Hepatitis E associated determinants and diagnostic biomarkers during pregnancy and its prenatal consequences in Multan, Punjab tertiary care setting (Pakistan).

OBJECTIVE: Hepatitis E virus (HEV) is the most common cause of acute viral hepatitis in the world. Hepatitis E infection is commonly widespread by the fecal oral routes and contaminated water. This study was designed to explore the prevalence and risk factors of hepatitis E infection in pregnant women of the Multan district, Pakistan. METHODS: The study comprised of a total of 500 enrolled patients, among which, 105 pregnant females with hepatitis E infection fulfilled the criteria for anti-HEV antibodies. Pregnant women without significant complications and without hepatitis E infection were excluded from this study. Hepatic profile, complete blood count, coagulation markers, and standard protocol were also assessed for fetal maternal hemorrhage. RESULTS: Our results showed that 105 patients (66.66%, CI 95%) had HEV infection with mean age 25±5 years. Serum bilirubin levels were increased in 74 patients (70.47%), aspartate transaminase was elevated > 200 IU/L in 71 patients (67.61%), alanine transaminase was above the 100 IU/L in 65 patients (245 IU/L), and low platelet counts were found in 45 patients (42.85%). Moreover, fetal distress cases were 9 (10.84%) and maternal distress cases were about 11 (13.25%). Fetal mortality cases were 39 (37.14%), and maternal mortality cases were about 22 (20.95%) due to hepatic comma, intravascular coagulation, and hepatic failure. CONCLUSION: It was concluded that the prevalence of Hepatitis E during pregnancy is associated with high risk factors of unhygienic practices, blood transfusion, and noncompliance with universal infection control techniques. Maternal fatalities and fetal consequences were exacerbated by HEV infection.

Genotypes of Hepatitis C Virus and Efficacy of Direct-Acting Antiviral Drugs among Chronic Hepatitis C Patients in a Tertiary Care Hospital.

Hepatitis C virus (HCV) chronic infection is a major causative factor for several chronic liver diseases, including liver cirrhosis, liver cell failure, and hepatocellular carcinoma. The HCV has seven major genotypes. Genotype 4 is the most prevalent genotype in the Middle East, including Saudi Arabia, followed by genotype 1. The HCV genotype affects the response to different HCV treatments and the progression of liver disease. Currently, combinations of direct-acting antiviral drugs (DAAs) approved for the treatment of HCV achieve high cure rates with minimal adverse effects. Because real-world data from Saudi Arabia about the efficacy of DAAs are still limited, this study was conducted to assess the effectiveness of DAAs in treating patients with chronic hepatitis C and to identify the variables related to a sustained virologic response (SVR) in a real-world setting in Saudi Arabia. This prospective cohort study included 200 Saudi patients with chronic HCV who were 18 years of age or older and had been treated with DAAs at King Abdul-Aziz Specialized Hospital in Taif, Saudi Arabia, between September 2018 and March 2021. The response to treatment was assessed by whether or not an SVR had been achieved at week 12 post treatment (SVR12). An SVR12 was reached in 97.5% of patients. SVR12 rates were comparable for patients of different ages, between men and women, and between patients with and without cirrhosis. In addition, the SVR12 rates did not differ according to the infecting HCV genotype. In this study, the presence of cirrhosis and the patient's gender were independent predictors of who would not reach an SVR12 (known here as the non-SVR12 group) according to the results of univariate and multivariate binary logistic regression analyses based on the determinants of SVR12. In this population of patients with chronic HCV infection, all DAA regimens achieved very high SVR12 rates. The patients' gender and the presence of cirrhosis were independent factors of a poor response.

Toll- like receptor 2 polymorphism and IL-6 profile in relation to disease progression in chronic HBV infection: a case control study in Egyptian patients.

