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Introduction: The adenomatoid odontogenic tumor is a relatively uncommon odontogenic neoplasm representing about 4.7% of all odontogenic tumors. Objective: The aim of this study was to determine the demographic and clinical profile of the adenomatoid odontogenic tumors in a Sri Lankan population. Methods: Data gathered from the cases received for a period of 38 years from the Department of Oral Pathology, Faculty of Dental Sciences, University of Peradeniya. Request forms, biopsy reports and electronic data base of the department were used to obtain relevant information. Demographic data including age, gender and location of the tumor were included in the analysis. Results: Out of 116 cases of adenomatoid odontogenic tumor, the mean age was 21.02 ± 11.24. It occurs more fre quently in the second decade of life, more prevalent in females, most often associated with the maxilla, predominantly affecting anterior jaw bones and presenting mostly in the right side of the jaw bone. The results from the present study showed the statistically significant relationship with site of occurrence (maxilla/mandible) and age (p 0.05). Conclusion: Adenomatoid odontogenic tumor occurs more frequently in the second decade of life with a significant female predominance and the commonest site is anterior maxilla. This study revealed few differences on demographic and clinical presentations of adenomatoid odontogenic tumor from some regions of the world.

Understanding variations in host-parasite relationships with urbanization is vital for both, public health management and conservation of endemic animals with high anthropogenic interactions. Toque macaques (Macaca sinica) are such endemic old-world monkeys in Sri Lanka. Three macaque sub species inhabit the main climatic zones of the island; M. s. sinica, M. s. aurifrons and M. s. opisthomelas inhabit the dry zone, wet zone, and montane regions of the island, respectively. This study aimed to examine parasite prevalence in this host in association with urbanization. A total of 180 fecal samples were collected from the three sub species of toque macaques inhabiting the main climatic zones (dry, wet, and montane) in Sri Lanka; 20 samples each were collected from urban, suburban, and wild populations representing each climatic zone. Twenty gastrointestinal (GI) parasite genera types i.e. five types of protozoan cysts, two types of trematode ova, four types of cestode ova, eight types of nematode ova, and a single type of acanthocephalan ova were identified. The overall prevalence of parasites was 62% (112/180) with the highest prevalence of Strongyloides infection. In all three sub species, toque macaque populations with proximity to human settlements, including urban and suburban populations, manifested a greater GI parasitic prevalence, mean ova/cyst counts and species richness, compared to their wild counterparts. Importantly, records of five parasite types in toques in Sri Lanka are reported for the first time, while Moniliformis type was reported as a first record in free ranging macaques, globally. This study clearly demonstrated that human contact and habitat modification may influence patterns of parasitic infections in macaques. As most of the parasite types identified manifest zoonotic potential, with public health implications, close associations of macaques may cause a threat to human well-being.

Gastrointestinal (GI) parasites may impose detrimental consequences on wildlife populations due to their capacity to cause mortality and reduce fitness. Additionally, wild animals play an important role in the transmission of zoonoses. Despite this importance, information on GI parasites of tropical wild mammals is critically lacking. The present study aimed to document GI parasites of six wild-dwelling large mammal taxa in Sri Lanka: Asian elephant (Elephas maximus), Sloth bear (Melursus ursinus), civet (Paradoxurus sp.), Leopard (Panthera pardus), Grey langur (Semnopithecus priam) and buffalo (Bubalus sp). Fresh faecal samples (n = 56) collected from the Wasgomuwa National Park, Sri Lanka were subjected to coprological examination using faecal smears, and the brine floatation technique followed by microscopic identification; quantitative data were accrued using the formol-ether method. The survey revealed a high prevalence of GI parasites, where 86% (48/56) of faecal samples screened positive for parasitic infections. Faecal samples of the civet, buffalo and Leopard recorded 100% prevalence, while the lowest (40%) was recorded for the Grey langur. Eight types of GI parasites were documented: protozoan cysts, platyhelminth ova (three types of digenean and a single cyclophillidean type), nematode ova (strongyle, strongyloid, ascarid, and trichuroid types) and rhabditiform larvae. The buffaloes and civets had a comparatively high number and diversity of GI parasites (buffalo: 7 types, H’ = 1.02; civet: 6 types, H’ = 1.52), whilst only a single type (digenean) was detected in the Grey langur. Likewise, parasite loads were also highly variable; highest in the bear (486 per g faeces) and lowest in the monkey (10 per g faeces). The outcome of this survey is important on two accounts; i) to fill the knowledge gap on GI parasites of tropical wild mammals, and ii) the revelation of many first-time parasite-host records for some of the threatened wild-dwelling large mammals in Sri Lanka.

