International Workshop on Molecular Biology Techniques Related to Infectious/Genetic Diseases & Human Identification

Permanent URI for this collectionhttp://repository.kln.ac.lk/handle/123456789/8773

Browse

Search Results

Now showing 1 - 10 of 22
  • Item
    Drug discovery research, Sudan
    (Faculty of Medicine, University of Kelaniya, 2015) Mustafa, S.A.
    Introduction: Malaria and tuberculosis are important diseases in Sudan. Therefore, discovery of drugs for these diseases is very important to control these diseases. Methodology: Lead compounds for the treatment of many infectious and endemic diseases such as malaria and tuberculosis were prepared. Diversed compounds from natural and synthetic sources against drug targets related to the causative agents of these diseases were discovered. Broad biochemical and molecular biology techniques such as gene expression and protein purification, enzyme inhibition and kinetics studies, lead optimization and computational chemistry were used. Results: Our research has resulted in a battery of promising drug leads against many infectious and endemic diseases. Discussion: Promising drugs are needed extensive field trials.
  • Item
    Single nucleotide polymorphisms of ESR1 gene and the risk of breast cancer in Sudanese women
    (Faculty of Medicine, University of Kelaniya, 2015) Adam, K.M.
    Introduction: The prolonged exposure to estrogen is well established risk factor for breast cancer which is the most common malignancy among Sudanese women. The effect of estrogen hormone on target tissues is mediated by its alpha receptor, through the binding of the hormone to the receptor which in turn promote the proliferation and differentiation of mammary tissues. The estrogen receptor alpha is coded by polymorphic ESR1 gene. Objective: To evaluate the association between rs2234693, rs9340799 and rs1801132 single nucleotide polymorphisms on ESR1 gene and breast cancer risk, type and the receptor status. Methodology: Three single nucleotide polymorphisms were genotyped in 139 breast cancer cases and 139 age-matched cancer-free controls. Leucocytes DNA was extracted and genotyped by PCR-RFLP method. The estrogen receptor status for 65 cases were determined on a freshly obtained breast tissues according to standard histological procedures. Results: The results of breast cancer cases showed a young mean age 46.5±10.4SD years, high frequencies of married women 97.1%, and parity 89.9%. The family history of breast or other type of female cancers in Sudanese women, proved to be the strongest risk factor, with 12 folds increase in the risk of developing breast cancer (OR=11.8, 95% CI:4-34.2, P-0). Of all the three studied SNPs, rs9340799 (XbaI) was the only one to show statistically significant association with the risk of breast cancer (P-0.03), where unadjusted logistic regression of the genotypic variants of this SNP showed that Xx genotype had (OR=1.6, 95% CI:0.85-3.04, P-0), and xx genotype (OR=2.6, 95% CI:1.23-5.46, P-0.01). The association of genotypic variants of different SNPs with estrogen receptor status, showed a statistically significant association between Xx variant of (XbaI) with positive receptor status (OR= 5.3, 95% CI:1.8-15.8, P-0.002). The other two SNPs (PvuII and HinfI) showed statistically significant association only with mammographic density (P-0.0 and 0.01 respectively). Discussion: The family history of female cancers and SNPs, rs9340799 (XbaI) are possible risk factors affecting breast cancer in Sudanese women.
  • Item
    Genomic and transmission electron microscopic characterization of coliphage lytic to Eschericia coli strains
    (Faculty of Medicine, University of Kelaniya, 2015) Kaikabo, A.A.; Abdulkarim, S.M.; Abas, F.; Sieo, C.C.
    Introduction: Avian Pathogenic Escherichia coli (APEC) and human Uropathogenic E. coli (UPEC) share similar genetic relatedness and causing human and poultry diseases. The two showed great overlap based on their serophylogenetic groups and virulence genotypes. The ability of APEC to spread to human, and its capability to act as human UPEC or its potential to act as reservoir of virulence genes for UPEC has been discerned. Recent reports showed that APEC were incriminated in human Urinary Tract Infections (UTI) which is most common bacterial infections causing significant morbidity and mortality and huge economic burden on the healthcare system worldwide. Also, Extended Spectrum Beta Lactamase (ESBL) producing E. coli resistant to third generation cephalosporin has been isolated from poultry products. These pathogens are a great threat to public health and food safety. Poultry is believed to be the source of these pathogens causing diseases in human and avian species. Objective: To isolate and characterize bacteriophage lytic to susceptible E. coli strains. Methodology: Bacteriophage lytic to E. coli strains from chicken was isolated by simple enrichment, soft agar overlay and incubation at 37ºC for 24 hours. The identity of the phage was determined by transmission electron microscopy and partial sequencing of the capsid gene (gp23). Results: Morphologically the phage possesses icosahedral head and contractile tail, and detection of gp23 gene revealed the phage as T4 like coliphage and a member of the family myooviridae. Discussion: Due to its lytic activity, the isolated bacteriophage may offer useful application for biocontrolling susceptible E. coli strains.
