The Prevalence of Blood Borne Diseases in Blood Donors of Peshawar, Khyber Pakhtunkhwa, Pakistan.

BACKGROUND: The goal of the study was to investigate the burden of transfusion- transmitted infections (TTIs) hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), syphilis, and malarial parasite (MP) in ABO Blood Groups and Rh Type System among voluntarily blood donors in Khyber Pakhtunkhwa (KPK), Pakistan. It is a retrospective single center cross sectional study. This study was conducted from June 2020 to September 2021 (16 months) at the frontier foundation thalassemia center Peshawar KPK. Donors were physically healthy and fit for donation. Donors with physical disabilities and/or having co-morbid conditions were excluded from the report. METHODS: All the samples were screened for anti-HIV, anti-HCV, HBsAg, Syphilis, and Malarial Parasite via ELISA kit and Immune Chromatographic Technique (ICT), respectively. A total of 6311 blood donations were evaluated. The majority of the donations (92%) were from (VNRBD) voluntary non-remunerated blood donation, while only 8% came from replacement donors. RESULTS: Amongst 6311 blood donations, 1.50 % (n = 95) were infected at least with one pathogen, HBV positive cases were 0.855 % (n = 54), HCV positive cases were 0.316% (n = 20), syphilis positive were 0.30% (n = 19) and MP positive cases were only 0.031% (n = 2). HBV, HCV, syphilis and malaria infections rates were found to be low as compared to the previous data published, while no case was reported for HIV. The study also revealed the distribution pattern of the aforementioned pathogens in blood groups and the Rh type system of the reactive samples. CONCLUSION: The lower reported in our study indicates the awareness among the people of Peshawar about TTIs and their precautions. The prevalence rate that we are reporting is less than previously published articles in the same domain.

Molecular prevalence, characterization and associated risk factors of Anaplasma spp. and Theileria spp. in small ruminants in Northern Pakistan.

This study was conducted in four districts (Malakand, Swat, Bajaur and Shangla) of Northern Pakistan to investigate the prevalence, associated risk factors and phylogenetic analyses of Theileria and Anaplasma species in small ruminants. A total of 800 blood samples, 200 from each district, were collected from apparently healthy animals. PCR assays were performed using generic primers for Anaplasma spp. and Theileria spp. as well as species specific primers for A. ovis and T. ovis. Overall infection prevalence was 361/800 (45.1%). Theileria spp. infection prevalence (187/800, 23.3%) was higher than Anaplasma spp. (174/800, 21.7%). Amplified partial 18S rRNA genes were sequenced and enrolled animals were found to be infected by T. ovis (115/800, 14.3%), and at least two more Theileria species (72/800, 9%) were present (T. lestoquardi and T. annulata). All blood samples that were found to be positive for Anaplasma spp. were also positive for A. ovis. Infection prevalence was higher in sheep (227/361, 28.3%) compared to goats (134/361, 16.6%) (p < 0.005). Univariable analysis of risk factors showed that host, age, grazing system and acaricide treatment were significant determinants (p < 0.05) for both Theileria and Anaplasma infections. Multivariable analysis revealed that host, sex, age, tick infestation and grazing system were significant risk factors (p < 0.005) for both pathogens. Phylogenetic analysis revealed variants among the A. ovis and T. annulata samples analysed, indicating that different genotypes are circulating in the field while T. ovis presented the same genotype for the samples analysed.

