Seasonal Trends with the Comparison of Venous and Capillary Blood for Diagnosis of Malaria among Children in Karachi, Pakistan.

Background: More than 250 million people are infected by malaria parasites annually while around one million children less than 5 years of age die every year due to malaria. We aimed to assess the seasonal trends and usefulness of capillary and venous blood for rapid diagnosis of malaria. Methods: This cross-sectional study of 18 months duration was conducted at the National Institute of Child Health (NICH), Karachi. All patients reporting fever as chief complaint were recruited as study subjects. A semi-structured questionnaire was used to collect demographic information, presenting complaints, awareness of caregivers regarding malaria, preventive measures and history of malaria fever. Three ml Venous (2-3ml) as well as peripheral blood (3-4 drops) samples of all patients were collected for microscopy and rapid diagnostic tests (RDTs). Results: Out of total 477 patients with fever Venous and Capillary Blood RDTs methods detected 33(6.9%) and 30(6.3%) as the malaria positive while Venous and Capillary Blood Microscopy detected 30(6.1%) and 32(6.7%) cases respectively. Plasmodium Vivax infection was the most prevalent (87.9%) and majority (39.39%) of the cases occurred in the quarter, July to September. Conclusion: July to September is the peak season for malaria and P. Vivax (87.9%) is the predominant strain in Karachi. Venous and capillary blood are equally useful for malaria diagnosis however, convenience and less invasiveness may justify the preference of capillary blood over venous blood for early diagnosis of malaria.

Regulation of quackery amid unprecedented HIV outbreak in Sindh, Pakistan.

OBJECTIVE: To assess the problem of unlicensed practitioners and quacks in Sindh, Pakistan. METHODS: A cross-sectional study was conducted in 29 districts of Sindh province in Pakistan from December 2019 to January 2020. Initial data available with Sindh Health Department about locations where quacks were practicing was used to identify unlicensed practitioners. A structured questionnaire was developed which contained information about certification of practitioners and an observational checklist was developed to assess infection prevention and control practices (IPC) and injection safety. RESULTS: A total of 4315 private practitioners were inspected out of which 3022 (70%) were unlicensed health practitioners belonging to different categories. Within the six surveyed divisions of Sindh, the highest proportion of unlicensed practitioners were documented in Shaheed Benazirabad division (n=739; 24.5%) followed by Hyderabad (n=599; 19.8%). In Mirpur Khas, there were 510 (16.9%), in Karachi 310 (10.3%), in Sukkur 484 (16%) and in Larkana there were 380 (12.6%) unlicensed practitioners. Poor IPC was observed in 89.4% (3861/4315) of all health providers. Reuse of syringes and intravenous drip sets was observed among 78.7% (1916/2432) of the untrained providers across the province. It was also found that 155 MBBS doctors had given their names on rent to be used as a signboard outside the clinics of some of the unlicensed practitioners. CONCLUSION: The problem of quackery is widespread in the Sindh province. It can be proactively addressed by shutting down all unlicensed practitioners and educating the community to avoid visiting them in order to reduce the probability of exposure to unsafe healthcare practices.

Exposure to aflatoxin B1 and associated risk factors in hepatitis C patients in cosmopolitan city of Pakistan: facility-based study.

INTRODUCTION: population-based follow-up study has been designed to investigate the contributing factors to high exposure to Aflatoxin B1 (AFB1) and the subsequent associated risk factors among hepatitis C-infected patients at a referral centre, Karachi. Pakistan. Hepatitis C infection affects millions of individuals worldwide and confers high morbidity and mortality, especially in lower middle-income countries (LMICs) including Pakistan. A literature review of recent studies has revealed that a number of hepatocellular carcinomas (HCC) cases are markedly increased in Pakistan, where one of the potential causes of HCC is hepatitis C virus. The objectives of this study were to determine frequency of Aflatoxin B1 (AFB1) exposure and other associated characteristics among hepatitis C patients at a referral centre, Karachi, Pakistan. METHODS: a semi-structured pre-coded pro forma designed to collect socio-demographic, Pharmacological, biochemical and clinical information from patients and hospital records. Patient´s pre and post polymerase chain reaction (PCR), serum alanine aminotransferase (ALT) levels and other blood parameters were analysed. AFB1 exposure was determined using an ELISA kit and validated through high-performance liquid chromatography (HPLC). RESULTS: AFB1 exposure was found in 30 (34%) patients. Post treatment responders were 49 (55.6%). More than 37% of study participants had a family history of hepatitis C. About 74% had a history of surgical procedure, and around 36% of study participants had a blood transfusion history. Up to 36% participants were fond of spicy food and around 25% study participants were eating roadside food on daily basis. CONCLUSION: high frequency of AFB1 exposure due to risky dietary habits, low level of formal education and awareness are contributing factors may be responsible for high exposure of AFB1. Effective and multidimensional strategies are needed to prevent advance stage progression of disease and associated complications.

