37 research outputs found

    Analysis of heat-induced DNA damage during PCR and verification, validation and comparative analysis of two PCR megaplexes

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    Biological evidence collected at crime scenes are often subjected to forensic deoxyribonucleic acid (DNA) testing. During forensic DNA testing the DNA from the evidence and known samples are extracted, purified, amplified using Polymerase Chain Reaction (PCR), and analyzed using capillary electrophoresis (CE). In order to appropriately compare the profile of the suspect to the evidence, it is essential that interpretation parameters and optimized processing schemes are established. This study endeavors to accomplish this by: first, evaluating whether the PCR temperature cycling is detrimental to the amplification process; and second, by establishing and comparing interpretation parameters for two commonly employed short tandem repeat (STR) megaplexes. To evaluate the effects of temperature cycling on downstream signal, a dynamic systems model was developed, validated, and used to test the effects of temperature on DNA damage and the subsequent fluorescence signal. Though DNA is generally thought to be a stable molecule, heat-induced damage does occur. Specifically, this model assesses the damage to the guanine and cytosine bases during temperature cycling. The model conducts the amplification of a single locus during PCR and generates the peak height observed after capillary electrophoresis. The model was designed to assess not only the effects of heat-induced DNA damage but to also incorporate variability in PCR efficiency. The simulated data indicate that heat-induced DNA damage does not significantly reduce the allelic signal. Also, although changes in PCR efficiency introduce variability in the peak heights at all targets, the peak heights observed with and without heat-induced DNA damage are not significantly different. In fact, the variation in PCR efficiency has a larger effect on the number of amplicons produced than does the heat-induced DNA damage. The second part of this study compares two PCR amplification megaplexes, PowerPlex® Fusion and GlobalFiler®, by evaluating their sensitivities, limits of detection, presence of artifacts, heterozygous peak balance, and ability to amplify minor contributors in DNA mixtures. Analysis of single source samples using weighted least squares regression analysis indicates that PowerPlex® Fusion has greater analytical sensitivities and lower limits of detection at comparable dye channels, and both kits display similar heterozygous balance. However, the GlobalFiler® processing scheme produced fewer artifacts for the various single source samples analyzed, particularly at higher target amounts. Also, analysis of two and three person DNA mixtures indicates that both megaplexes perform equally well when detection of the minor contributor is the criterion

    Immunohistochemical Expression of Nkx3.1 in Prostatic Adenocarcinoma and Benign Prostatic Hyperplasia at a Tertiary Care Hospital in Karachi

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    OBJECTIVES To determine NKX3.1 expression in prostatic adenocarcinoma and benign prostate hyperplasia on Immunohistochemistry at a tertiary care hospital in Karachi. METHODOLOGY 74 prostatic specimens were recruited in this comparative cross-sectional study at the Department of Pathology, Pakistan Navy Station Shifa Hospital, Karachi, from January 2018 to February 2019. Of these, 37 specimens were of prostatic adenocarcinoma, and 37 were benign prostate hyperplasia. All specimens were subjected to immunohistochemical staining with NKX3.1. Statistical analysis was done by using SPSS version 23.0. The association of the extent of NKX3.1 staining between the adenocarcinoma and hyperplasia group was assessed using the Chi-square test x2. RESULTSOf the 37 cases of benign prostatic hyperplasia subjected to NKX3.1 staining, 32 showed positive staining with strong to moderate intensity. No staining was observed in 5 cases. 37 cases of adenocarcinoma prostate stained for NKX3.1 revealed positive staining in 30 cases with strong to moderate intensity. Negative staining was seen in 7 cases. The prostatic adenocarcinoma showed a statistically significant association of NKX3.1 positivity compared to benign prostatic hyperplasia cases. The p-value was found to be 0.03. CONCLUSION NKX3.1 staining was highly specific for prostate epithelium, as it was positive in most cases. This immune marker was useful for distinguishing prostatic origin in the context of metastatic lesions. Adding NKX3.1 protein staining to a panel of features may add value to the diagnosis if applied in the appropriate clinicopathologic context