Abstract Prevention of re-establishment (POR) refers to the prevention of malaria outbreak/epidemic occurrence or preventing re-establishment of indigenous malaria in a malaria-free country. Understanding the effectiveness of the various strategies used for POR is, therefore, of vital importance to countries certified as “malaria-free” or to the countries to be thus certified in the near future. This review is based on extensive review of literature on both the POR strategies and elimination schemes of countries, (i) that have reached malaria-free status (e.g. Armenia, Mauritius, Sri Lanka), (ii) those that are reaching pre-elimination stage (e.g. South Korea), and (iii) countries at the control phase (e.g. India). History has clearly shown that poorly implemented POR programmes can result in deadly consequences (e.g. Sri Lanka); conversely, there are examples of robust POR programmes that have sustained malaria free status that can serve as examples to countries working toward elimination. Countries awaiting malaria elimination status should pre-plan their POR strategies. Malaria-free countries face the risk of resurgence mostly due to imported malaria cases; thus, a robust passenger screening programme and cross border collaborations are crucial in a POR setting. In addition, sustained vigilance, and continued funding for the national anti-malarial campaign programme and for related research is of vital importance for POR. With distinct intrinsic potential for malaria in each country, tailor-made POR programmes are built through continuous and robust epidemiological and entomological surveillance, particularly in countries such as Sri Lanka with increased receptivity and vulnerability for malaria transmission. In summary, across all five countries under scrutiny, common strengths of the POR programmes are (i) a multipronged approach, (ii) strong passive, active, and activated passive case detection, (iii) Indoor residual spraying (IRS), and (iv) health education/awareness programmes.

Abstract Background Dopaminergic neuronal loss begins years before motor symptoms appear in Parkinson disease (PD). Thus, reliable biomarkers for early diagnosis and prognosis of PD are an essential pre-requisite to develop disease modifying therapies. Inflammation-derived oxidative stress is postulated to contribute to nigrostriatal degeneration. We evaluated the role of selected serum immune mediators (IFNγ, TNFα, IL-10, and NOx) in PD progression and estimated their usefulness in preclinical diagnosis. Methods This case-control study recruited 72 PD patients with varying disease durations ( 3 years, n = 30) and 56 age- and gender-matched controls (26 with other neurological disorders as disease controls, and 30 healthy controls). Serum cytokine levels and NOx quantified using Sandwich Enzyme Linked Immunosorbent Assay kits, and the Griess test, respectively, were evaluated for diagnostic accuracy of optimal marker combinations by the CombiROC method. PD patients were clinically evaluated for motor and non-motor symptoms, and staged based on Hoehn and Yahr (H-Y) scale. Results A significant increase in serum IFNγ and IL-10 was observed in PD compared to healthy controls (p

Inflammation contributes to knee osteoarthritis (KOA) where many immunological mediators participate in its initiation and progression. Most clinicians manage primary (pKOA) and secondary osteoarthritis (sKOA) alike. Whether immunological profiles of pKOA and sKOA differ remains obscure. Hence, we aimed to differentially identify potential serum immunologic diagnostic markers of pKOA and of sKOA. This case control study used 46 KOA patients (pKOA, n = 30; sKOA, n = 16), and 60 age, gender matched controls (normal healthy, n = 30; systemic lupus erythematosus [SLE] disease controls, n = 30) where serum was assayed for cytokines (TNF-α, IL-1β, IL-6, IL-10) and nitric oxide derivatives (NOx). Sandwich ELISA assessed cytokine levels, while the 'Griess assay' quantified NOx levels. The diagnostic accuracy of optimal marker combinations was evaluated by the CombiROC web tool. Compared with pKOA, sKOA serum displayed significantly elevated levels of pro inflammatory cytokines (TNF-α, IL-1β, IL-6) with a concurrent decrease in the anti-inflammatory cytokine, IL-10 (P

