silk fibroin to improve mechanical properties and CM11 peptide as an antimicrobial peptide. The peptide was coated on the scaffold using the soaking method. The fabricated scaffold was characterised by SEM and FTIR, and their mechanical strength, biodegradation, peptide release, and cell cytotoxicity analyses were performed. Then, their antimicrobial activity was measured against antibiotic-resistant strains of Pseudomonas aeruginosa and Staphylococcus aureus. The in vivo biocompatibility of this scaffold was evaluated by subcutaneously implanting it under the skin of the mouse and counting lymphocytes and macrophages in the implanted area. Finally, the regenerative ability of the scaffold was analyzed in the mouse full-thickness wound model by measuring the wound diameter, H&E staining, and examining the expression rate of genes involved in the wound healing process. The developed scaffolds exerted an inhibiting effect on the bacteria growth, indicating their proper antimicrobial property. In vivo biocompatibility results showed no significant count of macrophages and lymphocytes between the test and control groups. The wound closure rate was significantly higher in the wound covered with fibroin electrospun-amniotic membrane loaded with 32 µg/mL CM11, where the relative expression rates of collagen I, collagen III, TGF-ß1 and TGF-ß3 were higher compared with the other groups.

The effect of bergamot (KoksalGarry) supplementation on lipid profiles: A systematic review and meta-analysis of randomized controlled trials.

This systematic review and meta-analysis were conducted to evaluate the impact of bergamot (KoksalGarry) and its nutraceutical compounds on lipid profiles. PubMed, Web of Knowledge, Scopus, and Google Scholar searched for relevant articles. Trials investigating the effect of oral bergamot supplementation on serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) in adults were included. The mean differences and standard deviations were pooled using a random-effects model. Fourteen trials were included in this systematic review and meta-analysis. Bergamot supplementation significantly decreased serum levels of TC (weighted mean difference (WMD): -63.60 mg/dL; 95% CI: -78.03 to -49.18; p < .001), TG (WMD: -74.72 mg/dL; 95% CI: -83.58 to -65.87; p < .001), LDL-C (WMD: -55.43 mg/dL; 95% CI: -67.26 to -43.60; p < .001), and increased HDL-C (WMD: 5.78 mg/dL; 95% CI: 3.27 to 8.28; p < .001), respectively. Our systematic review of the effects of nutraceuticals containing bergamot on lipid markers showed inconsistent results. The results showed that bergamot supplementation might improve lipid profiles. The findings for nutraceutical compounds containing bergamot were inconsistent. However, the clinical efficacy of bergamot on lipid profiles needs to be further established through higher-quality studies.

Twelve weeks of treadmill exercise training with green tea extract reduces myocardial oxidative stress and alleviates cardiomyocyte apoptosis in aging rat: The emerging role of BNIP3 and HIF-1α/IGFBP3 pathway.

In this study, we consider the effect of treadmill exercise training, green tea extract, and combination of exercise training with green tea extract, in aging rat cardiac myocytes apoptosis markers (i.e., HIF-1α, BNIP3, Bax, IGFBP3, Bcl-2, caspase-3, MDA, GPx, Bax/Bcl-2 ratio, and hematoxylin and eosin). Twenty-four rats (male, Wistar) were divided into four groups: (I) control (n = 6), (II) green tea extract (n = 6), (III) exercise (n = 6), and (IV) exercise + green tea extract (n = 6). Exercise groups performed 12 weeks of running on a rodent treadmill at 17-27 m.min-1 (60-75% vo2peak) for 5 days per week. Green tea extract involved 300 mg.kg-1 , 5 days per week for 12 weeks. After being euthanized, the blood and heart were collected for glutathione peroxidase (GPx) activity, malondialdehyde (MDA), HIF-1α, BNIP3, insulin-like growth factor-binding protein-3 (IGFBP3), Bax, Bcl-2, caspase-3, Bax/Bcl-2 ratio, and hematoxylin and eosin level measurements. Compared to control, the ANOVA demonstrated significant effects of green tea extract (F = 14.646 to 32.453, p = .009 to .001, η = 0.295 to 0.715) and exercise training (F = 9.213 to 133.828, p = .007 to .001, η = 0.315 to η = 0.870) on HIF-1a, BNIP3, Bax, IGFBP3, Bcl-2, caspase-3, MDA, GPx, and Bax/Bcl-2 ratio. However, the combination of green tea extract and exercise had no effect on the aforementioned apoptosis markers when compared to isolated green tea extract or isolated exercise (F = 0.002 to 4.068, p = .057 to .968, and η = 0.001 to 0.169). PRACTICAL APPLICATIONS: Isolated exercise training and green tea extract may provide a cardioprotective effect on aging-induced apoptosis through the downregulation of HIF-1α, BNIP3, and IGFBP3 in the heart muscle. However, further research is needed to clarify the effects of combining exercise and green tea.

