Analytical assessment of clinical sensitivity and specificities of pharmaceutical rapid SARS-CoV-2 detection nasopharyngeal swab testing kits in Pakistan.

Despite of the global unity against COVID-19 pandemic, the threat of SARS-CoV-2 variants on the lives of human being is still not over. SARS-CoV-2 pandemic has urged the need of rapid viral detection at earliest. To cope with gradually expanding scenario of SARS-CoV-2, accurate diagnosis is extremely crucial factor which should be noticed by international health organizations. Limited research followed by sporadic marketing of SARS-CoV-2 rapid pharmaceutical detection kits raises critical questions against quality assurance and quality control measures. Herein we aimed to interrogate effectivity and specificity analysis of SARS-CoV-2 pharmaceutical rapid detection kits (nasopharyngeal swab based) using conventional gold standard triple target real-time polymerase chain reaction (USFDA approved). A cross-sectional study was conducted over 1500 suspected SARS-CoV-2 patients. 100 real time-PCR confirmed patients were evaluated for pharmaceutical RDT kits based upon nasopharyngeal swab based kits. The SARS-CoV-2 nasopharyngeal swab based rapid diagnostic kit (NSP RDTs) analysis showed 78% reactivity. Among real time PCR confirmed negative subjects, 49.3% represented false positivity. The positive predictive analysis revealed 67.82%, while negative predictive values were 64.40%. The NSP RDTs showed limited sensitivities and specificities as compared to gold standard real time PCR. Valid and authentic detection of SARS-CoV-2 is deemed necessary for accurate COVID-19 surveillance across the globe. Current study highlights the potential consequences of inadequate detection of SARS-CoV-2 and emerging novel mutants, compromising vaccine preventable diseases. Current study emphasizes need to wake higher authorities including strategic organizations for designing adequate measures to prevent future SARS-CoV-2 epidemics.

Hepatitis C virus associated ALT, AST, GGT, Bili T, HB, HBA1C, CREAT, PT, aPPT, AFP, CEA, CA 125, CA 19-9, iPTH biomarkers, computed tomography and HCV burden of disease during pre COVID-19 era (2018-2019) and post COVID-19 era (2020-2022) in Pakistan.

The national burden of HCV has significantly mounted over the period of last few decades placing Pakistan at the worst placement of second largest burden of HCV globally. Herein for the first time from Pakistan, we examined clinical correlation of potential biomarkers with HCV. Nation-wide study was conducted on 13,348 suspected HCV patients during 2018-2022. During pre-COVID-19 era of 2018-2019, prevalence of HCV remained 30%. During 2018, among HCV positive patients, 91% of ALT, 63% of AST, 67% of GGT, 28% of Bili T, 62% of HB, 15% of HBA1C, 25% of CREAT, 15% of PT, 15% of aPTT and 64% of AFP were abnormal. During 2019, among HCV infected 74.47% of ALT, 63.54% of AST, 70.24% of GGT, 24.71% of Bili T, 8.77% of HB and 75% of AFP were raised. CT/CAT scan revealed 4.65% liver complications (mild 13.04%, moderate 30.43% and severe 56.52%). During 2020, HCV prevalence remained 25%. 65.17% of ALT, 64.20% of AST, 68.75% of GGT, 31.25% of Bili T, 20.97% of HB, 4.65% of CREAT and 73.68% of AFP levels were raised. CAT analysis revealed liver complications among 4.41% (14.81% mild, 40.74% moderate, and 44.44% sever). 85.71% of participants diabetes was out of control. During 2021, HCV prevalence remained 27.1%. ALT (73.86%), AST (50.6%), GGT (67.95%), Bili T (28.21%), HB (20%), CREAT (5.8%) and AFP (82.14%) levels were abnormal. During 2022, the levels of ALT (56.06%), AST (56.36%), GGT (56.6%), Bili T (19.23%), HB (43.48%), HBA1C (14.81), CREAT (18.92%), AFP (93.75%) were abnormal. CAT analysis revealed 7.46% liver complications (25% mild, 30.36% moderate, and 42.86% sever). During 2021-2022, 83.33% of subject's diabetes was not controlled.

