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

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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
Could food be our medicine?
(Faculty of Medicine, University of Kelaniya, 2015) Suresh, T.S.
Introduction: Non communicable diseases are becoming an alarming threat to humans at present. The high prevalence of diabetes mellitus in Asia is a challenge to health care professionals. With numerous limitations attributed to existing allopathic drugs, more attention is being paid to introduce functional foods which exert beneficial health effects. A few studies were conducted with the aim of identifying suitable functional foods that can be popularized among the community. The hypoglycaemic effects of the hot water extract of dried flowers of Aegle marmelos and the culinary mushrooms; American oyster (Pleurotus ostreatus) and abalone (P. cystidiosus) in Type II diabetic patients on diet control were studied. Methodology: The study was conducted at the Faculty of Medical Sciences, University of Sri Jayewardenepura, Sri Lanka. Following ethical clearance, ninety, volunteer patients were recruited (n=30/group). A glucose challenge test was performed on Day 1 and the concentration of serum glucose and insulin were measured. Same procedure was repeated on a subsequent day where patients received a single dose of respective extracts ½h prior to glucose and serum glucose and insulin were measured. The extracts were given daily for 2 weeks and liver enzymes and creatinine levels were measured. Results: Consumption of A. marmelos, P. autreatus and P. cystidiosus extracts signifcantly reduced (p<0.05) the concentrations of fasting as well as 2h serum glucose in the teste subjects. The serum insulin concnetrations were signifcantly increased (p<0.05) following treatment. The serum concentrations of liver enzymes and creatinine were not affected. Discussion: The drink of A. marmelos is a very popular beverage in Sri Lanka. The findings validadte its use as a functional food which can be safely consumed daily. Similarly the study confirms that the American oyster and abalone mushrooms which are consumed fresh as well as dried, are capable of improving glucose tolerance in diabetic patients.
Deletion analysis of Azoospermic Factor c (AZFc) region of Y chromosome and its effect on spermatogenic impairment in infertile and normozoospermic in South India
(Faculty of Medicine, University of Kelaniya, 2015) Shivaprasad, H.S.; Suttur, S.M.
Introduction: The Azoospermia Factor c (AZFc) region of Y chromosome undergoes de novo mutation and considered as a significant risk factor for spermatogenic impairment which has been well distinguished in infertile and also in fertile individuals with varied effect. However, the impact of these deletions on etiology of male infertility remains poorly understood Indian heterogeneous population. This AZFc sub deletions increase the risk factor of spermatogenic impairment that results in infertile/sub fertile condition. Objective: To investigate the correlation between the AZFc subdeletions and spermatogenic phenotypes in admixed population of South India. Methodology: We recruited the fertile normozoospermic (n= 250) and infertile (n=250) individuals with age group ranging 20-45 years. Semen analysis with sperm function test was performed as per the WHO protocol and five AZFc specific sequence tagged site markers were employed to detect deletions in various gene clusters in AZFc regions namely, DAZ (sY254,sY255), gr/gr (sY1291), b1/b3 (sY1291, sY1197) and b2/b3 deletions (sY1191) by means of standard PCR. Results: In comparison with other AZFc sub deletions we recorded a high occurrence of gr/gr sub deletion (36.3%) in infertile (n=16) and b2/b3 deletion in fertile (n=12) individuals. Interestingly, we have not observed any changes in sperm count, motility, morphology and vitality among the fertile normozoospermic group. In contrast, azoospermic and oligozoospermic subconditions were predominant in infertile individuals with impaired sperm motility, vitality, and volume. Our current investigation is in accordance with the Han Chinese population study in China/East Asians and also among Japanese population where they have recorded the positive association of gr/gr sub deletions with spermatogenic impairment among infertile. Discussion: There is a significant differences in the frequency of gr/gr deletions between fertile and infertile groups but not for other AZFc subdeletions. Thus, the Y chromosome AZFc gr/gr deletions result in spermatogenic impairment by reducing the sperm quality. Further, screening of single nucleotide variant and loss of specific gene copy in AZFc region and its effect on male infertility using RFLP approach is in progress.
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.
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.
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.
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.
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.
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.
Involvement of control of Lymphatic Filarisis in Sri Lanka: Laboratory, Anti-Filariasis Campaign (AFC)
(Faculty of Medicine, University of Kelaniya, 2015) Gamini, P.D.