Factors contributing to the public proneness towards quacks in Sindh.

The present study is to explore the factors and reasons behind public proneness towards quacks in the rural areas of Sindh, Pakistan and to manifest the public on how these quacks are duping vulnerable and quackery-prone peoples for financial gain which may induce human lives in life-threatening health conditions. The study also interprets a better understanding of the public needs, especially in the rural areas of the Sindh that may give a hope for deliverance from quacks.

Vitamin D status in under five children in diverse communities of Karachi.

BACKGROUND & OBJECTIVE: Vitamin D deficiency (VDD) prevalence is very high in pediatric population in developing countries including Pakistan. VDD contribute significantly to morbidity and mortality among children under five years. Therefore, it is vital to study vitamin D levels in population for future interventions and disease control. Our objective was to determine 25(OH)D levels in children of one to 59 months of age in socio-economically diverse communities of Karachi, Pakistan. METHODS: The cross- sectional analytical survey was carried out over 6 months from January -June 2017. Following proportionate sampling technique four clusters were randomly selected from Korangi, Saddar, Sindhi para and Manzoor colony, Karachi. Blood samples for 25(OH)D and zinc levels were carried out using ELISA and colorimetry. VD level <20 ng/ml was defined as VDD and serum zinc <65ug/dl as low zinc levels. Data including area of residency, gender, ethnicity, parent's education, family income, house status, duration of sun exposure and history of VD and zinc intake in last 3 months was collected from parents / care seekers on pretested and pre-coded semi-structured questionnaire. Data was analyzed using SPSS version-20. Frequencies and percentages were computed for categorical variables like gender, type of malnutrition whereas mean with standard deviation was used for VD and zinc levels and vitamin D status was compared in three different residential categories according to nutritional status. RESULTS: Out of 120 children, 67 (56%) were boys and 53 (44%) girls. Mean VD level was 22.8±14.8 ng/ml. Around 60% (70) children were VD deficient, whereas 15 % (18) had insufficient 25(OH)D levels. VDD was more prevalent among low socio-economic group with no obvious difference in age category. Mean zinc level was 123.8±47.45 ug/dl and it was either normal or high (42%) rather than low. Malnutrition was observed in 65% children and majority (82%) of them were undernourished. Optimal sun exposure was reported in 24% children only. CONCLUSION: Vitamin D deficiency was highly prevalent in our study population. Children of low socio-economic strata and with sub-optimal sun light exposure are at high risk of vitamin D deficiency. Unexpectedly, high zinc levels in majority of our children with low VD status needs further evidence to substantiate this inverse relation.

Malnutrition and poor oral health status are major risks among primary school children at Lasbela, Balochistan, Pakistan.

BACKGROUND: This survey was focusing on health and oral hygiene status of primary school children at Lasbela district considering the comparatively less developed and socio demographically deprived part of the Country. METHODS: A cross sectional survey was conducted to determine the health status of primary school children in seven tehseels of district Lasbela, Balochistan after applying proportionate sampling technique from March 2015 to July 2015. Field teams visited assigned schools to screen children and collect health related data on predesigned and pre coded proforma. RESULTS: Out of 200 schools, 196 schools found opened, while 2% of schools (04) remained closed. A total of 6363 students were clinically screened. About 45% of the school children had normal body mass index (BMI) and rest were falling in different categories of malnutrition. More than 19% had ear, nose and throat (ENT) problems and around 19% presented with clinical anemia. Less than 50% of children had scar of BCG vaccination and 4% informed about use of gutka/supari chewing (smokeless tobacco use). CONCLUSION: In conclusion, we estimated high prevalence of malnutrition, poor oral health including smokeless tobacco use, and low BCG coverage among primary school children at Lasbela. Current scenario suggests immediate and contextually focused interventions to confine existing public health risks and avoid future burden of disease.