Natural secondary metabolites of sponges of the genus Haliclona are associated with an array of biological activity with therapeutic usage. We investigated the immunopharmacological properties of a presumably novel marine sponge species from Sri Lanka, Haliclona (Soestella) sp. Sponge material was collected from southern Sri Lanka by scuba diving. Sponge identification was based on spicule and skeleton morphology using light microscopy. Selected in vivo and ex vivo tests investigated nonfunctional and functional immunomodulatory activity of the Haliclona (Soestella) sp. crude extract (HSCE) in the Wistar rat model. Compared to the controls, rats orally gavaged daily for 14 consecutive days with 15 mg/kg dose of the HSCE manifested a significant reduction of immune cell counts of total WBCs (by 17%; p

Abstract Background The standard dose of rituximab used in rheumatoid arthritis (RA) is 1000 mg but recent studies have shown that low dose (500 mg) is also effective. Efficacy of low dose rituximab in rheumatoid arthritis (RA) refractory to first-line non-biologic Disease Modifying Anti Rheumatic Drugs (DMARDs), compared to leflunomide is unknown. In a tertiary care referral setting, we conducted a randomized, double blind controlled clinical trial comparing the efficacy and safety of low-dose rituximab-methotrexate combination with leflunomide-methotrexate combination. Methods Patients on methotrexate (10-20 mg/week) with a Disease Activity Score (DAS) > 3.2 were randomly assigned to rituximab (500 mg on days 1 and 15) or leflunomide (10-20 mg/day). The primary end-point was ACR20 at 24 weeks. Sample of 40 had 70% power to detect a 30% difference. ACR50, ACR70, DAS, EULAR good response, CD3 + (T cell), CD19 + (B cell) and CD19 + CD27+ (memory B cell) counts, tetanus and pneumococcal antibody levels were secondary end points. Results Baseline characteristics were comparable in the two groups. At week 24, ACR20 was 85% vs 84% (p = 0.93), ACR50 was 60% vs. 64% (p = 0.79) and ACR70 was 35% vs 32% (P = 0.84), in rituximab and in leflunomide groups respectively. Serious adverse events were similar. With rituximab there was significant reduction in B cells (p 0.05), T cells (P > 0.05) or tetanus antibody levels (P > 0.05). Conclusions Leflunomide-methotrexate combination is as efficacious as low-dose rituximab-methotrexate combination at 24 weeks, in RA patient’s refractory to initial DMARDs. The high responses seen in both groups have favorable cost implications for patients in developing countries. Changes in immune parameters with leflunomide are novel and need further characterization. Trial registration The trial was registered with the Sri Lanka Clinical Trials Registry (SLCTR), a publicly accessible primary registry linked to the registry network of the International Clinical Trials Registry Platform of the WHO (WHO-ICTRP) (registration number: SLCTR/2008/008 dated 16th May 2008).

Abstract Background The leaf concentrate of Carica papaya is a traditionally acclaimed immunomodulatory remedy against numerous diseases; nonetheless comprehensive scientific validation of this claim is limited. The present study thus investigated the immunomodulatory potential of Carica papaya mature leaf concentrate (MLCC) of the Sri Lankan wild type cultivar using nonfunctional and functional immunological assays. Methods Wistar rats (N = 6/ group) were orally gavaged with 3 doses (0.18, 0.36 and 0.72 ml/100g body weight) of the MLCC once daily for 3 consecutive days. Selected nonfunctional (enumeration of immune cells and cytokine levels) and functional (cell proliferation and phagocytic activity) immunological parameters, and acute toxic effects were determined using standard methods. Effect of the MLCC (31.25, 62.5, 125, 250, 500 and 1000 μg/ml) on ex vivo proliferation of bone marrow cells (BMC) and splenocytes (SC), and in vitro phagocytic activity of peritoneal macrophages (PMs), and their corresponding cytokine responses were evaluated. The phytochemical profile of the MLCC was established using liquid chromatography-mass spectrometry (LS-MS) and Gas chromatography-mass spectrometry (GC-MS). Results Counts of rat platelets, total leukocytes, lymphocyte and monocyte sub populations, and BMCs were significantly augmented by oral gavage of the MLCC (p

