Conference Papers

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This collection contains abstracts of conference papers, presented at local and international conferences by the staff of the Faculty of Medicine

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    Reconstruction of Metabolic Pathways for the Setaria digitata Whole Genome
    (Sri Lanka Medical Association, 2020) Rashanthy, N.; Kothalawala, M.S.A.; Mugunamalwaththa, T.S.; Darshika, W.A.S.; Lakmali, G.L.Y.; de Zoysa, K.; Chandrasekharan, N.V.; Gunawardene, Y.I.N.S.; Suravajhala, P.; Dassanayake, R.S.
    INTRODUCTION AND OBJECTIVES: Setaria digitata is a Wolbachia-free filarial parasite that resides in the abdominal cavity of ungulates. It can cause cerebrospinal nematodiasis (CNS) in unnatural hosts such as sheep, goats, which causes a serious threat to livestock farming. Furthermore, S. digitata can also infect humans causing several conditions showing a gradual adaption to humans. METHODS: Despite, to date, complete a metabolic pathway reconstruction of S. digitata has not been undertaken and therefore, in this study the latter analyses were carried out using BLAST2GO software. RESULTS: Metabolic pathway analysis based on Kyoto Encyclopedia of Genes and Genomes (KEGG) database identified 111 enzymes found in total of 246 contigs that involve in 95 metabolic pathways, in which the most over-represented pathways are Biosynthesis of antibiotics, Phosphatidylinositol signaling system and Purine metabolism. Since S. digitata does not harbor Wolbachia endosymbiont, it was theorized that the S. digitata genome must encodes genes to carryout haem, riboflavin and nucleotides pathways, otherwise encoded by Wolbachia genome, potentially through lateral transfer of Wolbachia to an ancestor of S.digitata. Here, KEGG analysis identified 16 enzyme coding genes involve in nucleotide biosynthesis and one enzyme involve in riboflavin biosynthesis pathway. Although studies have revealed that FAD and glutathione pathways are complete in all nematode genomes, the genes encoding FAD and glutathione pathways were not found in the S. digitata. Moreover, complete nucleotide synthesis pathway and haem synthesis pathway were not found. CONCLUSION: This suggests that S. digitata may have evolved its own sequences to encode those biosynthetic pathways and hence calling for investigations to undertake characterization of genes involved in these pathways.
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    Development of a quantitative PCR assay to evaluate HER2 status of Gastric carcinoma in a cohort of Sri Lankan patients
    (Sri Lanka Medical Association, 2016) Kannangara, D.K.S.; Subasinghe, D.; Lokuhetti, M.D.S.; Dassanayake, R.S.; Gunawardene, Y.I.N.S.
    INTRODUCTION AND OBJECTIVES: Human epidermal growth factor receptor2(HER2) protein overexpression and/or HER2gene amplification is linked to dismal outcome of Gastric carcinoma(GCa). Immunohistochemistry(IHC) and fluorescence in situ hybridization(FISH) are key-methods to identify patients for HER2 targeted therapy. Drawbacks of both methods warrant novel tests. The study aimed to determine whether quantitative Polymerase Chain Reaction (qPCR) could serve as a supplementary-method to evaluate HER2 status of GCa in a cohort of Sri Lankan patients and investigate correlation between HER2 assessed by different methods and clinic-pathological features. METHOD: Twenty GCa-patients with known IHC-HER2 scores were evaluated. qPCR was performed for HER2gene and Ameloid precursor protein (reference gene) in Formalin fixed paraffin embedded GCa tissue. Threshold values(Ct) were analyzed using Pfaffl-method to detect HER2gene amplification. RESULTS: HER2positivity by IHC(protein) and qPCR(gene) were 20% and 35% respectively. Sensitivity and specificity of qPCR was 67% and 76% respectively and results were reproducible. HER2protein positivity was correlated with Tumour TNM-stage and Lauren-histological types(P<0.05). Positive expression of HER2gene was correlated with depth of tumour invasion, differentiation and Lymph node-status(P<0.05). Diagnostic consistency between IHC and qPCR(κ=0.146) was slightly agreeable(0.01
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    Development of modified mismatch PCR-RFLP to screen mutations in codon 12 and 13 of K- ras gene of colorectal (CRC) patients in Sri Lanka
    (Sri lanka Medical Association, 2015) Dhilhani, M.F.F.; de Zoysa, M.I.M.; Chandrasekharan, N.V.; Gunawardene, Y.I.N.S.; Lokuhetti, M.D.S.; Dassanayake, R.S.
