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Malnourishment is one of the major issues faced by people of mountain areas of Pakistan. Common bean, being a rich source of protein, minerals, and nutraceutical compounds, is considered as a potential food to address the malnutrition problems in these areas. Azad Jammu and Kashmir is widely distributed with the indigenous unexplored landraces of common bean. In the present work, the nutritional profiling of 36 common bean ecotypes collected from the wide range of altitude, based on protein percentage, lysine contents, antioxidants activity, and total phenolic contents was carried out. Significant protein and lysine contents were found in the range of 13 - 24.5% and 4.4 - 7.4%, respectively. The DPPH free radical assay revealed very high antioxidants capacity of 67.8 - 97.8% with significant variation in the selected common bean germplasm. Total phenolic contents were determined using Folin-Ciocalteu reagent method and ranged from 68.9 to 110.3 mg GAE/100 g. Principal Component Analysis (PCA) revealed significant variation, based on hierarchical clustering, within germplasm in all studied attributes. Two main factors were elucidated which imparted 67.36% variation based on PCA. Lysine gene was identified, and its presence was confirmed through primer's amplification. E20 and E34 were found comparatively more nutritious based on biochemical characterisation. The present work will help in selection of high nutrient ecotypes for further substantial research including breeding of nutritionally promising ecotypes for farming community of Azad Jammu and Kashmir. [ABSTRACT FROM AUTHOR]

Zanthoxylum alatum Roxb. is a medicinal plant which abundantly grows in the hilly areas of Pakistan. The dried fruit of the plant is used as condiment and has excellent spice value. S-carvone was isolated from Zanthoxylum alatum, purified using chromatographic methods, and studied by GC-MC, FT-IR, and NMR techniques. The in vitro antioxidant activity of carvone was evaluated by different in vitro systems, including lipid peroxidation, 2,2-dipenyl-1-picrylhydrazyl (DPPH) and phosphomolybdenum assay. The isolated carvone showed inhibition activity against thiobarbituric acid reactive species (TBARS) induced by different pro-oxidants (10 M FeSO and 5 μM sodium nitroprusside) in rat liver and brain homogenates. The extract also caused the scavenging of DPPH radical and the reduction of molybdenum, Mo(VI) to Mo(V). The results indicated that S-carvone possesses high antioxidant activity. [ABSTRACT FROM AUTHOR]

Sea buckthorn (Hippophae rhamnoides L. ssp. turkestanica) is a very important multipurpose plant in areas of northern Pakistan. The berry of sea buckthorn is rich in nutrients and medicinal compounds such as vitamins, carotenoids, flavonoids, oil, carbohydrates, organic acids, amino acids and minerals. The chemical composition and nutritive value of sea buckthorn berries from eight populations from different areas of northern Pakistan were compared. The concentration of vitamin C in the berries was 250-333 mg/100 g; the seed oil was 7.69-13.7%, oil in dried pulp was 19.2-29.1% and the pytosterol content of the seed oil was 3.3-5.5%; there were 0.5-25 mg/L anthocyanins in the fruit juice. The potassium (2.8-7.2 g/kg), sodium (0.065-0.8 g/kg), calcium (0.7-1.25 g/ kg), magnesium (139-240 mg/kg), iron (40-225 mg/kg) and phosphorus (110-133 mg/kg) contents were high in dried berries. Sea buckthorn berries from Pakistan are a good source of phytonutrients and minerals. [ABSTRACT FROM AUTHOR]

Brucellosis is a disease that poses a higher risk of transmission to animals and people who have close interactions with them, such as farmers. Therefore, the present study was designed to determine the seroprevalence of bovine brucellosis and associated risk factors in Kasur, Punjab, Pakistan. One hundred pooled milk samples from 100 smallholder mixed herds of cows and buffaloes with a total study population of 425 animals (277 cattle and 148 buffalos) with a range of up to seven animals in each herd were collected and tested through a milk ring test (MRT). Blood samples were collected from the animals of MRT-positive herds and then subjected to an indirect enzyme-linked immunosorbent assay (i-ELISA) for the presence of antibodies against Brucella abortus. Of the 100 herds, 16 % (n = 16) were MRT-positive. In MRT-positive herds, 77 animals (43 female cattle, three male cattle, 21 female buffaloes, and ten male buffaloes) were present; 32 (41.6 %) were i-ELISA seropositive, with the highest seropositivity (54.8 %) observed in buffaloes as compared to cows (32.6 %). Almost 38 % of the farmers were unaware of brucellosis. Herd-level risk factors indicated Brucella seropositivity increased in herds with recent abortion history and bull exchange during breeding. Conversely, herds with veterinary assistance exhibited a decreased Brucella seropositivity. The findings highlight the need for a government-led awareness campaign emphasizing routine screenings, especially for breeding bulls, to stop further disease spread. Early detection and management can enhance animal health, milk production, and livestock holders' economic well-being., Competing Interests: Declaration of competing interest The authors declare that they have no competing interests, and this study was conducted without any commercial or financial relationships., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Mapping tick distribution and pathogens in unexplored areas sheds light on their importance in zoonotic and veterinary contexts. In this study, we performed a comprehensive investigation of the genetic diversity of tick and tick-borne pathogens (TBPs) detection infesting/infecting small ruminants across northern Pakistan. We collected 1587 ixodid ticks from 600 goats and sheep, an overall tick infestation rate of 50.2 %. Notably, gender-based infestation rates were higher in female goats and sheep compared to their male counterparts. Age-wise analysis showed that the tick infestation rate was higher in older animals. This study identified 11 ixodid tick species within three genera: Hyalomma, Haemaphysalis, and Rhipicephalus, which were taxonomically classified using 16S rRNA and cytochrome oxidase I (cox1) molecular markers. Sequence analysis indicated that reported ticks are similar to ixodid species found across various Asian and African countries. Tick-borne pathogens were detected by amplifying 16S rRNA and citrate synthase (gltA) for bacterial pathogens and 18S rRNA for apicomplexan parasites. The present study reported a diverse array of TBPs in ticks from the study area, with Rickettsia massiliae (24.5 %) and Theleria ovis (16.4 %) as the most prevalent bacterial and apicomplexan pathogens. Phylogenetically, detected TBPs shared evolutionary relatedness with identical TBPs from old and new world countries. These findings highlight the presence of zoonotic TBPs in ixodid ticks from Pakistan. In addition, it also provides a foundation for future epidemiological research on ticks and TBPs, emphasizing their relevance in both zoonotic and veterinary contexts., Competing Interests: Declaration of competing interest There is no competing interest among the authors., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

