Medicine
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This repository contains the published and unpublished research of the Faculty of Medicine by the staff members of the faculty
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Item Proteome profiling of cutaneous leishmaniasis lesions due to dermotropic Leishmania donovani in Sri Lanka(BioMed Central, 2024) Manamperi, N.H.; Edirisinghe, N.M.; Wijesinghe, H.; Pathiraja, L.; Pathirana, N.; Wanasinghe, V.S.; De Silva, C.G.; Abeyewickreme, W.; Karunaweera, N.D.BACKGROUND Characterization of the host response in cutaneous leishmaniasis (CL) through proteome profiling has gained limited insights into leishmaniasis research compared to that of the parasite. The primary objective of this study was to comprehensively analyze the proteomic profile of the skin lesions tissues in patients with CL, by mass spectrometry, and subsequent validation of these findings through immunohistochemical methods.METHODS Eight lesion specimens from leishmaniasis-confirmed patients and eight control skin biopsies were processed for proteomic profiling by mass spectrometry. Formalin-fixed paraffin-embedded lesion specimens from thirty patients and six control skin specimens were used for Immunohistochemistry (IHC) staining. Statistical analyses were carried out using SPSS software. The chi-square test was used to assess the association between the degree of staining for each marker and the clinical and pathological features.RESULTS Sixty-seven proteins exhibited significant differential expression between tissues of CL lesions and healthy controls (p < 0.01), representing numerous enriched biological processes within the lesion tissue, as evident by both the Kyoto Encyclopedia of Genes and Genomes (KEGG) and Reactome databases. Among these, the integrated endoplasmic reticulum stress response (IERSR) emerges as a pathway characterized by the up-regulated proteins in CL tissues compared to healthy skin. Expression of endoplasmic reticulum (ER) stress sensors, inositol-requiring enzyme-1 (IRE1), protein kinase RNA-like ER kinase (PERK) and activating transcription factor 6 (ATF6) in lesion tissue was validated by immunohistochemistry.CONCLUSIONS In conclusion, proteomic profiling of skin lesions carried out as a discovery phase study revealed a multitude of probable immunological and pathological mechanisms operating in patients with CL in Sri Lanka, which needs to be further elaborated using more in-depth and targeted investigations. Further research exploring the intricate interplay between ER stress and CL pathophysiology may offer promising avenues for the development of novel diagnostic tools and therapeutic strategies in combating this disease.Item Diversity of Wolbachia infections in Sri Lankan mosquitoes with a new record of Wolbachia supergroup B infecting Aedes aegypti vector populations(Nature Publishing Group, 2024) Wijegunawardana, N.D.A.D.; Gunawardene, Y.I.N.S; Abeyewickreme, W.; Chandrasena, T.G.A.N.; Thayanukul, P.; Kittayapong, P.Wolbachia bacteria are common endosymbionts of insects and have recently been applied for controlling arboviral vectors, especially Aedes aegypti mosquito populations. However, several medically important mosquito species in Sri Lanka were present with limited information for the Wolbachia infection status. Therefore, the screening of Wolbachia in indigenous mosquitoes is required prior to a successful application of Wolbachia-based vector control strategy. In this study, screening of 78 mosquito species collected from various parts of the country revealed that 13 species were positive for Wolbachia infection, giving ~ 17% infection frequency of Wolbachia among the Sri Lankan mosquitoes. Twelve Wolbachia-positive mosquito species were selected for downstream Wolbachia strain genotyping using Multi Locus Sequencing Type (MLST), wsp gene, and 16S rRNA gene-based approaches. Results showed that these Wolbachia strains clustered together with the present Wolbachia phylogeny of world mosquito populations with some variations. Almost 90% of the mosquito populations were infected with supergroup B while the remaining were infected with supergroup A. A new record of Wolbachia supergroup B infection in Ae. aegypti, the main vectors of dengue, was highlighted. This finding was further confirmed by real-time qPCR, revealing Wolbachia density variations between Ae. aegypti and Ae. albopictus (p = 0.001), and between males and females (p < 0.05). The evidence of natural Wolbachia infections in Ae. aegypti populations in Sri