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Objective: To identify the types and frequency of pre-analytical errors in the haematology laboratory and to reduce these preanalytical errors by taking corrective measures. Study Design: Cross-sectional study. Place and Duration of Study: Haematology Laboratory of Pak Emirates Military Hospital, Rawalpindi Pakistan, from Mar to Apr 2022 after applying corrective measures, pre-analytical errors were re-analysed from Jun to Jul 2022. Methodology: The study included all samples received in the haematology laboratory for complete blood count, peripheral blood smear, coagulation profile, and blood malarial parasite. Blood samples were checked for clotted samples, insufficient quantity, diluted samples, hemolysed samples, lipemic samples, and degenerated samples. After that, corrective measures were taken to reduce the pre-analytical errors. Results: The total number of samples received in the first session was 30,169, and pre-analytical errors were present in 508(0.016%) samples. The most frequent pre-analytical errors were of clotted samples 300(59.1%) followed by quantity not sufficient 168(33.1%), hemolysed samples 68(13.4%), diluted samples 49(9.6%), degenerated samples 22(4.3%) and lipemic samples 18(3.5%). After applying corrective measures, pre-analytical errors were reduced in clotted samples 78(52.7%), quantity not sufficient 39(26.4%), hemolysed samples 14(9.5%), diluted samples 11(7.4%), degenerated samples 5(3.4%) and lipemic samples 4(2.7%) respectively. Conclusion: This study showed the relative frequency of various pre-analytical errors in a haematology laboratory. It also signifies the implementation of corrective measures for reducing these pre-analytical errors.

BACKGROUND: This study aimed to determine the association of single-nucleotide polymorphisms (SNPs) of interleukin-6 (IL-6) gene with the survival and disease status of patients after allogeneic hematopoietic stem cell transplantation. MATERIALS AND METHODS: It was a prospective cohort study of total 102 participants, 51 patients along with their donors who had human leukocyte antigen-identical-matched allogeneic hematopoietic stem cell transplantation (aHSCT). Their pretransplant and posttransplant blood samples were collected for extraction of DNA for genotyping by sequence-specific primers polymerase chain reaction and gel electrophoresis followed by sequencing. The pre- and posttransplant levels of IL-6 were measured using enzyme-linked immunosorbent assay technique. RESULTS: The mean age of our patients was 19.83 ± 12.5 years having males 65 (63.7%) and females 37 (36.3%). The frequency of −174G/C SNP among acute graft-versus-host disease (aGVHD) group was GG = 53.8%, GC = 34.6%, and CC = 11.5%, and for −597G/A SNP, it was GG = 69.2%, GA = 15.4%, and AA = 15.4%. Our results showed that the presence of G allele in both homozygous and heterozygous forms was associated with increased aGVHD incidence, while the homozygous CC and AA mutant genotypes correlated with the lowest number of cases of aGVHD. The polymorphism −597G/A was significantly associated with the incidence of aGVHD with P = 0.04. The GG genotype in −174G/C and −597G/A was found to be significantly associated with aGVHD with P < 0.0001. High serum levels of IL-6 pre- and posttransplant were found to be significantly associated with the incidence of aGVHD with P < 0.0001. CONCLUSION: We concluded that the two SNPs − 174G/C and − 597G/A in the promoter region of IL-6 gene present either in donors or recipients predisposed to increase the risk of development of aGVHD following aHSCT. Their evaluation in our transplant setting may help in the risk stratification of transplant recipients early in time and predict the onset of aGVHD and mortality.

Objective: To determine the genotypic frequency of RHC antigen in females of reproductive age. Study Design: Cross-sectional study. Place and Duration of Study: Armed Forces Institute of Transfusion (AFIT) and Pak Emirates Military Hospital, (PEMH) in collaboration with NUMS Pakistan, from Dec 2020 to Dec 2021. Methodology: Females aged 16 to 45 years were recruited in the study. Demographic data, including age, parity, ethnicity etc.,was recorded on a proforma. Venous blood samples were collected in EDTA tubes, and DNA was extracted using 5% Chelex TM. A conventional Polymerase chain reaction and amplified products were subjected to Polyacrylamide Gel electrophoresis. Results: Amongst 200 females, 172(86%) had the expression of RHC antigen, while 28(14%) were negative for the respective antigen. These results were valuable in predicting the risk of hemolytic disease in fetuses and newborns and alloimmunization in our population. Conclusion: Females in the reproductive age group had RHC positivity of 86%. This data will help predict the risk of HDFN in future pregnancies and individuals at risk of alloimmunization in RHC-negative women.