    INTRODUCTION AND OBJECTIVES: Mutations in K-ras codon 12, 13 of exon 2 are known to affect prognosis and impart resistance to anti EGFR monoclonal antibody therapy in CRC. Although several diagnostic tools have been developed for K-ras mutation testing, these procedures are too expensive or time consuming. Oufaim was to develop an effective, reliable and inexpensive method for the detection of K-ras mutations in codons 12 and 13 of exon 2 in CRC patients in Sri Lanka, and to relate the mutational status to liver metastasis, METHOD: The mismatch PCR-RFLP was developed and used to screen mutations in codon 12 and 13 for DMA isolated from paraffinized tumour tissue of 30 CRC patients followed up for 5 year after surgery to detect liver metastasis. Cross-tabulations were generated between K-ras mutations and the metastatic status. The Chi Square test was used to indicate statistical significance of the association. RESULTS: Analysis of banding pattern obtained from restriction digestion of PCR amplified region containing codon 12 and/or 13 of KRAS gene of 14(46.6%) CRC patients revealed the presence of mutations. Of the 30 patients, 13(43.3%) had developed liver metastases. There was a significant association between the presence of a K-ros mutation and the occurrence of liver metastasis (X2=4.693, p=0.003). CONCLUSION: This mismatch PCR-RFLP protocol is a suitable method to screen codon 12 and 13 mutation of K-ros gene to predict liver metastasis. Presence of these mutations is associated with the occurrence of liver metastasis during the first 5 years after surgery.
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    Potential use of IGR Pyriproxifen for control of dengue vector Aedes albopictus
    (Sri Lanka College of Microbiologists, 1999) Gunawardene, Y.I.N.S.; de Silva, N.R.; Abeyewickreme, W.
    Dengue Fever (DF) Dengue Haernorrhagic Fever (DHF) is now a significant problem in Sri Lanka. The incidence of DHF has increased cyclically since the first recognized outbreak in 1989. Of the 203 suspected DHF cases recorded in 1989, 87 were confirmed and 26 deaths were reported. By 1997, 5882 clinical cases, 1558 serologically confirmed cases and over 300 deaths have been reported. Without an effective vaccine against dengue, and considering the clinical difficulty in managing DHF cases, vector control has become an important com¬ponent in the integrated attempts for dengue con¬trol. Aedes aegypti and Aedes albopictus not only transmit dengue but are also a nuisance and cause annoyance by their day biting behaviour. In this study an attempt was made to control Ae. albopictus by exploiting its oviposition behaviour, us¬ing an Insect Growth Regulator (IGR), Pyriproxifen. The IGR is known to interfere with the synthesis and deposition of chitin in insects and thereby prevent growth and development. Different concentrations of the IGR (0.01 g/1 -0.08g/1) were tested against Ae.albopictus, in laboratory experiments. The most effective larvisidal concentration of Pyriproxifen for Ae.albopictus was determined to be 0.03g/1. Field trials were also carried out to determine the feasi¬bility of employing used automobile tyres contain¬ing Pyriproxifen as ovitraps against Ae.albopictus. Tyres containing water were treated with Pyriproxifen (0.03g/1) and the emergence of adults was recorded. Pyriproxifen at a concentration of 0.03g/1 was found to inhibit the emergence of adult Aedes mosquitoes up to 6 weeks.
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    Dengue as-a public health problem in Sri Lanka
    (La Fondation pour l’Université de Lyon, 2009) Hapangama, H.A.D.C.; Gunawardene, Y.I.N.S.; Hapugoda, M.D.; Premaratna, R.; Manamperi, A.; Gunasena, S.; Abeyewickreme, W.
    Dengue infection is an important global public health problem and an increasing number of persons from the South Asian region have been directly or indirectly affected by the disease. In Sri Lanka, dengue has become a major threat to public health in many urban and sub-urban' areas during past three decades. Rapid unplanned urbanization and increasing human population has increase the rate of infection and the frequency. The study area, Gampaha District is the second most populous district in the country having a population density of 1 539 persons per km2 and was the district reporting the second highest incidence of dengue in 2008. Therefore, current research efforts are focused on dengue transmission, examining the presence of sub-clinical infections, role of vector mosquitoes and Knowledge, Attitude and Practices (KAP) of the community on dengue infection in an effort to contain the disease. In the present study, dengue antibodies were detected in samples collected from clinically suspected patients and as well as in samples collected from volunteers. Volunteer sera collected around the confirmed cases had a 23.6% sero-positive rate for dengue IgM antibodies. The rate of asymptomatic recent infections was calculated to be 16.9%. In present study we have serologically confirmed the presence of subclinical infections and according to the published data this is the first confirmation of asymptomatic dengue infections in Sri Lanka. According to the entomological investigations carried out, the common breeding places for Aedes vectors were found to be discarded small containers. Even though Ae. Aegypti has been considered as the principal vector transmitting dengue fever, current studies highlighted the predominant ro!e of Ae. albopictus in the disease transmission. A previous study in Sri Lanka also suggested that prevalence and .presence of high-density of Ae. albopictus should be considered as a risk factor for endemic/epidemic dengue. In view of the above, the spread of dengue by Ae. albopictus should be a matter of great concern. Findings of KAP survey revealed that the community possessed substantially higher knowledge on the spread of dengue, vectors, vector breeding and also seriousness of the infection. However it was observed that good knowledge does not necessarily lead to good practices. Since the attitudes of the respondents were found to be good and most of them were supportive of control measures; next effort of the present study is to see how a novel community mobilized solid waste management system will be effective in dengue vector control.