TITLE: Prévalence moléculaire, caractérisation et facteurs de risque associés d'Anaplasma spp. et Theileria spp. chez les petits ruminants du nord du Pakistan. ABSTRACT: Cette étude a été menée dans quatre districts (Malakand, Swat, Bajaur et Shangla) du nord du Pakistan pour étudier la prévalence, les facteurs de risque associés et les analyses phylogénétiques des espèces de Theileria et Anaplasma chez les petits ruminants. Au total, 800 échantillons de sang, 200 de chaque district, ont été prélevés sur des animaux apparemment sains. Les tests PCR ont été réalisés en utilisant des amorces génériques pour Anaplasma spp. et Theileria spp. ainsi que des amorces spécifiques à l'espèce pour A. ovis et T. ovis. La prévalence globale de l'infection était de 361/800 (45,1 %). La prévalence de l'infection à Theileria spp. (187/800, 23,3 %) était plus élevée que celle d'Anaplasma spp. (174/800, 21,7 %). Le gène de l'ARNr partiel 18S amplifié a été séquencé et les animaux concernés se sont révélés infectés par T. ovis (115/800, 14,3 %) et au moins deux autres espèces de Theileria (72/800, 9 %) étaient présentes (T. lestoquardi et T. annulata). Tous les échantillons de sang trouvés positifs pour Anaplasma spp. ont également été trouvés positifs pour A. ovis. La prévalence de l'infection était plus élevée chez les moutons (227/361, 28,3 %) que chez les chèvres (134/361, 16,6 %) (p < 0,005). Une analyse univariée des facteurs de risque a montré que l'hôte, l'âge, le système de pâturage et le traitement acaricide étaient des déterminants significatifs (p < 0,05) pour les infections à Theileria et Anaplasma. L'analyse multivariée des facteurs de risque a révélé que l'hôte, le sexe, l'âge, l'infestation par les tiques et le système de pâturage étaient des éléments de facteurs de risque importants (p < 0,005) pour les deux agents pathogènes. L'analyse phylogénétique a révélé des variantes parmi les échantillons d'A. ovis et de T. annulata analysés indiquant que différents génotypes circulent sur le terrain tandis que T. ovis présentait le même génotype pour tous les échantillons analysés.

Inhibition of G-protein signalling in cardiac dysfunction of intellectual developmental disorder with cardiac arrhythmia (IDDCA) syndrome.

BACKGROUND: Pathogenic variants of GNB5 encoding the ß5 subunit of the guanine nucleotide-binding protein cause IDDCA syndrome, an autosomal recessive neurodevelopmental disorder associated with cognitive disability and cardiac arrhythmia, particularly severe bradycardia. METHODS: We used echocardiography and telemetric ECG recordings to investigate consequences of Gnb5 loss in mouse. RESULTS: We delineated a key role of Gnb5 in heart sinus conduction and showed that Gnb5-inhibitory signalling is essential for parasympathetic control of heart rate (HR) and maintenance of the sympathovagal balance. Gnb5-/- mice were smaller and had a smaller heart than Gnb5+/+ and Gnb5+/- , but exhibited better cardiac function. Lower autonomic nervous system modulation through diminished parasympathetic control and greater sympathetic regulation resulted in a higher baseline HR in Gnb5-/- mice. In contrast, Gnb5-/- mice exhibited profound bradycardia on treatment with carbachol, while sympathetic modulation of the cardiac stimulation was not altered. Concordantly, transcriptome study pinpointed altered expression of genes involved in cardiac muscle contractility in atria and ventricles of knocked-out mice. Homozygous Gnb5 loss resulted in significantly higher frequencies of sinus arrhythmias. Moreover, we described 13 affected individuals, increasing the IDDCA cohort to 44 patients. CONCLUSIONS: Our data demonstrate that loss of negative regulation of the inhibitory G-protein signalling causes HR perturbations in Gnb5-/- mice, an effect mainly driven by impaired parasympathetic activity. We anticipate that unravelling the mechanism of Gnb5 signalling in the autonomic control of the heart will pave the way for future drug screening.

Reduction of cardiac iron overload by optimising iron chelation therapy in transfusion dependent thalassaemia using cardiac T2* MRI: a quality improvement project from Pakistan.

OBJECTIVES: Cardiac T2* MRI (T2*CMR), for accurate estimation of myocardial siderosis, was introduced as part of a QI collaborative to optimise chelation therapy in order to improve cardiac morbidity in transfusion dependent thalassaemia (TDT) patients. We report the impact of this QI initiative from two thalassaemia centres from this collaborative. DESIGN AND SETTING: A key driver based quality initiative was implemented to improve chelation in TDT patients registered at these two centres in Karachi, Pakistan. Protocol optimisation and compliance to treatment through training, communication and feedback were used as the drivers for QI intervention. Preintervention variables (demographics, chelation history, T2*CMR, echocardiography and holters) were collected from January 2015 to December 2016) and compared with variables in the post implementation phase (January to December 2019). A standardised adverse event severity for chelators and its management was devised for safe drug therapy as well as ensuring compliance to the regimen. Preintervention and postintervention variables were compared using non-parametric test. P value<0.05 was statistically significant. RESULTS: 100 patients with TDT, median age 17 (9-34) years, were included. An increase or stabilisation of T2*CMR was documented in 82% patients in the postintervention phase especially in patients with severe myocardial iron overload (5.5 vs 5.3 ms, p <0.01). Significantly fewer patients had abnormal echocardiographic findings (3.5% vs 26%, p <0.05) in the postintervention versus preintervention period. CONCLUSION: This QI initiative improved the chelation therapy leading to improved cardiac status in TDT patients at the participating centres.