Chronic hepatitis B (CHB) has a wide range of outcomes depending on host immune responses mainly Toll-like receptors (TLRs) signaling and released cytokines. Toll-like receptor 2 (TLR2) single nucleotide polymorphisms (SNPs) and interleukin 6 (IL-6) may influence the course of CHB. We aimed to elucidate the relation between TLR-2 polymorphism, IL-6 profile, and CHB progression. We analyzed TLR-2 polymorphism (SNP; rs3804099) in 185 CHB patients and 60 controls using TaqMan allelic discrimination assay. Serum IL-6 levels were assessed by ELISA. IL-6 levels were considerably higher in active CHB and cirrhotic patients compared with inactive carriers and controls (P < 0.001). IL-6 showed positive correlation with ALT and advanced fibrosis in active CHB patients (r = 0.31, P = 0.02). A significant positive correlation was noticed between IL-6 and HBV DNA PCR in all CHB groups. TT genotype of rs3804099/TLR-2 was significantly more prevalent in inactive carriers compared to active hepatitis patients (P = 0.04, OR = 0.39 and 95% CI: 0.16-0.95). Both heterozygous CT and mutant TT genotypes were significantly more frequent among inactive carriers compared to cirrhotic patients (P = 0.01, OR = 0.33, 95% CI: 0.13-0.81 and P = 0.009, OR = 0.32, 95% CI: 0.13-0.77). TT genotype was significantly related to lower IL-6 levels in active hepatitis and cirrhotic groups (P = 0.005 and P = 0.001, respectively) showing that TLR mutations would be associated with milder hepatitis activity and lower possibility for disease progression. There may be a positive association between TLR2 rs3804099 polymorphism and hepatitis B activity. IL-6 is a good indicator of CHB disease progression.

Exploring the effect of Jasmonic Acid for Aphids control for improving the yield of Triticum aestivum varieties.

Many biotic and abiotic factors influence the production of wheat (Triticum aestivum L.). Among biological agents, aphids are destructive pests effecting wheat yield drastically. This study was designed to evaluate the impact of foliar Jasmonic acid spray on aphid population as well as on plant growth during aphid infestation in two wheat varieties i.e., Borlaug-2015 and Zincol-2015. Plants are cultivated in pots and treated with jasmonic acid at concentrations of 0.1 and 1 mM (JA). The results revealed that length of shoot and roots decreased after aphid stress and was improved (21-24%) by JA spray. Photosynthetic pigments increased after applying the jasmonic acid spray compared to control plants. Jasmonic acid spray helped the plants to recover from aphid stress by enhanced production of antioxidant enzymes CAT (Catalase) (65-71%), SOD (Superoxide dismutase) (71-74%) and POD (Peroxidase) (61-65%). Consequent to improved defence system, plants treated with JA had fewer aphids as compared to control (60-73% reduction), 24 h after spray. The higher concentration of JA (1 mM) proved more effective as compared to 0.1 mM jasmonic acid. Moreover, Zincol-2015 appeared tolerant as compared to Borlaug-2015 against aphid infestation. The application of jasmonic acid as an exogenous foliar application showed an overall positive impact on the physiological and biochemical attributes of both varieties. It helps the plants to enhance resistance against the biotic stress and can be adopted as future alternative for aphid management. However, detailed studies regarding understanding of underlying molecular mechanisms are needed to optimize the mode for field application.

Innovative Use of Biodex Balance System to Improve Dynamic Stabilization and Function of Upper Quarter in Recreational Weightlifters: A Randomized Controlled Trial.

Background and Objectives: Following an injury, upper-body strength and proprioception training is typically suggested. To our understanding, no prior research has looked into the impact of balance training on upper-body strength and stability. So, this study investigated the effects of Biodex balance training on enhancing the dynamic stability, strength, and function of the upper quarter (UQ) in recreational weightlifters. Materials and Methods: Fifty male weightlifters were randomly assigned into two groups. The experimental group received an upper-extremity Biodex balance training program three times/week for eight weeks, while the control group underwent a regular weightlifting training routine. Pre- and post-test scores of the upper-quarter dynamic stability, strength, and function were measured for both groups using the shoulder active repositioning accuracy test, two-minute push-up test, and the upper-quarter Y-balance test (UQ-YBT) and one-arm hop test, respectively. Results: Post-test values were significantly greater for the normalized UQ-YBT test than pre-test values in both groups (p < 0.05). Post-test values of the experimental group were significantly greater than the control group (p < 0.05). Regarding the shoulder active repositioning accuracy test and the time of the one-arm hop test, post-test values were significantly lower than pre-test values for both groups (p < 0.05), and post-test values of the experimental group were significantly lower than those of the control group (p < 0.05). The post-test value of the two-minute push-up test of each group was significantly higher than the pre-test value (p < 0.05), without any significant difference between both groups (p > 0.05). Conclusions: Adding upper-body Biodex balance training to a regular weightlifting training routine was effective in enhancing the upper quarter's dynamic stability and function.