Trajectory of cause of death among brought dead neonates in tertiary care public facilities of Pakistan: A multicenter study.

BACKGROUND: Considering the fact that Pakistan is amongst the countries with very high neonatal mortality rates, we conducted a research study to determine the possible causes and characteristics of neonates presenting dead to the emergency department of tertiary public health care facilities of Pakistan using verbal autopsies. METHODS: A descriptive case series study was conducted in emergency department/pediatrics ward/neonatal ward/nursery unit of ten tertiary care public health facilities, situated in seven major cities of Pakistan from November, 2011 to June, 2013. Precoded verbal autopsy proforma was used to collect information regarding cause of death, family narratives and other associated risks accountable for pathway to mortality. RESULTS: We identified 431 neonates presenting dead to the emergency department (238 males and 193 females). Sepsis (26.7%), birth asphyxia (18.8%) and persistent pulmonary arrest (17.2%) were main primary causes of brought death. Around 72% brought dead neonates were referred from doctors/health care facilities and more than 28% caregivers mentioned that they were not informed about the diagnosis/ailment of their deceased newborn. CONCLUSIONS: Findings of our study suggest that infectious disease remains the main primary cause of neonatal mortality. Underweight in newborns (64%) was estimated as a leading associated risk. Delays in referrals to respective health care facility enlightened the concern of sub-standard prerequisites of neonatal care that could be one of the major contributing risk factor of high mortality rates.

Synthesis and bio-molecular study of (+)-N-Acetyl-α-amino acid dehydroabietylamine derivative for the selective therapy of hepatocellular carcinoma.

BACKGROUND: The purpose of present work is to synthesize novel (+)-Dehydroabietylamine derivatives (DAAD) using N-acetyl-α-amino acid conjugates and determine its cytotoxic effects on hepatocellular carcinoma cells. METHODS: An analytical study was conducted to explore cytotoxic activity of DAAD on hepatocellular carcinoma cell lines. The cytotoxicity effect was recorded using sulforhodamine B technique. Cell cycle analysis was performed using Propidium Iodide (PI) staining. Based on cell morphology, anti growth activity and microarray findings of DAAD2 treatment, Comet assay, Annexin V/PI staining, Immunoperoxidase assay and western blots were performed accoringly. RESULTS: Hep3B cells were found to be the most sensitive with IC50 of 2.00 ± 0.4 µM against (+)-N-(N-Acetyl-L-Cysteine)-dehydroabietylamine as DAAD2. In compliance to time dependent morphological changes of low cellular confluence, detachment and rounding of DAAD2 treated cells; noticeable changes in G2/M phase were recorded may be leading to cell cycle cessation. Up-regulation (5folds) of TUBA1A gene in Hep3B cells was determined in microarray experiments. Apoptotic mode of cell death was evaluated using standardized staining procedures including comet assay and annexin V/PI staining, Immuno-peroxidase assay. Using western blotting technique, caspase dependant apoptotic mode of cell death was recorded against Hep3B cell line. CONCLUSION: It is concluded that a novel DAAD2 with IC50 values less than 8 µM can induce massive cell attenuation following caspase dependent apoptotic cell death in Hep3B cells. Moreover, the corelation study indicated that DAAD2 may have vital influence on cell prolifration properties.

Functional and structural abnormalities of the kidney and urinary tract in severely malnourished children - A hospital based study.

