Mutations in Plasmodium knowlesi Kelch protein 13 and the dihydropteroate synthase gene in clinical samples

Objective:To determine the genetic diversity,natural selection and mutations in Plasmodium(P.)knowlesi drug resistant molecular markers Kelch 13 and dhps gene in clinical samples of Malaysia.Methods:P.knowlesi full-length gene sequences Kelch 13 gene(PkK13)from 40 samples and dhps gene from 30 samples originating from Malaysian Borneo were retrieved from public databases.Genetic diversity,natural selection,and phylogenetic analysis of gene sequences were analysed using DNAsp v5.10 and MEGA v5.2.Results:Seventy-two single nucleotide polymorphic sites(SNPs)across the full-length PkK13 gene(63 synonymous substitutions and 9 non-synonymous substitutions)with nucleotide diversity ofπ~0.005 was observed.Analysis of the full-length Pkdhps gene revealed 73 SNPs andπ~0.006(44 synonymous substitutions and 29 non-synonymous substitutions).A high number of haplotypes(PkK13;H=37 and Pkdhps;H=29)with haplotype diversity of Hd~0.99 were found in both genes,indicating population expansion.Nine mutant alleles were identified in PkK13 amino acid alignment of which,7(Asp3Glu,Lys50Gln,Lys53Glu,Ser123Thr,Ser127Pro,Ser149Thr and Ala169Thr)were within the Plasmodium specific domain,2(Val372Ile and Lys424Asn)were in the BTB/POZ domain and no mutation was observed within the kelch propeller domain.The 29 non-synonymous mutations in the Pkdhps gene were novel and only presented in exon 1 and 2.Conclusions:Monitoring the mutations from clinical samples collected from all states of Malaysia along with clinical efficacy studies will be necessary to determine the drug resistance in P.knowlesi.

Development of micro/nanostructured‒based biomaterials with biomedical applications

Natural biomaterials are now frequently used to build biocarrier systems,which can carry medications and biomolecules to a target region and achieve a desired therapeutic effect.Biomaterials and polymers are of great importance in the synthesis of nanomaterials.The recent studies have tended to use these materials because they are easily obtained from natural sources such as fungi,algae,bacteria,and medicinal plants.They are also biodegradable,compatible with neighborhoods,and non-toxic.Natural biomaterials and polymers are chemically changed when they are linked by cross linking agents with other polymers to create scaffolds,matrices,composites,and interpenetrating polymer networks employing microtechnology and nanotechnology.This review highlights how microengineered and nanoengineered biomaterials are utilized to produce efficient drug-delivery systems and biomedical and biological therapies and how innovative sources of biomaterials have been identified.

The potency of N,N′-diphenyl-1,4-phenylenediamine and adiposederived stem cell co-administration in alleviating hepatorenal dysfunction complications associated with type 1 diabetes mellitus in rats

Background:The increasing occurrence of diabetes mellitus(DM)noted worldwide has considerably elicited concern in the recent past.DM is associated with elevated vascular complications,morbidity,mortality,and poor quality of life.In this context,mesenchymal stem cells(MSCs)have shown significant therapeutic potentialities in managing and curing type 1 DM owing to their self-renewable,immunosuppressive,and differentiation capacities.We investigated the potential action of N,N′-diphenyl-1,4-phenylenediamine(DPPD),a well-known synthetic antioxidant to enhance the therapeutic ability of the adipose-derived stem cells(AD-MSCs)in alleviating kidney and liver complications in diabetic rats.Methods:Over the four weeks of experiments,albino male rats(n=36)were split into six test groups:control,DPPD(250 mg/kg,i.p.),STZ-diabetic(D),D+DPPD,D+AD-MSCs(1×10^(6)cell/rat,i.v.),and D+AD-MSCs+DPPD treated groups.Results:Significant declines in the renal and hepatic oxidative stress markers(MDA,ROS,and AGEs)were observed coupled with a significant elevation in many antioxidant marker levels(GSH,SOD,CAT,GPx,HO-1,and TAC)in the diabetic rats treated with either DPPD or AD-MSCs or their co-administered injection compared to the diabetic untreated rats.This was suggested to be the leading cause of amelioration of the kidney functions(as measured by urea,uric acid,and creatine levels)and liver functions(as evidenced by the levels of AST,ALT,ALP,bilirubin,total proteins,albumin,and globulins).Conclusion:DPPD and AD-MSCs co-administration showed superior results in terms of the enhancement of the relative hepato-renal function,indicating the beneficial role of DPPD supplementation in increasing the therapeutic potential of AD-MSCs.

