In Silico Screening and Experimental Validation of Novel MexAB-OprM Efflux Pump Inhibitors of Pseudomonas aeruginosa.

The emergence of multidrug-resistant Pseudomonas aeruginosa possesses a significant public health concern. Constitutively expressed MexAB-OprM efflux pumps in P. aeruginosa significantly contribute to its resistance to a variety of antibiotics. The development of efflux pump inhibitors (EPIs) has emerged as an attractive strategy in reversing antibiotic resistance. In this study, structure-based virtual screening techniques were used for the identification of new MexAB-OprM efflux inhibitors. The predicted poses were thoroughly filtered by induced fit docking procedures followed by in vitro microbiological assays for the validation of in silico results. Two compounds, NSC-147850 and NSC-112703, were able to restore tetracycline susceptibility in MexAB-OprM overexpressing Pseudomonas aeruginosa ATCC® 27853™ strain. This correlation observed between in silico screening and positive efflux inhibitory activity in vitro suggests that NSC-147850 and NSC-112703 have potential as EPIs and may be effective in combination therapy against drug-resistant strains of P. aeruginosa.

Identification of Drug Combination Therapies for SARS-CoV-2: A Molecular Dynamics Simulations Approach.

Purpose: The development of effective treatments for coronavirus infectious disease 19 (COVID-19) caused by SARS-Coronavirus-2 was hindered by the little data available about this virus at the start of the pandemic. Drug repurposing provides a good strategy to explore approved drugs' possible SARS-CoV-2 antiviral activity. Moreover, drug synergism is essential in antiviral treatment due to improved efficacy and reduced toxicity. In this work, we studied the effect of approved and investigational drugs on one of SARS-CoV-2 essential proteins, the main protease (Mpro), in search of antiviral treatments and/or drug combinations. Methods: Different possible druggable sites of Mpro were identified and screened against an in-house library of more than 4000 chemical compounds. Molecular dynamics simulations were carried out to explore conformational changes induced by different ligands' binding. Subsequently, the inhibitory effect of the identified compounds and the suggested drug combinations on the Mpro were established using a 3CL protease (SARS-CoV-2) assay kit. Results: Three potential inhibitors in three different binding sites were identified; favipiravir, cefixime, and carvedilol. Molecular dynamics simulations predicted the synergistic effect of two drug combinations: favipiravir/cefixime, and favipiravir/carvedilol. The in vitro inhibitory effect of the predicted drug combinations was established on this enzyme. Conclusion: In this work, we could study one of the promising SARS-CoV-2 viral protein targets in searching for treatments for COVID-19. The inhibitory effect of several drugs on Mpro was established in silico and in vitro assays. Molecular dynamics simulations showed promising results in predicting the synergistic effect of drug combinations.

Azithromycin Misuse During the COVID-19 Pandemic: A Cross-Sectional Study from Jordan.

Objective: Since coronavirus disease 2019 (COVID-19) became a global pandemic, repurposing known drugs was the quickest way to combat the disease. The initial screening revealed that azithromycin (AZM) might have potential against COVID-19. Although clinical trials did not prove such efficacy, many countries have put AZM within their guidelines for treating COVID-19. Therefore, the present study was designed to assess the misuse of AZM in Jordan during the COVID-19 pandemic. Methods and Results: A cross-sectional study was conducted among community pharmacies in Jordan from March 27 to May 8, 2021, and 184 pharmacies data were collected from the Google forms. During COVID-19, 42.9% of pharmacies sold more than 20 packs of prescribed AZM per month compared to 46.7% of pharmacies used to sell 0-5 packs AZM prescriptions per month before the pandemic. During COVID-19, pharmacists significantly dispensed AZM with and without prescriptions 107% and 127%, respectively, more than before the pandemic (p < 0.0001). Overall, pharmacists stocked 121% more AZM packs during COVID-19 than before the pandemic (p < 0.0001). Additionally, most pharmacists (59.7%) believed that AZM could cure COVID-19 patients. However, using multinomial logistic regression analysis, low-experienced pharmacists were unsure if AZM could positively affect COVID-19 patients (p < 0.05, OR = 3.76, 95% CI = 1.23-11.52). Furthermore, low-experienced pharmacists believed that increased use of AZM for the treatment of viral infections could lead to negative consequences (p < 0.001, OR = 0.161, 95% CI 0.063-0.414). Conclusion: This study demonstrated that AZM is misused by physicians, pharmacists, and the public in Jordan. Since AZM efficacy on SARS-CoV-2 is scarce, there is a need for new guidelines by governmental health authorities to implement strict enforcement of AZM dispensing during COVID-19 to avoid negative consequences of bacterial resistance.

The effect of cigarette smoke condensate (CSC) on Pseudomonas aeruginosa virulence and antibiotic sensitivity.