    Self-medication amongst university students of Karachi: prevalence, knowledge and attitudes

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    OBJECTIVE: To determine the prevalence, attitude and knowledge of self-medication amongst university students of Karachi, Pakistan. METHODS: This cross-sectional, study was conducted from Jan-Feb 2007. A convenience sample was taken from 2 medical and 2 non-medical universities of the city of Karachi, Pakistan. Data was analyzed using SPSS v 14 and associations were tested using the Chi square test. RESULTS: Of the 572 participants (mean age=21 +/- 1.8 years, Male: Female ratio=1:1.5), 295 were medical and 277 were non-medical students. The prevalence of self-medication was 76%. Forty three percent students stated that they alter the regimen of prescribed medicines while 61.9% stated that they stop taking a prescribed medicine without consulting a doctor. The most common reason for self-medication was previous experience (50.1%) and the most common symptoms were headache (72.4%), flu (65.5%), and fever (55.2%). Commonly used medicines were analgesics (88.3%), antipyretics (65.1%) and antibiotics (35.2%). Eighty seven percent of students thought self-medication could be harmful and 82.5% students thought that it was necessary to consult a doctor before taking a new medicine. There was no significant difference between the self medication practices of medical and non medical students (p=0.8) CONCLUSION: Prevalence of self-medication is high in the educated youth, despite majority being aware of its harmful effects. There is a need to educate the youth to ensure safe practices. Strict policies need to be implemented on the advertising and selling of medications to prevent this problem from escalating

    Estimation of Chromium in Soil-Plant-Animal Continuum: A Case Study in Ruminants of Punjab, Pakistan

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    The increased use of waste water for agriculture purposes has increased around the globe. There are toxic metals present in waste water which affect plants, animals and human health. But at the same time it also contains useful nutrients which increase growth of plants. Different indices were applied to evaluate the metals present in water soil plant and milk. Various and correlation were determined with the help of SPSS, mean significance was found at the probability levels of 0.05, 0.001 and 0.01. The highest value of Cr was found in Avena sativa (0.7872 mg/kg) collected from site 5 while lower concentration of Cr was observed in Brassica campestris (0.0743 mg/kg) at site 4. In soil samples, Trifolium alexandrinum showed highest value of Cr (0.9887 mg/kg) at site 1 while lowered concentration was observed in Zea mays (0.1862 mg/kg) at site 3. Milk samples of site 5 had higher value of Cr (0.2898 mg/kg) and lowest at (0.1540 mg/kg) site 2. Water samples of site 3 had high value of Cr in them (1.849 mg/kg) and lowered concentration was found in water samples of site 5 (0.219 mg/kg). Cr concentration in fodders ranges from 0.0743 to 0.7872 mg/kg, soil 0.1862 to 0.9887 mg/kg, milk 0.1540 to 0.2898 mg/kg and water 0.219 to 1.849 mg/kg. In water samples, Cr level were above than permissible limit. In milk samples, concentration of Cr was greater than permissible limit which shows that it is hazardous to human health, may be due to pollution of environment like air, water and soil. This study concluded that if animals are allowed to graze on contaminated fodders and drink wastewater then metals were accumulated in their tissues and milk which causes toxicity to human health

    Global, regional, and national cancer incidence, mortality, years of life lost, years lived with disability, and disability-Adjusted life-years for 29 cancer groups, 1990 to 2017 : A systematic analysis for the global burden of disease study