High-throughput computational screening and in vitro evaluation identifies 5-(4-oxo-4H-3,1-benzoxazin-2-yl)-2-[3-(4-oxo-4H-3,1-benzoxazin-2-yl)phenyl]-1H-isoindole-1,3(2H)-dione(C3),as a novel EGFR—HER2 dual inhibitor in gastric tumors

Gastric cancers are caused primarily due to the activation and amplification of the EGFR or HER2 kinases resulting in cell proliferation,adhesion,angiogenesis,and metastasis.Conventional therapies are ineffective due to the intra-tumoral heterogeneity and concomitant genetic mutations.Hence,dual inhibition strategies are recommended to increase potency and reduce cytotoxicity.In this study,we have conducted computational high-throughput screening of the ChemBridge library followed by in vitro assays and identified novel selective inhibitors that have a dual impediment of EGFR/HER2 kinase activities.Diversity-based High-throughput Virtual Screening(D-HTVS)was used to screen the whole ChemBridge small molecular library against EGFR and HER2.The atomistic molecular dynamic simulation was conducted to understand the dynamics and stability of the protein-ligand complexes.EGFR/HER2 kinase enzymes,KATOIII,and Snu-5 cells were used for in vitro validations.The atomistic Molecular Dynamics simulations followed by solvent-based Gibbs binding free energy calculation of top molecules,identified compound C3(5-(4-oxo-4H-3,1-benzoxazin-2-yl)-2-[3-(4-oxo-4H-3,1-benzoxazin-2-yl)phenyl]-1H-isoindole-1,3(2H)-dione)to have a good affinity for both EGFR and HER2.The predicted compound,C3,was promising with better binding energy,good binding pose,and optimum interactions with the EGFR and HER2 residues.C3 inhibited EGFR and HER2 kinases with IC50 values of 37.24 and 45.83 nM,respectively.The GI50 values of C3 to inhibit KATOIII and Snu-5 cells were 84.76 and 48.26 nM,respectively.Based on these findings,we conclude that the identified compound C3 showed a conceivable dual inhibitory activity on EGFR/HER2 kinase,and therefore can be considered as a plausible lead-like molecule for treating gastric cancers with minimal side effects,though testing in higher models with pharmacokinetic approach is required.

Effect of Lonicerae Flos extracts on reflux esophagitis with antioxidant activity

AIM:To observe the effects of traditional anti-inflammatory medicine Lonicerae Flos (LF) on rat reflux esophagitis (RE) induced by pylorus and forestomach ligation compared with the well-known proton antioxidant, α-tocopherol.METHODS:Rats were pretreated with three different dosages of LF (500, 250 and 125 mg/kg) orally, once a day for 14 d before pylorus and forestomach ligation. Nine hours after pylorus and forestomach ligation, changes to the stomach and esophagus lesion areas, gastric volumes, acid and pepsin outputs, antioxidant effects, esophageal lipid peroxidation, superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), myeloperoxidase and glutathione (GSH) levels, and collagen contents (marker of flexibility) were observed on the esophageal and fundic histopathology. The results were compared with an α-tocopherol (once orally, 1 h before operation, 30 mg/kg) treated group in which the effects on RE were already conf irmed.RESULTS: Pylorus and forestomach ligations caused marked increases of gross esophageal and gastric mucosa lesion areas, which corresponded with histopathological changes. In addition, increases of esophageal lipid peroxidation, decreases of SOD, CAT, and GSH-free radical scavengers, increases of collagen were observed. However, these pylorus and forestomach ligation induced RE were dose-dependently inhibited by treatment of 500, 250 and 125 mg/kg of LF extract, mediated by antioxidant effects. RE at 250 mg/kg showed similar effects α-tocopherol.CONCLUSION: The results suggest that antioxidant effects of LF could attenuate the severity of RE and prevent the esophageal mucosal damage, and validate its therapeutic use in esophageal reflux disease.