The livestock sector contributes almost 11% of Pakistan's GDP and is crucial to 35 million people's livelihoods. Ticks are a major economic threat, as over 80% of livestock, such as bovines, are tick-infested with Hyalomma and Rhipicephalus tick species. Hyalomma anatolicum and Rhipicephalus microplus are the most common tick species collected from livestock, transmitting primarily anaplasmosis, babesiosis, and theileriosis. We aimed to identify the geographical distribution of these two tick species and hot spot areas where the risk of these diseases being transmitted by these ticks is high. Following the PRISMA guideline, two authors conducted an independent review of literature sourced from various databases. We screened 326 research articles published between January 1, 1990, and December 31, 2023, focused on identifying the tick species at the district level. Thirty studies from 75 districts, representing 49.3% of the country's total area, detected at least one tick species through collection from animals. R. microplus was present in 81% (n = 61) and H. anatolicum in 82% (n = 62) of these sampled districts. We employed spatial and conventional statistical methods with Geographic Information Systems (GIS) after mapping the weighted distribution of both ticks (the number of ticks per standard unit of sampling effort). We identified northwestern and northcentral regions of the country as hotspots with the highest tick distribution, which aligned with the documented high prevalence of anaplasmosis, babesiosis, Crimean-Congo hemorrhagic fever (CCHF), and theileriosis in these regions. This underscores the urgent need for robust tick control measures in these districts to safeguard animal health and boost the livestock economy., Competing Interests: The authors declare that there were no conflicts of interest., (Copyright: © 2024 Hussain et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The escalating issue of air pollution has become a significant concern in urban regions, including Islamabad, Pakistan, due to the rise in air pollutant emissions driven by economic and industrial expansion. To gain a deeper understanding of air pollution, a study was conducted during winter 2022-2023, assessing physical, chemical, and biological factors in Islamabad. The findings revealed that the average concentration of fine particulate matter (PM2.5) was notably greater than the World Health Organization (WHO) guidelines, reaching 133.39 μg/m³. Additionally, the average concentration of bacteria (308.64 CFU/m³) was notably greater than that of fungi (203.55 CFU/m³) throughout the study. Analytical analyses, including SEM-EDS and FTIR, showed that the PM2.5 in Islamabad is composed of various particles such as soot aggregates, coal fly ash, minerals, bio-particles, and some unidentified particles. EF analysis distinguished PM2.5 sources, enhancing understanding of pollutants origin, whereas Spearman's correlation analysis elucidated constituent interactions, further explaining air quality impact. The results from the Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-OES) indicated a gradual increase in the total elemental composition of PM2.5 from autumn to winter, maintaining high levels throughout the winter season. Furthermore, a significant variation was found in the mass concentration of PM2.5 when comparing samples collected in the morning and evening. The study also identified the presence of semi-volatile organic compounds (SVOCs) in PM2.5 samples, including polycyclic aromatic hydrocarbons (PAHs) and phenolic compounds, with notable variations in their concentrations. Utilizing health risk assessment models developed by the US EPA, we estimated the potential health risks associated with PM2.5 exposure, highlighting the urgency of addressing air quality issues. These findings provide valuable insights into the sources and composition of PM2.5 in Islamabad, contributing to a comprehensive understanding of air quality and its potential environmental and health implications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Vector-/tick-borne pathogens (V/TBPs) pose a potential threat to human and animal health globally. Information regarding canine V/TBPs is scarce and no specific study has been conducted so far to explore the microbial diversity within ticks infesting dogs from Pakistan. Herein, this knowledge gap is addressed by assessing the genetic diversity and prevalence pattern of V/TBPs in ixodid ticks with special implications for public and canine health., Methods: A total of 1150 hard ticks were collected from 300 dogs across central Khyber Pakhtunkhwa (KP), Pakistan. After morpho-molecular identification, 120 tick samples were screened for the presence of V/TBPs by amplifying 16S rRNA/gltA (Rickettsia/Ehrlichia and Wolbachia sp.), 18S rRNA (Theileria sp.) and cox1 (Dirofilaria sp.) genes through PCR followed by sequencing and phylogenetic study., Results: In toto, 50 ixodid ticks (50/120, 41.7%) were found positive for V/TBPs DNA. The detected V/TBPs were categorized into five genera and eight species, viz. Ehrlichia (E. canis and Ehrlichia sp.), Rickettsia (R. massiliae, R. raoultii and Rickettsia sp.), Theileria (T. annulata), Dirofilaria (D. immitis) and Wolbachia (Wolbachia sp.). The pathogen prevalence patterns showed that R. massiliae was the most prevalent zoonotic V/TBP (19.5%), followed by E. canis (10.8%), Rickettsia sp. (7.5%), R. raoultii (6.7%), T. annulata (5.8%), D. immitis (5.8%), Wolbachia sp. (4.2%) and Ehrlichia sp. (3.3%), respectively. Among the screened tick species, most Rhipicephalus sanguineus sensu lato samples were found positive for V/TBP DNA (20/20,100%) followed by Rh. turanicus sensu stricto (13/20, 65%), Hyalomma dromedarii (8/20, 40%), Rh. haemaphysaloides (6/20, 30%), Hy. excavatum (2/20, 10%) and Rh. microplus (1/20, 5%). Co-occurrence of V/TBP was also detected in tick specimens (single V/TBP infection: 32 ticks; double and triple: 13 and 5 tick samples). The detected pathogens shared a phylogenetic relationship with similar isolates published in NCBI GenBank from Old and New World countries., Conclusion: Ixodid ticks infesting dogs harbor a diverse array of V/TBPs including zoonotic agents from Pakistan. Furthermore, the presence of D. immitis in ticks that infest dogs raises the possibility that this parasite has either attained its dead-end host (i.e. the tick) while feeding on dogs or has expanded its range of intermediate/paratenic hosts. Further research work is needed to investigate the epidemiology and confirm the vector competence of screened tick species for these pathogens from Pakistan., (© 2023. The Author(s).)