Anti-Filariasis Campaign (AFC) is actively involved in control of lymphatic filarisis in Sri Lanka. Laboratories situated in the Head Office are actively involved in control of lymphatic filarisis in Sri Lanka. Microscopic identification of filarial parasite in Night Blood Films (NBF) and vector mosquitoes is conducted as a routine work. Quality assurance for NBF samples received from the regional centres of the Anti-Filarisis Campaign is also conducted. Immune Chromatographic Test (ICT) for the detection of filarial antigens in suspected individuals and school children is also conducted. Field samples are tested by CPCR, qPCR and C-ELISA.
Molecular diagnosis of Down syndrome cases
(Faculty of Medicine, University of Kelaniya, 2015) Podder, G.; Madhusnata, D.
Introduction: Down Syndrome (DS), the most frequent form of mental retardation, is characterized by well-defined and distinctive phenotypic features. Research DS-related genes is based on studying the genes located on chromosome 21. Understanding the genes involved may help to target medical treatment to individuals with DS. It is estimated that chromosome 21 contains 200 to 250 genes. It has been identified major genes involved in DS characteristics are normally in the region 21q21–21q22.3. Methodology: Blood samples from 85 DS cases and 30 normal cases were analyzed for D21S11 marker gene. Fluorescent dye-labeled primers were used in PCR amplification of this marker. The PCR amplified product was analyzed by GeneScan software version 4.1 automatically. Results: The short tandem repeat marker frequency was estimated in triallelic, diallelic, and monoallelic patterns. Seventy nine (92.94%) of 85 DS cases showed 3 distinct peaks for D21S11 marker and 5 (5.88%) DS cases showed two uneven peaks. One DS (1.17%) sample showed only one large peak. For healthy individuals, two peaks were observed in all cases. Discussion: Polymorphic STR DNA markeris useful for determining the numbers of chromosome 21. The high sensitivity and automation of the procedures suggest a good prospect for use of this method in detection of DS. However, this is a preliminary investigation and a large-scale study is necessary to validate the clinical application of this protocol.
Nutritive value of traditional foods used for breakfast
(Faculty of Medicine, University of Kelaniya, 2015) Senavirathna, I.
Introduction: The nutritive value of traditional breakfast foods or foods prepared with indigenous raw materials available in Sri Lanka are a neglected area of research. These foods have been consumed by Sri Lankans for centuries but have been slowly replaced with wheat based food products. Objective: To study a few such foods/foods made with indigenous raw materials for their nutritional properties. Methodology: The foods studied included four traditional tubers and foods made with Cycas circinalis (madu) seeds (roti and pittu) and Vateria copallifera (hal) fruit (pittu) and Caryota urens (kithul) pith (roti, thalapa and muffin). The proximate compositions (ash, moisture, digestible carbohydrate, crude protein, soluble and insoluble dietary fibre and total fat contents) of these foods made according to standard recipes were determined by the AOAC methods. Antioxidant capacity was determined as Trolox Equivalents (TEAC) and phenolic/polyphenol content as Gallic Acid Equivalence (GAE/100g fresh weight). The glycaemic indices were determined with healthy volunteers according to WHO criteria. Results: Kithul thalapa and buthsarana were categorized as high GI foods with (± SEM) 128 ± 11 and 110 ± 8 respectively. Muffin prepared from kithul flour and hulankeeriya had medium GI values of 92 ± 9 and 82 ± 8 respectively. The madu roti and pittu, kithul roti, hal pittu and two raja ala varieties (raja ala violet and white) had low GI (66 ± 6, 72 ± 4, 57 ± 4, 67±7, and 64 ± 9 and 69 ± 4 respectively) Phenolic/polyphenolic contents of the raw flour ranged from 79-1162 (GAE/100 g FW) with the antioxidant capacity ranging from 3-225 TEAC. Discussion: Traditional breakfast foods could be recommended for both healthy and people with impaired glucose tolerance.
Production of industrially important enzymes and microbial lipids from biomass of Municipal Solid Wastes (MSW) in Bangladesh
(Faculty of Medicine, University of Kelaniya, 2015) Iqbal, A.; Sohag, M.H.; Ahmed, J.; Rana, M.; Rakibuzzaman, S.M.; Islam, K.; Azad, A.K.