The First Saudi Report of Novel and Common Mutations in the gyrA and parC Genes Among Pseudomonas Spp. Clinical Isolates Recovered from Taif Area.

Background and Aims: Reports examine quinolone resistance mechanisms among Pseudomonas spp. are sporadic in the Kingdom of Saudi Arabia (KSA). We previously examined the genetic bases of plasmid-mediated quinolone resistance among Pseudomonas spp. clinical isolates. This study investigated chromosomally mediated quinolone resistance mechanisms via investigation of the mutations in the gyrA and parC genes. Methods: The minimum inhibitory concentration (MIC) to different quinolones was determined. Twenty-nine quinolone resistant Pseudomonas spp. clinical isolates were included. The gyrA and parC genes were sequenced by Sanger capillary electrophoresis. Multiple sequence alignment for the translated gyrA and parC genes was performed to identify mutation sites. Results: Of the 29 isolates, 27 isolates were P. aeruginosa and two were P. putida. The cluster analysis of the quinolone susceptibility pattern revealed seven susceptibility phenotypes (A-G) based on susceptibility patterns rather than the MIC values. Also, 22 different susceptibility phenotypes were detected based on MIC values. All isolates exhibited a missense mutation at position 83 (S83I/T/F) of the gyrA gene in addition to six missense mutations at positions outside the QRDR of this gene. In addition, 82.8% (24/29) of the isolates harbored a missense mutation in the parC gene at position 87 (S87L), along with six novel mutations outside the QRDR of the parC gene. Haplotyping of the gyrA, parC, and the overall QRDR revealed six, 10, and 13 different haplotypes, respectively. Conclusion: This study documents the incidence of the commonly reported mutations in the gyrA and parC genes in addition to novel mutations in these genes among Pseudomonas spp. clinical isolates recovered from KSA. Together with our previous findings, these data provide an insight into the genetic background of quinolone resistance among Pseudomonas spp. clinical isolates in KSA.

In vitro and in vivo study of inhibitory potentials of α-glucosidase and acetylcholinesterase and biochemical profiling of M. charantia in alloxan-induced diabetic rat models.

OBJECTIVES: Diabetes mellitus is a multifactorial chronic disease that affects the human population and it is the third most common cause of death worldwide. Momordica charantia is used as popular folk medicine and its action against diabetes mellitus remains unclear. We investigated the inhibitory potentials of α-glucosidase, acetylcholinesterase, and biochemical profiling of M. charantia in alloxan-induced diabetic rat models. METHODS: An In vivo study was carried out by using twenty male albino Wistar rats randomly divided into five groups each comprising four rats. Diabetes mellitus was induced by single intraperitoneal administration of 80 mg/kg body weight of alloxan and treatment with plant extract was conducted for a period of thirty days to check their impact on body weight and differentblood values. Biochemical profiling and characterization were performed by in vitro assays and HPLC, and FTIR. Histopathologic effects of M. charantia were examined through automated image analysis. Results were analyzed through Tukey's test, a complete randomized design and two factorial designs under CRD. RESULTS: Methanolic extract demonstrated potent alpha-glucosidase (72.30 ± 1.17%) and acetylcholinesterase (50.12 ± 0.82%) inhibitory activities. HPLC analysis confirmed the existence of vital flavonoids, antioxidants, and saponins. FTIR revealed the presence of hydroxyl groups, esters, alkanes, alkenes, alkynes, ketones, alcohols, amines and carboxylic acids as major functional groups. Results of in vivo study demonstrated that co-administration of alloxan and methanolic extract of M. charantia significantly improved the levels of fasting blood glucose, glycated hemoglobin and insulin in diabetic rats. CONCLUSION: M. charantia can be recommended as a therapeutic adjunct for diabetic patients as it can provide favorable remedial action in the context of the diabetes continuum of metabolic syndrome.

Histomorphometric study of placental blood vessels of chorion and chorionic villi vascular area among women with preeclampsia.