AIMS: Despite a decline in tobacco smoking in the developed world, the developing world has witnessed an increase in such activity over recent years. An increase in antibiotic resistance has accompanied this increase in tobacco use, and we suggest that the two may be linked. This study aims to investigate the effect of cigarette smoke exposure on bacterial virulence and susceptibility to antibiotics. METHODS AND RESULTS: Pseudomonas aeruginosa passaged in the presence of Cigarette Smoke Condensate (CSC) exhibited reduced susceptibility towards Amikacin (p = 0.02), Tobramycin (p = 0.03) and Aztreonam (p = 0.007) and was accompanied by changes in growth dynamics as exposure to CSC increased. These observed changes persisted after passaging bacteria in CSC-free medium for 10 days. The genotoxicity of CSC on P. aeruginosa was evaluated by the standard Comet assay, which demonstrated DNA damage in the P. aeruginosa genome in Passage 15 compared to the CSC-unexposed cells. Gene expression analysis on selected virulence and quorum sensing genes showed that both flagellar (fliC and fleR) and quorum sensing (lasI/R and rhII) genes were significantly up-regulated in Passage 15. CONCLUSIONS: Results confirm the genotoxic effect of cigarette smoke manifested in an increased antibiotic resistance, coupled with increased bacterial virulence SIGNIFICANCE AND IMPACT OF STUDY: This study is the first to elucidate a clear link between tobacco smoke residues and both increases in antibiotic resistance and the up-regulation of bacterial virulence markers.

Methicillin-Resistant Staphylococci (MRS): Carriage and Antibiotic Resistance Patterns in College Students.

Asymptomatic carriage of methicillin-resistant Staphylococci (MRS) may allow for the unseen dissemination of antibiotic-resistant strains through the population. This study investigates the prevalence and epidemiological risk factors that contribute to the spread of MRS in a university setting in Amman, Jordan. A cross-sectional questionnaire-based study was performed in December 2019. Five hundred and four students enrolled in the study and provided skin and nasal swabs. Swabs were then processed to isolate MRS on Mannitol Salt Agar (MSA) + 4 µg/ml oxacillin. Isolates were tested for their antibiotic susceptibility using the disc diffusion assay. Epidemiological risk assessment was performed using the Chi-square test and univariate and multivariant analysis. The percentage carriage of MRS in the 504 students was 40.4%. The carriage rate of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus epidermidis (MRSE) from the skin and nasal areas was 13.5% and 26.9%, respectively. The percentage of male carriers was significantly higher than females, and the only identified epidemiological risk factor related to the carriage was attendance at a fitness club. All MRS isolates were resistant to oxacillin (100%), cefoxitin (45.5%), erythromycin (35.2%), gentamycin (10.2%), ciprofloxacin (12.7%), nitrofurantoin (12.2%), linezolid (7.8%), amikacin (1.47%), and Vancomycin (0.49%). 42% of MRS expressed a multiple antibiotic resistance (MAR) index above 0.2. Three isolates expressed a MAR index of 0.8. MRS has been exhibited to be present in an otherwise healthy population of students, which may then act as a reservoir for MAR strains.

Changes in public knowledge and perceptions about antibiotic use and resistance in Jordan: a cross-sectional eight-year comparative study.

BACKGROUND: Resistance to antibiotics is a growing problem, worldwide and particularly in developing countries like Jordan. Raising public awareness on appropriate antibiotic use is crucial to combat this problem. The current study describes the change in public Knowledge and attitudes towards the use of antibiotics over a period of 8 years. METHODS: Two cross-sectional studies were performed 8 years apart on Jordanians of different age groups, and social settings, residing in Amman, Jordan. Convenience non-probability sampling techniques were used. In 2010, a questionnaire was distributed in paper form, whereas in 2018 snowball sampling was used to disseminate an identical electronic questionnaire. Chi-square test and post hoc analysis were done using the z-test to compare column proportions, adjustment for multiple testing using the Bonferroni method. Multiple logistic regression was used to adjust for case mix for each survey. Comparisons were made across the two studies and within each study. RESULTS: A total of 711 participants in 2010 and 436 participants in 2018 were surveyed. Over the 8-year period, there was a significant improvement in the beliefs regarding the use of antibiotics such as disagreeing to keeping left over antibiotics for later use from 57 to 70% (p < 0.05) and disagreeing to buying antibiotics without physicians' consent increased from 80 to 89% (P value < 0.001). There was no significant change in the beliefs that support self-medication such as: using antibiotics from a friend (72 to 77%) buying antibiotics without a prescription (42 to 45%), and getting information about medication use from leaflet without referring to a health care professional (60 to 63%). There were some areas of confusion regarding antibiotic range of effectiveness, and origin of resistance. Agreement about antibiotic resistance being a problem in Jordan increased significantly from 44 to 60% (p < 0.001). In addition, there was a significant increase in the percentage of participants who said that they don't request antibiotics from physicians (56 to 75% (P ≤ 0.001) and who said they would trust physicians' decisions about the necessity of antibiotics (70 to 83% P < 0.05). CONCLUSION: Findings indicate the need for better suited, and more inclusive, public educational campaigns.