Introduction: Municipal Solid Wastes (MSW) in developing countries is usually dumped in open field resulting environmental pollution and public health hazards. Methane, the second prevalent greenhouse gas having impact on climate change over 20 times greater than carbon dioxide is emitted from the rotten MSW. Organic part which is almost 70-80% of MSW can be converted to bioresources to mitigate the climate change and environmental pollution. Objective: To produce industrially important enzymes and microbial lipids from biomass of MSW. Methodology: We have isolated and identified proteolytic and cellulolytic bateria and fungi from MSW, cowdung and sea sands. We have successfully used MSW as raw materials in the bioreactor for production of industrially important protease and cellulase enzymes from bacterial and fungal isolates and microbial lipids from oliagenous yeast Lipomyces starkeyi. The enzymes were purified and characterized. Results: Amount of accumulated lipids of L. starkeyi grown in MSW hydrolysate media under optimal conditions was ~45% (v/w) of dry biomass of the yeast cells. Gas chromatographic analysis revealed that this lipid was composed of saturated and unsaturated fatty acids. The fatty acid composition of this microbial lipid is similar to that of vegetable oils. Discussion: Our study indicates that organic MSW might be a valuable alternative feedstock for production of microbial lipids and industrially important enzymes.
Review: Thermotherapy for treatment of cutaneous leishmaniasis in Sri Lanka
(Faculty of Medicine, University of Kelaniya, 2015) Silva, H.; Karunaweera, N.D.
Leishmaniasis is a neglected tropical disease caused by a protozoan parasite and transmitted by phlebotomine sand flies. In Sri Lanka, Cutaneous Leishmaniasis (CL) is caused by Leishmania donovani, a usually virsceralizing strain in other parts of the world. Thus it is important to study the treatment methods available for treatment of CL. Commonly used treatment for leishmaniasis in Sri Lanka is Intra-Lesional (IL), intramuscular or intravenous sodium stibogluconate. Thermotherapy is currently studied as an alternative treatment. Several clinical trials have been carried out in the world to see the efficacy and safety of thermotherapy versus intralesional SSG for treatment of CL. A device generating heat using radio waves had been used in these studies. This device is costly and is not freely available in Sri Lanka. A device with a lower cost would be better for Sri Lanka. A pilot study conducted in Peru looked into the use of a novel low-cost thermotherapy for CL using a hand Held Exothermic Crystallization Thermotherapy (HECT) device. 25 patients enrolled in the pilot study and 13 patients treated on compassionate grounds showed a definitive clinical cure rate of 68.4%. Another study conducted in Pakistan using a HECT device had a 83% cure rate by Day180. Aforementioned studies show the efficacy and safety of thermotherapy with HECT device is comparable with SSG, well tolerated by patients and the only side effect experienced was reversible burns which healed without a scar. On the contrary, conventional treatment with IL-SSG is painful, requires many hospital visits, have adverse effects, high treatment cost and poor patient compliance. Thus thermotherapy using HECT device might be considered as a better alternative treatment method for CL patients in a developing country like Sri Lanka.
Role of genetic and biochemical factors associated with non syndromic hearing loss
(University of Kelaniya, 2015) Santra, B.
Introduction: Hearing impairment is one of the most common widespread birth defects and one of every 1000 new born has bilateral permanent Sensorineural Hearing Loss (SNHL). About 50% of SNHL involves genetic factors and 46 genes have been identified as causally related to nonsyndromic SNHL. In spite of this large genetic heterogeneity, mutations in GJB2 and GJB6 genes are primarily responsible for most of the genetic hearing loss. A free radical is an unstable cluster of molecules can cause errors in genetic “messages” by altering DNA. This can, among other things, lead to a reduced blood supply to organs such as the inner ear and brain, there by damaging hearing. Our bodies produce enzymes known as antioxidants, such as Superoxide Dismutase (SOD) and catalase to counteract the damage. Objectives: To study to investigate the genetic cause of deafness by genetic screening of the GJB2 genes and enzyme and catalase in deaf patients. Methodology: Patients with sensorineural congenital HI were selected after obtaining their consents. Clinical samples were tested using PCR/RFLP for 35delG & 167delT mutations. PCR was used to amplify two regions of the exon 2 of connexin26 and PCR products were analyzed using Bsl I and Pst I. Blood level of SOD and catalase was estimated by spectrophotometric analysis. Results: 87.8% had normal genotype for the studied mutations, 4.88% were compound heterozygotes, 7.32% homozygotic for the 35delG mutation, 9.76% heterozygotic for 35delG/GJB2. The 167delT mutation was not detected in any of the deaf individuals. Discussion: Our data confirmed the presence of the 35delG mutation in the GJB2 gene in cases of non-syndromic bilateral moderate to profound sensorineural hearing loss in West Bengal. For the 167delT mutation the allele frequency was zero for the familial and sporadic groups. These findings underline the importance of a genetic diagnosis that may clarify the etiology and provide early treatment for children. Genetic counseling for their family members will help in early detection and treatment.