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    Ovi-trap based surveillance for Aedes in dengue endemic Gampaha District, Sri Lanka during rainy season
    (Faculty of Tropical Medicine, Mahidol University, 2008) Hapangama, H.A.D.C.; Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Benjamin, S.; Abeyewickreme, W.
    Dengue is endemic in Gampaha District, Sri Lanka. Aedes aegypti and Aedes albopictus have been incriminated as vectors in urban and suburban environments. An ovitrap-based weekly surveillance was carried out to determine the distribution and abundance of Dengue vectors, in six dengue endemic areas; Galudupita, Gongitota, Wanawasala, Kerangapokuna, Mabole and Welikadamulla in the Gampaha District during May and June 2008 in both indoors and outdoors. Galudupita and Gongitota gave the highest ovitrap indices, both indoors and outdoors as 97% and 73% respectively. Of the total positive ovitraps(1320), positivity for Ae. albopictus, Ae. aegypti and both were 63.5%, 32.9% and 3.6% respectively. Highest mean number of Ae. aegypti and Ae. albopictus larvae per ovitrap in both indoor and outdoor ovitraps were observed Galudupita and Gongitota respectively. However, statistical analysis showed that ovitrap index of Ae. aegypti in six study was not significantly different from each other (P > 0.05). Outdoor mean number of Ae. aegypti larvae per ovitrap in Galudupita, was significantly higher than other sites (F=3.587, P=0.012). Indoor Ae. albopictus ovitrap index of Gongitota was significantly higher than of Welikadamula (F=2.59, P=0.046) while outdoor Ae. albopictus ovitrap index of Gongitota and Wanawasala was significantly higher than of Welikadamula F=3.19 P=.0.02). Results irnplicate that Ae. albopictus prefers more vegetation as observed in Gongitota and Wanawasala. In highly populated and urbanized Galudupita, area with less vegetation, Ae. albopictus population was less than Ae. aegypti even outdoors. Study implicates that Ae. albopictus is the predominant indoor and outdoor breeder during rainy season.
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    Serotype distribution of dengue in Gampaha District, Sri Lanka
    (Faculty of Tropical Medicine, Mahidol University, 2008) Jayasooriya, D.H.S.W.; Gunawardene, Y.I.N.S.; Manamperi, A.; Hapugoda, M.D.; Premaratna, R.
    Dengue Virus (DENV) affecting millions of people mainly in the tropical and subtropical regions is a positive sense RNA virus which exists in four different serotypes (DENV1-4) and several subtypes. All four serotypes have been found in Sri Lanka. The objective of the study was to determine the DENV serotype distribution in patients and mosquitoes of Gampaha district. Serum was isolated from acute phase blood samples collected from dengue suspected patients during the febrile period. Mosquitoes were collected from different locations where dengue confirmed cases were reported. Pooled and crushed mosquito heads and serum samples were separately subjected to RNA extraction, RT-PCR, semi nested PCR with Dengue serotype specific primers and solution hybridization with Phosphorous 32 labeled probes respectively. Out of 93 patients, 34 were positive for DENV out of which 19 corresponded to DENV2, 5 to DENV3 and 10 were co-infected with DENV2 and DENV3. Of the positive infections corresponding to serotypes 2, 3, 2-3 co-infected; 7/19 (36.8%), 3/5(60%), 0/10 (0%) had developed dengue haernorrhagic fever respectively Among 14 mosquito pools, 5 were positive of which 3 were infected with DENV2 and 2 with DENV3. It was observed that although both DENV2 and DENV3 circulate among patients and in mosquitoes in Garnpaha District, DENV2 was predominant in both. Only Aedes albopictus 'presence in all localities was significant. A greater percentage of severe form of the disease among DENV3 infected patients was observed while a less severe form of the disease was observed in DENV2 and DENV3 co-infected patients
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    Entomological assesment of Wuchereria bancrofti transmission following mass treatment in Gampaha District, Sri Lanka.
    (Faculty of Tropical Medicine, Mahidol University, 2007) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Abeyewickreme, W.