Design of two immunotoxins based rovalpituzumab antibody against DLL3 receptor; a promising potential opportunity.

Background and purpose: The lack of a new effective treatment for small cell lung cancer (SCLC) is an unresolved problem. Due to the new identification of delta-like ligand 3 (DLL3) and its high expression in SCLC patients, the use of DLL3 in target therapy can be effective. The use of bacterial toxins belonging to the ADP-ribosyl transferase toxins family and human enzymes to remove cancerous cells has been effective in the structure of immunotoxins. In this study, single-chain fragment variable of rovalpituzumab antibody fused to granzyme B (Rova-GrB) and PltA of typhoid toxin (Rova-Typh) as immunotoxins were designed, and bioinformatics analysis was done. Experimental approach: In silico analysis including the physicochemical properties, evaluation of the secondary and tertiary structure, refinement and validation of 3D models, and docking were performed. Immunotoxin genes were cloned and expressed in the Escherichia coli BL21 (DE3) host, purified, subsequently confirmed by western blotting and their secondary structure was evaluated by the circular dichroism method. Findings/Results: The bioinformatics analysis showed that Rova-GrB and Rova-Typh had hydrophilic properties, their codon optimization parameters were standard, validation parameters were improved after immunotoxin refinement, and docking analysis showed that the binding domain of immunotoxins could bind the N-terminal region of DLL3. immunotoxins had high expression and after purification under denaturing condition by Ni-NTA column, the immunotoxins were dialyzed against PBS buffer. Conclusion and implications: The immunotoxins had the right structure and can be produced in a prokaryotic host. The recombinant immunotoxins against DLL3 can be promising therapeutic agents for SCLC cancer.

Rescuing humanity by antimicrobial peptides against colistin-resistant bacteria.

It has been about a century since the discovery of the first antibiotic, and during this period, several antibiotics were produced and marketed. The production of high-potency antibiotics against infections led to victories, but these victories were temporary. Overuse and misuse of antibiotics have continued to the point that humanity today is almost helpless in the fight against infection. Researchers have predicted that by the middle of the new century, there will be a dark period after the production of antibiotics that doctors will encounter antibiotic-resistant infections for which there is no cure. Accordingly, researchers are looking for new materials with antimicrobial properties that will strengthen their ammunition to fight antibiotic-resistant infections. One of the most important alternatives to antibiotics introduced in the last three decades is antimicrobial peptides (AMPs), which affect a wide range of microbes. Due to their different antimicrobial properties from antibiotics, AMPs can fight and kill MDR, XDR, and colistin-resistant bacteria through a variety of mechanisms. Therefore, in this study, we intend to use the latest studies to give a complete description of AMPs, the importance of colistin-resistant bacteria, and their resistance mechanisms, and represent impact of AMPs on colistin-resistant bacteria. KEY POINTS: • AMPs as limited options to kill colistin-resistant bacteria. • Challenge of antibiotics resistance, colistin resistance, and mechanisms. • What is AMPs in the war with colistin-resistant bacteria?