INTRODUCTION: The direction of blood movement in normal and abnormal placenta is curious from a morphometric point of view. Once pregnancy is compromised by an illness like hypertension, maternal and foetal distress can lead to negative outcomes. The quantitative variations in the blood vessels within the chorion and the chorionic villi in placentas from pregnancies are complicated by preeclampsia (PE) and are poorly defined. The purpose of this study was to calculate and explore the morphometric measurement of blood vessels involved in the progress of hypertension through pregnancy within the chorion and the chorionic villi among normotensive women (n = 39) versus a preeclamptic group (n = 35). METHODS: Measurements used a computerized morphometry system and a Vascular Medicine Institute (VMI) calculator. RESULTS: Our data showed a significant decrease in vessel area (VA), wall area (WA), lumen area (LA), mean wall thickness-boundary (MWTB), mean wall thickness-rosette (MWTR), mean diameter-rosette (MDR), mean wall thickness-skeleton (MWTS), and external diameter-skeleton (EDS) in preeclampsia women compared to normotensive women. There were no significant differences between preeclampsia and control group in lumen area. DISCUSSION: We concluded that preeclamptic chorion and chorionic villi vessels are linked with significant structural discrepancies; future studies should address morphological events that occur throughout pregnancy including associations between arterial elastic properties-mainly collagen and structural proteins in hypertensive patients. A more integrated approach involving parallel analysis of the effects of potential vasoactive factors on the morphology of foetal vessel alteration is also needed.

Biological control potential of entomopathogenic fungal strains against peach Fruit fly, Bactrocera zonata (Saunders) (Diptera: Tephritidae).

The peachfruit fly, Bactrocera zonata (Saunders) is a polyphagous pest in nature, belonging to order, Diptera and their respective family is Tephritidae. It mostly feeds on different crops, vegetables and fruits. Different traditional chemical insecticides have been used to control this notorious pest. Excessive consumption of pesticides has become a major threat to the fresh fruits trade since many importing countries refused to accept the shipments due to public health and environmental concerns. There is a growing trend to control these pests using the most effective biological control methods and other preventive measures have been adopted for reducing their attacks. Fungal agents have been used as biological agents to manage the attack of different insects pest through biological means. The present study was conducted to assess the virulence of three entomopathogenic fungi, Metarhizium anisopliae, Beauveria bassiana and Verticillium lecanii, against Bactrocera zonata stages under different laboratory conditions. The results showed that B. bassiana and M. anisopliae were more effective in pathogenicity and potentially kill at all stages of B. zonata as compared to V. lecanii. The highest mortality rate for the third larval instar and the pupal stage were recorded after exposure to the 1 × 1010 conidia/ml concentrations, B. bassiana, with 68.67% and 89.67%, respectively. Adult B. zonata flies were the most susceptible to all entomopathogenic fungi. However, M. anisopliae was more virulent against B. zonata adult flies than B. bassiana and V. lecanii at 1 × 1010 conidial concentration. Therefore, the entomopathogenic fungi B. bassiana and M. anisopliae can be used as an cost effective bio-insecticide in the integrated pest management programs to control B. zonata. This study will be helpful to overcome this pest through biological control means.

Incidence of Epstein-Barr Virus Among Women With Breast Cancer Using Monoclonal Antibodies for Latent Membrane Protein 1 (LMP1).

BACKGROUND/AIM: Breast cancer is a common type of cancer in Sudan. Numerous studies propose viral oncogenesis as an etiological factor for breast cancer. The aim of the study was to analyze the presence of the Epstein-Barr virus (EBV) using monoclonal antibodies against latent membrane protein 1 (LAMP1) and determine the correlation between the presence of EBV and clinicopathological characteristics. PATIENTS AND METHODS: This study used immunohistochemistry to analyze the presence of EBV in 202 samples from Sudanese women diagnosed with breast cancer. Clinicopathological data were collected from patient records from the Radiation and Isotopes Centre in Khartoum State, Republic of Sudan. RESULTS: This study included 202 patients 168 (83.2%), 16 (7.9%), and 18 (8.9%), diagnosed with invasive ductal carcinoma, invasive lobular carcinoma, and papillary carcinoma, respectively. Axillary lymph node metastasis was present in 57 (28.2%) of cases, while 11 patients (5.4%) tested positive for EBV. The mean age of patients was 48.14±14.4 years. EBV infection was more frequently detected in invasive ductal carcinoma cases, and EBV positivity was not associated with cancer type, grade, progesterone levels, and HER2 expression. On the other hand, a statistically significant association was found between EBV presence and lymph node involvement, estrogen receptor status, and age group. CONCLUSION: EBV may not play a vital role in the pathogenesis of breast carcinoma in Sudanese women.