خلاصة: أهداف البحث: تؤدي المضاعفات المرتبطة بـ كوفيد-١٩ إلى تخثر منتشر داخل الأوعية، الأمر الذي تم الإبلاغ عنه على أنه أحد أسباب الوفيات لدى هذه الفئة من المرضى. هدفت هذه الدراسة إلى تحليل حدوث التخثر المنتشر داخل الأوعية لدى غير الناجين ولتقييم العلاقة بين التخثر المنتشر داخل الأوعية والأمراض المصاحبة. طرق البحث: تم تحليل السجلات الطبية لـ ١٥٤ من غير الناجين من كوفيد-١٩، الذين تم إدخالهم إلى المستشفى بين أبريل ٢٠٢٠ ويوليو ٢٠٢٠ بأثر رجعي. تم تطبيق معايير الجمعية الدولية للتخثر والإرقاء لتحديد حدوث اعتلال التخثر. وتم استخدام تحليل خواص التشغيل المتلقي لتقييم الارتباط بين التخثر المنتشر داخل الأوعية والأمراض المصاحبة. النتائج: من بين ١٥٤ من غير الناجين، لوحظ وجود تخثر منتشر داخل الأوعية غير العلني في ٩٤.٨٪ من المرضى بينما استوفى ٥.٢٪ فقط المعايير العلنية للتخثر المنتشر داخل الأوعية بمتوسط عمر ٦٤.٦سنة. وكان معدل الوفيات ٤.٥مرات أعلى بين الذكور مقارنة بالإناث. وكان مستوى د-دايمر>٢٥٠نانوغرام / مل في ٦٨.٨٪ من المرضى بما في ذلك ٨٨.٩٪ من المرضى غير الصريحين و١٠٠٪ من مرضى التخثر المنتشر داخل الأوعية العلني. وكان زمن البروثرومبين في حالات التخثر المنتشر داخل الأوعية غير العلنية والعلنية ١٧.٣ثانية و٢٤.٤ ثانية على التوالي. كما تم العثور على الحدث التخثري وأمراض الكلى المزمنة لتكون من العوامل الرئيسية للتنبؤ ويليها داء السكري وارتفاع ضغط الدم، دون دلالة إحصائية. الاستنتاجات: نستنتج أن درجة معايير الجمعية الدولية للتخثر والإرقاء لتحديد حدوث اعتلال التخثر لا يمكن أن تتنبأ بالوفيات لأن التخثر المنتشر داخل الأوعية المرتبط بـ كوفيد-١٩ تختلف عن التخثر المنتشر داخل الأوعية الذي يسببه الإنتان. من بين المرضى الذين يعانون من أمراض خطيرة، فإن المرضى الأكبر سنا الذين يعانون من أمراض مصاحبة، وزيادة مستويات د-دايمر وزمن البروثرومبين المطول هي معايير أكثر موثوقية بين غير الناجين من كوفيد-١٩. Abstract: Objectives: Complications related to coronavirus disease 2019 (COVID-19) may lead to disseminated intravascular coagulation (DIC), which has been reported to be among the known causes of mortality in such patients. This study aims to analyse the incidence of DIC in COVID-19 non-survivors and to assess the association between DIC and its comorbidities. Methods: The medical records of 154 non-survivors of COVID-19, hospitalised between April 2020 and July 2020, were retrospectively analysed. The International Society on Thrombosis and Haemostasis (ISTH) criteria for DIC were applied to identify the occurrence of coagulopathy. The receiver-operating characteristic (ROC) analysis was used to assess the association between DIC and its comorbidities. Results: Out of 154 non-survivors, non-overt DIC was observed in 94.8% of the patients, whereas only 5.2% fulfilled the overt criteria of DIC with a mean age 64.6 years. The mortality rate was 4.5 times higher among men than women. The D-dimer level was >250 ng/ml in 68.8% of the patients including 88.9% of the non-overt and 100% of the overt DIC patients. Prothrombin time (PT) in non-overt and overt DIC cases was 17.3 s and 24.4 s, respectively. Thrombotic event and chronic kidney disease were found to be the main predictors of DIC (p