Epigenetic states and expression of imprinted genes in human embryonic stem cells

AIM: To investigate the epigenetic states and expres- sion of imprinted genes in five human embryonic stem cell (hESC) lines derived in Taiwan. METHODS: The heterozygous alleles of single nucleo- tide polymorphisms (SNPs) at imprinted genes were analyzed by sequencing genomic DNAs of hESC lines and the monoallelic expression of the imprinted genes were confirmed by sequencing the cDNAs. The expres- sion profiles of 32 known imprinted genes of five hESC lines were determined using Affymetrix human genome U133 plus 2.0 DNA microarray. RESULTS: The heterozygous alleles of SNPs at seven imprinted genes, IPW , PEG10 , NESP55 , KCNQ1 , ATP10A ,TCEB3C and IGF2 , were identified and the monoallelic expression of these imprinted genes except IGF2 were confirmed. The IGF2 gene was found to be imprinted in hESC line T2 but partially imprinted in line T3 and not imprinted in line T4 embryoid bodies. Ten imprinted genes, namely GRB10 , PEG10 , SGCE, MEST , SDHD , SN- RPN , SNURF , NDN , IPW and NESP55 , were found to be highly expressed in the undifferentiated hESC lines and down-regulated in differentiated derivatives. The UBE3A gene abundantly expressed in undifferentiated hESC lines and further up-regulated in differentiated tissues. The expression levels of other 21 imprinted genes were relatively low in undifferentiated hESC lines and five of these genes (TP73 , COPG2 , OSBPL5 , IGF2 and ATP10A ) were found to be up-regulated in differentiated tissues. CONCLUSION: The epigenetic states and expression of imprinted genes in hESC lines should be thoroughly studied after extended culture and upon differentiation in order to understand epigenetic stability in hESC lines before their clinical applications.

Prevalence and antimicrobial resistance of Salmonella spp.in raw retail frozen imported freshwater fish to Eastern Province of Saudi Arabia

Objective:To determine the proportion of imported frozen fish contaminated with Salmonella among retail food stores and supermarkets in the Eastern Province of Saudi Arabia.Methods:A total of 223 frozen freshwater fish purchased from different supermarkets and grocery stores were analyzed for the presence of foodborne pathogen Salmonella.The isolation of Salmonella was determined and confirmed by using the methods of US Food and Drug Administration's Bacteriological Analytical Manual.CHROMagar Salmonella plus,biochemical tests and API 20E strips.Antimicrobial susceptibilities of Salmonella isolates were determined by the disk diffusion method on Muller-Hinton agar,as described by Kirby-Bauer.in accordance with the guidelines of the Clinical and Laboratory Standards Institute.Results:Out of the total 223 fish samples(20 of catfish,18 of carfu,20 of mirgal,25 of milkfish,35 of mackerel,75 of tilapia,and 30 of rohu),89(39.9%)were tested positive for Salmonella.The prevalence of positive samples were reported for the freshwater fish of pangas(60.0%,n=12),carfu(27.7%,n=5),mirgal(35.0%,n=7),milkfish(52.0%,n=13),mackerel(31.4%,n=11),tilapia imported from Thailand(64.0%,n=16),tilapia imported from India(28.0%,n=14),rohu imported from Thailand(26.6%,n=4)and rohu imported from Myanmar(46.6%,n=7).A total of 140 isolates of Salmonella spp.were yielded from at least seven different types of frozen freshwater fish imported from 5 different countries and were tested for their susceptibility to 16 selected antimicrobial agents.The highest antibiotic resistance was observed to tetracycline(90.71%)followed by ampicillin(70%)and amoxicillin-clavulanic acid(45%).Conclusions:The obtained results of this study shows that these raw retail imported frozen freshwater fish are contaminated with potentially pathogenic Salmonella spp.And the study recommend and suggest that there is a need for adequate consumer measures.

