Molecular Analyses of Phenylketonuria in The Intellectually Disabled Children from Faisalabad, Punjab, Pakistan

ABSTRACT Background and Objective: Phenylketonuria (PKU) is a rare inherited metabolic disorder, caused by mutations in the phenylalanine hydroxylase (PAH). It is a treatable disorder if diagnosed earlier in life. The objective was to identify PKU patient(s) amongst the intellectually disabled children. Methods: Blood samples (n=100) were collected from intellectually disabled children from Faisalabad, Pakistan. Screening was performed on plasma samples through High Performance Liquid Chromatography (HPLC), and DNA samples were examined for mutation analysis of PAH through direct PCR and SSCP analyses. Results: In the current study, 85% consanguinity rate was observed, with the average BMI (16.15 kg/m2) and head circumference(50.21 cm) was observed and the age range of the patients were 8-14 years. Moreover, through biochemical and genetic analyses, not a single PKU patient was identified. Conclusion: Based on just one previous report and our small dataset it is concluded that either mutations are not common in the hotspot regions or chances of occurrence of PKU might be rare in Pakistan. Moreover, there is a need of more research on large scale to find the incidence of PKU in Pakistan

Accuracy of MRI in Diagnosis of Invasive Placenta by Taking Per Operative Findings as Gold Standard

Objective: To assess the diagnostic accuracy of magnetic resonance imaging (MRI) in detecting invasive placentas using per-operative findings as the gold standard.Methodology: A prospective cross-sectional study was conducted in the diagnostic radiology department of KRL General Hospital Islamabad during Oct 2019 to Sep 2021. Sixty prenatal individuals were identified as having a high risk of invasive placenta and underwent MRI (Phillips 1.5 T) to confirm the diagnosis. A trainee radiologist and a consultant radiologist reviewed the images. The MRI's sensitivity, specificity, positive predictive value, negative predictive value, and accuracy was calculated using a 2×2 contingency tables. Results: Ten cases of invasive placenta were detected postoperatively (gold standard). The MRI had a sensitivity of 90%, a specificity of 93%, a positive predictive value of 90%, a negative predictive value of 90%, and an accuracy of 92.3 percent, respectively. Conclusion: The study concluded that magnetic resonance imaging (MRI) offers a good diagnostic accuracy and is a reproducible technology for prenatal identification of invasive placentas

Comparison of the virulence markers of helicobacter pylori and their associated diseases in patients from Pakistan and Afghanistan

BACKGROUND/AIM: Helicobacter pylori is a Gram-negative bacteria, which is associated with development of gastroduodenal diseases. The prevalence of H. pylori and the virulence markers cytotoxin-associated gene A and E (cagA, cagE) and vacuolating-associated cytotoxin gene (vacA) alleles varies in different parts of the world. H. pylori virulence markers cagA, cagE, and vacA alleles in local and Afghan nationals with H. pylori-associated gastroduodenal diseases were studied. PATIENTS AND METHODS:Two hundred and ten patients with upper gastrointestinal symptoms and positive for H. pylori by the urease test and histology were included. One hundred and nineteen were local nationals and 91 were Afghans. The cagA, cagE, and vacA allelic status was determined by polymerase chain reaction. RESULTS:The nonulcer dyspepsia (NUD) was common in the Afghan patients (P = 0.025). In Afghan H. pylori strains, cagA was positive in 14 (82%) with gastric carcinoma (GC) compared with 29 (45%) with NUD (P = 0.006), whereas cagE was positive in 11 (65%) with GC and 4 (67%) with duodenal ulcer (DU) compared with 12 (18%) with NUD (P \u3c 0.001 and 0.021, respectively). The vacA s1a/b1was positive in 10 (59%) of GC compared with 20 (31%) in NUD (P = 0.033). In Pakistani strains, cagE was positive in 12 (60%) with GC, 7 (58%) with GU, 12 (60%) with DU compared with 11 (16%) with NUD (P \u3c 0.001, 0.004, and \u3c 0.001, respectively). In Pakistani strains, cagA/s1a/m1 was 39 (33%) compared with Afghans in 17 (19%) (P = 0.022). Moderate to severe mucosal inflammation was present in 51 (43%) Pakistani patients compared with 26 (28%) (P = 0.033) in Afghans. It was also associated with grade 1 lymphoid aggregate development in Pakistani patients 67 (56%) compared with 36 (40%) (P = 0.016) in Afghans. CONCLUSION: Distribution of H. pylori virulence marker cagE with DU was similar in Afghan and Pakistan H. pylori strains. Chronic active inflammation was significantly associated with Pakistani H. pylori strains