Weligama coconut leaf wilt disease (WCLWD) causes heavy losses in the coconut cultivations of southern Sri Lanka. The in-house developed and validated indirect ELISA was based on specific polyclonal antibodies raised in female New Zealand White rabbits, against partially purified WCLWD associated phytoplasma. This ELISA has the potential to distinguish secA PCR confirmed, WCLWD associated phytoplasma positive palms from phytoplasma free palms at high accuracy (93%) and sensitivity (92.7%), but with marginal specificity (79%). The calculated ELISA cross reactivity index (CRI) values were low for sugarcane white leaf (7%) and sugarcane grassy shoot (8%) infected leaves, but with marked highCRIfor both Bermuda grass white leaf (69%) and areca nut yellow leaf (70%) infected leaves. SecA gene based phylogenetic relationships of the WCLWD associated phytoplasma with these other locally prevalent phytoplasma strains elucidated this immunological cross reactivity, which was further reiterated by virtual restriction fragment length polymorphism analysis. Based on scanning electron microscopy, this study provides additional visual evidence, for the presence of phytoplasmas in WCLWD infected tissues.

Background & objectives: Artemisinin isolated from Artemisia annua is the most potent antimalarial against chloroquine resistant Plasmodium falciparum malaria. We previously reported that the ethanolic leaf extract of Artemisia vulgaris, an invasive weed and the only Artemisia species in Sri Lanka, possess both potent and safe antimalarial activity (in terms of antiparasitic properties) in a P. berghei murine malaria model. We report here a prototype study that investigated antidisease activities of A. vulgaris ethanolic leaf extract (AVELE) in a P. berghei ANKA murine malaria model that elicit pathogenesis similar to falciparum malaria. Profound thrombocytosis and thrombocytopenia in mice were detected in early-stage (Day 3), and at a later stage of infection (Day 6), respectively. Plasmodium berghei infected mice, 7 or 8 days post-infection reached end-stage disease with rapid drop in body temperature and usually die within 24 h, as a consequence of cerebral malaria. Methods: Three doses of the AVELE (500, 750 and 1000 mg/kg) were used to assess antidisease activity of A. vulgaris in terms of survival, effects on thrombocyte related pathology and end-stage disease, antipyretic activity, and antinociception, using standard methodology. Results: The 1000 mg/kg dose of AVELE significantly increased survival, reversed the profound thrombocytopenia/ thrombocytosis (p ≤0.01), altered the end-stage disease (p ≤0.05), and manifested significant antipyretic and antinociceptive (p ≤0.05) activities. Interpretation & conclusion: We conclude that a crude ethanolic leaf extract of A. vulgaris, showed potent antimalarial properties, in terms of antidisease activities; antipyretic activity, peripheral and central antinociception, increased survival, averted end-stage disease and reversed hrombocytopenia/thrombocytosis.