Synthesis and biological evaluation of ^(18)F-FB-NGA as a hepatic asialoglycoprotein receptor PET imaging agent

Asialoglycoprotein receptor(ASGP-R)is a hepatic membrane receptor that uniquely exists on the surface of mammalian hepatocytes,and has been used as target of liver functional imaging agents for many years.We labeled the Galactosyl-neoglycoalbumin(NGA)with 18F to get a PET molecular probe 18F-FB-NGA and evaluated its ability as a liver functional PET imaging agent.The 18F-FB-NGA was prepared with NGA by conjugation with Nsuccinimidyl-4-18F-fluorobenzoate(18F-SFB)and purified with PD-10 desalting column.The radiolabeling yield and radiochemical purity of 18F-FB-NGA were determined by radio-HPLC.Starting with 18F-F–,the total time for 18F-FB-NGA was about 120±10 min.The decay-corrected radiochemical yield is about 25–30%.The radiochemical purity of purified 18F-FB-NGA was more than 98%.Labeled with 185–1850 MBq 18F-SFB,the specific activity of 18F-FBNGA was estimated to be 7.83–78.3 TBq/mmol.Biodistribution of 18F-FB-NGA in normal mice was investigated after injection through the tail vein.The results showed that the liver accumulated 39.47±3.42 and 12.12±6.11%ID/g at 10 and 30 min after injection,respectively.Dynamic MicroPET images in mice were acquired with and without block after injection of the radiotracer,respectively.High liver activity accumulation was observed at 5 min after injection in normal group.On the contrary,the liver accumulation was significantly lower after block,indicating the specific binding to ASGP-R.18F-FB-NGA is probably a potential PET liver imaging agent.

Prevalence of intestinal parasite infections among patients in local public hospitals of Hail,Northwestern Saudi Arabia

Objective:To evaluate the prevalence of intestinal parasites among patients in Hail,Northwestern Saudi Arabia.Methods:Stool samples were collected from 130 patients(69females and 61 males) in Hail General Hospital.Each sample was examined by direct wet mount microscopic examination using both normal saline and Lugol's iodine preparation and concentration techniques using salt and formol-ether solutions.Permanent stained smears were performed for intestinal coccidian using modified Ziehl-Neelsen technique.Results:The overall prevalence of intestinal parasitic infection was 45.38%(59 cases).Forty-four(33.84%)were found to be infected with one or more intestinal protozoa.5(3.84%) were infected with helminthes and 10(7.69%) had mixed infection with both helminthes and protozoa.The most common intestinal helminth detected was Ancyloxtoma duvdenale(n=5.3.84%),followed by Ascaris lumlmcnidex.Taenia sp.and Trichuris trichiura(n=2 for each species,1.5%).For intestinal protozoa,the coccidian Cryptosporidium parvum(n=25,19.23%) was the most common followed by Entamoeba histolytica/dispur(n=21,16.15%),Giurdia lamblia(n=15,11.54%),Entamoeba coli(n=5.3.85%) and Blastocysts hominis(n=3,2.30%).The prevalence of intestinal parasitic infections in females was significantly higher than in males(P<0.05).Conclusions:This is the first study highlighting that intestinal parasites are still an important public health problem in Northwestern Saudi Arabia Therefore,health education would be the best way to prevent from intestinal parasite infections which are mainly food borne diseases.

Prediction of COVID-19 Cases Using Machine Learning for Effective Public Health Management

