Chitosan-Aspirin Combination Inhibits Quorum-Sensing Synthases (lasI and rhlI) in Pseudomonas aeruginosa.

BACKGROUND: Quorum sensing (QS) controls the virulence of P. aeruginosa. This study aims to determine the anti-QS activity of aspirin alone and in combination with chitosan to reach maximum inhibition. We tested ten virulent Pseudomonas aeruginosa (P. aeruginosa) isolates and screened for N-acyl homoserine lactone (AHL) production using Agrobacterium tumefaciens as a biosensor. P. aeruginosa isolates were treated with sub-minimum inhibitory concentrations (MICs) of aspirin and chitosan-aspirin. We used broth microdilution and checkerboard titration methods to determine the MICs and the synergistic effect of these two compounds, respectively. Real-time polymerase chain reaction (PCR) was used to estimate the anti-QS activity of the aspirin-chitosan combination on the expression of lasI and rhlI genes. RESULTS: Aspirin decreased the motility and production of AHLs, pyocyanin, and biofilm. Chitosan potentiated the inhibitory effect of aspirin. The chitosan-aspirin combination inhibited lasI and rhlI gene expression in PAO1 (ATCC 15692) by 7.12- and 0.92-fold, respectively. In clinical isolates, the expression of lasI and rhlI was decreased by 1.76 × 102- and 1.63 × 104-fold, respectively. Molecular docking analysis revealed that aspirin could fit into the active sites of the QS synthases lasI and rhlI with a high binding affinity, causing conformational changes that resulted in their inhibition. CONCLUSIONS: The chitosan-aspirin combination provides new insights into treating virulent and resistant P. aeruginosa.

The role of family and culture in the disclosure of bad news: A multicentre cross-sectional study in Pakistan.

Objectives: Disclosure of bad news is distressing for patients and family members. Our aim was to assess patients' perceptions and preferences regarding bad news in the health setting. Methods: Cross-sectional, multi-centered study supported by an external grant in 15 Government and Private Hospitals across Pakistan. A sample size of 1673 patients and family members was used. Ethics permission/consent was taken from each participating hospital and participant. Responses were compared across provinces, gender, age, education and income. Results: >80% patients preferred their relatives to know the diagnosis first and they wanted the news to be disclosed to them by doctors. Significant association between education level, income and preference for wanting to know the diagnosis was found. Reasons for wanting to know the diagnosis included treatment, prognosis and prevention options whereas reasons for not wanting to know included fear of emotions and God's will. Conclusion: The majority of Pakistani patients want to be informed and want the family to know first. Preferences for disclosure vary across, age, education and income level. Innovation: First countrywide study on this topic. Identifies need for culturally sensitive guidelines that include the family's role in disclosure of bad news.

Teaching Clinical Skills Using Online Modality through Modified Peyton's Framework: An Experience from a Medical University in Pakistan.

Introduction: Medical institutions worldwide faced an unprecedented situation during COVID-19 of identifying alternative strategies to sustain the continuum of learning process. This led to several innovations in the traditional medical curriculum. This study explored the effectiveness and feasibility of using the Peyton's framework modified for a virtual platform (Microsoft Teams) for teaching clinical skills to first and second-year medical students at The Aga Khan University, Karachi, Pakistan. Methods: In 2020-2021, the modified Peyton's framework was integrated in the clinical skills (CS) curriculum for all first- and second-year students (N=200). For evaluation, a mixed-method design was used, with pre-and post-session questionnaires. Students' satisfaction was obtained through the standard session evaluation tool of the university. For the qualitative arm, to explore the instructors' experiences, purposive sampling was used (n=8) and a focused group discussion (FGD) was conducted. Finally, performance of the students at the end of year summative Objective Structured Clinical Examination (OSCE) was compared with the students of previous year. Quantitative data were analysed using STATA® version 15.1, using paired t-test to compare the differences in OSCE scores in selected CS stations. A p-value of <0.05 was considered significant for all tests. The FGD was transcribed and analysed through manual content analysis. Results: Nine clinical skills (that included history and examination skills) were taught using the virtual platform. There was a significant improvement in post-session questionnaires in seven of these skills (p value <0.01). Session evaluations showed that most students were satisfied with the learning experience. The instructors showed that the online teaching offered a promising platform for teaching history taking skills. The OSCE scores showed mixed results, with significant improvement in two out of four repeated stations by using paired t-test [abdominal exam (87.33 ± 8.99, <0.001); and precordial examination (88.45 ± 8.36, 0.001)]. Conclusions: Modifying Peyton's framework to a virtual platform allowed us to sustain the continuum of clinical education during the COVID-19 pandemic. The results support the use of a blended learning environment for teaching clinical skills.