The antimicrobial effect of quorum sensing autoinducers of Pseudomonas aeruginosa, C12-HSL and C4-HSL, against MDR Staphylococcus aureus isolates.

In the current study, we investigated the antibacterial activity of main quorum sensing autoinducers of Pseudomonas aeruginosa, C12-HSL and C4-HSL, against MDR Staphylococcus aureus isolates and their synergistic effects with some common antibiotics. Forty clinical isolates of S. aureus were collected and their antibiotic susceptibility pattern was evaluated. Then, 10 resistant isolates were selected for further studies. In the following, the antibacterial activity of quorum sensing C12-HSL and C4-HSL inducers of P. aeruginosa was evaluated against selected isolates based on the microdilution method and Time Killing assay as well as their synergistic activity with selected antibiotics. The ability of inductors to hemolysis and their cytotoxicity on CHO and HeLa cell lines was also assessed. For the assessment of antibacterial activity, Acinetobacter baumannii was used as negative control. The results demonstrated that C12 and C4 have antibacterial activity against MDR S. aureus isolates but had no effect on A. baumannii. Time Killing test showed that at 2X MIC concentration, the maximum inhibition (100%) is observed after 120 min for C12 and 240 min for C4. The IC50 of inducers was about 512 µg/ml. In addition, no synergistic effects were observed.

A contemporary review on the important role of in silico approaches for managing different aspects of COVID-19 crisis.

In the last century, the emergence of in silico tools has improved the quality of healthcare studies by providing high quality predictions. In the case of COVID-19, these tools have been advantageous for bioinformatics analysis of SARS-CoV-2 structures, studying potential drugs and introducing drug targets, investigating the efficacy of potential natural product components at suppressing COVID-19 infection, designing peptide-mimetic and optimizing their structure to provide a better clinical outcome, and repurposing of the previously known therapeutics. These methods have also helped medical biotechnologists to design various vaccines; such as multi-epitope vaccines using reverse vaccinology and immunoinformatics methods, among which some of them have showed promising results through in vitro, in vivo and clinical trial studies. Moreover, emergence of artificial intelligence and machine learning algorithms have helped to classify the previously known data and use them to provide precise predictions and make plan for future of the pandemic condition. At this contemporary review, by collecting related information from the collected literature on valuable data sources; such as PubMed, Scopus, and Web of Science, we tried to provide a brief outlook regarding the importance of in silico tools in managing different aspects of COVID-19 pandemic infection and how these methods have been helpful to biomedical researchers.

In Silico Analysis of Inhibiting Papain-like Protease from SARS-CoV-2 by Using Plant-Derived Peptides.

SARS-CoV-2 is a corona virus that has been the cause for one of the deadliest pandemics of history, started since 2019. Suppressing the activity of the critical enzymes in the SARS-CoV-2 could potentially inhibit a vital step in viral life cycle. Papain-like protease (PLpro) could be regarded as a critical enzyme in viral replication of SARS-CoV-2. In this research, it was aimed to suppress the activity of PLpro enzyme by using potential plant-derived protease inhibitor peptides. For this purpose, 11 plant derived peptides that could potentially inhibit protease activity were selected from literature. The structures of the PLpro and the peptide ligands were acquired from PDB (protein data bank) and after structural optimization, were docked by using HADDOCK 2.4 program. Analyzing the results indicated that VcTI from Veronica hederifolia provides effective molecular interactions at both liable Zn site and classic active site of PLpro, making it a potential inhibitory ligand for this enzyme that could be used for halting the replication of SARS-CoV-2. Molecular dynamic assay confirmed that the selected receptor and ligand complex was stable. Future in vitro and in vivo investigations are required to verify the efficiency of this compound as a potential therapeutic against SARS-CoV-2 infection. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s10989-021-10331-8.

Supplementation of zebrafish (Danio rerio) diet using a short antimicrobial peptide: Evaluation of growth performance, immunomodulatory function, antioxidant activity, and disease resistance.