Correction to: Simple protoplast isolation system for gene expression and protein interaction studies in pineapple (Ananas comosus L.).

[This corrects the article DOI: 10.1186/s13007-018-0365-9.].

Simple protoplast isolation system for gene expression and protein interaction studies in pineapple (Ananas comosus L.).

Background: An efficient transformation protocol is a primary requisite to study and utilize the genetic potential of any plant species. A quick transformation system is also crucial for the functional analysis of genes along with the study of proteins and their interactions in vivo. Presently, however, quick and effective transformation systems are still lacking for many plant species including pineapple. This has limited the full exploration of the genetic repository of pineapple as well as the study of its genes, protein localization and protein interactions. Results: To address the above limitations, we have developed an efficient system for protoplast isolation and subcellular localization of desired proteins using pineapple plants derived from tissue culture. A cocktail of 1.5% (W/V) Cellulase R-10 and 0.5% (W/V) Macerozyme R-10 resulted in 51% viable protoplasts with 3 h digestion. Compared to previously reported protocols, our protoplast isolation method is markedly faster (saving 4.5 h), requires only a small quantity of tissue sample (1 g of leaves) and has high yield (6.5 × 105). The quality of the isolated protoplasts was verified using organelle localization in protoplasts with different organelle markers. Additionally, colocalization analysis of two pineapple Mg2+ transporter genes in pineapple protoplasts was consistent with the results in a tobacco transient expression system, confirming that the protoplast isolation method can be used to study subcellular localization. Further findings showed that the system is also suitable for protein-protein interaction studies. Conclusion: Based on our findings, the presently described method is an efficient and effective strategy for pineapple protoplast isolation and transformation; it is convenient and time saving and provides a greater platform for transformation studies.

Diagnostic Value of Noninvasive Computed Tomography Perfusion Imaging and Coronary Computed Tomography Angiography for Assessing Hemodynamically Significant Native Coronary Artery Lesions.

BACKGROUND: The objective of this study is to determine the diagnostic performance of computed tomography perfusion (CTP) with and without computed tomography angiography (CTA) in assessment of hemodynamically significant coronary artery lesions in comparison to invasive fractional flow reserve (FFR). MATERIALS AND METHODS: PubMed and Cochrane Center Register of Controlled Trials from January 2010 searched through December 2014. Nine original studies were selected evaluating the diagnostic performance of CTP with and without CTA to invasive coronary angiography in evaluation of hemodynamic significance of coronary lesions (n = 951). RESULTS: The sensitivity, specificity, LR+ and LR- and DOR of CTA+CTP were 0.85 [95% confidence interval (CI: 0.79-0.89)] 0.94 (CI: 0.91-0.97), 15.8 (CI: 7.99-31.39), 0.146 (CI: 0.08-0.26), and 147.2 (CI: 69.77-310.66). Summary Receiver Operating Characteristics (SROC) results showed area under the curve (AUC) of 0.97 indicating that CTA+CTP may detect hemodynamically significant coronary artery lesions with high accuracy. The sensitivity, specificity, LR+ and LR- and DOR of CTP were 0.83 (CI: 0.78-0.87), 0.84 (CI: 0.80-0.87) 5.26 (CI: 2.93-9.43), 0.209 (CI: 0.12-0.36), and 31.97 (CI: 11.59-88.20). CONCLUSIONS: This result suggests that CTP with CTA significantly improves diagnostic performance of coronary artery lesions compared to CTA alone and closely comparable with invasive FFR.

Hepatocellular Carcinoma in Nonalcoholic Fatty Liver Disease.