Silencing of tumor-suppressor genes (TSGs) by DNA promoter hypermethylation is an early event in carcinogenesis; hence, TSGs may serve as early tumor biomarkers. We determined the promoter methylation levels of p16INK4a, RASSF1A, TIMP3, and PCQAP/MED15 TSGs in salivary DNA from oral cancer (OC) and oropharyngeal cancer (OPC) patients, using methylation-specific PCR coupled with densitometry analysis. We assessed the association between DNA methylation of individual TSGs with OC and OPC risk factors. The performance and the clinical validity of this quadruple-methylation marker panel were evaluated in discriminating OC and OPC patients from healthy controls using the CombiROC web tool. Our study reports that RASSF1A, TIMP3, and PCQAP/MED15 TSGs were significantly hypermethylated in OC and OPC cases compared to healthy controls. DNA methylation levels of TSGs were significantly augmented by smoking, alcohol use, and betel quid chewing, indicating the fact that frequent exposure to risk factors may drive oral and oropharyngeal carcinogenesis through TSG promoter hypermethylation. Also, this quadruple-methylation marker panel of p16INK4a, RASSF1A, TIMP3, and PCQAP/MED15 TSGs demonstrated excellent diagnostic accuracy in the early detection of OC at 91.7% sensitivity and 92.3% specificity and of OPC at 99.8% sensitivity and 92.1% specificity from healthy controls.

Background & objectives: Artemisinin isolated from Artemisia annua is the most potent antimalarial drug against chloroquine-resistant Plasmodium falciparum malaria. Artemisia vulgaris, an invasive weed, is the only Artemisia species available in Sri Lanka. A pilot study was undertaken to investigate the antiparasitic activity of an A. vulgaris ethanolic leaf extract (AVELE) in a P. berghei ANKA murine malaria model that elicits pathogenesis similar to falciparum malaria. Methods: A 4-day suppressive and the curative assays determined the antiparasitic activity of AVELE using four doses (250, 500, 750 and 1000 mg/kg), Coartem® as the positive control and 5% ethanol as the negative control in male ICR mice infected with P. berghei. Results: The 500, 750 and 1000 mg/kg doses of AVELE significantly (p ≤0.01) inhibited parasitaemia by 79.3, 79.6 and 87.3% respectively, in the 4-day suppressive assay, but not in the curative assay. Chronic administration of the high dose of AVELE ruled out overt signs of toxicity and stress as well as hepatotoxicity, renotoxicity and haematotoxicity. Interpretation & conclusion: The oral administration of a crude ethonolic leaf extract of A. vulgaris is non-toxic and possesses potent antimalarial properties in terms of antiparasitic activity.

Parasitic investigations of the subspecies Mus mayori pococki were carried out in four rainforests that included two man and biosphere reserves and two forest reserves of Sri Lanka from October 2006 to August 2007. Rodents were live-trapped using Shermans traps. Of the 117 individuals of M. m. pococki captured 73% were infested with four types of ectoparasites mites of the genus Echinolaelaps, a louse Polyplax serrata, a larval stage of hard tick Ixodes and pseudoscorpions of the genus Megachernes. Mites were the most abundant ectoparasite of this rodent host. Faecal examination revealed the presence of a nematode larva of the Order Strongylida and five types of parasitic ova; three nematode ova types i.e. strongyle, strongyloides, ascarid types, and cestode and mite ova. In comparison to the non-infested hosts, those infested did not show a significant difference in body weight and size. Both sexes had an equal probability of being exposed to ectoparasites. The present study on the parasitic investigations of M. m. pococki reports four new host-ectoparasite and six new endoparasitic records for the Sri Lankan rodent host.

Understanding variations in host-parasite relationships with urbanization is vital for both, public health management and conservation of endemic animals with high anthropogenic interactions. Toque macaques (Macaca sinica) are such endemic old-world monkeys in Sri Lanka. Three macaque sub species inhabit the main climatic zones of the island; M. s. sinica , M. s. aurifrons and M. s. opisthomelas inhabit the dry zone, wet zone, and montane regions of the island, respectively. This study aimed to examine parasite prevalence in this host in association with urbanization. A total of 180 fecal samples were collected from the three sub species of toque macaques inhabiting the main climatic zones (dry, wet, and montane) in Sri Lanka; 20 samples each were collected from urban, suburban, and wild populations representing each climatic zone. Twenty gastrointestinal (GI) parasite genera types i.e. five types of protozoan cysts, two types of trematode ova, four types of cestode ova, eight types of nematode ova, and a single type of acanthocephalan ova were identified. The overall prevalence of parasites was 62% (112/180) with the highest prevalence of Strongyloides infection. In all three sub species, toque macaque populations with proximity to human settlements, including urban and suburban populations, manifested a greater GI parasitic prevalence, mean ova/cyst counts and species richness, compared to their wild counterparts. Importantly, records of five parasite types in toques in Sri Lanka are reported for the first time, while Moniliformis type was reported as a first record in free ranging macaques, globally. This study clearly demonstrated that human contact and habitat modification may influence patterns of parasitic infections in macaques. As most of the parasite types identified manifest zoonotic potential, with public health implications, close associations of macaques may cause a threat to human well-being. [Display omitted] • GI parasites of the 3 endemic, endangered Sri Lankan macaque subspecies recorded. • Parasitic infections were examined in realtion to urbanization of macaque habitats. • 20 genera types of protozoan, helminth and acanthocephalan groups identified. • Urban/sub-urban habitats led to significantly higher parasite prevalence than in the wild. • Moniliformis sp. in macaques was recorded for the first time, globally, along with five novel local records of parasite-toque macaque associations. [ABSTRACT FROM AUTHOR]