Anomalous Right Coronary Artery Originating From the Pulmonary Artery (ARCAPA): A Rare Presentation in the Fifth Decade.

Presented is a case report of a 54-year-old male Jehovah's Witness diagnosed with an anomalous right coronary artery originating from the main pulmonary artery with the initial presentation with a history of episodic fatigue, chest pain, palpitation, and bodily weakness. The patient was managed with reimplantation of the anomalous coronary artery onto the ascending aorta.

Hypertension in the United States Fire Service.

Hypertension is a major risk factor for atherosclerotic cardiovascular disease and cardiac remodeling and is associated with an increased risk of sudden cardiac events, the leading cause of duty-related death in the fire service. We assessed systemic blood pressures and prevalence of hypertension among US firefighters by decade of life. Medical records of career firefighters (5063 males and 274 females) from four geographically diverse occupational health clinics were assessed. Hypertension was defined as systolic blood pressure ≥130 mmHg or diastolic blood pressure ≥80 mmHg, or taking antihypertensive medication. Results from the firefighter sample were compared to the US general population (2015-2016 and 2017-2018 National Health and Nutrition Examination Surveys). Among the total sample, 69% of firefighters met the criteria for hypertension and 17% were taking antihypertensive medications. Percentages of hypertensive male and female firefighters were 45% and 11% among 20-29 years old, respectively, and increased to 78% and 79% among 50-59 years old, respectively. Compared to the general population, male firefighters had a higher prevalence of hypertension (p < 0.05) across all age groups (11-16% higher). In order to improve firefighter health and protect against sudden incapacitation in this public safety occupational group, increased efforts are necessary to screen for and manage high blood pressure.

Identification of potential lockdown areas during COVID-19 transmission in Punjab, Pakistan.

OBJECTIVES: Real-time COVID-19 spread mapping and monitoring to identify lockdown and semi-lockdown areas using hotspot analysis and geographic information systems and also near future prediction modeling for risk of COVID-19 in Punjab, Pakistan. STUDY DESIGN: Data for all COVID-19 cases were collected until 20 October 2020 in Punjab Province. METHODS: The methodology included geotagging COVID-19 cases to understand the trans-mobility areas for COVID-19 and characterize risk. The hotspot analysis technique was used to identify the number of areas in danger zones and the number of people affected by COVID-19. The complete lockdown areas were marked down geographically to be selected by the government of Pakistan based on increased numbers of cases. RESULTS: As per predictive model estimates, almost 9.2 million people are COVID-19 infected by 20 October 2020 in Punjab Province. The compound growth rate of COVID-19 decreased to 0.012% per day and doubling rate increased to 364.5 days in Punjab Province. Based on Pueyo model predictions from past temporal data, it is more likely that Punjab and Pakistan entered into peak around the first week of July 2020, and the decline of growth rate (and doubling rate) of reported cases started afterward. Hospital load was also measured through the Pueyo model, and mostly, people in the 60+ years age group are expected to dominate the hospitalized population. CONCLUSIONS: Pakistan is experiencing a high number of COVID-19 cases, with the maximum share from Punjab, Pakistan. Statistical modeling and compound growth estimation formulation were done through the Pueyo model, which was applied in Pakistan to identify the compound growth of COVID-19 patients and predicting numbers of patients shortly by slightly modifying it as per the local context.

Hepatitis C virus induces oxidative stress and DNA damage by regulating DNAPKCs, ATM, ATR and PARP mediated signaling and guards cell from cancerous condition by upregulating RB, P53 and downregulating VEGF.