Objective: To identify the association between acute infection and acute coronary syndrome, and to evaluate the outcomes in such cases., Methods: The cross-sectional, retrospective, analytical study was conducted at the Aga Khan University Hospital, Karachi, from July to December 2020, and comprised data from January to December 2019 of acute coronary syndrome patients aged >18 years. Data related to demographics, comorbidities, smoking status and history of dyslipidaemia. Binary logistic regression was used to explore the association of infections with acute coronary syndrome. Data was analysed using SPSS 26., Results: Of the 1202 patients with acute coronary syndrome, 189(15.7%) had infection before the coronary event. The mean age of the patients was 68.5±12.4 years, and 97(51.3%) of them were females. Community-acquired pneumonia was found in 105(55.6%) patients, followed by urinary tract infection 64(33.9%) and cellulitis 8(4.2%). For pneumonia, the odds of having non-ST elevated myocardial infarction was 1.1 (95% confidence interval: 0.4-3.0). With urinary tract infections, unstable angina was associated with an odd ratio of 4.2 (95% confidence interval: 1-17.4), and ST elevated myocardial infarction was associated with odd ratio of 3.7 (95% confidence interval: 0.4-31)., Conclusions: Bacterial infections were found to be associated with acute coronary syndrome. Bacterial infections with pneumonia and urinary tract infections showed a higher risk of myocardial ischaemia..

Natural pristine environments including cold habitats are thought to be the potent reservoirs of antibiotic-resistant genes and have been recurrently reported in polar glaciers' native bacteria, nevertheless, their abundance among the non-polar glaciers' inhabitant bacteria is mostly uncharted. Herein we evaluated antibiotic resistance profile, abundance of antibiotic-resistant genes plus class 1, 2, and 3 integron integrases in 65 culturable bacterial isolates retrieved from a non-polar glacier. The 16S rRNA gene sequencing analysis identified predominantly Gram-negative 43 (66.15%) and Gram-positive 22 (33.84%) isolates. Among the Gram-negative bacteria, Gammaproteobacteria were dominant (62.79%), followed by Betaproteobacteria (18.60%) and Alphaproteobacteria (9.30%), whereas Phyla Actinobacteria (50%) and Firmicutes (40.90%) were predominant among Gram-positive. The Kirby Bauer disc diffusion method evaluated significant antibiotic resistance among the isolates. PCR amplification revealed phylum Proteobacteria predominantly carrying 21 disparate antibiotic-resistant genes like; bla AmpC 6 (100%), bla VIM-1, bla SHV and bla DHA 5 (100%) each, bla OXA-1 1 (100%), bla CMY-4 4 (100%), followed by Actinobacteria 14, Firmicutes 13 and Bacteroidetes 11. Tested isolates were negative for bla KPC, qnrA, vanA, ermA, ermB, intl2, and intl3. Predominant Gram-negative isolates had higher MAR index values, compared to Gram-positive. Alignment of protein homology sequences of antibiotic-resistant genes with references revealed amino acid variations in bla NDM-1, bla OXA-1, bla SHV, mecA, aac(6)-Ib3, tetA, tetB, sul2, qnrB, gyrA, and intI1. Promising antibiotic-resistant bacteria, harbored with numerous antibiotic-resistant genes and class 1 integron integrase with some amino acid variations detected, accentuating the mandatory focus to evaluate the intricate transcriptome analysis of glaciated bacteria conferring antibiotic resistance., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

Coxiellosis, also known as Q fever, is a zoonotic disease caused by Coxiella burnetii, a gram-negative bacterium that exerts a significant deleterious impact on the productive and reproductive capabilities of livestock, severely effecting the economics of this sector. In this study, 448 sera samples from cattle (n = 224) and buffalo (n = 224) were collected from 112 farms in Pakistan and examined for antibodies against C. burnetii using an indirect ELISA. Ticks were also collected from these animals. Serological analysis revealed a 23.66% and 27.23% seroprevalence of Q fever in cattle and buffalo, respectively. Odds ratio (OR) analysis of the factors associated with C. burnetii seropositivity was performed, and a multivariable logistic model identified five main variables associated with the seropositivity for coxiellosis. These were: (i) the absence of acaricide use (OR 5.61; 95% CI 2.97-10.94); (ii) the presence of ticks (OR 3.23; 95% CI 1.87-5.69); (iii) the abortion history during the preceding year on the farm (OR 14.96; 95% CI 8.09-29.34); (iv) the presence of sheep and goats (OR 2.47; 95% CI 1.20-5.35); and (v) the absence of a separate parturition area (OR 3.17; 95% CI 1.76-5.86). This study provides new insights into the seroprevalence of Q fever in large ruminants across seven studied districts of Punjab, Pakistan, also providing baseline data to inform improved herd management and on-farm practices for the prevention and control of Q fever in large ruminants in the region. Results of this work suggest that further molecular investigation of coxiellosis is warranted to provide a more thorough evaluation of C. burnetii epidemiology in Pakistan., (© 2022. The Author(s).)