The impact of COVID-19 safety interventions on creating a controlled environment on campus.

Objectives: During COVID-19 the re-opening of educational institutes was frequently debated, however with the decline in the number of COVID-19 cases, The Aga Khan University (AKU) in Karachi, Pakistan opened its campus for medical and nursing students after more than 6 months of closure. To ensure gradual resumption of activities on-campus, a combination of interventions was diligently deployed to minimize student infection rates. Scarce literature exists on students' perceptions regarding decisions implemented by university leadership. The aim of the study was to determine the efficacy of these interventions. Methods: We conducted a convergent, parallel, mixed-methods observational study targeting medical and nursing students. An online questionnaire was disseminated to elicit students' degree of (dis)agreement on a four-point Likert scale. Focused group discussions (FGDs) were conducted to comprehend reasons for (dis)agreement. Results: Total of 183 students responded to questionnaire (59.0% nursing, 67.8% female), 11 FGDs were conducted with 85 students. Interventions with highest agreement were mandatory face masks policy (94.54%), weekly mandated COVID-testing (92.35%) and students' Academic Bubble (91.26%); highest disagreement was for Sehat Check application (41.53%); and stay strong campaign (40.44%). Four themes emerged from FGDs: Effective safety interventions, Safety interventions with limited effectiveness, Utility of Sehat Check Application and Future recommendations for informing policy. Conclusion: It is paramount to seek student-feedback at forefront of university re-opening strategy. Clear communication channels are as important as an administrative response system's robustness. Bidirectional communication channels are fundamental and requisite during ever-changing policies and regulations. Engaging student representatives in decision making or implementation processes (such as "pilot" before "roll-out") would allow any potential issues to be managed early on. Gather real-time anonymous feedback and identify key areas that need further promulgation and those that need to be replaced with more effective ones.

Use of simulation based technology in pre-clinical years improves confidence and satisfaction among medical students.

OBJECTIVE: To determine perception of medical students about learning from integrated simulated clinical skill sessions as part of the undergraduate curriculum. METHODS: The cross-sectional study was conducted at the Centre for Innovation in Medical Education, Aga Khan University (AKU), Karachi, from July 2018 to February 2019, and comprised first year medical students undertaking the Respiration and Circulation module of the curriculum. Quantitative data was collected using a questionnaire and the responses were assessed on a five-point Likert scale. Data was analysed using SPSS 21. Qualitative data was gathered through focused group discussion with students and an in-depth interview with the facilitator conducting the sessions. The data was subjected to thematic analyses. RESULTS: Of the 161 subjects, 71(44%) participated in the session I and 90(56%) in the session II. Altogether 68(96%) students in session I and 81(90%) in session II believed integrated sessions to be effective in achieving learning objectives, and 65(92%) in session I and 79(88 %) in session II found them motivating, while 61(86%) in session I and 76(84%) in session II expressed the confidence that they had accomplished learning objectives and felt they had learned practical clinical skills; session I, 59(84%), session II, 73(81%). Qualitative analysis revealed that these sessions enhanced understanding of the subject matter and student engagement. CONCLUSIONS: Integrated clinical skills sessions improved students' interest, engagement and confidence. It should be implemented in undergraduate medical teaching curriculum.

Chitosan and chitosan-zinc oxide nanocomposite inhibit expression of LasI and RhlI genes and quorum sensing dependent virulence factors of Pseudomonas aeruginosa.

The great ability of Pseudomonas aeruginosa to cause chronic infection is attributed to several virulence factors, biofilm formation, intrinsic and acquired resistance to many antibiotics. Anti-quorum sensing (QS) and anti-virulence therapy are promising alternatives to the existing antibiotic therapy. In this study, the effect of chitosan and the prepared chitosan-zinc oxide (CH/ZnO) nanocomposite on QS-dependent virulence factors and acyl homoserine lactone "AHL" production was studied. The chemical structure of the prepared CH/ZnO nanocomposite was characterized by FT-IR spectrum and XRD. The thermal stability and particle size were determined. Chitosan causes a significant decrease in AHL, biofilm, pyocyanin production and motility of P. aeruginosa. CH/ZnO nanocomposite augments the inhibitory activity of chitosan in both phenotypic and genotypic levels. Both chitosan and CH/ZnO nanocomposite downregulate the expression of LasI and RhlI genes using quantitative real-time PCR. The expression of RhlI gene in PAO1 is reduced by 1240 folds after treatment with CH/ZnO nanocomposite. The expression of LasI and RhlI genes in clinical isolates is reduced by 1778.07 and 627.29 folds upon treatment with CH/ZnO. These promising results may find a rescue in the battle of fighting P. aeruginosa by repressing its QS-dependent virulence factors.