Significance of induced hybridized metallic and non-metallic nanoparticles in single-phase nano liquid flow between permeable disks by analyzing shape factor.

The current communication is designed by keeping in the mind high heat transfer capabilities of nanoliquids with the dispersion of diversified-natured nanoparticles in poorly conducting base liquids. Here, an amalgamation of metallic (Cu) and hybridization of metallic and non-metallic oxide (Cu-TiO2) nanoparticles to uplift thermophysical attributes of water is deliberated. The magnetically affected flow between rotating disks under the impact and permeability aspect is assumed. Empirical relations for effective dynamic viscosity, density, and heat capacitance to show mesmerizing features of obliged nanoparticles are also expressed. In addition, mathematical relations also depend on morphological factors like shape, size, and diameter of inducted nanoparticles. The mathematical formulation of the problem is conceded in the form of a system of ODEs after using similarity transformation on dimensional PDEs. Simulations of the complex coupled differential structure are solved by using a numerical approach by employing shooting and Runge-Kutta procedures jointly. The impact of flow concerning variables on associated distributions is revealed through tabular and graphical manner. Quantities of engineering interest associated with work like wall friction and thermal flux coefficients at walls of the disk are also calculated. It is deduced from an examination that the addition of metallic particles raises heat transfer more than non-metallic particles. A significant impression of magnetic field on shear stress is executed by hybrid nanoparticles along the surface of disks. In addition, elevation in Nusselt number and depreciation in skin friction coefficient is revealed against increasing magnitude of nanoparticle volume fraction. A positive trend in skin friction coefficient is manifested against the increasing magnitude of Reynold number. It is also observed that by increasing the size and shape of hybrid nanoparticles thermal conductivity and viscosity of the base fluid increases.

Value of Serum miRNA-96-5p and miRNA-99a-5p as Diagnostic Biomarkers for Hepatocellular Carcinoma.

Purpose: Circulatory microRNAs (miRNAs) have the potential to be employed as markers for cancer detection and as prognostic tools for disease management. As a result, our goal was to explore the effectiveness of serum miRNA-96-5p and miRNA-99a-5p as diagnostic tools in hepatocellular carcinoma (HCC). Patients and methods: Blood samples were collected from 55 patients with HCV-induced HCC, 55 patients with HCV-induced liver cirrhosis, and 55 healthy controls. The expression levels of miRNA-96-5p and miRNA-99a-5p were measured using quantitative RT-PCR. Results: miRNA-96-5p expression levels were increased in HCC patient sera, while miRNA-99a-5p levels were reduced. According to ROC curve analysis, using a combination of circulating miRNA-96-5p, miRNA-99a-5-, and alpha-fetoprotein (AFP) improves the accuracy of diagnoses for HCC, with an area under the curve (AUC) of 0.97, compared to AUCs of 0.82, 0.86, and 0.73, respectively, for the individual biomarkers. Furthermore, the present data suggested that higher serum miRNA-96-5p levels were linked to larger tumors and metastasis, whereas lower serum miRNA-99a-5p levels were exclusively linked to HCC metastasis. Conclusion: Using miRNA-96-5p and miRNA-99a-5p in combination with AFP increased both sensitivity and specificity for the diagnosis of HCC. Furthermore, serum levels were linked to tumor size and metastasis. These findings suggested that serum miRNA-96-5p and miRNA-99a-5p could be used as non-invasive biomarkers for the diagnosis of HCC.

Biogenic Synthesis of Ag Nanoparticles of 18.27 nm by Zanthozylum armatum and Determination of Biological Potentials.