Tick infestation is a major public and animal health concern causing significant financial losses, especially in tropical and subtropical regions of the world. This study aimed at investigating the epidemiologic profile of ticks infesting cattle and molecular identification of R. microplus in the centrally ignored part of Khyber Pakhtunkhwa, Pakistan. A total of 600 cattle from 20 farms were examined for the tick infestation, among them 358 (59.7%) cattle were infested with ticks. A total of 2118 nymph, larvae and adult tick stages were collected and morphologically identified followed by molecular confirmation of Rhipicephalus microplus. Host-based demographic and ecological parameter analysis revealed significantly higher tick infestation in adult, female, exotic, freely grazing, and with irregular/no acaricides treated cattle. The univariate logistic analysis showed that host age, gender, breed, acaricides use, and feeding method were significantly (P < 0.05) associated, whereas multivariate analysis revealed only host breed and feeding method were potential risk factors (P < 0.05) for tick infestation. Microscopy-based examination identified four different species of ticks including R. microplus (44.5%), Hyalomma anatolicum (38.5%), and Hyalomma marginatum (10.5%) and Hyalomma excavatum (6.5%). Tick infestation pattern showed that 55.9% of cattle was found co-infested with R. microplus and H. anatolicum followed by R. microplus and H. anatolicum and H. marginatum (29.3%) then R. microplus, H. anatolicum, H. marginatum, and H. excavatum (11.2%). Sequencing of the second internal transcribed spacer (ITS2-) and 16S rRNA gene fragments also confirmed the molecular identification of Rhipicephalus microplus. Phylogenetic analysis of ITS-2 revealed all sequences clustered in single clade of the R. microplus while the 16S rRNA nucleotide sequences showed that R. microplus in this study was clustered together in clade A along with other isolates from Pakistan, China, and India. The high tick infestation suggests the need for designing strategic and integrated control measures for ticks in order to ensure good health of domestic animals in this region of Pakistan., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: In the initial stage of infectious diseases such as COVID-19 creates a prodigious uncertainty not only in general population but also in health care professionals. This often leads to emotional distress in general public and particularly in health care professionals., Objectives: During COVID-19 pandemic in Pakistan, the health care professionals experienced unusual stressors. This study aimed to examine the coping responses, optimism, pessimism and psychiatric morbidity of health professionals serving the COVID-19 patients., Participants: Total 87, health care professionals take part in study, whom 36 were physicians and 51 nursing staff. Among these 44 were male, and 43 were female., Method: Researchers used cross sectional research design in this study. Physicians and nurses completed self-reported questionnaires. Participants provided demographics data and recorded their responses to self-administered questionnaires. Researchers administered Brief coping orientation to problems experiences (COPE) for assessing the coping strategies, while they assessed psychiatric morbidity through general health questionnaires. Similarly, future expectancy of health care professional was assessed by using life orientation scale. Participants were recruited from quarantine words in two federal government hospitals providing health care services to COVID-19 patients in Pakistan., Result: The result showed a significant relationship in optimism and problem focus coping style and avoidance coping style. Moreover, male health professionals score high on optimism as compared to female health professionals. While, avoidance coping style were seen higher in female health professional as compare to male. The result revealed that optimism and psychiatric morbidity were significantly positive in health professionals having problem focus and avoidance coping style., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background: A healthy diet in the adolescence period is essential for physical, mental, and immunological development. We aimed to assess macronutrient consumption in the diet of adolescent school children using 24 h recalls in four seasons of the year., Method: This was a longitudinal study conducted from February 2014 to June 2015. The study population included 155 school children aged 7-14 years from an urban school in Karachi. 24HR recall was conducted on 4 random days of the 4 main seasons. A food composition table was developed where the weight, calories, carbohydrate, fat, and protein content of the food items were listed. Macronutrients quantification was calculated by using proportional weight from the food composition table. Food groups were also assigned to each food item including vegetables, fruits, grains, protein foods, dairy products, and oils., Results: A total of 155 adolescent children aged between 7 and 14 years were approached. Out of the 155 preadolescents and adolescents, 150 (96.7%) agreed to participate. The mean (SD) age of the children was 11.31 (1.6) years, and 59% of all the children were males. Overall mean (SD) daily intake for all seasons was 195.31 (86.87) grams of carbohydrates, 94.77 (71.87) grams of proteins, and 55.87 (30.79) grams of fats. Carbohydrates formed 48.16%, protein 21.92%, and fat 29.93% of the total caloric intake. The mean (SD) daily caloric intake was 1517 (644) grams. Overall, the highest source of calories was from carbohydrate 781 (347) Kilocalories (Kcal), followed by fat 502 (277) Kcal and protein 379 (287). The Carbohydrate intake in 24 h was highest in the autumn; 212.81 (85.37), and there was a significant difference in carbohydrate intake in all seasons (p value 0.003). Consumption of discretionary food group was high (31.3%), and consumption of fruits and vegetables was low (29%)., Conclusion: The study reports a suboptimal caloric intake of fewer than 2000 cal/day among the adolescents from school. The highest source of calories was from carbohydrates.The highest consumption of food was in autumn and the least in summer. Fruits and vegetable intake was low, and discretionary food intake was high., (© 2021. The Author(s).)