Hepatitis C virus is responsible for liver damage and various metabolic disorders. HCV infections promote oxidative stress and cause damage to macromolecules. The aim of our study was to design a preliminary study with establishment of HCV genotype 3a infectivity assay in order to determine DNA damage in Huh-7 cell line at 72 hours post inoculation. Quantitative expression levels of COX-2 and GSR (oxidants and antioxidants), DNAPKCs, ATM, ATR and PARP (DNA damage and repair genes), RB and P53 (tumor suppressor genes) and VEGF (angiogenesis marker) were observed via real time PCR. Our findings revealed 1.533 fold upregulated expression of COX-2. The expression level of GSR was increased by1.27 fold and VEGF expression decreased by 0.367 fold. Thus, preventing cells to enter cancerous phase.

Characteristics of Aerococcus viridans isolated from bovine subclinical mastitis and its effect on milk SCC, yield, and composition.

Aerococcus viridians (A. viridans), an environmental Gram-positive bacterium, has been documented to be associated with bovine mastitis. However, its exact role in bovine mastitis and the changes it brings about in milk characteristics are not yet known. The objectives of the current study were to describe the antibiotic resistance of A. viridans from bovine mastitis as well as the correlation between existence of this pathogen in udders and the somatic cell counts (SCC), daily milk yield, and composition of individual cow. One-year sampling for subclinical mastitis composite milk was conducted based on monthly DHI data from September 2013 to August 2014, in a commercial herd located in Beijing, China. All samples were cultured and pathogens were identified using microbiology method. A. viridians isolates were further identified by API identification system and 16S ribosomal RNA (rRNA) sequencing method. Kirby-Bauer disk diffusion method was used to test the antibiotic resistance of A. viridians against kinds of antimicrobial substance. SCC, milk yield, and milk composition data were from monthly Dairy Herd Improvement (DHI) results. Results showed that a total of 279 (16.67%) A. viridans isolates were identified from among 1674 bacterial isolates cultured from milk samples with high SCC. The incidence of mastitis caused by A. viridans was the highest (48-53%) during the summer season. Majority of the isolates were susceptible to most of antimicrobial compounds tested, especially to ß-lactams, but were found to be resistant (50-90%) to aminoglycosides, sulfonamides, and tetracycline. The average SCC of the A. viridans infected cows was significantly higher (1000.0 × 103 cells/mL) (P < 0.01) as compared to healthy cows (72.4 × 103 cells/mL) and daily milk yield decreased (P > 0.05) by 1.86 kg/day. Reductions were also observed in fat content (P > 0.05), lactose (P < 0.01), and total solids (P > 0.05), whereas protein content increased significantly (P < 0.01) in milk samples of cows infected with A. viridans. The results of this study suggest that A. viridans could be considered as an emerging aetiological agent of bovine subclinical mastitis wherein it exerts an effect on SCC, milk yield, and composition.

Diagonal Earlobe Crease as a Significant Marker for Coronary Artery Disease: A Case-control Study.

OBJECTIVES: To investigate the association between diagonal earlobe crease (DELC) and coronary artery disease (CAD). Limited data exists in South Asia and no prior studies have been performed in Pakistan to assess this relationship. METHODS: In this case-control study, 200 participants from December 2015 to March 2016 at Shifa International Hospital, Islamabad, Pakistan were enrolled. Consecutive non-probability sampling was used to recruit patients. Cases were enrolled from cardiac care unit (CCU) of the hospital with angiography-proven CAD. Controls were selected from surgical, medical and neurology units of the hospital if they had no previously established evidence or symptoms of CAD. Patients were evaluated in terms of age and any history of hypertension, diabetes and/or smoking. Cases and controls were examined separately by two investigators for the unilateral or bilateral presence of DELC of the lobular portion of either auricle. Patients with ear piercings were excluded from the study. The data was analyzed in statistical product and service solutions (SPSS) (IBM, Delaware, Chicago), and an online statistical software. RESULTS: Out of the 200 patients, 126 (63%) were males and 74 (37%) were females. In the 100 cases, 76 had DELC and 24 had no crease whereas, among the 100 controls, 36 had DELC and 64 had no DELC (p <0.001, OR = 5.63, CI = 2.91-10.93). The prevalence of diseases such as hypertension, diabetes, smoking among the cases and controls were 66%, 53%, 27% and 27%, 18%, 25% respectively. The effect of hypertension and diabetes on the presence of DELC was statistically significant (p <0.05) but the impact of smoking on DELC presence was insignificant (p >0.05). CONCLUSION: There is a significant association between DELC and CAD. This is the first case-control study from South Asia disclosing this important correlation. Our study also reports a high frequency of DELC in patients suffering from hypertension and diabetes mellitus. No association between smoking and DELC was found.