Objective: To see the effect of severe acute respiratory syndrome coronavirus 2 (SARS CoV- 2) on red cell indices in patients with COVID-19 disease. Study Design: Cross sectional study. Place and Duration of Study: The study was conducted in the department of Pathology, Pak Emirates Military Hospital, Rawalpindi, from Apr 2020 to Apr 2020. Methodology: The study includes 100 confirmed cases of COVID-19 infection admitted in PEMH. Cases was confirmed by real time polymerase chain reaction (RT-PCR) performed on pharyngeal swabs. After obtaining the informed consent and approval from the ethical committee patient’s complete blood counts (CBC) were performed. Haemoglobin and red cell indices values were collected and analyzed using SPSS version 22. Results: Mean age of the study population was 43 years ± 14.4 SD with minimum of 20 years and maximum of 75 years. Out of 100 samples 96 were males and 4 females. Mean haemoglobin was 14.4 g/dl ± 1.93 g/dl. Red cell indices shows normochromic normocytic anemia. Among these 100 patients with moderate to severe disease admitted in the hospital 96 survived and 4 deceased. Conclusion: COVID-19 patients show normocytic normochromic values. Therefore it is really important to evaluate the laboratory indices at baseline and during the evolution of disease which can help clinicians in making a structured treatment approach and urgently provide good intensive care to those who are in vital need.

Objective: To investigate the incidence and epidemiology of Dengue fever, and to evaluate the utility of NS1 antigen test for the rapid diagnosis of disease at Combined Military Hospital Malir, Karachi. Study Design: Cross sectional prospective study. Place and Duration of Study: Study was carried out at Combined Military Hospital Malir Cantt Karachi, form Jan to Dec 2018. Methodology: Patients with acute febrile illness that reported to Combined Military Hospital Malir Cantt Karachi, Pakistan were enrolled in our study. The diagnosis was made on the basis of Dengue Serology by one or more of the following tests NS1 antigen, IgM or both by using ICT methods and a compatible clinical picture. The demographic characteristics, clinical features were noted. Results: A total of 993 sera were collected from patients with suspected dengue infection out of which 89 were laboratory-confirmed dengue cases suggesting the incidence of dengue fever of 8.96%. Positive NS1 antigen alone was observed in majority of the cases with the frequency of 78 males and 07 females. NS1 antigen along with IgM antibody was observed in 1 male and 1 female only. Peak incidence was seen in the age group of 21-40 years i.e. 71%. Conclusion: Diagnosis of Dengue Virus infection can be established early by keeping high suspicion for dengue in patients who presents with an acute febrile illness by using rapid diagnostic test that is dengue specific NS1 antigen along with IgM and IgG antibodies.

Objective: To determine the association of low maternal serum vitamin-D levels with serum vitamin-D levels of their term neonates and higher risk of early onset neonatal sepsis (EOS). Study Design: Prospective observational study. Place and Duration of Study: Neonatal intensive care unit (NICU) of Department of Pediatrics; Combined Military Hospital (CMH) Malir Cantt Karachi, from Apr 2016 to Apr 2017. Material and Methods: Ninety three term neonates with EOS formed the “sepsis group” and equal number (93) of healthy term neonates without sepsis were included in the control group selected as per inclusion criteria. Blood samples of mothers and their neonates were drawn during the first 72 hours after delivery for measure-ment of serum vitamin-D levels. Results: Mean serum vitamin-D levels were 18.16 ± 8.21 ng/ml and 13.93 ± 13.29 ng/ml in the mothers and their neonates respectively in the sepsis group and were significantly lower (p-value