OBJECTIVES: The association of malnutrition and systemic diseases like chronic kidney disease (CKD) is well known. Various urinary tract abnormalities may be associated with malnutrition. So objective of current study was to determine the frequency of functional and structural urinary tract abnormalities in severely malnourished children admitted in Nutritional Rehabilitation Unit (NRU) of a tertiary care facility, Karachi. METHODS: This descriptive cases series of 78 children was conducted in NRU from October 2014 - March 2015. All newly admitted children aged 2-60 months, diagnosed as Severe Acute Malnutrition (SAM) were studied and children with known kidney and urinary tract disorders were excluded. Detailed history, examination and investigations like serum creatinine, ultrasound kidney and urinary tract in addition to routine tests for SAM, were done. A proforma was used to collect demographic data, clinical history, physical findings, and radio-imaging and biochemical investigations. Glomerular filtration rate (GFR) was calculated using Schwartz equation. Data was analyzed using descriptive statistics. RESULTS: Among 78 children, male to female ratio was equal. Mean age was 18±15.53 months and majority (79.48%) of children were below 24 months. Majority (82%) of children with SAM had marasmus whereas 18% had edematous malnutrition. Out of 78, 57 (73%) children had either functional (80.7%) and or structural (19.3%) abnormalities whereas 21(36.84%) had normal functional and structural status. Most common functional abnormality was subnormal GFR (<90ml/min/1.73 m2) found in all 46 children. Functional abnormities were more common in children below 24 months. Other functional disorders were Bartter syndrome, renal tubular acidosis and urinary tract infection (UTI) found in two cases each. Common structural abnormalities were echogenic kidneys (n=4, 36%), hydronephrosis (n=3, 27%), hypoplastic kidneys (n=3, 27%) and calculi (n=1, 9%). Subnormal GFR was also found in all cases with structural abnormalities. UTI was observed exclusively in two children among 11 with structural abnormalities. CONCLUSION: A high frequency of functional abnormalities and noticeable proportion of structural abnormalities of urinary tract were detected in children with SAM. Current finding suggest that multicenter study at national level may be undertaken to generate better data about prevalence of renal diseases in SAM.

Frequency of Stillbirths in a Tertiary Care Hospital, Karachi.

BACKGROUND AND OBJECTIVE: Pakistan accounts for the highest stillbirth rate in the world. Therefore, this observational study was planned to determine the prevalence of stillbirths and its associated demographic characteristics in the given context. Hence our objective included: To determine the frequency of stillbirths with reference to parity and gestational age in a tertiary care public hospital, Karachi. To determine the socio-demographic characteristics of families with stillbirths. METHODS: All pregnant mothers who delivered stillbirth babies at Gynaecology and Obstetrics ward of Jinnah Postgraduate Medical Center, Karachi a tertiary care facility were prospectively enrolled from October 2012 to September 2013. Deliveries occurred before 28 weeks of gestational age were excluded. Gestational age was confirmed from hospital record and attending physicians. Data was collected on predesigned proforma and analyzed using descriptive statistics. RESULTS: Among 7708 registered deliveries, 137 were stillbirths. A total of 84 mothers were primiparous and 12% of mothers were below 20 years at the time of delivery. Majority of stillbirths were macerated type (80.3%) and 20% were fresh stillbirth. About 55% of still births occurred between 33-37 weeks and 20% between 28-32 weeks. Almost 80% (109) of stillbirths were low birth weight and only 20% (28) were normal birth weight. CONCLUSION: This study shows that stillbirths are more common in primiparous mothers in a given context. Conducting awareness sessions with special focus on antenatal and obstetrical care of primiparous may be helpful to reduce still births.

High prevalence of typhoidal Salmonella enterica serovars excreting food handlers in Karachi-Pakistan: a probable factor for regional typhoid endemicity.

BACKGROUND: Typhoid fever is the persistent cause of morbidity worldwide. Salmonella enterica serovar's carriers among food handlers have the potential to disseminate this infection on large scale in the community. The purpose of this study was to determine the prevalence of typhoidal S. enterica serovars among food handlers of Karachi. METHODS: This cross-sectional study was conducted in Karachi metropolis. A total of 220 food handlers were recruited on the basis of inclusion criteria from famous food streets of randomly selected five towns of Karachi. Three consecutive stool samples were collected from each food handler in Carry Blair transport media. Culture, biochemical identification, serotyping, and antimicrobial susceptibility tests for S. enterica serovars were done. RESULTS: Out of 220 food handlers, 209 consented to participate, and among them, 19 (9.1%) were positive for S. enterica serovars. Serotyping of these isolates showed that 9 (4.3%) were typhoidal S. serovars while 10 (4.7%) were non-typhoidal S. serovars. Of the typhoidal S. serovars, 7 were S. enterica serovar Typhi and 1 each of S. enterica serovar Paratyphi A and B. The resistance pattern of these isolates showed that 77.7% were resistant to ampicillin and 11.1% to cotrimoxazole. All typhoidal S. enterica serovar isolates were sensitive to chloramphenicol, ceftriaxone, cefixime, nalidixic acid, and ofloxacin. CONCLUSIONS: Carrier rate of typhoidal S. enterica serovars in food handlers working in different food streets of Karachi is very high. These food handlers might be contributing to the high endemicity of typhoid fever in Karachi, Pakistan.