When pure is not so pure: chloramine-related hemolytic anemia in home hemodialysis patients.

Worldwide, chloramines are used as the preferred disinfectant for city water supplies. Although they have distinct advantages compared with chlorine and are deemed harmless to the general population, hemodialysis (HD) patients are at risk from chloramine-induced hemolytic anemia. In recent years, this has been highlighted in regional dialysis units but not as frequently in the home HD group. We report on 2 home HD patients who succumbed to severe oxidative hemolysis due to high mains water chloramine concentrations. Both patients were extensively investigated for other cause of anemia before a definitive diagnosis was reached. Delays in diagnosing this uncommon condition can be costly in terms of significant morbidity and excessive usage of recombinant erythropoietin and blood transfusion. Prevention primarily involves enforcing strict water quality control and establishing regular communication with water supply boards and home HD patients. Double (inline) carbon filters should be installed in patient's homes as an effective means for removing high incoming chloramine concentrations.

Engineering of chaperone systems and of the unfolded protein response.

Production of recombinant proteins in mammalian cells is a successful technology that delivers protein pharmaceuticals for therapies and for diagnosis of human disorders. Cost effective production of protein biopharmaceuticals requires extensive optimization through cell and fermentation process engineering at the upstream and chemical engineering of purification processes at the downstream side of the production process. The majority of protein pharmaceuticals are secreted proteins. Accumulating evidence suggests that the folding and processing of these proteins in the endoplasmic reticulum (ER) is a general rate- and yield limiting step for their production. We will summarize our knowledge of protein folding in the ER and of signal transduction pathways activated by accumulation of unfolded proteins in the ER, collectively called the unfolded protein response (UPR). On the basis of this knowledge we will evaluate engineering approaches to increase cell specific productivities through engineering of the ER-resident protein folding machinery and of the UPR.

Report: prevalence and resistance pattern of Pseudomonas aeruginosa against various antibiotics.

A prospective study on various clinical isolates from patients admitted from various parts of NWFP and Afghanistan at Post Graduate Medical Institute (PGMI) Hayatabad Medical Complex, Peshawar was conducted from January 2000 to December 2004 to ascertain the prevalence and antimicrobial susceptibility of Pseudomonas aeruginosa infections. Among 4709 positive isolates, 314 (6.67%) were Pseudomonas aeruginosa. The highest rate of infection due to Pseudomonas aeruginosa was observed in orthopedic ward (24.61%) and OPD (20%), in other wards the infection was between 13% to 1.5%. Gender-wise prevalence showed 61.78% male and 38.22% females were infected by Pseudomonas aeruginosa. The highest percentage of Pseudomonas aeruginosa isolates were observed in pus (57.64%) and urine (24.2%) samples. Maximum Pseudomonas aeruginosa isolates were found between March to August and the highest percentage 13.846% was observed in June. Using the disc diffusion method, the resistance patterns of 314 isolates against 14 antimicrobial agents were determined. The highest resistance was observed against ampicillin (> or = 98.4%), ampicillin/sulbactam (85.3%), co-amoxiclave (83.8%) and ofloxacin (68.4%) and least resistance was observed against amikacin (24%). Similarly the MIC for ampicillin (4 to >2048 microg/ml), ampicillin/sulbactam (1 to 2048 microg/ml) and co-amoxiclave (1 to >2048 microg/ml) against clinical isolates of Pseudomonas aeruginosa was also high. High resistance of Pseudomonas aeruginosa against various commonly used antibiotics showed the alarming situation. The control of drug resistant Pseudomonas aeruginosa required rational prescribing and proper use of antibiotics.