Gastrointestinal (GI) parasites may impose detrimental consequences on wildlife populations due to their capacity to cause mortality and reduce fitness. Additionally, wild animals play an important role in the transmission of zoonoses. Despite this importance, information on GI parasites of tropical wild mammals is critically lacking. The present study aimed to document GI parasites of six wild-dwelling large mammal taxa in Sri Lanka: Asian elephant (Elephas maximus), Sloth bear (Melursus ursinus), civet (Paradoxurus sp.), Leopard (Panthera pardus), Grey langur (Semnopithecus priam) and buffalo (Bubalus sp). Fresh faecal samples (n = 56) collected from the Wasgomuwa National Park, Sri Lanka were subjected to coprological examination using faecal smears, and the brine floatation technique followed by microscopic identification; quantitative data were accrued using the formol-ether method. The survey revealed a high prevalence of GI parasites, where 86% (48/56) of faecal samples screened positive for parasitic infections. Faecal samples of the civet, buffalo and Leopard recorded 100% prevalence, while the lowest (40%) was recorded for the Grey langur. Eight types of GI parasites were documented: protozoan cysts, platyhelminth ova (three types of digenean and a single cyclophillidean type), nematode ova (strongyle, strongyloid, ascarid, and trichuroid types) and rhabditiform larvae. The buffaloes and civets had a comparatively high number and diversity of GI parasites (buffalo: 7 types, H' = 1.02; civet: 6 types, H' = 1.52), whilst only a single type (digenean) was detected in the Grey langur. Likewise, parasite loads were also highly variable; highest in the bear (486 per g faeces) and lowest in the monkey (10 per g faeces). The outcome of this survey is important on two accounts; i) to fill the knowledge gap on GI parasites of tropical wild mammals, and ii) the revelation of many first-time parasite-host records for some of the threatened wild-dwelling large mammals in Sri Lanka. [Display omitted] • GI parasites of six native large mammals from the wild recorded from Sri Lanka. • Novel GI parasite records of locally and globally endangered large mammals reported. • High prevalence (86%) of infected fecal samples; eight GI parasite types identified. • Buffaloes and civets had high parasite diversity; highest parasite load in the Sloth bear. • GI parasite diversity and prevalence differed between wild and captive mammals. [ABSTRACT FROM AUTHOR]