COVID-19 is a pandemic that has affected nearly every country in the world.At present,sustainable development in the area of public health is considered vital to securing a promising and prosperous future for humans.However,widespread diseases,such as COVID-19,create numerous challenges to this goal,and some of those challenges are not yet defined.In this study,a Shallow Single-Layer Perceptron Neural Network(SSLPNN)and Gaussian Process Regression(GPR)model were used for the classification and prediction of confirmed COVID-19 cases in five geographically distributed regions of Asia with diverse settings and environmental conditions:namely,China,South Korea,Japan,Saudi Arabia,and Pakistan.Significant environmental and non-environmental features were taken as the input dataset,and confirmed COVID-19 cases were taken as the output dataset.A correlation analysis was done to identify patterns in the cases related to fluctuations in the associated variables.The results of this study established that the population and air quality index of a region had a statistically significant influence on the cases.However,age and the human development index had a negative influence on the cases.The proposed SSLPNN-based classification model performed well when predicting the classes of confirmed cases.During training,the binary classification model was highly accurate,with a Root Mean Square Error(RMSE)of 0.91.Likewise,the results of the regression analysis using the GPR technique with Matern 5/2 were highly accurate(RMSE=0.95239)when predicting the number of confirmed COVID-19 cases in an area.However,dynamic management has occupied a core place in studies on the sustainable development of public health but dynamic management depends on proactive strategies based on statistically verified approaches,like Artificial Intelligence(AI).In this study,an SSLPNN model has been trained to fit public health associated data into an appropriate class,allowing GPR to predict the number of confirmed COVID-19 cases in an area based on the given values of selected parameters. Therefore, this tool can help authorities in different ecological settingseffectively manage COVID-19.

Hepcidin-orchestrated Hemogram and Iron Homeostatic Patterns in Two Models of Subchronic Hepatic injury

Objective This study was designed to evaluate hematological disorders and the orchestrating roles of hepcidin and IL-6 in rat models of thioacetamide(TAA) and carbon tetrachloride(CCl_4) hepatotoxicity. Methods Rats were intraperitoneally injected with TAA(10 mg/100 g rat weight dissolved in isosaline) or CCl_4(100 μL/100 g rat weight diluted as 1:4 in corn oil) twice weekly for eight consecutive weeks to induce subchronic liver fibrosis. Blood and tissue samples were collected and analyzed. Results CCl_4 but not TAA significantly decreased the RBCs, Hb, PCV, and MCV values with minimal alterations in other erythrocytic indices. Both hepatotoxins showed leukocytosis, granulocytosis, and thrombocytopenia. By the end of the experiment, the erythropoietin level increased in the CCl_4 model. The serum iron, UIBC, TIBC, transferrin saturation%, and serum transferrin concentration values significantly decreased, whereas that of ferritin increased in the CCl_4 model. TAA increased the iron parameters toward iron overload. RT-PCR analysis revealed increased expression of hepatic hepcidin and IL-6 m RNAs in the CCl_4 model and suppressed hepcidin expression without significant effect on IL-6 in the TAA model. Conclusion These data suggest differences driven by hepcidin and IL-6 expression between CCl_4 and TAA liver fibrosis models and are of clinical importance for diagnosis and therapeutics of liver diseases.

Evaluation of NFC-Guidable System to Manage Polypharmacy in Elderly Patients

A complete and thorough understanding by patients of their prescriptions is one of the most critical components of a successful treatment journey.Being unfamiliar with the intricacies of prescribed medication can cause serious health risks due to not adhering to prescription instructions or noting potential drug interactions,which can lead to life-threatening injuries.Pharmacists face communication barriers(including non-English speaking patients),lack of time,lack of knowledge,workload,and frequent interruption when dispensing medicines often preventing them from providing the necessary guidance to their patients.To minimize this risk,an NFC-Guidable polypharmacy system was developed integrating Near Field Technology(NFC)into prescription packages.The intention being,to automate prescription identification for elderly patients in hospitals and primary health care(PHC)centers where Arabic is their native language and patients have five or more prescriptions at a time.The system provides prescription information such as name,expiration date based on the Islamic calendar,appropriate dosage,the correct time for a dose,contraindication,and other relevant instructions in Arabic.The objectives of the system are to improve patient safety and enhance the quality of prescription dispensing for elderly and non-English speaking patients.An evaluation study with polypharmacy elderly patients and pharmacists is conducted to understand the perception and acceptance of the system by both groups.The strengths and weaknesses of the system as well as suggestions for improvement from participants were also gathered.The study results indicate that the system helps patients to avoid misuse of sensitive medication and authenticates their prescribed prescription drugs before taking them.The system also assists pharmacists in improving the efficiency of dispensing medication and streamline processes.Overall,the application was well received by both patients and pharmacists.Thus,NFC technology should be integrated into the healthcare sector to overcome language barriers,the lack of specialized medication labels for illiterate or patients with sight constraints as well as unreasonable expectations placed on pharmacists.