Infection remains a leading cause of neonatal mortality among infants delivered at a tertiary hospital in Karachi, Pakistan.

INTRODUCTION: The current cohort study was conducted to determine the frequency and compare the mortality rate with associated characteristics among low birth weight and normal birth weight infants during the neonatal period at a tertiary healthcare facility, Karachi. METHODOLOGY: Close-ended structured questionnaires were used to collect information from the parents of 500 registered neonates at the time of birth. Follow-ups by phone on the 28th day of life were done to determine the mortality among low birth weight and normal birth weight babies during the neonatal period. RESULTS: The neonatal mortality rate ranged from as low as 2.4% in the normal birth weight and 16.4% in the low birth weight categories to as high as 96% in the very low birth weight category. Respiratory distress syndrome (24.2%) and sepsis (18.2%) were reported as the leading causes of neonatal deaths. The babies' lengths of stay ranged from 2 to 24 hours, and around 90% of neonatal deaths were reported in the first seven days of life. More than 6% of neonates died at home, and 7.6% of the deceased babies did not visit any healthcare facility or doctor before their death. In the 12-15 hours before their deaths, 13.6% of the deceased babies had been unattended. Around 90% of the deceased babies were referred from a doctor or healthcare facility. CONCLUSIONS: The present estimates of neonatal mortality are very high among low birth weight and very low birth weight categories. Infectious diseases, including respiratory distress syndrome (24.2%) and sepsis (18.2%), were leading causes of neonatal deaths.

Comparative evaluation of in vitro cytotoxic effects among parent abietyl alcohol and novel fatty acid ester derivatives against MCF7 and hepatocellular carcinoma cell lines.

Synthesis of twelve hitherto unreported esters of abietyl alcohol and screening of these esters against four cancer cell lines including one breast cancer line MCF7 and four hepatocellular carcinoma cell lines (HCC) Huh7, Hep3B, Snu449 and Plc has been determined using SRB assay. The Cell cycle progression showed changes in cellular behaviour after 48 and 72 hours in MCF7 and Huh7 cell lines. Abietyl alcohol was obtained from the reduction of abietic acid, a tricyclic diterpene, isolated from oleoresin of Pinus longifolia Roxberghii.

Synthesis and characterization of amino acid conjugates of oleanolic acid and their in vitro cytotoxic effect on HCC cell lines.

Oleanolic acid (3ß-hydroxy-olean-12-en-28-oic acid; OA-01), a pentacyclic triterpene, exhibit a wide range of pharmacological and biological activities. We have isolated oleanolic acid from methanolic extract of Periploca aphylla, collected from surroundings of Karachi in the month of February. Furthermore, four known and two new C-28 amino acid conjugates of oleanolic acid were prepared to explore potential of these compounds on HCCs and one breast cancer cell line. Cytotoxic effects revealed that as compare to parent compound (OA-01), two derivatives OA-04 (p<0.0001) and OA-06 (p<0.01) showed significantly increased/higher inhibition rates.

Comparative cost and efficacy trial of Pakistani versus Indian anti snake venom.