Objective: To study the relationship between dysfunctional schema modes and post-traumatic stress disorder among trauma survivors., Methods: The cross-sectional study was conducted at the Pakistan Institute of Medical Sciences, Islamabad, Pakistan, from March to August 2019, and comprised patients of traumatic brain injury and orthopaedic trauma. Data was collected using Schema Mode Inventory and the Clinician Administered Post-Traumatic Stress Disorder Scale version 5. Data was analysed using SPSS 23., Results: Of the 281 patients, 203(72.2%) were males, 78(27.8%) were females, and 157 (55.9%) were aged 18-40 years. Besides, 137(48.8%) had orthopaedic trauma, 96(34.2%) traumatic brain injuries, and 48(17%) had multiple injuries. Post-traumatic stress disorder symptoms and dysfunctional schema modes were higher in patients with moderate injuries with a mean of 42.14±7.36. Intentionally injured patients 42.70±6.92 and female trauma survivors 42.05±8.26 had more symptoms., Conclusions: Maladaptive schema mode could lead to post-traumatic stress disorder symptoms among trauma survivors with history of orthopaedic injury, traumatic brain injury and multiple injuries.

Objective: To compare the knowledge, attitude and practice regarding diabetes mellitus among diabetics and non-diabetics., Methods: The cross-sectional study was conducted at the Government College University, Faisalabad, Pakistan, from December 2017 to April 2018, and comprised subjects recruited randomly from different cities of Punjab, Pakistan. Data was collected using a predesigned structured questionnaire regarding socio-demographic characteristics, general knowledge about diabetes, perception regarding indication, risk factors, diagnosis, and complications, and practices followed for treatment and management of diabetes., Results: Of the 2,000 subjects, 972(48.6%) had family history of diabetes, 1338(66.9%) were living in urban areas, 1068(53.4%) were university graduates, 804(40.2%) were employed and 1152(57.6%) belonged to socio-economically balanced families. Composite knowledge score was significantly associated with age and socio-economic status (p<0.05). A highly significant association was observed regarding family history (p<0.001), level of education (p<0.0001) and occupation (p<0.001) with composite knowledge score., Conclusions: The knowledge level about diabetes was seen to be average.

Purpose: Patients with acquired brain injury are a vulnerable group in Pakistan with limited psychosocial services available to them. The current study sought to determine the incidence of post-traumatic stress disorder (PTSD) in ABI patients with traumatic and non-traumatic injury., Patients and Methods: A cross-sectional research design was used. The assessment of PTSD was carried out by using the Clinician Administered PTSD Scale (CAPS-5). In the study, there were 132 adults older than 18 years who have ABI-either traumatic or non-traumatic. The study was conducted in a public hospital in Islamabad, Pakistan, and the data was collected one-month post brain injury., Results: The results showed that traumatic brain injury (TBI) patients have more PTSD symptoms as compared to non-TBI patients (p = 0.00). Intrusive (p = 0.00) and hyper arousal symptoms (p = 0.04) of PTSD were higher in TBI than in non-TBI patients. There was statically non-significant mean difference in symptoms of PTSD among male and female patients (p = 0.34). The moderately brain injury patients have high PTSD symptoms as compared to mild brain injury patients (p = 0.04). Further, the frequency of severe and extreme PTSD symptoms was high in TBI patient as compared to non TBI patients (p = .05) CONCLUSION: PTSD severity and PTSD symptoms such as intrusive and avoidance symptoms were higher in TBI patients than in non-TBI patients. This study contributed to the literature by assessing the onset of PTSD in patients with acquired brain injury. This study also provided knowledge to plan psychosocial interventions for patients with acquired brain injury having PTSD symptoms., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: This study aimed to assess the extent of knowledge and understanding of rabies disease in rural and urban communities of Pakistan. It also identified malpractices after suspected dog bite that might pose a risk for humans contracting rabies., Methods: A cross-sectional study was conducted (n = 1466) on people having different age groups and educational levels in four different geographic regions of Punjab and Khyber Pakhtunkhwa provinces in Pakistan. Knowledge, attitude, and practices of people were assessed using a structured questionnaire. We used a bivariate and multivariate analysis to study the association between rabies related mortalities in near or extended family members and different risk behaviors., Results: Our results demonstrate that the majority of the juvenile population (less than 18 years of age) were not aware of the clinical signs of rabies in animals. 75% of the total respondents were not vaccinated against rabies, 60% did not seek a doctor's advice after a suspected animal bite, and 55% had inadequate health care facilities for rabies patients in local hospitals. Respondents that had pets at home had not vaccinated (38%; p < 0.05; odds ratio 1.58) themselves against rabies due to lack of knowledge and awareness of pre-exposure prophylaxis for rabies (51%; p < 0.05; odds ratio 1.25). They also tend to not visit doctor after suspected bite (52%; p < 0.05; odds ratio 1.97), which may had resulted in more deaths (65%; p < 0.05; odds ratio 1.73) of someone in their near or extended family due to rabies., Conclusions: Lack of knowledge about the nature of rabies disease and prophylaxis has contributed to increase of rabies related deaths. Inadequate health care facilities and poor attitude of not seeking medical attention after suspected dog bite are the major reasons of rabies related deaths. These findings could help in devising a targeted management strategy and awareness program to control and reduce the incidence of human rabies related deaths in Pakistan.