Frequency and Factors Associated with Adult Immunization in Patients Visiting Family Medicine Clinics at a Tertiary Care Hospital, Karachi.

Objective The goal of this study was to determine the frequency and factors associated with adult immunization in patients visiting family medicine clinics at a tertiary care hospital in Karachi. Methods A cross-sectional study was conducted from March 2014 to March 2015 in a tertiary care hospital in Karachi, Pakistan. Participants more than 18 years were invited to participate in the study. A pretested questionnaire was used to collect information. Data were entered and analyzed using IBM SPSS Statistics for Windows, version 19.0 (Armonk, NY: IBM Corp). Results A total of 340 patients were surveyed. The majority of patients were female (69.5%) with a mean age of 35.47 years. The majority were married (61.1%), and 30% of the participants had completed graduation or postgraduate education (20%). Most of the patients believed that vaccines can be used in adults to prevent disease (62.2%). Patients believed that the hepatitis B vaccine, influenza vaccine, and hepatitis A vaccine can be administered to adults (58.1%, 29.9%, 33.8%, respectively). The major sources of their information regarding vaccination in adults were friends or relatives (25%) and media (23.2%). Regarding availability of vaccines, 71.3% thought a hepatitis B vaccine is available, 54.9% thought a tuberculosis vaccine is available, and 49.3% thought a tetanus toxoid vaccine is available. Only 36.4% respondents received any vaccine in adulthood. The majority of patients (62.2%) received the hepatitis B vaccine in adulthood. The major reason given for not receiving vaccines was lack of awareness (62.4%). Conclusion Low adult vaccination coverage rates and awareness, as highlighted by the results of this study, show the dire need to address this major preventive strategy. This information can be utilized to conduct larger community-based surveys, to conduct health awareness sessions in the community, and to educate our doctors regarding the availability and benefits of adult vaccines.

Energy drinks consumption practices among medical students of a Private sector University of Karachi, Pakistan.

Consumption of energy drinks has become popular among students and athletes over the past few years. To explore the phenomenon, a cross-sectional survey was conducted through a self-administered pilot-tested questionnaire. Frequency of energy drinks consumption was found to be 121(52%) in a sample of 233 medical students. Red bull was the most common brand consumed 101(43%). The major reasons reported for its usage were to gain/replenish energy by 36(15.4%), and studying for examination by 34(14.6%). Television was reported as the major source of information 153(66%) followed by friends 113(48%). There was a high frequency of energy drinks' consumption among medical students of a private university. There is a strong need to create awareness regarding these drinks, especially among adolescents and teenagers.

Importance of serum IL-18 and RANTES as markers for breast carcinoma progression.

BACKGROUND: Interleukin-18 (IL-18), a cytokine that plays an important role in the T-cell-helper response, acts as an angiogenic factor and a tumor suppressor. RANTES (regulated upon activation normal T-cells expressed and secreted) is a member of the C-C chemokine family with chemoattractant activity for a variety of cell types. High incidence and intensity of RANTES were noted in advanced breast carcinoma. AIM OF THE STUDY: To correlate the levels of RANTES and IL-18 in serum of breast cancer patients with bone or other organ metastasis compared to breast cancer patients without metastasis and healthy controls and to estimate the role of each of them as a prognostic marker for the progression of the disease. PATIENTS AND METHODS: The study was conducted on 60 breast cancer patients (25 cases with no metastasis and 35 cases with metastasis) who were admitted to the outpatient clinic of the NCI, Cairo University during the period from March 2004 to September 2004 and 30 apparently healthy controls who were volunteers at the blood bank of the NCI, Cairo University. RESULTS: Showed that there was a statistically significant difference between the level of IL-18 in breast cancer patients without metastasis and the control group (p<0.05) while there was a highly significant difference between the metastatic group and the control group (p<0.001). There was a significant increase in IL-18 levels between metastatic and non-metastatic cases (p<0.01). RANTES showed a significant increase in breast cancer cases with no metastasis and the control group (p<0.05) and it showed a highly significant increase in metastatic patients compared to controls (p<0.001). There was no significant increase in the level of RANTES in metastatic compared to non-metastatic patients (p>0.05). CONCLUSIONS: IL-18 is an important non invasive marker suspecting metastasis. Even though RANTES levels were higher in cancer patients compared to controls, its role in staging of breast cancer was not clear in this study.