Objective: To find out the pattern of Liver Function tests (LFTs) in different spectrums of Acute Viral Hepatitis (AVH). Study Design: Cross sectional. Place and Duration of Study: The study was conducted at Pathology Department of Combined Military Hospital, Malir Karachi from March 2017 to August 2017. Materials and Methods: A total of 360 subjects of all age groups, both male and female, with biochemical features of acute viral hepatitis (AVH) were included. After clinical history and examination, liver function tests (LFTs) were performed that included total bilirubin, alanine amino transferases (ALT), alkaline phosphatase (ALP), followed by viral markers for hepatitis A virus (HAV), hepatitis B virus (HBV) hepatitis C virus (HCV) and hepatitis E virus (HEV). Results: Among the 360 patients studied, males (n=308, 86%) outnumbered females (n=52, 14%). Biochemical pattern showed ALT value reaching >75 times the upper limit of normal (ULN). Similarly rise of serum total bilirubin was up to 30 times of ULN while rise of serum ALP was not more than 5 times of ULN. HEV was found to be the most common (28%) in seropositive AVH in the study group. Highest number (24.4%) of HEV infection was found among males in the age group of 20-30 years. ALT and bilirubin were significantly raised in patients with HEV as compared to other causes of AVH. Conclusion: Pattern of LFTs varies in spectrum of AVH. Highest rise of ALT indicates extensive liver damage in HEV infection. How to cite this: Yousaf S, Younas M, Murtaza M, Akhtar F, Jaffar SR, Ijaz A. Pattern of Liver Function Test in Spectrum of Acute Viral Hepatitis. Life and Science. 2020; 1(1): 29-33. doi: http://doi.org/10.37185/LnS.1.1.38

To investigate the accumulation of lead (Pb) by radish (Raphanus sativus) cultivars a study was carried out at Department of Soil and Environmental Sciences, Gomal University, Dera Ismail Khan (Pakistan), during 2012. Two radish varieties i.e., exotic and local, were used. The treatments included sewage water and different concentrations of Pb @ 25, 100, 200 and 400 mg L-1. The results showed that the total biomass of both the radish varieties were nonsignificantly influenced by the applied Pb concentrations and sewage water, except for root diameter which were significantly greater in the local cultivar (3.261 cm).Pb treatments significantly reduced the growth and yield of both the cultivars. While the Pb uptake by the root and leaf of radish plants was increased by the increasing the applied Pb levels, with the highest value for root (19.008 mg kg-1) and leaf (16.134 mg kg-1) in the treatment receiving the highest applied Pb concentrations. The total biomass, fresh weight of root and root diameter was found significantly higher except for Pb @ 400 mg L-1, in the plants receiving sewage water as compared to the control and different levels of Pb. The interaction amongst the varieties and treatments were found significantly different for various parameters. Thus, it can be concluded, that the use of sewage water and Pb contaminated wastewater results in higher metal concentration in the radish root and may lead to different types of health problems to consumers.

Steroids remain an essential part of immunosuppressive therapy for renal transplant patients since the start of transplant era. Different immunosuppressive regimens are prescribed so as to minimize the side effects. The purpose of our study is to compare the outcome of early steroid withdrawal with steroid maintenance protocol. It is a prospective nonrandomized study. All patients that received renal transplants from January 2011 to December 2013 were included in the study. Early steroid withdrawal at day 5 was done in low immunological risk patients, and the results were compared with the steroid maintenance group, at one-year, posttransplant. Outcome measures included acute rejection (AR), slow graft function and delayed graft function (SGF and DGF), patient and graft survival, and new-onset diabetes after transplant (NODAT), dyslipidemia, hypertension, and obesity. A total of 249 patients were divided into two groups - 105 patients had early steroid withdrawal and 144 patients were maintained on steroid therapy. Outcome measures were compared one-year posttransplant. There was no significant difference in AR, patient and graft survival, creatinine level, and weight gain. However, a significant difference in systolic and diastolic blood pressure, lipid profile, NODAT, SGF, and DGF was found in the steroid group. Our study shows that early steroid withdrawal is a safe standard of care in low immunological risk patients.