Bisphenol A (BPA) is considered as xenoestrogen, a crucial component utilized for the manufacturing of plastic products. It has a potential to disrupt the endocrine system and induces endocrine-related metabolic disorders. We aimed to investigate the exposure of BPA in Pakistani population and its association with sociodemographic features, dietary habits, and risk factors of diabetes mellitus (DM). This cross-sectional study was conducted on 400 participants among which 61.75% participants were diabetic and 38.25% were non-diabetic. We developed a structured questionnaire, gathered sociodemographic data, and collected their urine and blood samples for the estimation of BPA and various biomarkers as risk factors of DM, respectively. Pearson correlation coefficient was determined for urinary BPA levels and DM risk factors. Urinary BPA values were adjusted for confounders. Sociodemographic data shown that urinary BPA level was significantly higher (p < 0.05) in obese people (BMI > 27) living in semi-urban and industrial areas. BPA was detectable in 75% of study participants. Urinary BPA level was found to be higher in diabetic participants compared with that of non-diabetics. A significant correlation is observed between BPA exposure and DM risk factors. We found that urinary BPA level was correlated with elevated levels of HbA1c (r = 0.6028), HOMA-IR (r = 0.5356), CRP (r = 0.6946), BUN (r = 0.6077), AST (r = 0.5151), FFA (r = 0.5759), TGs (r = 0.5608), and MDA (r = 0.6908). Hence, our study adds to the growing body of evidence supporting the role of BPA exposure as a risk factor for DM and may be associated with higher glycemic index, increased pro-inflammatory and oxidative stress biomarkers, dyslipidemia, and impaired functioning of the liver and kidney. Heating food in plastic containers and consumption of packed food items are the main sources of BPA exposure which are positively associated with DM.

Crimean-Congo Haemorrhagic Fever (CCHF) is a deadly viral zoonotic disease, which is endemic in Pakistan. We report a case study analysis of three cases of CCHF which occurred in Chakwal, Pakistan in 2016. The disease was suspected in three patients exhibiting clinical symptoms suggestive of CCHF; two of the three patients died. The presence of CCHF was detected by seroconversion (IgG) from the surviving patient, while the antigen was detected in Hyalomma ticks originating from animals in the vicinity. This report indicates increase threat emergence of CCHF in Pakistan and highlights its zoonotic implications requiring immediate interventions under the "One-Health" concept., (© 2019 Blackwell Verlag GmbH.)

Background: Genetic polymorphisms in the human arginase-1 (ARG1) gene locus and their effects on cardiovascular disease have not been thoroughly elucidated. The aim of the present study was to investigate the association of the variant ARG1 alleles rs2781666 and rs2781667 with coronary artery disease (CAD)., Methods: ARG1 rs2781666G/T and rs2781667C/T polymorphisms were characterized in a case-control study consisting of 200 complex Pakistani families with CAD history. Heritability of susceptibility/variant alleles was investigated from parent-offspring trios in these families. Determination of serum liped levels was performed spectrophotometrically, while serum arginase-1 activity and the concentrations of nitric oxide metabolites were detected by enzyme colorimetric assay. Genotyping of the two polymorphic sites in the ARG1 gene was performed using polymerase chain reaction and restriction analysis., Results: A significant increase in arginase-1 activity was observed in CAD patients compared with controls (p < 0.0001). Arginase-1 was negatively correlated with serum nitrite and nitrate (r = -0.8137 and r = -0.8444, respectively). There was a significant difference in distribution of genotypes for rs2781666 and rs2781667 polymorphisms between patients and controls (p < 0.001 for each). Similarly, the variant T allele at both loci showed a significant association with the disease compared with subjects free of CAD (p < 0.0001 for each). The transmission-disequilibrium test revealed a significant association of rs2781666 and rs2781667 polymorphisms with CAD (p < 0.0001 for each)., Conclusion: This report is the first to describe arginase-1 activity and an association between ARG1 gene polymorphisms and familial CAD from Pakistan.

Background: Evidence-based decision making is essential for appropriate prioritization and service provision by healthcare systems. Despite higher demands, data needs for this practice are not met in many cases in low- and middle-income countries because of underdeveloped sources, among other reasons. Emergency departments (EDs) provide an important channel for such information because of their strategic position within healthcare systems. This paper describes the design and pilot test of a national ED based surveillance system suitable for the Pakistani context., Methods: The Pakistan National Emergency Department Surveillance Study (Pak-NEDS) was pilot tested in the emergency departments of seven major tertiary healthcare centres across the country. The Aga Khan University, Karachi, served as the coordinating centre. Key stakeholders and experts from all study institutes were involved in outlining data needs, development of the study questionnaire, and identification of appropriate surveillance mechanisms such as methods for data collection, monitoring, and quality assurance procedures. The surveillance system was operational between November 2010 and March 2011. Active surveillance was done 24 hours a day by data collectors hired and trained specifically for the study. All patients presenting to the study EDs were eligible participants. Over 270,000 cases were registered in the surveillance system over a period of four months. Coverage levels in the final month ranged from 91-100% and were highest in centres with the least volume of patients. Overall the coverage for the four months was 79% and crude operational costs were less than $0.20 per patient., Conclusions: Pak-NEDS is the first multi-centre ED based surveillance system successfully piloted in a sample of major EDs having some of the highest patient volumes in Pakistan. Despite the challenges identified, our pilot shows that the system is flexible and scalable, and could potentially be adapted for many other low- and middle-income settings.