    Global program to eliminate Lymphatic filariasis (Lf) relies mainly on Mass Drug Administration (MDA). Success of filariasis control programs depends on careful monitoring of infection levels in human populations as well as vectors following the introduction of the drug intervention. This study was conducted to assess the current level of Lf transmission following mass distribution of Diethylcarbamazine-Albendazole in the Gampaha district, Sri Lanka. Field study was conducted in 45 sites in all Medical Officer of Health (MOH) areas of the Gampaha district identified by the Anti Filariasis Campaign (AFC) as high risk for bancroftian filariasis. Investigation revealed 42.22% (19/45) of the sites were infested with mosquitoes positive for Wuchereria bancrofti. Only the sites in urban and semi urban areas were positively infested while rural areas were free of infection. An infection rate of 5.26% was observed among the mosquitoes caught from households and the larval density was 8.7 per positive mosquito. According to a study conducted by the AFC in a sample population (14 sites), the prevalence of Lf was 0.038%. Data recorded by the AFC in 1994 indicated that the infective rate, positivity of mosquitoes and microfilaria density were 90%, 3.05% and 23 respectively. The present study confirms that the level of transmission of W. bancrofti has not decreased in the Gampaha district, despite the MDA Programme been implemented since 2002. Therefore, a proper screening-programme combined with anti filarial treatment and. vector control programme is urgently required to minimize filarial morbidity and interrupt filarial transmission within the country.
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    Transmission study of dengue fever in Gampaha District, Sri Lanka
    (Faculty of Tropical Medicine, Mahidol University, 2007) Hapangama, H.A.D.C.; Gunawardene, Y.I.N.S.; Hapugoda, M.D.; Dayanath, M.Y.D.; Abeyewickreme, W.
    RATIONALE: Dengue is an important flaviviral infection in Sri Lanka.OBJECTIVES; To determine the effect of possible contributing factors on transmission of Dengue METHODS: A total of 40 households situated around 7 selected dengue cases confirmed by both IgM-IgG ELISA and HAI assay were taken. A minimum of 3 inhabitants from each household were tested for anti-dengue antibodies. At each household, mosquito surveillance was carried out. RESULTS: Of the 40 households, 26(65%) were positive for dengue viral infection. Of the 148 inhabitants, 41 had evidence of exposure to dengue. Of the 41. asymptomatic dengue infection was observed in 25(61%). Paired sera were collected from 36/41 volunteers and tested by HAI assay which confirmed dengue virus infection in 4(11.1%) and were confirmed secondary. Three (75%) of them were asymptomatic. Balance 32(88.9%) was diagnosed as recent dengue infections. Of that 21(65.6%) were asymptomatic probable secondary 7(33.3%), recent dengue 14(66.6%)]. Clustering of cases was observed in all localities varying from 56.5%. There was a significant statistical association between the presence of a definitive dengue infection with the presence of Aedes vectors (x2=3.1, p=0.1). Ae. albopictus were found in all localities 'while Ae. aegypti was found only in one. Breteau index for Ae. albopictus varied from 0-260 while- container index varied from 0-84%. Highest clustering of dengue cases were observed in localities with the highest indices for' mosquitoes. CONCLUSIONS: Study confirms that the presence of asymptomatic infections and clustering of cases. The predominant vector was Ae. albopictus.
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    Patterns of monthly Culex mosquito density variation in Gampaha district, Sri Lanka.
    (Faculty of Tropical Medicine, Mahidol University, 2009) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S.; Manamperi, A.; Abeyewickreme, W.
    BACKGROUND: The ecology, development, behavior, and survival of mosquitoes and the transmission dynamics of the diseases they transmit are strongly influenced by climatic factors. OBJECTIVE: The objective of this study was to identify the population density variation of Culex mosquitoes within a period of one year in Gampaha district, Sri Lanka together with potential climatic factors that influenced the Culex population density variation. METHODOLOGY: Culex mosquitoes were routinely collected on monthly basis from 9 sites in Gampaha district. Climate data was obtained from the Department of Meteorology. RESULTS: An exponential growth of Culex population densities was observed in all sites starting in December to February during the study period. The maximum density occurred in January and decreased from March until July. It again increased during August and thereafter decreased until December. Among the study sites the maximum Culex density (mosquitoes/man-hour) was observed in Hekiththa ranging between 89 to 22 and the minimum was from Kurukulawa ranging from 6 to 1. Climatic data suggest that temperature is a limiting factor for the Culex population growth while it was strongly influenced by the rain fall pattern. DISCUSSION: Similar Culex population density variation pattern was observed in all sites but exhibited enormous variation between sites, probably due to different local conditions. Also it was suggested that estimation of W, bancrofti transmission levels in Culex mosquitoes should be practiced in field settings where high mosquito density was observed. Since mosquito density appears difficult to be analyzed by individual dissection use of pool-screen PCR-ELISA would be a better method.