OBJECTIVES: To compare the efficacy, safety and cost of Pakistani anti-snake venom with that imported from India METHODS: The comparative cross-sectional study was conducted from June to September 2010 and comprised patients hospitalised following Krait snake bite in Mithi and Umerkot hospitals of Tharparker district who had incoagulable blood test on admission (20-minute whole blood clotting time). Basic demographies of patients, the site of bite and swelling around the bite and joints were entered in a proforma. For blinding, the liquid anti-snake venoms were packed in opaque polythene bags and marked as "A" and "B", and refrigerated. Four ampoules of the venom were mixed in a drip and given over one hour while looking for adverse reactions. In case of milder reactions, the venom was temporarily stopped and anti-histamines and analgesics were given. For more severe reactions intramuscular adrenaline was recommended. Coagulation was again checked after 6 hours and, if blood was still incoagulable, the dose of the venom was repeated after 6 hours. Once coagulation was confirmed on two occasions, the patient was discharged. Total doses of the venom given to achieve coagulation, the reactions and the cost of the venom were analysed at the end to see the response and cost-effectiveness. RESULTS: Of the 80 cases - 40 (50%) from each hospital - 6 (7.5%) had to be excluded for lack of data. Out of the remaining 74 (92.5%) patients, 38 (52.35%) received Pakistani anti-snake venom (A), and 36 (48.64%) received Indian anti-snake venom (B). Immediate reaction to the venom was seen in 23 (60.5%) cases with "A" and 25 (69.4%) with "B". In terms of evenomation, 23 (60.5%) cases with "A" attained restoration of coagulation with the first dose, compared to 13 (36.11%) with "B", showing a significantly better response with "A" (p < 0.035). Mean of 1.66 doses of "A" and 1.94 of "B" were used to neutralise venom, again showing lesser doses of Pakistani anti-snake venom. Cost-wise "A" was 2.5 times cheaper than the imported "B". CONCLUSION: Pakistani anti-snake venom was significantly quicker and better as well as cheaper than the Indian anti-snake venom.

Biosafety perspective of clinical laboratory workers: a profile of Pakistan.

INTRODUCTION: Biosafety during lab work is an important concern in developing countries. Some critical issues concerning biosafety are lack of training, exceeding workload, working too fast, deciding not to follow safe practices, and skepticism about biohazards. This study aimed to determine biosafety perception and practices of laboratory technicians during routine work in clinical laboratories of Pakistan. METHODOLOGY: A total of 1,782 laboratory technicians were interviewed from major public sector hospitals and a few private hospital laboratories throughout Pakistan. RESULTS: A total of 1,647 (92.4%) males and 135 (7.6%) females participated in the study, with over half (59.7%) having more than five years of work experience. Results showed that 28.4% of the laboratory technicians from Punjab, 35.7% from Sindh, 32% from Balochistan and 38.4% from Khyber Pakhtoon Khawa (KPK) did not use any personal protective equipment. Almost 46% of the respondents (34.2% from Punjab, 61.9% from Sindh, 25.2% from Balochistan and 85% from KPK) said they reused syringes either occasionally or regularly. Furthermore, 30.7% of the respondents said they discard used syringes directly into municipal dustbins. The majority (66.7%) claimed there are no separate bins for sharps, so they throw these in municipal dustbins. Mouth pipetting was reported by 28.3% technicians. Standard operating procedures were not available in 67.2% labs, and accident records were not maintained in 83.4%. No formal biosafety training had been provided to 84.2% of the respondents. CONCLUSION: Laboratory technicians in Pakistan lack awareness of good laboratory practices and biosafety measures, and also face a lack of resources.

Exposure to risk factors for hepatitis B and C viruses among primary school teachers in Karachi.

INTRODUCTION: The study aimed to determine hepatitis B vaccination status and assess the exposure of risk factors for hepatitis B and C among primary schoolteachers in Karachi, Pakistan. METHODOLOGY: In two hundred school teachers from 30 primary schools in Karachi participated in the study between January and June 2008 by completing an anonymous, self-administered questionnaire. Exposure to and knowledge of hepatitis B and C were assessed, as well as mode of transmission and prevention. The percentage of vaccinated and non-vaccinated teachers was also estimated. RESULTS: Only 73 (36.5%) respondents were vaccinated against HBV. Nine percent (17) of the teachers had received more than 10 therapeutic injections while about 56% (101) took between 5-10 injections per annum. Fifteen (8%) of the teachers confirmed they had been injected with re-used syringes. More than 8% (17) of participants' family members were suffering from hepatitis B or C, while 10% (20) of family members had died of liver diseases without any known history. More than 13% (27) of participants shared razors, brushes, cigarettes and hukahs. Statistically significant difference was also observed in risk factors of hepatitis B and C among male and female respondents. CONCLUSION: Hepatitis B vaccination among school teachers of Karachi was around 37% with a high use of therapeutic injections and syringe reuse. Health awareness programs and educational workshops are needed for teachers, who can later educate the children.