Our objective was to study nonalcoholic fatty liver disease (NAFLD) as a relevant risk factor associated with hepatocellular carcinoma (HCC) in patients with and without cirrhosis. HCC is a common cancer worldwide that predominantly involves patients with hepatic cirrhosis. HCC has recently been linked to NAFLD, the hepatic manifestation of obesity and related metabolic disorders. This association is alarming due to the high prevalence of NAFLD globally, which may contribute to the rising incidence of HCC. A 31-year-old female with a history of dyslipidemia, hypertension, and diabetes mellitus presented with abdominal pain that persisted for six months. The pain was associated with gastrointestinal symptoms and weight loss. She was drug-free and a nonalcoholic and a nonsmoker. The physical examination was unremarkable. The abdominal exam showed a soft and non-tender abdomen, with no organomegaly or ascites. The laboratory evaluation was unremarkable. The imaging studies showed a hypodense lesion in the right hepatic lobe with strong arterial enhancement. Subsequently, the patient underwent a liver biopsy. The histopathology results were consistent with HCC. The patient underwent an uneventful segment VI liver resection and tumor-free margins were achieved. In our patient, NAFLD was designated as an independent etiology for HCC, without cirrhosis. Our patient recovered well and has been disease free for over a year. HCC may complicate non-cirrhotic NAFLD with mild or absent fibrosis, greatly expanding the population potentially at higher risk of HCC. These results provide new targets for surveillance, prevention, early recognition, and effective treatment of HCC associated with NAFLD.

Electrochemical characterization and in-vitro bio-assessment of AZ31B and AZ91E alloys as biodegradable implant materials.

The degradation of magnesium alloys, AZ31B and AZ91E, are under review due to a their ability to degrade under physiological conditions and successively yield an oxidized biocompatible by-product which can safely be absorbed by the body. By exploiting the biodegradability of magnesium alloys, the prospects of developing an unprecedented class of implant are at hand. To do so however, the rate of corrosion of the alloys must be modified in order to better suit physiological conditions. Therefore, anodization was carried out on AZ31B and AZ91E specimens to alter the surface chemistry to reduce the corrosion rates and improve biocompatibility. Scanning electron microscopy, energy dispersive spectroscopy, atomic force microscopy and contact angle meter, were used to characterize and compare the surfaces of untreated and anodized magnesium alloys. Corrosion behavior was evaluated by electrochemical tests using potentiodynamic polarization and electrochemical impedance spectroscopy, to verify changes in corrosion rates as a result of anodization. Finally, a bio-assessment using MTS assays and fluorescent microscopy were carried out to ensure that the anodization process had no compromise on the biocompatibility of the magnesium alloys. The study indicated that the anodization process did alter the surface chemistry of the alloys, yielding slower corrosion rates, while causing no adverse effects in regards to biocompatibility.

Effect of oxytocin on serum biochemistry, liver enzymes, and metabolic hormones in lactating Nili Ravi buffaloes.

Studies reporting the effects of oxytocin on the health of lactating animals are lacking and still no such data is available on Nili Ravi buffalo, the most prominent Asian buffalo breed. The present study was conducted to investigate the effect of oxytocin on physiological and metabolic parameters of lactating Nili Ravi buffaloes. Healthy lactating buffaloes (n = 40) of recent calving were selected from a commercial dairy farm situated in the peri-urban area of district Faisalabad, Pakistan. These buffaloes were randomly allocated to two equal groups viz experimental and control, comprising 20 animals each. Twice-a-day (morning and evening) milking practice was followed. The experimental and control buffaloes were administered subcutaneously with 3 mL of oxytocin (10 IU/mL) and normal saline respectively, prior to each milking. Serum biochemical profile including glucose, total cholesterol (tChol), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), total proteins (TP), C-reactive protein (CRP), liver enzymes aspartate transaminase (AST), alanine transaminase (ALT), and metabolic hormones triiodothyronine (T3) and thyroxine (T4) were studied. Results revealed significantly higher (P ≤ 0.01) levels of glucose, total cholesterol, LDL-C, triglycerides, total proteins, and C-reactive protein in experimental (oxytocin-injected) lactating buffaloes compared to control group. Liver enzymes AST and ALT as well as serum T4 concentration was significantly higher (P ≤ 0.01) in oxytocin-injected lactating buffaloes as compared to control animals. It was concluded that oxytocin had the key role in increasing the metabolic parameters and hormones, resulting in the optimization of production. But, at the same time, it may pose a threat to the animal health.

In situ synthesis of MIL-100(Fe) in the capillary column for capillary electrochromatographic separation of small organic molecules.