Short-chain bioactive peptides are new and promising antimicrobial, immune moderating, and antioxidant agents. Therefore, the present study was conducted to evaluate in vitro antibacterial activity of CM11, a short antimicrobial peptide (AMP), against Streptococcus iniae and Yersinia ruckeri as fish pathogenic bacteria using standard disk diffusion and microdilution assays. In addition, in vivo effects of CM11 on fish growth, immunity, antioxidant activity, and disease resistance were evaluated using zebrafish (Danio rerio) as an animal model. For in vivo study, based on in vitro susceptibility results, four diets were designed to include zero (as control), 10, 20, and 50 µg of CM11 per g diet referred to as control, P1, P2, and P3 treatments, respectively. After eight weeks of dietary trial, fish were challenged with Streptococcus iniae, and the survival rate was calculated for a period of two weeks. Results showed that CM11 effectively inhibited the growth of S. iniae and Y. ruckeri on agar plates at a concentration of eight µg/ml. Minimum inhibitory and minimum bactericidal concentrations of CM11 were measured at 8 and 32 µg/ml for S. iniae and 16 and 64 µg/ml Y. ruckeri, respectively. In vivo results showed no noticeable effects on fish growth parameters, however, feed conversion ratio (FCR) was found lower in P3 and P2 compared to control (P < 0.05). Immunological and antioxidant responses were found strongly affected by CM11 in all treatment groups in which the highest values were found in the P3 treated group. Key immune and antioxidant genes were up-regulated particularly in fish receiving the highest level of CM11 (P3). Fish receiving the CM11 peptide showed better survival when challenged with S. iniae. These findings suggest the potential of CM11 for use in aquaculture as an antibacterial and immunostimulant agent.

Conjugation of imipenem to silver nanoparticles for enhancement of its antibacterial activity against multi-drugresistant isolates of Pseudomonas aeruginosa.

Due to the broad-spectrum of antibiotic resistance, herein we investigated the possibility of using imipenemconjugated silver nanoparticles (IMP-AgNPs) against multidrug-resistant isolates of Pseudomonas aeruginosa. For this purpose, 200 clinical isolates were tested against different antibiotics to determine the antimicrobial susceptibility. To identify blaVIM and blaIMP resistance genes, PCR was used. The synthesized AgNPs and conjugants were characterized using UV-vis spectroscopy, XRD, SEM, TEM, DLS, and FTIR. The stability, drug release kinetics, cytotoxicity, hemolytic and apoptotic effects of NPs were also investigated. MIC of the imipenem, AgNPs, and conjugants were evaluated versus P. aeruginosa isolates. Finally, the effects of the IMP-AgNPs to heal burn wounds in rats was evaluated. According to the results, about 68% of isolates showed resistance to imipenem (MIC ≥ 64 µg/ml to ≥ 512 µg/ml). Analytical results verified the synthesis of AgNPs and IMP-AgNPs. A Dose-dependent decrease happened in terms of the MIC values of IMP-AgNPs were also affected by the existence of resistant genes. Low cytotoxic was observed regarding AgNPs which lead to apoptosis. The histopathological results showed a considerable epithelization in treated groups with IMPAgNPs. Accordingly, IMP-AgNPs can be considered as a powerful antibacterial agent to treat the infections caused by multidrug-resistant P. aeruginosa.

Antiviral peptides against Coronaviridae family: A review.

The Coronaviridae family comprises large enveloped single-stranded RNA viruses. The known human-infecting coronaviruses; severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), novel SARS-CoV-2, human coronavirus (HCoV)-NL63, HCoV-229E, HCoV-OC43 and HKU1 cause mild to severe respiratory infections. The viral diseases induced by mammalian and avian viruses from Coronaviridae family pose significant economic and public health burdens. Due to increasing reports of viral resistance, co-infections and the emergence of viral epidemics such as COVID-19, available antiviral drugs show low or no efficacy, and the production of new treatments or vaccines are also challenging. Therefore, demand for the development of novel antivirals has considerably increased. In recent years, antiviral peptides have generated increasing interest as they are from natural and computational sources, are highly specific and effective, and possess the broad-spectrum activity with minimum side effects. Here, we have made an effort to compile and review the antiviral peptides with activity against Coronaviridae family viruses. They were divided into different categories according to their action mechanisms, including binding/attachment inhibitors, fusion and entry inhibitors, viral enzyme inhibitors, replication inhibitors and the peptides with direct and indirect effects on the viruses. Reported studies suggest optimism with regard to the design and production of therapeutically promising antiviral drugs. This review aims to summarize data relating to antiviral peptides particularly with respect to their applicability for development as novel treatments.