Lanka is an extremely rare incident that has the potential to be used for arboviral vector control.Item Immuno-dominant dengue NS1 peptides as antigens for production of monoclonal antibodies(Frontiers Media S.A, 2022) Munasinghe, E.; Athapaththu, M.; Abeyewickreme, W.Synthetic peptides have recently become common as antigens for antibody production. Peptides are short chains of amino acids that can be used to elicit an immune response. The immunogenicity of the peptide antigens varies depending on the length, charge, solubility, and amino acids contained in the peptide sequence. Dengue NS1 protein is an important target antigen in the early detection of dengue infection. In this study, peptides corresponding to a highly conserved region from the dengue NS1 region were designed and synthesized. Balb/C mice were immunized against each peptide and spleen cells extracted from the immunized mice were fused with NS0 murine myeloma cells. Hybridoma clones obtained from the fusions were tested against peptides using ELISA. Out of 1,830 growing clones, 28 clones produced antibodies reacting with dengue NS1 peptides. A purified monoclonal antibody reacting with all four peptides was tested for reactivity with dengue NS1 native protein using dengue-confirmed serum and urine samples. The monoclonal antibody shows significant reactivity with both serum and urine. The findings of the current research can be used to detect dengue infection using urine, which ultimately results in the prevention of dengue epidemics through painless diagnosis, following treatment, and patient management to safeguard human and economic wellness.Item Occurrence of major and potential malaria vector immature stages in different breeding habitats and associated biotic and abiotic characters in the district of Trincomalee Sri Lanka(New Delhi : National Institute of Malaria Research, 2020) Ranathunge, R.M.T.B.; Kannangara, D.N.; Gunathilaka, P.A.D.H.N.; Abeyewickreme, W.; Hapugoda, M.D.BACKGROUND & OBJECTIVES: Understanding the effect of biotic and abiotic factors on the biology and ecology of immature stages of anopheline larvae is very important in controlling malaria vector mosquitoes. Therefore, this study was focused on the monitoring of ecological factors affecting the distribution, dynamics, and density of malaria vector mosquitoes in the District of Trincomalee, Sri Lanka. METHODS: Permanent and temporary breeding habitats were identified and selected from five possible malaria sensitive sites in the district of Trincomalee. Anopheles larvae and macro-invertebrates were collected using standard methods for 16 months (from October 2013 to January 2015) and they were identified microscopically. Eight physico-chemical parameters of the breeding habitats were measured. RESULTS: Overall, a total of 4815 anopheline larvae belonging to 13 species were collected from 3,12,764 dips from 18 permanent and temporary breeding habitats. The abundance of anopheline larvae showed a significant positive correlation (p <0.05) with physico-chemical parameters in breeding habitats, such as temperature, dissolved oxygen, and turbidity. A total of 35 macro-invertebrate taxa were collected from the anopheline mosquito breeding habitats. INTERPRETATION & CONCLUSION: This study represents the first systematic update of water quality parameters, macro-invertebrate communities associated with Anopheles mosquito oviposition sites in the District of Trincomalee, Sri Lanka. Rainfall intensity and wind speed are critical meteorological factors for the distribution and abundance of malaria vectors. Knowledge generated on the ecology of Anopheles mosquitoes will help to eliminate malaria vectors in the country. KEYWORDS: Macro-invertebrates; Sri Lanka; malaria vectors; physico-chemical parameters.Item Larval indices of vector mosquitoes as predictors of dengue epidemics: An approach to manage dengue outbreaks based on entomological parameters in the districts of Colombo and Kandy, Sri Lanka(Hindawi Pub. Co., 2020) Udayanga, L.; Aryaprema, S.; Gunathilaka, N.; Iqbal, M. C. M.; Fernando, T.; Abeyewickreme, W.BACKGROUND: Early detection of dengue epidemics is a vital aspect in control programmes. Predictions based on larval indices of disease vectors are widely used in dengue control, with defined threshold values. However, there is no set threshold in Sri Lanka at the national or regional levels for Aedes larval indices. Therefore, the current study aimed at developing threshold values for vector indices in two dengue high-risk districts in Sri Lanka. METHODS: Monthly vector indices (House Index [HI], Container Index [CI], Breteau Index for Aedes aegypti [BIagp], and Ae. albopictus [BIalb]), of ten selected dengue high-risk Medical Officer of Health (MOH) areas located in Colombo and Kandy districts, were collected from January 2010 to June 2019, along with monthly reported dengue cases. Receiver Operating Characteristic (ROC) curve analysis in SPSS (version 23) was used to assess the discriminative power of the larval indices in identifying dengue epidemics and to develop thresholds for the dengue epidemic management. RESULTS: Only HI and BIagp denoted significant associations with dengue epidemics at lag periods of one and two months. Based on Ae. aegypti, average threshold values were defined for Colombo as Low Risk (2.4 ≤ BIagp < 3.8), Moderate Risk (3.8 ≤ BIagp < 5), High Risk (BIagp ≥ 5), along with BIagp 2.9 ≤ BIagp < 4.2 (Low Risk), 4.2 ≤ BIagp < 5.3 (Moderate Risk), and BIagp ≥ 5.3 (High Risk) for Kandy. Further, 5.5 ≤ HI < 8.9, 8.9 ≤ HI < 11.9, and HI ≥ 11.9 were defined as Low Risk, Moderate Risk, and High Risk average thresholds for HI in Colombo, while 6.9 ≤ HI < 9.1 (Low Risk), 8.9 ≥ HI < 11.8 (Moderate Risk), and HI ≥ 11.8 (High Risk) were defined for Kandy. CONCLUSIONS: The defined threshold values for Ae. aegypti and HI could be recommended as indicators for early detection of dengue epidemics and to drive vector management activities, with the objective of managing dengue epidemics with optimal usage of financial, technical, and human resources in Sri Lanka.Item Climate change induced vulnerability and adaption for dengue incidence in Colombo and Kandy districts: the detailed investigation in Sri Lanka(BioMed Central., 2020) Udayanga, L.; Gunathilaka, N.; Iqbal, M. C. M.; Abeyewickreme, W.BACKGROUND: Assessing the vulnerability of an infectious disease such as dengue among endemic population is an important requirement to design proactive programmes in order to improve resilience capacity of vulnerable communities. The current study aimed to evaluate the climate change induced socio-economic vulnerability of local communities to dengue in Colombo and Kandy districts of Sri Lanka. METHODS: A total of 42 variables (entomological, epidemiological, meteorological parameters, land-use practices and socio-demographic data) of all the 38 Medical Officer of Health (MOH) areas in the districts of Colombo and Kandy were considered as candidate variables for a composite index based vulnerability assessment. The Principal Component Analysis (PCA) was used in selecting and setting the weight for each indicator. Exposure, Sensitivity, Adaptive Capacity and Vulnerability of all MOH areas for dengue were calculated using the composite index approach recommended by the Intergovernmental Panel on Climate Change. RESULTS: Out of 42 candidate variables, only 23 parameters (Exposure Index: six variables; Sensitivity Index: 11 variables; Adaptive Capacity Index: six variables) were selected as indicators to assess climate change vulnerability to dengue. Colombo Municipal Council (CMC) MOH area denoted the highest values for exposure (0.89: exceptionally high exposure), sensitivity (0.86: exceptionally high sensitivity) in Colombo, while Kandy Municipal Council (KMC) area reported the highest exposure (0.79: high exposure) and sensitivity (0.77: high sensitivity) in Kandy. Piliyandala MOH area denoted the highest level of adaptive capacity (0.66) in Colombo followed by Menikhinna (0.68) in Kandy. The highest vulnerability (0.45: moderate vulnerability) to dengue was indicated from CMC and the lowest indicated from Galaha MOH (0.15; very low vulnerability) in Kandy. Interestingly the KMC MOH area had a notable vulnerability of 0.41 (moderate vulnerability), which was the highest within Kandy. CONCLUSIONS: In general, vulnerability for dengue was relatively higher within the MOH areas of Colombo, than in Kandy, suggesting a higher degree of potential susceptibility to dengue within and among local communities of Colombo. Vector Controlling Entities are recommended to consider the spatial variations in vulnerability of local communities to dengue for decision making, especially in allocation of limited financial, human and mechanical resources for dengue epidemic management. KEYWORDS: Climate change; Dengue; Sri Lanka; Vulnerability.Item Field-based evaluation of Novaluron EC10 insect growth regulator, a chitin synthesis inhibitor against Dengue vector breeding in leaf axils of pineapple plantations in Gampaha district, Sri Lanka(BioMed Central, 