Role of MicroRNAs in B cell maturation and differentiation during human visceral leishmaniasis infection
(Faculty of Medicine, University of Kelaniya, 2015) Pandey, R.K.; Prajapati, V.K.
Introduction: Lymphocyte B cells play an important role to provide humoral protective immune response against Leishmania donovani by the producing polyclonal antibodies and the process of B cell maturation and differentiation depends on microRNA mediated gene regulation during Human Visceral Leishmaniasis (HVL) infection. Objective: To analyze the role of miRNA in B cell maturation and differentiation during HVL infection. Methodology: MicroRNAs are endogenously expressed, nonprotein encoding tiny RNA that negatively regulates gene expression at post transcriptional level. The present study depicts the identification of miRNA controlling expression of immune signaling molecule, genes and transcription factors using Target Scan web server, to control B-cell maturation and differentiation. Results: Bioinformatics analysis has shown that microRNAs such as miR-1178, miR-590-3p hinders the attachment of progenitor B cell to stromal cell in bone marrow by inhibiting VCAM-1, which is essential for the conversion of progenitor B cell to precursor B cell for maturation. Seed region present in miR-432 and miR- 3973 have the putative binding site in 3’-UTR region of cytokine IL-7 coding mRNA, suppress the specification and commitment of cell to the B cell lineage. Furthermore, we speculated that miR-4731-5p, miR-3678-3p, miR-4492 and miR762 may suppress the early stage B-cell maturation by suppression of various B-cell specific genes, including Pax-5, Vpre-B, λ5 and immunoglobulin heavy chain enhancer, results in aggravation of HVL pathogenesis. Moreover, miR-141 and miR-4698 can inhibit expression of RAG-1/RAG-2 and results in non specific antibody production. We also indicate that miR-132 and miR-212 are potential microRNAs that regulate Sox-4 specific response to stop the maturation of B cell, which enhances the progression of visceral leishmaniasis disease in human. Discussion: Our study highlights the importance of microRNAs in the B cell maturation and differentiation during L. donovani infection in human and provides the bioinformatics evidences to control HVL infection.
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.
Seroprevalence and associated factors for Parvovirus B19 among pregnant women presenting to antenatal clinics in a selected MOH area in Gampaha District
(Faculty of Medicine, University of Kelaniya, 2015) Thilakarathne, N.T.
Introduction: Up to 50% of pregnant women infected with Parvovirus B19 infection may be asymptomatic, but vertical transmission can still occur. If a pregnant woman gets Parvovirus B19 infection, there is a 30% chance of foetal transmission with a 5 - 9% risk of foetal loss. Foetus can be affected at any stage of the pregnancy with serious complications. Immunity to Parvovirus B19 has not being studied in Sri Lanka before. Detection of IgG antibodies to Parvovirus B19 will identify immunity to infection in pregnant mothers. Objective: To determine seroprevalence and describe associated factors for Parvovirus B19 among pregnant women presenting to antenatal clinics in Mahara MOH area in Gampaha district. Methodology: All immunocopmpetent pregnant women who presented to antenatal clinics in Mahara MOH area were included in the study until the calculated sample size (n = 318) was reached, during the 3 months study period. After obtaining informed written consent, interviewer administered structured questionnaires were used to gather data on socio-demographic and associated factors. Venous blood (5ml) was collected from and serum was separated and stored at -40C until processing. Parvovirus B19 IgG ELISA tests were done at Department of Virology, MRI to detect the immunity, using a validated commercial ELISA kit. Results of the test were made available to clinicians at the relevant antenatal clinics. Results: Results analysis is pending Discussion: Though in the past serodiagnostic tests were available at MRI, currently testing for Parvovirus B19 is not available at MRI / government laboratories. The results of this study may be useful in deciding re-establishment of sero-diagnostic tests for Parvovirus B19 infection in government laboratories in future.
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.
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.