Swimming exercise improves gene expression of PPAR-γ and downregulates the overexpression of TLR4, MyD88, IL-6, and TNF-α after high-fat diet in rat skeletal muscle cells.

Exercise training with anti-inflammatory effects can improve insulin sensitivity in muscle tissue. This study investigated the effects of eight-week swimming exercises on lipid profile, toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and peroxisome proliferator-activated receptor gamma (PPAR-γ) in gastrocnemius muscle of rats fed with high-fat diet (HFD). Thirty-two healthy male Wistar rats (8 weeks, 200 ± 20 g) were randomly divided into four groups (n = 8 each group): the control (C), aerobic exercise (E), HFD, and HFD + aerobic exercise (HFD & E). The exercise training protocol consisted of swimming 60 min/day, 5 days/week for eight weeks. Serum levels of glucose, insulin, and lipid profile were measured at end of the study. Protein expressions of TLR4, TNF-α, and IL-6 were determined by immunohistochemical method. Gene expression of TLR4/MyD88, TNF-α, IL-6, and PPAR-γ was evaluated by a real-time polymerase chain reaction in gastrocnemius muscle. HFD fed rats showed higher levels of cholesterol and LDL-c that were similar in weight gain. Meanwhile, the HFD group had a higher gene expression of TLR4, MyD88, TNF-α, IL-6, and lower gene expression of PPAR-γ compared to the control group (p < 0.05). Muscle protein expression of TLR4, TNF-α, IL-6 was lower in the E and HFD&E groups (especially when compared to HFD group, P < 0.05). We also showed a decrease in TLR4/MyD88 mRNA and an increase in PPAR-γ mRNA in gastrocnemius of E and HFD&E groups (compared to HFD group, p < 0.05). Insulin resistance in HFD&E groups show a significant decrease compared to the HFD group (p < 0.05). It seems that swimming aerobic exercise for eight weeks controlled the destructive effects of HFD on muscle inflammatory pathways along with the down-regulation of the TLR4/MyD88, inflammatory cytokine, and up-regulation PPAR-γ mRNA. It appears that the down-regulation in the expression of TLR4/MyD88 mRNA reduces the muscle pro-inflammatory cytokines, such as IL-6 and TNF-α, whose action may be caused by the adaptation of swimming aerobic exercise (an increase of PPAR-γ). Therefore, local and systemic inflammatory changes due to HFD and obesity may be affected by metabolic adaptations of aerobic exercise training, which requires further studies.

Molecular Detection of Sexually Transmitted Infections in Women with and without Human Papillomaviruses Infection Who Referred to Tehran West Hospitals in Iran.