  • Item
    Effect of Malus domestica and Moringa oleifera in Amelioration of thyroid Dysfunction in Cassava-cyanide consuming Female Rotus norvegicus
    (Faculty of Medicine, University of Kelaniya, 2015) Madukosiri, C.H.; Opara, D.C.; Unuaru, V.
    Introduction: Prolonged and frequent intake of residual cyanide through processed cassava (Manihot esculenta) foods can precipitate various types of toxicity syndromes including thyroid enlargement (Goitre) and other organ lesions. Since cassava is a staple food and a major energy source for the poor masses, not only in Nigeria but in many African countries and other developing worlds where it is considered to have great economic potentials, there is no intention to advocate discontinuation of the use of cassava as foods but to find ways of ameliorating the symptoms of toxicity in consuming animals due to hydrogen cyanide content. Objective: To study on cassava consuming Female Wistar Rats Methodology: About 24 albino wistar rats aged 8 to 9 weeks and weighing 150- 165g were divided into 5 groups and fed Ad libitum with heat-treated cassava cyanide diets for 28 days after seven days acclimatization in food and environment. Serum iodide/Thyroid Peroxidase (TPO) activity was measured spectrophotometrically at 353nm following active iodine formation from I- and H202; whereas thyroid hormone levels were determined using kit method. Serum bilirubin, malondialdehyde, minerals and antioxidant capacity as well as other phytochemical determinations in M. domestica and M. oleifera were carried out using standard laboratory methods. Results: Results showed that TPO activity (μL-1) were 5.055±0.119, 2.384±0.068, 3.627±0.634, and 3.449±0.070 for rats fed on the control, cassava-based feed diet, apple-cassava-based feed, and moringa-cassava-based feed respectively. This is an indication that the supplements might be helpful in treating thyroid dysfunction caused by reduced activity of the TPO enzyme. Antioxidant capacity of apple and moringa supplements determined was according to the decreasing order Duchess>Northwest greening>Golden delicious>Moringa seeds. Discussion: These supplements could be helpful in alleviating the symptoms of thyroid dysfunction mediated through hydrocyanide intoxication in cassavaconsuming animals.
  • Item
    Epstein-Barr virus (EBV) among Human Immunodeficiency Virus (HIV) infected individuals in Ogbomoso, South-Western Nigeria
    (Faculty of Medicine, University of Kelaniya, 2015) Oladipo, E.K.; Oloke, J.K.; Adewusi, A.P.; Afolabi, F.O.; Bambi, I.; Oke, I.C.; Oladimeji, W.O.
    Introduction: The Epstein - Barr virus (EBV), also called Human Herpes Virus (HHV-4), is a virus of the Herpes family, and is one of the most common viruses in humans. It is the best known cause of infectious mononucleosis. In AIDS patients, several distinct additional EBV-associated diseases may occur and some particular forms of malignancies. Objective: To determine the seroprevalence of EBV in Human Immunodeficient Virus (HIV) -infected individuals Methodology: Two hundred and seventy eight HIV-infected individuals in Ogbomoso, Oyo State were screened for EBV during the period of August to November, 2014 after obtaining their consent. The sera of the subjects were subjected to serological assay using Enzyme-Linked Immunosorbent Assay (ELISA). Results: The mean age of the subjects was 39.5 ±0.48 years and the mean CD4+ was 384.27 ±17.77 cells/μl. Out of 278 subjects tested, 11 (4%) were positive for anti-EBV IgM. Anti-EBV IgM was highest in the age group 41-50years, 5 (5.81%); females, 8 (4.85%); students (14.21%); and CD4+ 350-500 cells/μl, 4 (36.36%). The multivariate analysis using age and the CD4 count shows highest prevalence (27.27%) among the age range (41-50) years with the CD4+ count of 350- 500 cells/μl. Discussion: The result from this study has established that EBV IgM is present among individuals infected with HIV in Ogbomoso.
  • Item
    Significance of polymorphonuclear leucocytes with drumstic in malaria positive and normal patient in Ekiti State South Western, Nigeria
    (Faculty of Medicine, University of Kelaniya, 2015) Adejuyigbe, A.; Ologunde, A.C.