2020) Gunathilaka, N.; Ranathunga, T.; Hettiarachchi, D.; Udayanga, L.; Abeyewickreme, W.BACKGROUND: Insect growth regulators (IGRs) are considered a novel group of insecticides to control mosquitoes. Novaluron is an IGR with benzoylphenyl urea insecticide, which inhibits chitin synthesis in insects and can reduce insect population density; it is also known to have a high margin of safety for mammals. METHODS: The effective minimum concentration of novaluron formulation EC10 was tested. Six pineapple plantations [control (n = 3) and test (n = 3)] were selected from Meerigama Medical Officer of Health area in Gampaha District, Sri Lanka. Fifteen plots (10 × 10 m) were demarcated in each site with a 200 m distance apart. Leaf axils of 450 pineapple plants (30 plants × 15 plots) were screened for immature stages of Aedes mosquitoes weekly for 12 weeks. The required concentration (20 ppm) of novaluron was sprayed onto the selected pineapple plants (n = 1350) individually in 3 selected test sites for 5-10 s. The reduction in the vector population was interpreted as the percentage of reduction in immature stages of Aedes mosquitoes. RESULTS: The 100% mortality of the Ae. aegypti larvae within 24 h was observed at 20 ppm (0.05 ml of novaluron 100 g/l in 250 ml of water) as the minimum dose. Variation in the number of Aedes larvae present in the control and intervention sites was found to be significantly different throughout the entire observational period (χ2 = 128.29, df = 11, P < 0.001). The total elimination of Aedes larvae continued for up to 2 weeks and a 50% reduction was observed until the 8th week. CONCLUSIONS: The present study emphasizes that novaluron (10% EC) can be used as an effective larvicide at the treatment dose of 20 ppm. The residual effect of the IGR lasted for 12 weeks with a functional efficacy of 8 weeks. The 100% reduction of larval breeding was observed up to the 2nd week after application and the percentage reduction of immature stages remained > 50% until the 8th week. The lowest reduction (34.2%) was observed at 12 weeks after the initial treatment. Therefore, re-treatment may be recommended based on the reduction in the efficacy of the IGR. Keywords: Aedes; Field efficacy; Insect growth regulator; Novaluron; Pineapple plantation.Item Level of Awareness of Dengue Disease among School Children in Gampaha District, Sri Lanka, and Effect of School-Based Health Education Programmes on Improving Knowledge and Practices.(Hindawi Pub. Co., 2019) Radhika, N. M. L.; Gunathilaka, N.; Udayanga, L.; Kasturiratne, A.; Abeyewickreme, W.INTRODUCTION:Limited awareness and nonsystematized health education programmes have contributed adversely to the increase in dengue incidence at schools due to limited attention which has positively contributed to the increase in vector receptivity. The current study was conducted to evaluate the existing level of awareness of dengue infection among a selected group of school children and to assess the effectiveness of dengue awareness programmes to improve the existing knowledge and preventive practices on dengue.METHODS:A cohort of 2,194 students (13-15 years old) from 10 schools at Kelaniya educational zone, Gampaha District, Western Province of Sri Lanka, was enrolled for the current study, which was conducted during 2015-2016. A self-administered questionnaire, comprising 20 questions, was used to evaluate the present knowledge of the students on various aspects of dengue. A structured awareness programme (2 hours) was conducted for students, followed by a reassessment. General Linear Model (GLM) and chi-square test of independence were used to investigate the variations in knowledge levels.RESULTS:The majority of students were characterized by "Good" (46.31%, n= 1016) and "Moderate" (42.62%, n= 935) awareness, while only 2.92% (n= 64) of students fell into the "Excellent" (>80%) category prior to the awareness programme. Even though, existing knowledge of students about dengue ranged between "Moderate" and "Good" categories, awareness of "symptoms & patient care" and "control & prevention practices" were limited. After the programme, the awareness level reached the "Excellent" (41.84%, n=918) level indicating a significant increase by 38.92%, according to the chi-square test (p<0.05 at 95% level of confidence).CONCLUSION:We recommend implementing school-based educational programmes in order to raise the awareness and to translate knowledge into sound practice to control dengue disease epidemics in these areas.Item Predatory efficacy of five locally available copepods on Aedes larvae under laboratory settings: An approach towards bio-control of dengue in Sri Lanka.