Effect of <i>Ginkgo biloba</i>Extract Ingestion on Plasma Total Cortisol Levels during an Oral Glucose Tolerance Test in Normal Glucose Tolerant Individuals

Protracted periods of increased cortisol production, as may be seen in an acute illness, may lead to transient hyperglycemia. Increasing evidence suggests that cortisol may then mediate the development of insulin resistance and potentially lead to the development of overt diabetes. Evidence in animal studies also suggests that under conditions of stress Ginkgo biloba extract could reduce plasma cortisol production and so the primary aim of this study was to determine the effect of Ginkgo biloba extract ingestion on plasma cortisol production during an acute period of glucose challenge. Healthy non-diabetic, glucose tolerant volunteers (n = 30, (10/20, M/F);age, 45.7 ± 9.9 years old) completed a randomized, double-blind, placebo-controlled crossover study when they ingested Ginkgo biloba extract (120 mg/day as a single dose) and placebo during each 3-month arm. A standard 75 g oral glucose tolerance test was performed at the end of each cycle and blood was collected and used to measure plasma glucose, insulin, c-peptide and cortisol. Fasting plasma cortisol was significantly lower after the Ginkgo biloba cycle than the placebo cycle (326 ± 149 vs. 268 ± 121 nmol/L, respectively;p = 019). The plasma cortisol area under the curve during the 2-hour test (AUC0-2) was also significantly lower after ingestion of the Ginkgo biloba cycle compared to the placebo (668 ± 265 vs. 530 ± 213 nmol/L/h, respectively;p < 0.001). It is concluded that the ingestion of Ginkgo biloba extract has effect on the hypothalamic-pituitary-adrenal axis leading to reduced basal cortisol levels and reduced cortisol production in response to acute hyperglycemic challenge.

Assessment of Genetic and Immunological Factors Associated with Hepatitis C Infection in Saudi Hemodialysis Patients

The association of human leukocyte antigen (HLA) genotype and hepatitis C virus (HCV) infection has been reported in many populations. In addition, the HLA genotype distribution has been found to differ according to ethnicity. Currently, there are no published data concerning this relationship in the Saudi population. Therefore, the primary objective of this study was to determine the relationship between HLA genotype and HCV infection in a group of haemodialysis patients in a tertiary hospital in the Eastern Province of Saudi Arabia. A total of 152 haemodialysis patients and 160 controls were enrolled in the study. The patient group included 91 males and 61 females with a mean age of 48 years, who had been on haemodialysis for a period of 6 months to five years. The control group was randomly selected from donors attending the blood bank and included 155 males and five females with a mean age of 38 years. Of the patient group, 27 (17.8%) were found to have HCV antibodies by EIA. The seropositivity was confirmed by RIBA and by RT-PCR. The average viral load was 15 × 103 HCV RNA per 10 μl of plasma. The predominant HCV genotype was found to be genotype 4. HLA-class I and HLA class II were screened in 27 HCV-positive patients and in 30 age and sex matched controls by polymerase chain reaction/sequence specific primers (PCR/SSP) method. Measurements of cytokines Interleukin-2, 10, IFN-γ and TNFα showed that IL-2 level was 17.65 - 20.3 pg/l;IFN-γ was 22.15 - 41.1 pg/l;TNF-α was 2.5 - 5.6 pg/mL and IL-10 was 10.7 - 14.8 pg/mL. Measurement of CD markers showed the following: CD3 cells were measured. There was no correlation between infection with HCV and HLA type, cytokines levels and levels of different cell populations. CD markers are similar in the HCV-positive and HCV-negative hemodialysis patients. Further analysis and comparisons are needed, especially between the cytokines and CD markers in the hemodialysis patients and control groups.