Nanotechnology has become a dire need of the current era and the green synthesis of nanoparticles offers several advantages over other methods. Nanobiotechnology is an emerging field that contributes to many domains of human life, such as the formulation of nanoscale drug systems or nanomedicine for the diagnosis and treatment of diseases. Medicinal plants are the main sources of lead compounds, drug candidates and drugs. This work reports the green synthesis of Ag nanoparticles (AgNPs) using the aqueous bark extract of Zanthozylum armatum, which was confirmed by a UV absorption at 457 nm. XRD analysis revealed an average size of 18.27 nm and SEM showed the particles' spherical shape, with few irregularly shaped particles due to the aggregation of the AgNPs. FT-IR revealed the critical functional groups of phytochemicals which acted as reducing and stabilizing agents. The bark extract showed rich flavonoids (333 mg RE/g) and phenolic contents (82 mg GAE/g), which were plausibly responsible for its high antioxidant potency (IC50 = 14.61 µg/mL). Extract-loaded AgNPs exhibited the highest but equal inhibition against E. coli and P. aeruginosa (Z.I. 11.0 mm), whereas methanolic bark extract inhibited to a lesser extent, but equally to both pathogens (Z.I. 6.0 mm). The aqueous bark extract inhibited P. aeruginosa (Z.I. 9.0 mm) and (Z.I. 6.0 mm) E. coli. These findings-especially the biosynthesis of spherical AgNPs of 18.27 nm-provide promise for further investigation and for the development of commercializable biomedical products.

Heat flow saturate of Ag/MgO-water hybrid nanofluid in heated trigonal enclosure with rotate cylindrical cavity by using Galerkin finite element.

MHD Natural convection, which is one of the principal types of convective heat transfer in numerous research of heat exchangers and geothermal energy systems, as well as nanofluids and hybrid nanofluids. This work focuses on the investigation of Natural convective heat transfer evaluation inside a porous triangular cavity filled with silver-magnesium oxide/water hybrid nanofluid [H2O/Ag-MgO]hnf under a consistent magnetic field. The laminar and incompressible nanofluid flow is taken to account while Darcy-Forchheimer model takes account of the advection inertia effect in the porous sheet. Controlled equations of the work have been approached nondimensional and resolved by Galerkin finite element technique. The numerical analyses were carried out by varying the Darcy, Hartmann, and Rayleigh numbers, porosity, and characteristics of solid volume fraction and flow fields. Further, the findings are reported in streamlines, isotherms and Nusselt numbers. For this work, the parametric impact may be categorized into two groups. One of them has an effect on the structural factors such as triangular form and scale on the physical characteristics of the important outputs such as fluidity and thermal transfer rates. The significant findings are the parameters like Rayleigh and slightly supported by Hartmann along with Darcy number, minimally assists by solid-particle size and rotating factor as clockwise assists the cooler flow at the center and anticlockwise direction assists the warmer flow. Clear raise in heat transporting rate can be obtained for increasing solid-particle size.

Thyme Oil and Thymol Counter Doxorubicin-Induced Hepatotoxicity via Modulation of Inflammation, Apoptosis, and Oxidative Stress.

Doxorubicin (DOX) is an effective anticancer agent with a wide spectrum of activities. However, it has many adverse effects on various organs especially on the liver. Thymol, one of the major components of thyme oil, has biological properties that include anti-inflammatory and antioxidant activities. Thus, this study was designed to examine thyme oil and thymol for their ability to prevent doxorubicin-induced hepatotoxicity in Wistar rats. Hepatotoxicity was induced by an intraperitoneal injection of doxorubicin, at a dose of 2 mg/kg bw/week, for seven weeks. Doxorubicin-injected rats were supplemented with thyme oil and thymol at doses 250 and 100 mg/kg bw, respectively, four times/week by oral gavage for the same period. Treatment of rats with thyme oil and thymol reversed the high serum activities of AST, ALT, and ALP and total bilirubin, AFP, and CA19.9 levels, caused by doxorubicin. Thyme oil and thymol also reduced the high levels of TNF-α and the decreased levels of both albumin and IL-4. These agents ameliorated doxorubicin-induced elevation in hepatic lipid peroxidation and associated reduction in GSH content and GST and GPx activities. Further, the supplementation with thyme oil and thymol significantly augmented mRNA expression of the level of antiapoptotic protein Bcl-2 and significantly downregulated nuclear and cytoplasmic levels of the hepatic apoptotic mediator p53. Thus, thyme oil and thymol successfully counteracted doxorubicin-induced experimental hepatotoxicity via their anti-inflammatory, antioxidant, and antiapoptotic properties.