A case-control and trio-families study was performed to establish a potential association between TNF-alpha gene promoter SNPs at -308 and -238, and occurrence of CAD in a Pakistani population. In the first phase, 150 patients and 150 controls were enrolled in the case-control association study. In the second phase, heritability of susceptible alleles was investigated from 88 trio-families with CAD affected offspring. Biochemical analysis of lipids and hs-CRP was carried out spectrophotometrically, while serum TNF-alpha concentrations were determined by enzyme-linked immunosorbent assay. Genotyping of the TNF-alpha SNPs were determined by PCR-RFLP method. Elevated serum TNF-alpha and hs-CRP were observed from CAD vs. controls (P<0.0001; for both). The evaluation of TNF-alpha-308G>A polymorphism in case-control study revealed that the said SNP was significantly associated with the increased risk of CAD. The findings demonstrated a significant link between the TNF-alpha variant allele A at -308 and CAD (P=0.0035), whereas the -238 SNP was not associated with the disease. Haplotype A-G of the TNF-alpha gene at -308G>A and -238G>A showed higher frequency in the patient group compared with controls (P<0.05). Moreover, data showed preferential transmission of the disease susceptible allele A at TNF-alpha-308 from parent to affected offspring in a trio-family study (P<0.0001). The current research leads to conclusion that the TNF-alpha-308G>A polymorphism is associated with CAD in the study population. Furthermore, for the first time, we showed that the TNF-alpha-308A allele was significantly associated with the familial CAD in our high risk population., (Copyright © 2014. Published by Elsevier Inc.)

Background: We aimed to analyse the frequency and patterns of fall-related injuries presenting to the emergency departments (EDs) across Pakistan., Methods: Pakistan National Emergency Departments surveillance system collected data from November 2010 to March 2011 on a 24/7 basis using a standardized tool in seven major EDs (five public and two private hospitals) in six major cities of Pakistan. For all patients presenting with fall-related injuries, we analysed data by intent with focus on unintentional falls. Simple frequencies were run for basic patient demographics, mechanism of falls, outcomes of fall injuries, mode of arrival to ED, investigations, and procedures with outcomes., Results: There were 3335 fall-related injuries. In cases where intent was available, two-thirds (n = 1186, 65.3%) of fall injuries were unintentional. Among unintentional fall patients presenting to EDs, the majority (76.9%) were males and between 15-44 years of age (69%). The majority of the unintentional falls (n = 671, 56.6%) were due to slipping, followed by fall from height (n = 338, 28.5%). About two-thirds (n = 675, 66.6%) of fall injuries involved extremities, followed by head/neck (n = 257, 25.4%) and face (n = 99, 9.8%). Most of the patients were discharged from the hospital (n = 1059, 89.3%). There were 17 (1.3%) deaths among unintentional fall cases., Conclusion: Falls are an important cause of injury-related visits to EDs in Pakistan. Most of the fall injury patients were men and in a productive age group. Fall injuries pose a burden on the healthcare system, especially emergency services, and future studies should therefore focus on safety measures at home and in workplaces to reduce this burden.

In this study, we have performed autozygosity mapping on a large consanguineous Pakistani family segregating with intellectual disability. We identified two large regions of homozygosity-by-descent (HBD) on 16q12.2-q21 and 16q24.1-q24.3. Whole exome sequencing (WES) was performed on an affected individual from the family, but initially, no obvious mutation was detected. However, three genes within the HBD regions that were not fully captured during the WES were Sanger sequenced and we identified a five base pair deletion (actually six base pairs deleted plus one base pair inserted) in exon 7 of the gene FBXO31. The variant segregated completely in the family, in recessive fashion giving a LOD score of 3.95. This variant leads to a frameshift and a premature stop codon and truncation of the FBXO31 protein, p.(Cys283Asnfs*81). Quantification of mRNA and protein expression suggests that nonsense-mediated mRNA decay also contributes to the loss of FBXO31 protein in affected individuals. FBXO31 functions as a centrosomal E3 ubiquitin ligase, in association with SKP1 and Cullin-1, involved in ubiquitination of proteins targeted for degradation. The FBXO31/SKP1/Cullin1 complex is important for neuronal morphogenesis and axonal identity. FBXO31 also plays a role in dendrite growth and neuronal migration in developing cerebellar cortex. Our finding adds further evidence of the involvement of disruption of the protein ubiquitination pathway in intellectual disability.

Interleukin-6 (IL-6) is a well-known inflammatory cytokine and suggested to be involved in the development of coronary artery disease (CAD). IL-6 gene expression has been investigated with controversy in CAD patients. This study investigates the association of the IL-6 gene expression with CAD, the molecular basis for the regulation of interleukin-6 expression in a Pakistani population. Our data show that the serum IL-6 levels were increased in patients with CAD compared with healthy controls and that the IL-6 gene polymorphism at -174 was more prevalent in CAD cases. There was a statistically significant association between the IL-6 gene polymorphism and CAD, which may be associated with an increased risk for the disease. Moreover, circulating IL-6 and hs-CRP levels were significantly higher in patients with CC genotype (P < 0.0001 and P < 0.0001, resp.). In a binary logistic-regression model, an independent association was found between CAD and increased serum IL-6 and hs-CRP levels and -174G>C polymorphism. This is the first report on the IL-6 expression and the IL-6 gene polymorphism in patients with CAD from Pakistan, and hence it highlights a novel risk factor for the disease.

Background: To determine the accuracy, turnaround time and cost effectiveness of bedside monitoring of blood glucose levels by non-laboratory health care workers and centralized testing of blood glucose by automated analyzer in a tertiary care hospital., Methods: The study was conducted in Section of Chemical Pathology, Department of Pathology and Microbiology and Section of Endocrinology Department of Medicine, Aga Khan University and Hospital Karachi, from April 2005 to March 2006. One hundred and ten patients were included in the study. The blood glucose levels were analyzed on glucometer (Precision Abbott) by finger stick, using Biosensor Technology. At the same time venous blood was obtained to analyze glucose in clinical laboratory on automated analyzer (SYNCHRON CX7) by glucose oxidase method., Results: We observed good correlation between bed side glucometer and laboratory automated analyzer for glucose values between 3.3 mmol/L (60 mg/dl) and 16.7 (300 mg/dl). A significant difference was observed for glucose values less than 3.3 mmol/L (p = 0.002) and glucose values more than 16.67 mmol/l (p = 0.049). Mean Turnaround time for glucometer and automated analyzer were 0.08 hours and 2.49 hours respectively. The cost of glucose testing with glucometer was 48.8% lower than centralized lab based testing., Conclusion: Bedside glucometer testing, though less expensive does not have good accuracy in acutely ill patient with either very high or very low blood glucose levels.