BACKGROUND: According to the studies, many pathogens function as cofactors interacting with Human papillomavirus in the development of pre-cancer or cancer of the cervix. The aim of this study was to investigate the prevalence rate of Sexually Transmitted Infections (STIs) pathogens including Mycoplasma hominis, Ureaplasma urealyticum, Chlamydia trachomatis, Neisseria gonorrhoeae, Gardnerella vaginalis, and Streptococcus agalactiae in people with HPV and without HPV infection, and frequency rate of these pathogens in high and low risk of HPV. METHODS: Cervical samples of 280 women who referred to Tehran west hospitals in Iran, between 2019 and 2020, were collected. After DNA extraction of samples, identification of HPV and genotyping was performed, and then, to detect each microorganism, the PCR was carried out with specific primers. Finally, the results were analyzed using descriptive statistics tests. RESULTS: The mean age of patients was 37 years. Two groups of patients were identified based on positivity or negativity of HPV. In HPV-positive group (118 cases), the prevalence of U. urealyticum, M. hominis, N. gonorrhoeae, G. vaginalis, and S. agalactiae was 38 (13%), 7 (62%), 5.93%, 19.49%, 0.84% respectively. In HPV-negative group (162 cases), rate of infection with U. urealyticum, M. hominis, N. gonorrhoeae, G. vaginalis, and S. agalactiae was 29.62%, 6.17%, 3.08%, 16.04%, 0.61% respectively. Among the two groups, there was only 1 patient with C. trachomatis (0.84%), seen in HPV-positive group. CONCLUSION: In this study no significant association was found between HPV and bacteria such as G. vaginalis and S. agalactiae, and it was found that C. trachomatis, and especially N. gonorrhoeae are strongly associated with HPV infection.

Frequency of bla CTX-M and bla TEM Virulence Genes and Antibiotic Resistance Profiles among Klebsiella pneumoniae Isolates in Urinary Tract Infection (UTI) Samples from Hashtgerd, Iran.

BACKGROUND: Klebsiella pneumoniae (K. pneumoniae) is an opportunistic microorganism and one of the most important causes of urinary tract infection. This study aimed to evaluate the frequency of K. pneumoniae producing broad-spectrum beta-lactamase in urinary tract infection and to determine the pattern of drug resistance. METHODS: This study was performed on 50 samples of K. pneumoniae isolated from patients with urinary tract infection referred to the Medical Diagnostic Laboratory in Hashtgerd city. The isolates were first evaluated for antibiotic susceptibility by disk diffusion method according to the method proposed by the Clinical and Laboratory Standards Institute (CLSI). Then phenotypic detection of ESBLS was carried out by the DDST method. The frequency of gene bla TEM and bla CTX-M was determined by PCR. RESULTS: The highest resistance was observed to ampicillin (94%) and the highest sensitivity was observed to gentamicin (84%). 22 isolates (44%) were positive for ESBLs production. Of the 50 isolates studied, 34% had bla CTX-M and 28% had bla TEM and 11 (22%) had both genes simultaneously. Also, more than 77% of positive ESBLs isolates had the bla CTX-M gene and approximately 63.64% of positive ESBLs isolates had the bla TEM gene. CONCLUSION: Given the high prevalence of antibiotic-resistant and ESBL-producing isolates, early identification of these resistant isolates and their follow-up is essential to prevent further outbreaks. It is also important to use appropriate therapeutic strategies and proper and rational administration of antibiotics by physicians.

Molecular Detection and Evaluation of MLـ Resistance M. Pneumoniae Associated with Mutation in 23S RNA Gene among Iranian Patients with Respiratory Infections.

BACKGROUND: Mycoplasma pneumoniae is a common cause of community-acquired pneumonia. The global increased resistance of M. pneumoniae strains to macrolide (ML) has become a worrisome health problem. The widespread use of these medications has led to increased rate of reported ML-resistant M. pneumoniae (MRMP) throughout the world. This study was aimed to evaluate the resistance of M. pneumoniae against erythromycin due to mutations in the 23S rRNA gene of patients with respiratory infections in Iran. METHODS: In this study, 100 samples of throat swab from a patient with respiratory problems were collected. After the cultured of all samples in M. pneumonia-specific PPLO medium, PCR technique was performed with specific primers. Afterwards, the broth micro-dilution MIC assay was employed. Finally, the PCR product of the 23S rRNA gene was sequenced to detect mutations of domain V in 23S rRNA gene of MRMP. RESULTS: It was found that 17 cases (17%) were positive for mycoplasma genus and six cases (6%) positive for M. pneumoniae species. Also, analysis of the sequence of 23S rRNA gene, revealed that one of the samples had mutations at positions A2431G and G2491A. All positive samples M. pneumoniae with 23S rRNA gene were sensitive to erythromycin. CONCLUSION: These use of these antibiotics should be limited to prevent the emergence of MRMP in Iran.