Prognostic Significance of p16 Protein Expression in Breast Cancer.

BACKGROUND/AIM: Breast cancer is the most common cancer in Sudan. The p16 protein plays a vital role in the regulation of the cell cycle. PATIENTS AND METHODS: This study analysed the protein expression of p16 in 202 paraffin blocks from Sudanese women with breast cancer using immunohistochemistry. RESULTS: This study included 168 (83.2%), 16 (7.9%), and 18 (8.9%) patients with invasive ductal carcinoma, invasive lobular carcinoma, and papillary carcinoma, respectively. There were 95 cases (47.0%) with grade III, 70 cases (34.6%) with grade II, and 23 cases (11.4%) with grade I breast cancer. The hormone receptor status was available for 119 of the cases, and 31 (15.3%), 25 (12.4%), and 63 (31.2%) cases were positive for oestrogen, progesterone, and HER2 receptors, respectively. CONCLUSION: p16 protein expression was associated with high histologic grade, lymph node metastasis, and poor prognosis. p16 protein expression may potentially be used as a prognostic marker.

Biomacromolecules of chitosan - Bacopa saponin based LipL32 gene delivery system for leptospirosis therapy.

Leptospirosis is a severe bacterial infectious disease caused by the organisms belonging to the genus of Leptospira. The chitosan/Bacopa saponin/tripolyphosphate (CS/BS/TPP) nanoparticles conjugated with recombinant DNA vaccines were designed against Leptospirosis. Chitosan, a polysaccharide is suitable for delivery of drug, and gene due to its bio-compatible and biodegradable properties. Bacopa saponins are used for the induction of the immune response against microbial infections. The recombinant DNA vaccine construct was composed of the leptospiral outer membrane LipL32 gene tagged with EGFP and hGMCSF adjuvant in the pVAX1 mammalian expression vector along with the Cytomegalovirus (CMV) promoter. These recombinant DNA vaccine constructs was termed as pVAX1-EGFP-LipL32 and pVAX1-EGFP-hGMCSF-LipL32, and these constructs were conjugated with CS/BS/TPP nanoparticles by using the ionic gelation technique. Thus, CS/BS/TPP conjugated nanoparticle DNA vaccine was confirmed by functionality (FT-IR), crystalline nature (XRD) and surface charge (Zeta potential). The 90% encapsulation efficiency was observed in the conjugated nanoparticle DNA vaccine. In contrast, cell viability analysis validated that the synthesized DNA conjugated CS/BS/TPP nanoparticles showed low cytotoxicity up to 10 mg/mL. The results showed here are the initial establishment of DNA vaccine conjugated nanoparticles, which can be used as a potential anti-leptospiral vaccine.

Efficacy of the Newcastle Disease Virus Genotype VII.1.1-Matched Vaccines in Commercial Broilers.

Class II genotype VII Newcastle disease viruses (NDV) are predominant in the Middle East and Asia despite intensive vaccination programs using conventional live and inactivated NDV vaccines. In this study, the protective efficacies of three commercial vaccine regimes involving genotype II NDV, recombinant genotype VII NDV-matched, and an autogenous velogenic NDV genotype VII vaccine were evaluated against challenge with velogenic NDV genotype VII (accession number MG029120). Three vaccination regimes were applied as follows: group-1 received inactivated genotype II, group-2 received inactivated recombinant genotype VII NDV-matched, and group-3 received velogenic inactivated autogenous NDV genotype VII vaccines given on day 7; for the live vaccine doses, each group received the same live genotype II vaccine. The birds in all of the groups were challenged with NDV genotype VII, which was applied on day 28. Protection by the three regimes was evaluated after infection based on mortality rate, clinical signs, gross lesions, virus shedding, seroconversion, and microscopic changes. The results showed that these three vaccination regimes partially protected commercial broilers (73%, 86%, 97%, respectively, vs. 8.6% in non-vaccinated challenged and 0% in non-vaccinated non-challenged birds) against mortality at 10 days post-challenge (dpc). Using inactivated vaccines significantly reduced the virus shedding at the level of the number of shedders and the amount of virus that was shed in all vaccinated groups (G1-3) compared to in the non-vaccinated group (G-4). In conclusion, using closely genotype-matched vaccines (NDV-GVII) provided higher protection than using vaccines that were not closely genotype-matched and non-genotype-matched. The vaccine seeds that were closely related to genotype VII.1.1 provided higher protection against challenge against this genotype since it circulates in the Middle East region. Updating vaccine seeds with recent and closely related isolates provides higher protection.