Hepatitis B virus (HBV) and C virus (HCV) are major public health concerns in Pakistan. We conducted a baseline analysis of first-time replacement blood donors at the blood bank of Jinnah Postgraduate Medical Centre in the year 2000 with the view to developing low-cost sentinel surveillance for these infections. Among 7325 such donors, 264 (3.6%) were positive for anti-HCV and 344 (4.7%) for HBsAg. HCV seroprevalence was significantly positively associated with age and lower education. Those speaking Sindhi or other minor languages had a significantly higher prevalence of HBV infection. Blood bank data could provide reliable information to monitor trends in prevalence of these infections.

Autosomal recessive primary microcephaly (MCPH) is a rare human genetic disorder in which the head circumference is reduced because of abnormality in fetal brain growth. To date, six loci and four genes have been identified for this condition. Our study of primary MCPH led to the identification of 33 Pakistani families with different ethnic backgrounds. Most of these families showed linkage to MCPH5 locus on chromosome 1q31. Only one family with Pashtoon origin from a remote region in Pakistan linked to MCPH6 locus on chromosome 13q12.12-q12.13. Sequence analysis of exon 11 of CENPJ gene, located at MCPH6 locus, revealed a novel four base pair deletion mutation, which is predicted to be protein truncating.

Sea buckthorn (Hippophae rhamnoides ssp. turkestanica) is a very important medicinal plant in northern areas of Pakistan. The fruit of sea buckthorn is rich in nutrients and medicinal compounds such as vitamins, carotene, flavonoids, essential oil, carbohydrates, organic acids, amino acids, and minerals. In order to compare various populations of sea buckthorn for chemical composition, eight populations from different areas of northern Pakistan were compared using fruit characteristics. Phytochemical analysis of berries showed vitamin C (250-333 mg/100 g), seed oil (7.69-13.7%), oil in softer pulp (19.2-29.1%), phytosterol content of seed oil (3.3-5.5%), and anthocyanin (0.5-25 mg/L), while the mineral element composition analysis revealed high contents of potassium (140-360 ppm), sodium (20-80 ppm), calcium (70-98 ppm), magnesium (150-240 ppm), iron (40-150 ppm), and phosphorus (110-133 ppm). This study established sea buckthorn berries as a good source of biochemical and mineral elements. The high variation between different populations shows the potential of selecting and breeding of the raw material for various defined purposes.

Objective: To collect baseline information on congenital adrenal hyperplasia (CAH) and to identify relevant issues specific to this disease in Pakistan., Methods: A retrospective analysis of medical records of pediatric patients registered for serum 17 hydroxyprogesterone (17-OHP) measurement and documented to have CAH in the period 1987 to 1998 was carried out at The Aga Khan University, Karachi (AKU). The clinical notes were reviewed for documentation of CAH as the diagnosis., Results: Of the 152 children registered for 17-OHP testing, sixty-three were diagnosed with CAH. Salt wasting, simple virilization and non-classical CAH was found in 40 (63%), 18 (29.0%) and 5 (8.0%) patients respectively. Twenty-one (33.9%) patients were incorrectly assigned sex and of these, 20 (32.2%) patients were females who were either considered males or just not assigned gender. Parental consanguinity was found in 33 (52.3%) cases. No case had a history of similar features in either parent but in 19 (30.6%) cases similar features were present in siblings. Sixteen cases (25.4%) had a history of sibling death in the neonatal period and 7 had a history of sibling death in infancy. Maternal obstetric histories identified 3 (4.8%) cases with a history of still birth(s) and 4 (6.4%) with a history of abortion(s)., Conclusion: Children with CAH should be diagnosed early as a rational and judicious choice of sex assignment is a critical aspect of treatment. The high rate of consanguinity emphasized the need to establish the true incidence of the defect in Pakistani population.

Objective: To determine the response to i.v. anti-D and its comparison with splenectomy as second line therapy in refractory and relapsed cases of ITP, in the Aga Khan University Hospital, Karachi., Methods: A total of 23 patients with chronic ITP were treated with either anti-D or splenectomy as second line treatment. The patients were assessed for time to achieve a response to second line treatment, duration of response and adverse events., Results: There were 12 patients in the anti-D group and 11 in the splenectomy group. The mean platelet count at presentation was 9,000/cumm. The mean age was 8.9 years and 13.0 years and the male to female ratio was 1:1 and 1:1.2 in anti-D and splenectomy group respectively. 54.5% of the patient in the anti-D group responded compared to 81.8% in the splenectomy group. Median time to achieve a response was 7 days in the anti-D group and 1 day in the splenectomy group. Mean time to relapse was 87.8 days in the anti-D group and 55.4 days in the splenectomy group. No adverse events were recorded for any of the infusions of anti-D and none of the patients had more than 0.5 gm/dl fall in the hemoglobin level following anti-D infusion., Conclusion: It was thus concluded that Anti-D is a relatively safe, convenient and effective therapy for chronic ITP and can be used as a splenectomy sparing agent when treatment is clinically indicated.