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    Importance: Cancer and other noncommunicable diseases (NCDs) are now widely recognized as a threat to global development. The latest United Nations high-level meeting on NCDs reaffirmed this observation and also highlighted the slow progress in meeting the 2011 Political Declaration on the Prevention and Control of Noncommunicable Diseases and the third Sustainable Development Goal. Lack of situational analyses, priority setting, and budgeting have been identified as major obstacles in achieving these goals. All of these have in common that they require information on the local cancer epidemiology. The Global Burden of Disease (GBD) study is uniquely poised to provide these crucial data. Objective: To describe cancer burden for 29 cancer groups in 195 countries from 1990 through 2017 to provide data needed for cancer control planning. Evidence Review: We used the GBD study estimation methods to describe cancer incidence, mortality, years lived with disability, years of life lost, and disability-Adjusted life-years (DALYs). Results are presented at the national level as well as by Socio-demographic Index (SDI), a composite indicator of income, educational attainment, and total fertility rate. We also analyzed the influence of the epidemiological vs the demographic transition on cancer incidence. Findings: In 2017, there were 24.5 million incident cancer cases worldwide (16.8 million without nonmelanoma skin cancer [NMSC]) and 9.6 million cancer deaths. The majority of cancer DALYs came from years of life lost (97%), and only 3% came from years lived with disability. The odds of developing cancer were the lowest in the low SDI quintile (1 in 7) and the highest in the high SDI quintile (1 in 2) for both sexes. In 2017, the most common incident cancers in men were NMSC (4.3 million incident cases); tracheal, bronchus, and lung (TBL) cancer (1.5 million incident cases); and prostate cancer (1.3 million incident cases). The most common causes of cancer deaths and DALYs for men were TBL cancer (1.3 million deaths and 28.4 million DALYs), liver cancer (572000 deaths and 15.2 million DALYs), and stomach cancer (542000 deaths and 12.2 million DALYs). For women in 2017, the most common incident cancers were NMSC (3.3 million incident cases), breast cancer (1.9 million incident cases), and colorectal cancer (819000 incident cases). The leading causes of cancer deaths and DALYs for women were breast cancer (601000 deaths and 17.4 million DALYs), TBL cancer (596000 deaths and 12.6 million DALYs), and colorectal cancer (414000 deaths and 8.3 million DALYs). Conclusions and Relevance: The national epidemiological profiles of cancer burden in the GBD study show large heterogeneities, which are a reflection of different exposures to risk factors, economic settings, lifestyles, and access to care and screening. The GBD study can be used by policy makers and other stakeholders to develop and improve national and local cancer control in order to achieve the global targets and improve equity in cancer care. © 2019 American Medical Association. All rights reserved.Peer reviewe

    Measuring universal health coverage based on an index of effective coverage of health services in 204 countries and territories, 1990–2019 : A systematic analysis for the Global Burden of Disease Study 2019