Awareness of Folic Acid Intake among Women in the Childbearing Age in Hail Region—Saudi Arabia

This study evaluated the awareness of folic acid, timing of folic acid supplementation and source of knowledge of folic acid intake in Hail region-Saudi Arabia. A sample of 300 married women in the childbearing age (19 - 45 years) in Hail in the period between October/2011 and January/2012 participated in a survey study regarding awareness and use of folic acid before and during pregnancy. Participant women were asked to respond to different questions related to age, level of education, occupation, monthly income, gravidity, and other information concerning folic acid awareness, specific knowledge, supplementation practices, and source of knowledge about the preconceptional intake of folic acid. While 91.0% of the subjects were aware of folic acid, 81.0% knew that folic acid could prevent neural tube defects and 84.0% of the subjects took folic acid prior or during a certain stage of pregnancy. Nonetheless, only 10.0% of the subjects who proclaimed that they know the proper timing for folic acid supplementation recognized that folic acid should be taken prior and during the first trimester of pregnancy. University education was the strongest predictor of folic acid knowledge and university educated subjects had significantly (P P

Histochemical Patterns of Collagenic Fibers in the Benign and Malignant Breast Lesions

Cancer is one of the leading causes of death worldwide and breast cancer is the most commonly diagnosed cancer among women. An increased incidence of different types of breast cancer has been reported. This study was designed to evaluate the different types of breast cancer and its possible risk of neoplasmic transformation to an advanced malignant stage from a benign tumor. The histochemical patterns of collagen fibers in the benign and malignant breast lesions were evaluated. From the 50 tissue samples, 25 were malignant breast lesions and 25 were benign breast tumor. Hematoxylin and Eosin (HE), Van Gieson staining were performed to detect a benign and malignant tumor as well as collagen fibers. We found that significant cases after age of 35 were associated with ductal carcinoma while most of the cases within the age of 25 years were associated with fibrocystic changes. The intensity of collagen fiber was higher to Ductal Carcinoma while negative and less intense for Fibroblastic changes. Furthermore, a consistent association of other lesions, such as Lobular Carcinoma, Fibroadenoma, Papilloma and Fat necrosis and noticeable staining for collagen was observed for the different lesion. Our study suggested that women with age of 25 with benign lesion of fibrocystic change and ductal carcinoma are highly susceptible to develop advanced malignant tumor with age. Therefore, quantitative measurement of collagen fiber and regular follow-up are recommended to avoid the possible risk of developing advanced malignant lesions.

Studying the Importance of <i>VacA</i>Gene of <i>Helicobacter pylori</i>in Identifying the Pathogenicity of Strains by Comparing It with the Disease Status of the Subjects

Introduction: H. pylori is a primary pathogen isolated by Warren and Marshall in 1983. They called it as Campylobacter Pylori and in 1989 Goodwin et al. renamed it as Helicobacter pylori. Helicobacter pylorus is one of the responsible factors for causing gastritis, Peptic ulcer disease and is strongly associated with gastric carcinoma and gastric Malt lymphoma. Materials and Methods: In this study 300 biopsies were collected at Deccan College of Medical Sciences and Allied Hospitals, Hyderabad, India. Of these, 101 patients had peptic ulcer, 95 patients had gastritis and 4 had gastric carcinoma. A total of four gastric biopsy specimens were collected. One was used for culturing H. pylori, one for histological lesions and the remaining two one each from the antrum & corpus was collected in phosphate buffered saline for the DNA analysis. Results: Helicobacter pylori were isolated from many of the biopsies and the identification of Helicobacter pylori was confirmed in 200 biopsies with colony characteristics, Biochemical tests, and 16S rRNA amplification. The presence of VacA marker was detected by using appropriate primers. From the data obtained in our study, in a total of 169/200 isolates s1 region was found in 84.1% and s2 was found in 36/200 i.e. 18% of the isolates. In the middle region, m1awas found in 60/200 which was 30%, m1b was found in 60/200 i.e. 60% and m2 was found in 114/200 i.e. 57.1% of the isolates. Conclusion: Thus, VacA detection might be helpful for determination of which patients are at highest risk for severe clinical outcomes such as duodenal ulcer, gastric ulcer or gastric carcinoma and eventually, to define strategies for the treatment or prevention of H. pylori infection.

