Impact of COVID-19 Pandemic on Antimicrobial Resistance Pattern;Transition from Resistivity to Susceptibility

Objective: The study aimed to assess the antibiotic resistance pattern before and after the pandemic to provide the physicians with general guidance for antibiotic prescribing. Material and Methods: The yearly antibiograms of different tertiary care hospitals were extracted from Pakistan Antimicrobial Resistance (AMR) Network. The data timeline observed was from January 2016 to December 2020. The data were scrutinized to the most common organism studied with the most recurring antimicrobial used. Results: Among the Gram-positive organisms, increased resistivity against penicillin was observed against both the organism, while a good susceptibility was observed against vancomycin. Among the Gram-negative organisms, the highest resistance was observed in Ceftriaxone, Ciprofloxacin, and Cotrimoxazole. Staphylococcus aureus and Escherichia coli are the most prevalent organisms in tertiary care hospitals. Conclusion: While satisfactory susceptibility was observed in Amikacin and Piperacillin/Tazobactam. The post-pandemic era resulted in a decrease in AMR due to significant changes in antibiotic prescribing patterns. This report may guide future antibiotic prescribing.

Less disease severity of COVID-19 among smokers: an indication of investigating nicotine as preventive measure

The study was aimed to mention the epidemiology of smoking in Pakistani COVID-19 infected patients along with the disease severity, oxygen dependency and fatality rate. A cross sectional epidemiological study was carried out on 555 confirmed cases of COVID-19 infection. The median age was 47+or-16 years. 59% were male and 41% were female. Most of the patients (97.5%) survived, while only 2.5% expired. 25.6% patients required the oxygen. Total 17 (3%) COVID-19 patients with age 20-75 years were identified as smokers. No mortality was observed in smokers. The 1.4% smokers presented with mild disease, 1.2% with moderate disease and 0.4% had severe disease. According to Chi- Square test, there existed an insignificant difference (p- value: 0.38649) between smokers and non-smokers in disease severity levels. Smoking is a precursor for countless diseases, but it behaved differently in COVID-19 infected patients, as its prevalence was significantly low. We found no significant variation of the disease severity among the smokers and non-smokers. Profound experiments should be conducted to recommend whether nicotine can be used as a protective agent to negate COVID-19 infection.

Il-6, il-0, ifn gamma and crp in newly diagnosed covid 19 patients

The study was conducted in Baghdad for the period from July 1 to August 20 2020 on patients previously diagnosed with the emerging corona virus .. The study included 40 patients whose ages ranged from 20 to 80 years. The study also included taking 40 healthy people of the same age groups to detect interleukin 6 and 10 gamma interferon by using ELISA technique as well as measuring the level of the C-reactive protein. The study contained a withdrawal of five ml of venous blood from patients and patients and took all the necessary information from them such as gender, age, current living status, number of family members, as well as the history of infection and travel to countries affected by the virus to compare the two groups with regard to the immune response by measuring the variables above. The study showed that the highest mean level of IL-6 was found in newly diagnosed COVID-19 patients comparing with healthy control (60.76±8.46 v.s.39.03±5.26 pg/ml) (P:<0.01). The study also demonstrated that the level of IL-10 was significantly elevated in newly diagnosed COVID-19 patients as compared with healthy control (38.18±4.57 v.s.31.84±3.19 pg/ml). The study established that the level of IFN-Gamma was significantly increased in patients with new infection of COVID-19 as compared with healthy control (27.27±7.18 v.s. 13.81±3.83 pg/ml) (P:<0.01). The study proven that the level of CRP was increased significantly in patients with new infection of COVID-19 as compared with healthy control (33.18±3.19 v.s.3.±1.81 mg/dl) (P:<0.01). The study concluded that, There was a highly significant relation of IL-6, IL-0, IFN gamma, and CRP with COVID-19 disease in the first week of infection