Estimation of airship states and model uncertainties using nonlinear estimators

This Airships are lighter than air vehicles and due to their growing number of applications, they are becoming attractive for the research community. Most of the applications require an airship autonomous flight controller which needs an accurate model and state information. Usually, airship states are affected by noise and states information can be lost in the case of sensor's faults, while airship model is affected by model inaccuracies and model uncertainties. This paper presents the application of nonlinear and Bayesian estimators for estimating the states and model uncertainties of neutrally buoyant airship. It is considered that minimum sensor measurements are available, and data is corrupted with process and measurement noise. A novel lumped model uncertainty estimation approach is formulated where airship model is augmented with six extra state variables capturing the model uncertainty of the airship. The designed estimator estimates the airship model uncertainty along with its states. Nonlinear estimators, Extended Kalman Filter and Unscented Kalman Filter are designed for estimating airship attitude, linear velocities, angular velocities and model uncertainties. While Particle filter is designed for the estimation of airship attitude, linear velocities and angular velocities. Simulations have been performed using nonlinear 6-DOF simulation model of experimental airship for assessing the estimator performances. 1− uncertainty bound and error analysis have been performed for the validation. A comparative study of the estimator's performances is also carried out

Cytokine changes in colonic mucosa associated with Blastocystis spp. subtypes 1 and 3 in diarrhoea-predominant irritable bowel syndrome

We determined cytokines (e.g. interleukin-8, 10, 12 and TNF-α) expression by peripheral blood mononuclear cells (PBMCs) and in rectal mucosa in diarrhoea-predominant irritable bowel syndrome (D-IBS) with Blastocystis spp. Eighty patients with D-IBS and Blastocystis spp. infection were classified as \u27cases\u27 and 80 with D-IBS without Blastocystis spp. infection were classified as \u27control\u27. Cases were subdivided into D-IBS and Blastocystis sp. defined type 1 (subtype-specific primer SB83) and type 3 (SB227). Stool microscopy and culture were performed. Rectal biopsies were obtained for histology and cytokines by real-time PCR for mRNA expression of cytokines. PBMCs IL-8 was similar in different groups but in type 1, IL-8mRNA was increased compared with type 3 (P = 0·001) and control (P = 0·001). In type 1, IL-10 by PBMCs had a low mean value (14·5±1·6) compared with (16·7±1·5) type 3 and (16±2·3) in controls (

Glyphosate: cancerous or not? Perspectives from both ends of the debate

Glyphosate is non-selective herbicide. Studies published in the last decade, point towards glyphosate toxicity. Shikimic acid pathway for the biosynthesis of folates and aromatic amino acids is inhibited by glyphosate. Glyphosate carcinogenicity is still considered to be a controversial issue. The World Health Organizations’ International Agency recently concluded that glyphosate is “probably carcinogenic to humans.” Some researchers believed that glyphosate is not linked with carcinogenicity

Insight of Tp53 Mutations and their effect on Protein in Different Feline and Canine Neoplasms

Background: Mutations in the Tp53 gene, a tumor suppressor gene, may cause dysfunction in growing cells and hinder the phenomenon of apoptosis, an alleged cause of tumorigenesis. It is involved in conservation of the genome and DNA repair, mutations of this gene may cause the damaged cells to grow continuously.Methods: The type of molecular changes in Tp53 gene and their effects on physiochemical and structural properties of this protein in various Canine and Feline cancers were observed in this study by using online bioinformatics tools.Results: Our results indicated that lymphomas and perianal adenocarcinomas (PAC) have the same mutation at c. 104, while mammary tumors and canine transmissible venereal tumor (CTVT) contain different mutations. Referring to changes in protein, synonymous mutations in granulomas were observed while certain mutations in squamous cell carcinoma (SCC) and head & neck tumors were detected in Canis familiaris. In Felis catus, the mutant protein was similar to wild type protein with exception of mutant 5 of mammary tumor, which had a deletion at the 287 amino acid position.Conclusion: The insight gathered on the p53 mutant proteins in both species aided our understanding of the in-vivo fate of the p53 protein and its isoforms and the effects that morphological changes can have on the fate of cells. Furthermore, isolation of this protein may augment our understanding about the structural biology of these proteins

Anti-Helicobacter pylori activity and inhibition of Helicobacter pylori-induced release of IL-8 in AGS cells by plant extracts