    Introduction: Malaria is estimated to cause about millions cases with high death rate recorded annually. Sickle cell, B blood group and duff blood antigens have selective advantage against malaria infection caused by Plasmodium falciparum and vivax respectively. Objective: To study the effect of Polymorphonuclear Leucocytes with Drumstic (PMND) in immunity to malaria infection Methodology: Three selected local government areas in South West Nigeria were sampled. When the results obtained with the malaria negative female population compared with malaria positive population by subjected to Chi-square statistical analysis to verify whether the observed results has significant difference. Results: The percentage of normal individual female positive for PMND was 4% while none of the male patient screened was positive for PMND. When the results obtained with the malaria negative female population compared with malaria positive population by subjected to Chi-square statistical analysis to verify whether the observed results has significant difference. The result shown that there was no significant difference (X2 (α = 0.05) (1)< X2 cal) as the value positive for PMND in malaria positive compare with malaria negative female patient was only 5.26%. Discussion: This study shows that PMND have no immunity role to play in malaria infection.
  • Item
    Application of molecular genetic markers for genetic resource estimation in cattles in Ukrane
    (Faculty of Medicine, University of Kelaniya, 2015) Shkavro, N.
    Introduction: The unique Charolais herd of Ukrainian selection was begun from 1970 by cross breeding with Gray Ukrainian cattle breed. Such hybrids have better reproductive abilities with maintaining of high meat quality. Now, the problem of native and local farm animal breeds estimation and conservation must be solved by the whole complex of current DNA analysis methods. Genomic selection methods based on a Single Nucleotide Polymorphisms (SNPs) analysis is a widely used powerful new tool in an animal breeding program, especially some economically valuable traits genes identification (e.g. growth hormone and milk proteins genes (caseins and whey proteins) in addition and with the microsatellites markers investigated for individual identification and paternity testing, but only a few studies have explored their practical applications for meat production at the breed level. Objective: To apply microsatellites markers for meat production at the breed level. Methodology: DNA was isolated from the blood of animals by Kawasaki method. Polymorphism of microsatellite loci was estimated based on 10 markers panel, recommended by ISAG, using standard reagents Bovine PCR Typing Kit II by the capillary electrophoresis analysis on ABI PRISM 3110. The Growth Hormone (GH) and kapa-Casein (CSN3) gene polymorphism were identify by PCR-RFLP with the specific restriction ferments (AluI and HindII, respectively) in a 2% agarose gel. Results: 77 allelic variants of the microsatellite DNA markers were identified, the largest number of the alleles was observed for TGLA53 locus (12 alleles). All microsatellites, besides ETH10, showed a high polymorphism level. Sufficiently high B allele frequency (0,269) of kappa-casein gene was detected and a large number of animals (50.8%) have the heterozygous AB genotype variant. Insignificant advantage of V allele (0,511) frequency over L allele (0,489) of growth hormone gene was identified. Animals with heterozygous LV genotype prevailed (53.7%), frequencies of homozygous LL and VV genotypes were 0.221 and 0.242 respectively. Discussion: Results of this study will enhance meat production in the country
  • Item
    Role of the chromobox protein CBX7 in carcinogenesis
    (Faculty of Medicine, University of Kelaniya, 2015) Simair, A.A.; Lu, C.
    Introduction: Chromobox Protein Homolog 7 (CBX7) is a 251 amino acid nuclear protein that contains one N-terminal chromo domain and one C-terminal Pc box. Highly expressed in kidney, brain, heart and skeletal muscle, with weaker expression in peripheral blood leukocytes. The CBX7 functions as a component of the chromatin-associated Polycomb complex (PcG) and is involved in maintaining the transcriptionally repressed state of target genes. Additionally, CBX7 modifies chromatin and is thought to extend the cellular life span of epithelial cells by repressing p14 ARF expression, while simultaneously repressing telomerase activity. Due to its ability to repress the transcription of cellcycle related proteins, CBX7 is thought to play a role in tumorigenesis, specifically in the development of follicular lymphoma and thyroid cancer. Objective: To study on oncogenic activity of CBX7 in humans. Methodology: Expression pattern of CBX7 in a range of normal human tissues and tumor samples was tested to determine oncogenic activity of CBX7. Results: We found in different studies that CBX7 negatively or positively regulates the expression of several genes (SPP1, SPINK1, STEAP1, and FOS, FOSB, EGR1) associated to cancer progression, by interacting with their promoter regions and modulating their transcriptional activity. Furthermore, Cbx7 repressed transcription from the Ink4a/Arf locus and acted epistatically to the Arf-p53 pathway during tumorigenesis. Data identify CBX7 as a chromobox protein causally linked to cancer development and may help to explain the low frequency of INK4a/ARF mutations observed in human follicular lymphoma. Discussion: Mechanism by which the loss of CBX7 expression may contribute to the emergence of a more malignant phenotype and the loss of CBX7 expression might play a critical role in advanced stages of carcinogenesis by deregulating the expression of specific effector genes.