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    Background Achieving universal health coverage (UHC) involves all people receiving the health services they need, of high quality, without experiencing financial hardship. Making progress towards UHC is a policy priority for both countries and global institutions, as highlighted by the agenda of the UN Sustainable Development Goals (SDGs) and WHO's Thirteenth General Programme of Work (GPW13). Measuring effective coverage at the health-system level is important for understanding whether health services are aligned with countries' health profiles and are of sufficient quality to produce health gains for populations of all ages. Methods Based on the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019, we assessed UHC effective coverage for 204 countries and territories from 1990 to 2019. Drawing from a measurement framework developed through WHO's GPW13 consultation, we mapped 23 effective coverage indicators to a matrix representing health service types (eg, promotion, prevention, and treatment) and five population-age groups spanning from reproductive and newborn to older adults (≥65 years). Effective coverage indicators were based on intervention coverage or outcome-based measures such as mortality-to-incidence ratios to approximate access to quality care; outcome-based measures were transformed to values on a scale of 0–100 based on the 2·5th and 97·5th percentile of location-year values. We constructed the UHC effective coverage index by weighting each effective coverage indicator relative to its associated potential health gains, as measured by disability-adjusted life-years for each location-year and population-age group. For three tests of validity (content, known-groups, and convergent), UHC effective coverage index performance was generally better than that of other UHC service coverage indices from WHO (ie, the current metric for SDG indicator 3.8.1 on UHC service coverage), the World Bank, and GBD 2017. We quantified frontiers of UHC effective coverage performance on the basis of pooled health spending per capita, representing UHC effective coverage index levels achieved in 2019 relative to country-level government health spending, prepaid private expenditures, and development assistance for health. To assess current trajectories towards the GPW13 UHC billion target—1 billion more people benefiting from UHC by 2023—we estimated additional population equivalents with UHC effective coverage from 2018 to 2023. Findings Globally, performance on the UHC effective coverage index improved from 45·8 (95% uncertainty interval 44·2–47·5) in 1990 to 60·3 (58·7–61·9) in 2019, yet country-level UHC effective coverage in 2019 still spanned from 95 or higher in Japan and Iceland to lower than 25 in Somalia and the Central African Republic. Since 2010, sub-Saharan Africa showed accelerated gains on the UHC effective coverage index (at an average increase of 2·6% [1·9–3·3] per year up to 2019); by contrast, most other GBD super-regions had slowed rates of progress in 2010–2019 relative to 1990–2010. Many countries showed lagging performance on effective coverage indicators for non-communicable diseases relative to those for communicable diseases and maternal and child health, despite non-communicable diseases accounting for a greater proportion of potential health gains in 2019, suggesting that many health systems are not keeping pace with the rising non-communicable disease burden and associated population health needs. In 2019, the UHC effective coverage index was associated with pooled health spending per capita (r=0·79), although countries across the development spectrum had much lower UHC effective coverage than is potentially achievable relative to their health spending. Under maximum efficiency of translating health spending into UHC effective coverage performance, countries would need to reach 1398pooledhealthspendingpercapita(US1398 pooled health spending per capita (US adjusted for purchasing power parity) in order to achieve 80 on the UHC effective coverage index. From 2018 to 2023, an estimated 388·9 million (358·6–421·3) more population equivalents would have UHC effective coverage, falling well short of the GPW13 target of 1 billion more people benefiting from UHC during this time. Current projections point to an estimated 3·1 billion (3·0–3·2) population equivalents still lacking UHC effective coverage in 2023, with nearly a third (968·1 million [903·5–1040·3]) residing in south Asia. Interpretation The present study demonstrates the utility of measuring effective coverage and its role in supporting improved health outcomes for all people—the ultimate goal of UHC and its achievement. Global ambitions to accelerate progress on UHC service coverage are increasingly unlikely unless concerted action on non-communicable diseases occurs and countries can better translate health spending into improved performance. Focusing on effective coverage and accounting for the world's evolving health needs lays the groundwork for better understanding how close—or how far—all populations are in benefiting from UHC

    Measuring universal health coverage based on an index of effective coverage of health services in 204 countries and territories, 1990–2019: a systematic analysis for the Global Burden of Disease Study 2019