Plants used in popular diet were studied for anti-Helicobacter pylori activity and their effect on the expression of interleukin-8 (IL-8) from H. pylori infected gastric epithelial cells. Extracts were prepared of Allium sativum (A. sativum), Cuminum cyminum (C. cyminum),Piper nigrum (P. nigrum) and their mix in two different dilutions. AGS cell line and H. pyloristrains were used for co-culture experiments. Extracts bactericidal activity was determined by a viable colony count. ELISA (Enzyme linked immunosorbent assay) was used to determine IL-8 expression. DNA extracted from bacterial cells was used for polymerase chain reaction (PCR) of cytotoxin-associated gene (cagA) and E (cagE). Results revealedH. pylori strains sensitivity to A. sativum (5.5 mg/ml) was 57% (39/69) (p=0.06), and to 11 mg/ml was 65% (45/69) (p=0.02) compared to amoxicillin, respectively. CagE positive H. pylori, 37% (11/30) (p=0.02) were sensitive to plant mixture (23.5 mg/ml), 60% (18/30) (pC. cyminum (PP. nigrum (P=0.0046) and A. sativum(P=0.0021), respectively compared to positive and negative controls. Thus, dietary plants demonstrated an anti-H. pylori effect. They reduced IL-8 expression from the H. pyloriinfected AGS cells. Key words: Helicobacter pylori; Allium sativum, Cuminum cyminum, Piper nigrum, gastric epithelial cells, interleukin-8, ELISA, cagA, cagE

Prevalence of non Helicobacter pylori species in patients presenting with dyspepsia

<p>Abstract</p> <p>Background</p> <p>Helicobacter species associated with human infection include <it>Helicobacter pylori, Helicobacter heilmannii </it>and <it>Helicobacter felis </it>among others. In this study we determined the prevalence of <it>H. pylori </it>and non-<it>Helicobacter pylori </it>organisms <it>H. felis and H. heilmannii </it>and analyzed the association between coinfection with these organisms and gastric pathology in patients presenting with dyspepsia. Biopsy specimens were obtained from patients with dyspepsia on esophagogastroduodenoscopy (EGD) for rapid urease test, histology and PCR examination for Helicobacter genus specific 16S rDNA, <it>H. pylori </it>phosphoglucosamine mutase (<it>glmM</it>) and urease B (<it>ureB</it>) gene of <it>H. heilmannii </it>and <it>H. felis</it>. Sequencing of PCR products of <it>H. heilmannii </it>and <it>H. felis </it>was done.</p> <p>Results</p> <p>Two hundred-fifty patients with dyspepsia were enrolled in the study. The mean age was 39 ± 12 years with males 162(65%). Twenty-six percent (66 out of 250) were exposed to cats or dogs. PCR for Helicobacter genus specific 16S rDNA was positive in 167/250 (67%), <it>H. pylori glmM </it>in 142/250 (57%), <it>H. heilmannii </it>in 17/250 (6%) and <it>H. felis </it>in 10/250 (4%), respectively. All the <it>H. heilmannii </it>and <it>H. felis </it>PCR positive patients were also positive for <it>H. pylori </it>PCR amplification. The occurrence of coinfection of <it>H. pylori </it>and <it>H. heilmannii </it>was 17(6%) and with <it>H. felis </it>was 10(4%), respectively. Only one out of 66 exposed to pets were positive for <it>H. heilmannii </it>and two for <it>H. felis</it>. Histopathology was carried out in 160(64%) of 250 cases. Chronic active inflammation was observed in 53(56%) (p = 0.001) of the patients with <it>H. pylori </it>infection alone as compared to 3(37%) (p = 0.73) coinfected with <it>H. heilmannii </it>and <it>H. pylori </it>and 3(60%) coinfected with <it>H. felis </it>and <it>H. pylori </it>(p = 0.66). Intestinal metaplasia was observed in 3(3%)(p = 1.0) of the patients with <it>H. pylori </it>infection alone as compared to 2(25%) (p = 0.02) coinfected with <it>H. heilmannii </it>and <it>H. pylori </it>and 1(20%) coinfected with <it>H. felis </it>and <it>H. pylori </it>(p = 0.15).</p> <p>Conclusion</p> <p>The prevalence of <it>H. heilmannii </it>and <it>H. felis </it>was low in our patients with dyspepsia. Exposure to pets did not increase the risk of <it>H. heilmannii </it>or <it>H. felis </it>infection. The coinfection of <it>H. pylori </it>with <it>H. heilmannii </it>was seen associated with intestinal metaplasia, however this need further confirmation.</p