  • Item
    Detection of Salmonella at local poultry farms by rapid and reliable molecular diagnostic method
    (University of Kelaniya, 2015) Wajid, M.
    Introduction: Salmonella enterica having more than 2600 serovars causes moderate to severe gastrointestinal infections in humans and animals. Salmonella enterica serovar Gallinarum biovars Gallinarum and Pullorum (S. Gallinarum and S. Pullorum) are poultry specific bacterial pathogens causing fowl typhoid and pullorum disease respectively while S. Typhimurium and S. Enteritidis cause chicken infections of zoonotic importance. These infections are of considerable economic importance so early and precise diagnosis of the particular Salmonella is mandatory for effective treatment. Currently conventional diagnostic method practiced is based on postmortem guess which lack specificity and sensitivity. Once diagnosed, Salmonella infections are treated by excessive use to antibiotics in drinking water. The choice of antibiotics is usually based on previous treatment experience or just easy availability, while the sensitivity of the prevailing Salmonella serovars against particular antibiotic has not been investigated, thus not considered. This irrational use of antibiotics often results in treatment failure, so the veterinary doctors have no choice except to try other antibiotic. Such hit and trial approach causes huge economic loss to the poultry farmers as the cost of antibiotics used for larger controlled poultry sheds is very high. Objective: To detect Salmonella at local poultry farms by rapid and reliable molecular diagnostic method and to provide the base line data about sensitivity/resistance of the prevailing Salmonella serovars against commonly used antibiotics. Methodology: Suspected samples from local poultry farms were collected. These samples are being tested for Salmonella serovars by biochemical and molecular assays. The antimicrobial sensitivity/resistance will be checked phenotypically by disc diffusion assay followed by molecular detection of drug resistance genes. Results: Analysis of samples is ongoing. Discussion: This information will be helpful for doctors to select the effective antibiotic against which the resistance has not yet developed in local isolates of Salmonella.
  • Item
    Evaluation of lead toxicity among primary school children in Nepal
    (Faculty of Medicine, University of Kelaniya, 2015) Mehta, K.; Sherchand, O.; Rajendra, K.C.; Binod, D.; Madhab, L.; Baral, N.
    Introduction: Lead toxicity is one of the greatest environmental threats to children and primarily affects the developing brain. Objective: To estimate Blood Lead Level (BLL) and its associated risk factors and impact on health in primary school going children in Nepal. Methods: School children, 5-12 years (n=612) living near high traffic roads with air pollution were investigated from June 2012 to November 2013. Children (n=308) from 10 schools of industrial region eastern Nepal and others (n=304) from 8 schools of Kathmandu metropolitan city were enrolled in this study after taking consent from school teacher/guardians. Blood samples were collected from each child and BLL was determined using Flame Atomic Absorption Spectrophotometer (FAAS). Intelligent Quotient (IQ) was assessed using Seguin Form Board Performance Test. Results: We found 82% children (n=503) having detectable BLL and 64.7% children (n=396) had BLL more than 5 μg/dl. In addition 51.79% children (n=317) had above the WHO level of concern (10 μg/dl). Seven percent of children were found to be high risk of toxicity of lead exposure (>70 μg/dl). The BLL was found to be significantly negative correlated with blood hemoglobin (p=0.002), but only negative correlation with total calcium (-0.045) and IQ (-0.077). Using multivariate regression analysis, low socioeconomic status, increasing age, anemia, hypocalcaemia and living homes with chipped wall paints are potential factors associated with high BLL in children (P=0.001) of Nepal. Discussion: This first baseline study is a pointer to the perils of environmental lead exposure among children of Nepal. According to WHO lead level of concern (≥10 μg/dl), it reports more than 50% Nepalese children are the risk of lead poisoning and average mentally retarded. The negative correlation of BLL with IQ also explains that lead toxicity can disrupt the children’s brain development. The consequences of significant adversity early in life prompt an urgent call for innovative strategies to reduce lead toxicity within the context of a coordinated system of policies and services guided by childhood and early brain development.