Mucociliary Transport Deficiency in Syrian Hamsters with SARS-CoV-2

RATIONALE: SARS-CoV-2 causes COVID-19 disease and infects respiratory epithelial cells, but how it affects ciliated cell function and the mucociliary transport apparatus is unknown. Abnormal mucociliary function could predispose to COVID-19 progression and/or secondary infection. Micro-optical coherence tomography (μOCT) is a novel method to simultaneously visualize and quantify the functional microanatomy of airways. Here, we established a hamster model of COVID-19 and evaluated their tracheas by μOCT. METHODS: Adult golden Syrian hamsters were inoculated intranasally with 3.2 × 105 (high dose, HD, N=4) or 3.2 × 104 (low dose, LD, N=4) plaque-forming units of SARS-CoV-2 (WA/1 strain). Clinical signs were monitored daily, nasal brushes collected intermittently, and hamsters were euthanized seven days (D7) after inoculation. Tracheas were imaged by μOCT, nasal washes and bronchial alveolar lavage fluid (BALF) from right lung lobes were collected for quantitation of viral load by qRT-PCR, and left lungs were inflated with and fixed in 10% neutral buffered formalin for histological analysis. Age-matched hamsters were used as uninfected controls (N=5). RESULTS: SARS-CoV-2 hamsters lost weight through D7 in dose-dependent fashion (-11% in HD vs. -4% in LD, p=0.02) and HD hamsters showed moderate lethargy. Nasal brushes on D4 and nasal washes on D7 contain 105-106 genome copies of virus while BALF on D7 was less than 104 genome copies and intermittently detected in LD. Histology demonstrated patchy and multifocal interstitial pneumonia (type II pneumocyte hyperplasia and mononuclear cell infiltrate), with ∼20% area affected in HD that was more variable in LD. Functional microanatomy of tracheas revealed diminished area of active ciliary beating (control 18 ± 2 vs. LD 7 ± 1%, p=0.0002, control vs. HD 9 ± 1%, p=0.001), reduced ciliary beat frequency (control 10.88 ± 0.70 vs. LD 8.83 ± 0.34 Hz, p=0.01, control vs. HD 8.26 ± 0.33 Hz, p=0.001), and decreased periciliary liquid depth (control 6.41 ± 0.18 vs. HD 5.61± 0.12 μ m, p=0.027). Mucociliary transport rate was diminished (control 0.84 ± 0.19 vs. LD 0.48 ± 0.16 vs. HD 0.37 ± 0.13 mm/min) although not statistically significant. Additional cohorts are in progress. CONCLUSION: SARS-Cov-2 infected hamsters exhibit reduced body weight, high viral load, and histopathological injury through 7 days. SARS-CoV-2 caused functional deficits of the mucociliary transport apparatus, consistent with early findings in COVID-19 patients (see Vijaykumar et al.). Abnormal ciliated cell function is important to SARS-CoV-2 pathogenesis, and may help monitor progression and represent a treatment opportunity for COVID-19.

Cigarette smoke exposure increases ace-2 expression and sars-cov-2 infection severity in human and ferret airways and induces apoptotic cell injury in vitro

RATIONALE: Cigarette smoking is associated with COVID-19 prevalence and severity, but the mechanistic basis for how smoking alters SARS-CoV-2 pathogenesis is unknown. A potential explanation is that smoking could alter expression of angiotensin converting enzyme-2 (ACE2), which functions as the cellular receptor and point of entry. Here we investigated the severity of SARS CoV-2 infection ex vivo and in vitro and using tissue samples and primary ferret and human airway epithelial cells with and without antecedent exposure to cigarette smoke. Methods: ACE2 expression measured by Quantitative PCR (Q-PCR) of ferret lungs exposed to 6 months of cigarette smoke, and findings validated by immunofluorescence (IF). Primary airway cells isolated from airways of ferrets or human non-smokers or COPD subjects were grown until terminally differentiated at air liquid interface. Cells were then exposed to cigarette smoke extract (CSE) or vehicle control, then infected with SARS CoV-2 (3 MOI) or mock control. Viral copy was measured by Q-PCR. Viral infection was quantified by foci forming assay (FFU/mL) using VeroE6 cells. Results: Ferret lungs following 6 months of smoke exposure had increased ACE2 as compared to air controls by Q-PCR (>1.5 fold, P<0.05, N=6), and IF staining. Higher ACE2 expression was also observed in normal ferret airway cells exposed to CSE (>1.5 fold, P<0.05, N=3), normal Human Bronchial Epithelial (HBE) cells exposed to CSE (>2 fold, P<0.001, N=4), and HBE cells from COPD donors as compared to healthy controls (>2 fold, P<0.001, N=4). When ferret airway cells were inoculated with SARS-CoV-2, intracellular viral load of SARS-CoV-2 was increased in CSE exposed cells as compared to vehicle controls (103-104 vehicle Vs CSE 105 106 copy/μL). Viral infection was also increased >2 fold (P<0.01, N=5). Likewise, CSE exposed (105-107 copy/μL, P<0.0001, N=4) and COPD (106-108 copy/μL, P<0.0001, N=4) HBE had increased viral load as compared to controls (103-104 copy/μL, P <0.001, N=4), and 2 to 3-fold increase in viral infection, respectively. TUNEL staining was increased in infected cells, indicating apoptosis. Transcript analysis of HBE cells with and without SARS-CoV-2 by RNASeq to identify differentially expressed genes in CSE exposed cells as compared to controls is in progress. Conclusion: Cigarette smoke and CSE increased ACE2 expression in ferrets, and ferret and human cells respectively. CSE-induction increased viral replication and infection severity, resulting in increased apoptosis. Cigarette smoking likely influences the severity of SARS-CoV-2 infection by altering expression of ACE2, inducing airway cell apoptosis upon infection.

COVID-19 and a way forward for restaurants and street food vendors

COVID-19 has had a drastic impact on every field and walk of life. The long-lasting impacts of this pandemic have changed the way businesses used to be conducted and will have a strong impact on business models as well. The main objective of this qualitative study is to investigate the impact of COVID-19 on restaurants and small stalls of street food vendors in Pakistan and to suggest a way forward. A total of 30 interviews were conducted through conference calls. The findings proved that major issues faced by the restaurants are the massive decline in sales, massive layoffs, no economic activity, and no relief from the government. The major changes required in the existing business models highlighted by the interviewees are proper sanitization, changes in the sitting area, change in menus, and the need for innovative ideas to attract the customers back. The study is useful for the restaurants and street food vendors to help them out in this difficult phase and suggest a way forward to them.