Microsporidia infection in patients with autoimmune diseases.

Purpose: Microsporidium is a spore-forming intracellular parasite that affects a wide range of hosts including humans. The tumor necrosis factor alpha (TNF-α) plays a key role in the immunity to infection with microsporidia. Recently, the TNF-α antagonists have proven successful in treating variable autoimmune diseases. In the current study, we aimed to investigate the impact of using TNF-α antagonists as a therapeutic regimen in the prevalence of infections with microsporidia. Materials and Methods: Diarrheal patients with distinct autoimmune diseases (n = 100) were assigned to the study. Patients taking anti-TNF-α medications (n = 60) were allocated to Group 1A and those undergoing non-TNF-α inhibitor treatment (n = 40) to Group 1B. Furthermore, patients with diarrhea without autoimmune disorders (n = 20) were allocated as controls. Stool specimens, 3 per patient, were collected and microscopically examined for microsporidia spores. A microsporidia-specific stool polymerase chain reaction was used to confirm the microscopic findings. Results: Microsporidia infection was identified in 28.3% (17/60), 10% (4/40), and in 5% (1/20) of patients in Group 1A, Group 1B, and in the control group, respectively. Overall, infection was significantly high in cases compared to the controls and in patients receiving TNF-α antagonists compared to patients not given TNF-α inhibitors (P < 0.05). Finally, infection was significantly higher in cases treated with TNF-α antagonists for ≥2 months compared to cases treated for <2 months of duration (P < 0.05). Conclusion: There was a significant increase in microsporidia infection in autoimmune disease patients undergoing treatment with TNF-α antagonists, and the duration of treatment is one of the risk factors. The study highlights the importance of microsporidia testing in immunocompromised patients, particularly those undergoing treatment with anti-TNF-α drugs and emphasises the need for awareness among clinicians regarding this opportunistic parasite.

Prevalence, toxin gene profile, genotypes and antibiotic susceptibility of Clostridium difficile in a tertiary care hospital in Taif, Saudi Arabia.

Purpose: Clostridium difficile (C. difficile) is an important causative agent of nosocomial diarrhoea and has become a major worldwide public health concern. The current study was conducted to determine the prevalence of C. difficile infection (CDI) amongst patients with nosocomial diarrhoea in a large tertiary care hospital in Taif, Saudi Arabia, and to define molecular characteristics and antimicrobial sensitivity profiles of C. difficile strains isolated from those patients. Materials and Methods: Stool specimens were collected from 456 patients and were cultured for C. difficile isolation. The isolates were subjected to multiplex polymerase chain reaction (PCR) for detecting genes encoding the toxins (toxin A, toxin B and binary toxin [CDT]), genotyping by PCR ribotyping method and antimicrobial sensitivity testing using E test strips. Results: Seventy-four C. difficile strains were recovered, of which 44 (59.5%) were A+B+CDT-, 14 (18.9%) were A-B+CDT-, 4 (5.4%) were A+B+CDT+ and 12 (16.2%) were A-B-CDT-. Toxigenic strains, and hence CDI, were detected in 13.6% of the patients (62/456). Fourteen different ribotypes were distinguished amongst bacterial isolates, of which ribotypes 002, 001, 017, 014 and 020 were the most prevalent (20.3%, 18.9%, 18.9%, 9.5% and 8.1%, respectively). Four isolates (5.4%) belonged to ribotype 027. All bacterial isolates showed sensitivity to metronidazole, vancomycin and piperacillin-tazobactam. The isolates exhibited resistance to linezolid (2.7%), chloramphenicol (5.4%), rifampicin (13.5%), tetracycline (21.6%), moxifloxacin (48.6%), clindamycin (54%) and imipenem (83.8%). Multiple drug resistance was observed in 56.8% of the isolates. Conclusion: Further larger studies are required for an accurate understanding of CDI epidemiology in Saudi Arabia.