Because of the unusual properties of the structure, the metal organic frameworks (MOFs) have received great interest in separation science. However, the most existing methods for the applications of MOFs in separation science require an off-line procedure to prepare the materials. Here, we report an in situ, layer-by-layer self-assembly approach to fabricate MIL-100(Fe) coated open tubular (OT) capillary columns for capillary electrochromatography. By a controllable manner, the OT capillary columns with a tailored MIL-100(Fe) coating have been successfully synthesized. The results of SEM, XRD, FT-IR, and ICP-AES indicated that MIL-100(Fe) was successfully grafted on the inner wall of the capillary. Some neutral, acidic and basic analytes were used to evaluate the performance of the MIL-100(Fe) coating OT capillary column. Because of the size selectivity of lattice aperture and hydrophobicity of the organic ligands, three types of analytes were well separated with this novel MIL-100(Fe) coating OT capillary column. For three consecutive runs, the intraday relative standard deviations (RSDs) of migration time and peak areas were 0.4-4.6% and 1.2-6.6%, respectively. The interday RSDs of migration time and peak areas were 0.6-8.0% and 2.2-9.5%, respectively. The column-to-column reproducibility of retention time was in range of 0.6-9.2%. Additionally, the 10 cycles OT capillary column (10-LC) could be used for more than 150 runs with no observable changes on the separation efficiency.

Protein adsorption on citrate modified magnetic nanoparticles.

Magnetic iron oxide nanoparticles were prepared by chemical co-precipitation method and then modified with sodium citrate. These iron oxide nanoparticles were characterized by transmission electron microscopy (TEM), X-ray diffractometer (XRD), and vibrating sample magnetometer (VSM). BSA was adsorbed on these citrate modified nanoparticles using two types of buffers (acetate buffer, pH 4.0, 4.7 and phosphate buffer, pH 7.4). The results showed that the maximum adsorption of BSA was 83 mg/g at its isoelectric point (pH 4.7). Fourier-transform infrared spectroscopy (FTIR) was used to confirm the attachment of citrate groups and BSA on the prepared magnetic nanoparticles. BSA was desorbed from nanoparticles under alkaline conditions, which was confirmed by SDS-PAGE electrophoresis, UV-vis and fluorescence spectra. The desorbed BSA showed small changes in its structure. The adsorption results indicated that BSA adsorption on citrate modified iron oxide nanoparticles occurred mainly by electrostatic mechanism.

Empyema thoracis: management outcome.

BACKGROUND: Empyema thoracis results from postpneumonic effusion of bacterial origin or trauma. If untreated it may convert to fibro-purulent or an organising stage. METHODS: This study was conducted at cardiothoracic unit of Ayub Teaching Hospital Abbottabad from Jan 2008 to Aug 2009. Patients with diagnosis of empyema thoracic were studied. Their clinical features and investigations were recorded on a proforma. Antibiotics alone, aspiration, chest intubation, rib resection or decortication were performed where required. RESULTS: Out of 71 patients, males were 58 (81.69%) and females were 13 (18.3%). Majority of the patients (24, 33.8%), were below 20 years of age. Most of the patients (59, 83.09%), were from the poor socioeconomic class. Forty-one (57.76%) patients were smokers, 12 (16.9%) were diabetic, and 39 (54.92%) were hypertensive. Cough was the most common complain (62, 87.32%). Forty (56.33%) had more than 50% involvement of hemi thorax. In 44 (61.97%) cases, cause was tuberculosis. Fifty-one (71.83%) patients were treated with tube thoracostomy and 13 (18.3%) required decortication. CONCLUSION: Empyema thoracis commonly affects males. Apart from clinical assessment, x-ray chest is an important and simple investigation to detect empyema thoracic. Mostly tube drainage cures the diseases but in case of failure other options are also available.

Ankle brachial index measurement in primary care setting: how long does it take?

BACKGROUND: Peripheral arterial disease (PAD) affects over 8 million people in the United States and has been found to be associated with an increased incidence of coronary and carotid artery disease. The ankle brachial index (ABI) measurement is a highly specific noninvasive screening and diagnostic test for PAD, but is rarely performed in primary care office settings. This study sought to determine the actual performance time involved in completing an ABI in a primary care office. METHODS: Data were collected by one provider on a convenience sample of women who met the inclusion criteria. The time was recorded at the beginning and upon the completion of the ABI procedure for each patient. Analysis of the time data was completed and barriers to performing the ABI were recorded by the provider. RESULTS: The average time to complete an ABI was 5 minutes, with a range of 3-11 minutes. In 83.8% of patient encounters, the ABI procedure took less than 6 minutes to complete. Barriers identified by the provider included the additional time needed to explain the test and assist patients into the proper testing position. CONCLUSIONS: The actual performance time for an ABI in a primary care setting takes an average of 5 minutes, but additional time may be required for patient preparation and education. With proper scheduling and training, the ABI can be completed in a timely manner. The ABI is an important screening/diagnostic test that can be performed in primary care and potentially impact patient treatment plans.