(San Francisco, CA : Public Library of Science, 2019) Udayanga, L.; Ranathunge, T.; Iqbal, M. C. M.; Abeyewickreme, W.; Hapugoda, M.Many countries are in search of more effective and sustainable methods for controlling dengue vectors, due to undeniable inefficiencies in chemical and mechanical vector control methods. Bio-control of vectors by copepods is an ideal method of using interactions in the natural ecosystem for vector management, with minimum consequences on the environment. Current study determined the predatory efficacy of five locally abundant copepod species on, Aedes larvae under laboratory conditions. Copepods were collected from the pre-identified locations within the districts of Gampaha and Kandy, and identified morphologically. Individual species of copepods were maintained as separate colonies with Paramecium culture and wheat grain as supplementary food. Five adult copepods of each species was introduced into separate containers with 200 larvae (1st instar) of Aedes aegypti. Number of larvae survived in containers were enumerated at 3 hour intervals within a duration of 24 hours. Each experiment was repeated five times. The same procedure was followed for Ae. albopictus. Significance in the variations among predation rates was evaluated with General Linear Modelling (GLM) followed by Tukey's pair-wise comparison in SPSS (version 23). Significant variations in predation rates of studied copepod species were reported (p<0.05), whereby M. leuckarti indicated the highest followed by M. scrassus, while C. languides indicated the lowest predatory efficacy. The effect of different Aedes larval species on the predation rates of copepods remained significant (p<0.05), even though the effect on predatory efficiency was not significant. Based on the findings, both M. leuckarti and M. scrassus, with the highest predatory efficiencies, could be recommended as potential candidates for biological controlling of Aedes vectors in Sri Lanka.Item Use of mechanical and behavioural methods to eliminate female Aedes aegypti and Aedes albopictus for sterile insect technique and incompatible insect technique applications.(BioMed Central, 2019) Gunathilaka, N.; Ranathunge, T.; Udayanga, L.; Wijegunawardana, A.; Gilles, J. R. L.; Abeyewickreme, W.BACKGROUND:Sex separation of mosquitoes at different stages is currently being attempted to ensure the successful release of male mosquitoes in novel vector control approaches. Mechanical and behavioral techniques have been tried most frequently. METHODS: Batches of (n = 300) Aedes aegypti and Ae. albopictus pupae were used for standard sieving (using sieves with 1.12, 1.25, 1.40 and 1.60 mm mesh sizes) and the Fay-Morlan glass plate separation methods. Male and female separation by each method was calculated. For behavioral separation, a spiked blood meal with different concentrations (0, 2, 4, 6, 8 and 10 ppm) of ivermectin and spinosad (spinosyn, 12% w/v), were provided to a batch (n = 300) of adult Ae. aegypti and Ae. albopictus (1:1 sex ratio) followed by observation of mortality. An additional "double feeding method" involved provision of a further blood meal after 24 h, with the same concentrations of ivermectin and spinosad as the initial feeding, followed by a 48-h observation of mortality. All experiments were repeated five times. RESULTS: In the standard sieving method, the percentage of males and females separated at different pore sizes differed significantly (P < 0.05). The majority of the male pupae were collected in the 1.12 mm pore sized sieve for both Ae. aegypti (73%) and Ae. albopictus (69%) while females were retained mainly in the sieve with the pore size of 1.25 mm. In the Fay-Morlan glass plate separation, 99.0% of the Ae. aegypti and 99.2% of the Ae. albopictus introduced male pupae could be separated, but with female contaminations of 16 and 12%, respectively. Provision of a blood meal spiked with 8 ppm of ivermectin under the "double feeding" was identified as the most effective way of achieving 100% female elimination for both Aedes species. CONCLUSIONS: With 100% separation, use of a spiked blood meal is a more effective method of sex separation than the mechanical methods. Application of the spiked blood meal approach as a second separation level for sexes, after applying the Fay-Morlan glass plate method, could achieve 100% sex separation of sexes whilst allowing a reduction in the amount of toxicants required.