Antibiotic resistance assessment of Acinetobacter baumannii isolates from Tehran hospitals due to the presence of efflux pumps encoding genes (adeA and adeS genes) by molecular method.

OBJECTIVE: Acinetobacter baumannii (A. baumannii) has caused many problems in nosocomial infections. Efflux pumps are considered as one of the most important mechanisms of resistance in this bacterium and have the ability to excrete toxic substances such as antibiotics out of the cell. RESULTS: In this study, 60 isolates of A. baumannii were collected from patients in several hospitals in Tehran, Iran. After diagnosis using standard biochemical methods, the pattern of antibiotic susceptibility was determined using the disk diffusion method according to CLSI guidelines. The adeA and adeS genes were identified by PCR method. The highest resistance to Piperacillin and the lowest resistance to Gentamicin were observed (100% compared to 48.4%). 6.6% of the isolates had only adeA gene and adeS gene was observed in 8.4% of isolates and both genes were detected in 73.4% of the samples. Despite the high resistance of t A. baumannii o antibiotics and due to the high frequency of genes of adeA and adeS efflux pumps in A. baumannii isolates, it can be concluded that these efflux pumps may play an important role in resistance of this bacterium. By determining the pattern of antibiotic the resistance before treatment, the resistance of this pathogen can be prevented in societies.

The Efficacy of AgNO3 Nanoparticles Alone and Conjugated with Imipenem for Combating Extensively Drug-Resistant Pseudomonas aeruginosa.

INTRODUCTION: The extensive drug-resistant (XDR) Pseudomonas aeruginosa (P. aeruginosa) causes a range of infections with high mortality rate, which inflicts additional costs on treatment. The use of nano-biotechnology-based methods in medicine has opened a new perspective against drug-resistant bacteria. The aim of this study was to evaluate the effectiveness of the AgNO3 nanoparticles alone and conjugated with imipenem (IMI) to combat extensively drug-resistant P. aeruginosa. METHODS: Antibiotic susceptibility was carried out using disc diffusion method. Detection of different resistant genes was performed using standard polymerase chain reaction (PCR). The chemically synthesized AgNO3 particles were characterized using scanning electron microscope (SEM), dynamic light scattering (DLS) and X-ray diffraction (XRD) methods. Fourier transform infrared spectroscopy (FTIR) was accomplished to confirm the binding of AgNO3 with IMI. The microdilution broth method was used to obtain minimum inhibitory concentration (MIC) of AgNO3 and IMI-conjugated AgNO3. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was carried out on L929 cell line to study the cytotoxicity of nanoparticles. The data were analyzed by Eta correlation ratio and chi-square (X 2) test. RESULTS: Analysis of the antibiotic resistance pattern showed that 12 (24%) isolates were XDR, and MIC values of IMI were between 64 and 128 µg/mL. Frequency of SHV, TEM, CTX M, IMP, VIM, OPR, SIM, SPM, GIM, NDM, VEB, PER, KPC, OXA, intI, intII, and intIII genes were 29 (58%), 26 (52%), 26 (52%), 32 (64%), 23 (46%), 43 (86%), 3 (6%), 6 (12%), 3 (6%), 4 (8%), 7 (14%), 6 (12%), 18 (36%), 4 (8%), 19 (38%), 16 (32%), and 2 (4%), respectively. The XRD, SEM, DLS, and FTIR analysis confirmed the synthesis of AgNO3 nanoparticles and their conjugation with IMI. The AgNO3 nanoparticles had antimicrobial activity, and their conjugation with IMI showed enhanced effectiveness against XDR isolates. The synthesized AgNO3 showed no cytotoxic effects. CONCLUSION: The results suggest that IMI-conjugated AgNO3 has a strong potency as a powerful antibacterial agent against XDR P. aeruginosa.