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    Background Achieving universal health coverage (UHC) involves all people receiving the health services they need, of high quality, without experiencing financial hardship. Making progress towards UHC is a policy priority for both countries and global institutions, as highlighted by the agenda of the UN Sustainable Development Goals (SDGs) and WHO's Thirteenth General Programme of Work (GPW13). Measuring effective coverage at the health-system level is important for understanding whether health services are aligned with countries' health profiles and are of sufficient quality to produce health gains for populations of all ages. Methods Based on the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019, we assessed UHC effective coverage for 204 countries and territories from 1990 to 2019. Drawing from a measurement framework developed through WHO's GPW13 consultation, we mapped 23 effective coverage indicators to a matrix representing health service types (eg, promotion, prevention, and treatment) and five population-age groups spanning from reproductive and newborn to older adults (≥65 years). Effective coverage indicators were based on intervention coverage or outcome-based measures such as mortality-to-incidence ratios to approximate access to quality care; outcome-based measures were transformed to values on a scale of 0–100 based on the 2·5th and 97·5th percentile of location-year values. We constructed the UHC effective coverage index by weighting each effective coverage indicator relative to its associated potential health gains, as measured by disability-adjusted life-years for each location-year and population-age group. For three tests of validity (content, known-groups, and convergent), UHC effective coverage index performance was generally better than that of other UHC service coverage indices from WHO (ie, the current metric for SDG indicator 3.8.1 on UHC service coverage), the World Bank, and GBD 2017. We quantified frontiers of UHC effective coverage performance on the basis of pooled health spending per capita, representing UHC effective coverage index levels achieved in 2019 relative to country-level government health spending, prepaid private expenditures, and development assistance for health. To assess current trajectories towards the GPW13 UHC billion target—1 billion more people benefiting from UHC by 2023—we estimated additional population equivalents with UHC effective coverage from 2018 to 2023. Findings Globally, performance on the UHC effective coverage index improved from 45·8 (95% uncertainty interval 44·2–47·5) in 1990 to 60·3 (58·7–61·9) in 2019, yet country-level UHC effective coverage in 2019 still spanned from 95 or higher in Japan and Iceland to lower than 25 in Somalia and the Central African Republic. Since 2010, sub-Saharan Africa showed accelerated gains on the UHC effective coverage index (at an average increase of 2·6% [1·9–3·3] per year up to 2019); by contrast, most other GBD super-regions had slowed rates of progress in 2010–2019 relative to 1990–2010. Many countries showed lagging performance on effective coverage indicators for non-communicable diseases relative to those for communicable diseases and maternal and child health, despite non-communicable diseases accounting for a greater proportion of potential health gains in 2019, suggesting that many health systems are not keeping pace with the rising non-communicable disease burden and associated population health needs. In 2019, the UHC effective coverage index was associated with pooled health spending per capita (r=0·79), although countries across the development spectrum had much lower UHC effective coverage than is potentially achievable relative to their health spending. Under maximum efficiency of translating health spending into UHC effective coverage performance, countries would need to reach 1398pooledhealthspendingpercapita(US1398 pooled health spending per capita (US adjusted for purchasing power parity) in order to achieve 80 on the UHC effective coverage index. From 2018 to 2023, an estimated 388·9 million (358·6–421·3) more population equivalents would have UHC effective coverage, falling well short of the GPW13 target of 1 billion more people benefiting from UHC during this time. Current projections point to an estimated 3·1 billion (3·0–3·2) population equivalents still lacking UHC effective coverage in 2023, with nearly a third (968·1 million [903·5–1040·3]) residing in south Asia. Interpretation The present study demonstrates the utility of measuring effective coverage and its role in supporting improved health outcomes for all people—the ultimate goal of UHC and its achievement. Global ambitions to accelerate progress on UHC service coverage are increasingly unlikely unless concerted action on non-communicable diseases occurs and countries can better translate health spending into improved performance. Focusing on effective coverage and accounting for the world's evolving health needs lays the groundwork for better understanding how close—or how far—all populations are in benefiting from UHC

    Mapping local patterns of childhood overweight and wasting in low- and middle-income countries between 2000 and 2017

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    A double burden of malnutrition occurs when individuals, household members or communities experience both undernutrition and overweight. Here, we show geospatial estimates of overweight and wasting prevalence among children under 5 years of age in 105 low- and middle-income countries (LMICs) from 2000 to 2017 and aggregate these to policy-relevant administrative units. Wasting decreased overall across LMICs between 2000 and 2017, from 8.4% (62.3 (55.1–70.8) million) to 6.4% (58.3 (47.6–70.7) million), but is predicted to remain above the World Health Organization’s Global Nutrition Target of <5% in over half of LMICs by 2025. Prevalence of overweight increased from 5.2% (30 (22.8–38.5) million) in 2000 to 6.0% (55.5 (44.8–67.9) million) children aged under 5 years in 2017. Areas most affected by double burden of malnutrition were located in Indonesia, Thailand, southeastern China, Botswana, Cameroon and central Nigeria. Our estimates provide a new perspective to researchers, policy makers and public health agencies in their efforts to address this global childhood syndemic
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