Differential regulation of endogenous N- and P/Q-type Ca2+ channel inactivation by Ca2+/calmodulin impacts on their ability to support exocytosis in chromaffin cells.

P/Q-type (Ca(V)2.1) and N-type (Ca(V)2.2) Ca2+ channels are critical to stimulus-secretion coupling in the nervous system; feedback regulation of these channels by Ca2+ is therefore predicted to profoundly influence neurotransmission. Here we report divergent regulation of Ca2+-dependent inactivation (CDI) of native N- and P/Q-type Ca2+ channels by calmodulin (CaM) in adult chromaffin cells. Robust CDI of N-type channels was observed in response to prolonged step depolarizations, as well as repetitive stimulation with either brief step depolarizations or action potential-like voltage stimuli. Adenoviral expression of Ca2+-insensitive calmodulin mutants eliminated CDI of N-type channels. This is the first demonstration of CaM-dependent CDI of a native N-type channel. CDI of P/Q-type channels was by comparison modest and insensitive to expression of CaM mutants. Cloning of the C terminus of the Ca(V)2.1 alpha1 subunit from chromaffin cells revealed multiple splice variants lacking structural motifs required for CaM-dependent CDI. The physiological relevance of CDI on stimulus-coupled exocytosis was revealed by combining perforated-patch voltage-clamp recordings of pharmacologically isolated Ca2+ currents with membrane capacitance measurements of exocytosis. Increasing stimulus intensity to invoke CDI resulted in a significant decrease in the exocytotic efficiency of N-type channels compared with P/Q-type channels. Our results reveal unexpected diversity in CaM regulation of native Ca(V)2 channels and suggest that the ability of individual Ca2+ channel subtypes to undergo CDI may be tailored by alternative splicing to meet the specific requirements of a particular cellular function.

Standardization of Anaesthesia Ready Time and reasons of delay in induction of anaesthesia.

OBJECTIVE: Anaesthesia-Ready Time (ART) is the time taken by the anaesthetist to provide sufficient anaesthetic depth for start of surgery. Our aim was to set benchmark timings for ART and compare it with our current practice. METHODS: Benchmark ART time of 15 minutes was set for American Society of Anesthesiologists (ASA) class I and II patients, 30 minutes for ASA III and IV patients, 20 minutes for spinal and 30 minutes for epidural anaesthesia. An additional 15 minutes was added for each invasive procedure. RESULTS: Three hundred elective cases were audited. Seventy eight percent of the cases were within benchmark timings. The main causes of delay included undergraduate students performing procedures (24.6%), teaching invasive lines to postgraduates (21.3%) and paediatric patients (16.4%). CONCLUSION: The introduction of benchmark timings and its regular auditing can help standardize operating room booking time and reducing patient cost.

In-vitro antibacterial, antifungal and cytotoxic activities of some coumarins and their metal complexes.

A series of new antibacterial and antifungal coumarin-derived compounds and their transition metal complexes [cobalt (II), copper (II), nickel (II) and zinc (II)] have been synthesized, characterized and screened for their in vitro antibacterial activity against Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella typhi, Shigella dysenteriae, Bacillus cereus, Corynebacterium diphtheriae, Staphylococcus aureus and Streptococcus pyogenes bacterial strains and for in vitro antifungal activity against Trichophyton longifusus, Candida albicans, Aspergillus flavus, Microsporum canis, Fusarium solani, Candida glaberata. The results of these studies show the metal complexes to be more antibacterial and antifungal as compared to the uncomplexed coumarins. The brine shrimp bioassay was also carried out to study their in vitro cytotoxic properties.