Antibiotic resistant Cutibacterium acnes among acne patients in Jordan: a cross sectional study.

BACKGROUND: Antibiotics have been used for decades against Cutibacterium acnes (previously known as Propionibacterium acnes; C. acnes). Alarmingly, antibiotic resistance to this bacterium has become a worldwide problem in recent years. No studies are available on the antibiotic susceptibility patterns of C. acnes among Jordanian acne patients and how that is influenced by antibiotic use. This study aims to assess antibiotic resistance patterns of C. acnes clinical isolates and neighboring Gram-positive normal flora of the skin obtained from acne patients attending dermatology clinics in Amman -Jordan appraising the role of antibiotic consumption. METHODS: This is a cross-sectional study of acne patients presenting to selected dermatology outpatient clinics over a 6-month study period. Swabs obtained from inflamed lesions were cultured aerobically and anaerobically. Isolates were identified and screened for antibiotic susceptibility. In addition, all patients were asked to fill in a questionnaire that included questions about the history of antibiotic treatment. RESULTS: C. acnes was isolated from lesions of 100 patients out of 115 participants included in this study. 73% of the isolates were resistant to erythromycin and 59% to clindamycin 37% to doxycycline, 36% to tetracycline, 31% to trimethoprim / sulfamethoxazole, 15% to levofloxacin, and 3% to minocycline. Multi drug resistance (MDR) in C. acnes isolates as well as Staphylococcus aureus (S. aureus) and Staphylococcus epidermidis (S. epidermidis) with a similar pattern of resistance were detected from the same patient in most cases. A pattern of higher resistance towards variable antibiotic was observed in patients previously treated with antibiotics for acne management. CONCLUSIONS: The findings of this study demonstrate the distribution of antibiotic resistance of C. acnes towards used antibiotics and emphasizes the influence of antibiotic consumption on development of antibiotic resistance. The similar pattern of resistance between skin bacteria tested in this study highlights the genetic transfer of resistance between skin commensals including S. aureus and S. epidermidis hence promoting its circulation in the community.

The search of potential inhibitors of the AcrAB-TolC system of multidrug-resistant Escherichia coli: an in silico approach.

The number of infections caused by multidrug antibiotic resistance (MDR) species is increasing globally. The efflux pump system, AcrAB-TolC, confers Escherichia coli resistance to many antibiotics and results in poor treatment outcomes. Different rational drug design techniques were employed to search for a safe and effective AcrAB-TolC system inhibitor. Ligand docking was performed to analyze the binding of different ArcB substrates and/or inhibitors in the different AcrAB crystal structure binding sites. The validated docking site using the established docking preferences was used to perform virtual high-throughput screening on a large library of compounds. Domperidone, a known and safe over-the-counter antiemetic drug, was proposed as an effective ArcB inhibitor. Microbiological studies confirmed the computational results and domperidone reversed the resistance to the antibiotics: levofloxacin and ciprofloxacin in the MDR E. coli stains with an effect that surpassed the effect of the known efflux pump inhibitor, reserpine. In addition, it was able to increase both antibiotic effects on susceptible strains. This finding suggests that the antibiotic-domperidone combination can be used clinically to treat infections caused by multidrug-resistant E. coli strains.

Awareness of antibiotic use and antimicrobial resistance in the Iraqi community in Jordan.

INTRODUCTION: Antimicrobial resistance is a serious global health concern. It has considerable implications on societies' health and resources. In Jordan, there is a large Iraqi community due to the ongoing turmoil in Iraq. Unfortunately, health awareness and practices of this community are under-investigated due to scarcity of research. This paper assesses the awareness of antibiotic use and antimicrobial resistance in the Iraqi community residing in Amman, Jordan. Their level of interaction with health care professionals regarding antibiotics and differences in their antibiotic use between Iraq and Jordan are also discussed. METHODOLOGY: A cross-sectional questionnaire-based survey involving randomly selected Iraqis residing in Amman, Jordan was conducted. RESULTS: The study involved 508 participants. Sixty-two percent of participants agreed with buying antibiotics without a prescription, 29% agreed with obtaining antibiotics from friends or relatives, and 46% agreed with keeping leftover antibiotics for future use. Furthermore, 60% disagreed with not completing an antibiotic course and almost 90% of the sample listed viral diseases as an indication for antibiotics. Forty-four percent of participants abided by physicians' instructions on antibiotic use. Half of the participants believed that pharmacists provided instructions on antibiotics all the time, whereas physicians were perceived to do so by 29% of participants. CONCLUSIONS: Gaps exist in knowledge of antibiotic use and reasons for antimicrobial resistance among Iraqis residing in Jordan. These gaps should serve in planning educational campaigns to raise the community's awareness of responsible antibiotic use. Law enforcement to restrict access to antibiotics is also pivotal to tackle their misuse.