Differentiation of neuronal cells using a murine embryonic stem cell-based method

The differentiation and screening methodology proposed here is an efficient in vitro system to screen and study effects of small molecules and bioagents and is an alternative to studies that use live animals and embryos. The method is based on engineering a stable murine embryonic stem (ES) cell line expressing lineage-specific promoters that drive selection and reporter genes. Additionally, uniform embryoid bodies (EBs) are used for differentiation studies that allow synchronous differentiation. The reporter and selection marker genes are expressed only in lineages where the promoter is functional. The differentiated cell type can be identified by reporter gene expression and the selection marker can be used for selective enrichment of that particular cell population. The method described here is useful in screening small molecules or bioagents that can differentiate stem cells into particular lineages or cell types. Identified compounds are useful in areas such as stem cell-based regenerative medicine and therapeutics. The method described here has been applied to neuronal cell differentiation.

The Possibilities of Chronic Renal Failure Patients Contracting Occult Hepatitis B Virus Infection, Sudan

Introduction: Chronic kidney disease (CKD) is a major cause of death in sub-Saharan Africa. The effects of the CKD on the host and the continuous therapeutic measures increase the hypothesis of blood-borne diseases transmission. Objective: This study aimed to find the frequency of occult hepatitis B virus (OBI) in patients of chronic renal failure (CRF) and to study the possibilities of infection acquisition. Methods: During 2017 and 2019, two hundred CRF patients under regular haemodialysis and attending Gezira Hospital for Renal Diseases and Surgery were recruited. Plasma specimens were collected and used for detection of hepatitis B surface antigens (HbsAg), total hepatitis B core antibodies (anti-HBc) and hepatitis B virus DNA isolation. Nested PCR reaction was followed to identify HBV. Socio-clinical data for each participant was obtained. Results: Male patients represented 64% (128/200), most frequent age group was from 41 to 60 years with percentage of 56.5% (113/200), 86% (172/200) of CRF patients were received blood while 42% (84/200) get HBV vaccination. Hepatitis B core antibodies were found in 54% (108/200) of studied cases, and 22% (42/188) of tested DNA were positively amplified for target gene. Detection of Hepatitis B core antibodies was significantly associated with marital status while absence of vaccination significantly associated with the detection of both hepatitis B core antibodies and HBV DNA. Conclusion: This study found high frequency of OBI in CRF patients, to reduce the transmission of the disease, possible hypotheses should be studied, including blood transfusion, haemodialysis process and HBV vaccination status.

Sero-Prevalence of Hepatitis B Virus Infection among Blood Donors: A Retrospective Study in the Kintampo Municipal Hospital, Ghana

Background: Hepatitis B virus is one of the transfusion transmission infectious agents of public health relevance. Its prevalence varies across the globe. Establishing the sero-prevalence of the disease is critical to informing the direction of preventive and control strategies. Objective: The aim of this study therefore was to establish the hepatitis B surface antigen sero-prevalence among blood donors in the Kintampo municipality of Ghana. Methodology: This three-year hospital based retrospective study was conducted at the laboratory unit of the Kintampo Municipal Hospital. The laboratory uses one step immunochromatographic test kits to detect the presence of hepatitis B surface antigen in the sera of blood donors. Records on blood donors were reviewed for hepatitis B sero-positivity. Results: A total of 3402 blood donors were studied, out of which 3238 (95.2%) were males and 164 (4.8%) were females. The overall sero-prevalence of hepatitis B surface antigen was 9.6% (327/3402). Majority of the sero-positive donors were less than 40 years, with higher prevalence of 16.4% (35/214) in donors less than 20 years. The sero-prevalence in males and females was 9.7% (313/3238) and 8.5% (14/164) respectively. Conclusion: The Kintampo municipality has a relatively high prevalence of hepatitis B among blood donors. The probability of hepatitis B viral infection was age inclined as the youth seem to be at greater risk of contracting the disease.