Impacts of heavy metal toxicity on the immune system of the Indian green frog, Euphlyctis hexadactylus, in Bellanwila Attidiya, an urban wetland polluted with high levels of heavy metals, compared to the reference site in Bolgoda, in Sri Lanka was investigated. Significantly higher accumulation of selected heavy metals, copper (Cu), zinc (Zn), lead (Pb), and cadmium (Cd) were detected by AAS in frog liver and gastrocnemius muscle, in the polluted site than in the reference site. Non-functional immunotoxicity tests; total WBC, splenocyte and bone marrow cell counts, spleen weight/body weight ratio, neutrophil/lymphocyte ratio and basal immunoglobulin levels, and phagocytic capacity of peritoneal macrophages (immune functional test) were carried out using standard methodology. Test parameters recorded significantly lower values for frogs of the polluted site compared with their reference site counterparts, indicative of lowered immune response of frogs in the former site. In vitro phagocytic assay based on NBT dye reduction, measured the stimulation index (SI) of E. hexadactylus blood leukocytes, splenocytes and peritoneal macrophages, where SIs of frogs in the polluted site were significantly lower. Also, in vitro exposure of frog phagocytes to Cu, Zn, Pb and Cd at 10 −2 –10 −10 M, showed immunomodulation i.e. low concentrations stimulated phagocytosis while increased concentrations showed a trend towards immunosuppression. IC 50 values indicated Cd>Zn>Cu>Pb as the decreasing order of the potential of phagocytosis inhibition. In conclusion, this study clearly demonstrated immunomodulation of E. hexadactylus, stimulated by heavy metals. In-vitro studies evidently suggested the use of phagocytosis as a biomarker in Ecoimmunotoxicology to detect aquatic heavy metal pollution. [ABSTRACT FROM AUTHOR]

Objectives To do mapping and modeling of conformational B cell epitope regions of highly conserved and protective regions of three merozoitecandidate vaccine proteins of Plasmodium vivax ( P. vivax ), ie . merozoite purface protein-1 ( Pv MSP-1), apical membrane antigen −1 domain ∏ ( Pv AMA1-D∏) and region ∏ of the Duffy binding protein ( Pv DBP∏), and to analyze the immunogenic properties of these predicted epitopes. Methods 3-D structures of amino acid haplotypes from Sri Lanka (available in GeneBank) of Pv MSP-1 19 ( n =27), Pv AMA1-D∏ ( n =21) and Pv DBP∏ ( n =33) were modeled. SEPPA, selected as the best online server was used for conformational epitope predictions, while prediction and modeling of protein structure and properties related to immunogenicity was carried out with Geno3D server, SCRATCH Protein Server, NetSurfP Server and standalonesoftware, Genious 5.4.4. Results SEPPA revealed that regions of predicted conformational epitopes formed 4 clusters in Pv MSP-I 19 , and 3 clusters each in Pv AMA1-D∏ and Pv DBP∏, all of which displayed a high degree of hydrophilicity, contained solvent exposed residues, displayed high probability of antigenicity and showed positive antigenic propensity values, that indicated high degree of immunogenicity. Conclusions Findings of this study revealed and confirmed that different parts of the sequences of each of the conserved regions of the three selected potential vaccine candidate antigens of P. vivax are important with regard to conformational epitope prediction that warrants further laboratory experimental investigations in in vivo animal models. [ABSTRACT FROM AUTHOR]

The article discusses a study which presents the first systematic parasitic survey of reptiles of Sri Lanka that were acquired in the National Zoological Garden in the country. Accordingly, it serves as an addition to the existing knowledge database on reptilian parasites. The study documents the ectoparasites and intestinal parasites of 19 selected reptilian species including snakes, chelonians, and crocodilian. It cites that the adult intestinal parasites were gathered from the feces of hosts provided with anthelmintic. Results show that 62% of the reptiles were infected with parasites, and four new scientific host-parasite records were observed.

Abstract: Objective: To study the immunological profile, disease characteristics and socioeconomic status of a population of patients with rheumatoid arthritis (RA) in Sri Lanka. Methods: A case-control study was undertaken to characterize the immunoglobulin profiles of 105 RA and, age and gender matched osteoarthritis (OA) patients (n = 30) from the National Hospital, Sri Lanka. Healthy, non-arthritic individuals (n=30) served as controls. Sera were assayed for immunoglobulins [IgG, IgM, IgE and IgA isotypes] by establishing sandwich type ELISA. IgM, IgG and IgA rheumatoid factors (RFs) of 162RA patients were assayed by indirect ELISA. Disease characteristics and socioeconomic factors were accrued via an interviewer-administered questionnaire. Results: Higher IgG, IgM, IgE, IgA and lower IgG1, IgG2 levels were observed in RA sera compared with controls (P < 0.05). Novel correlations between disease characteristics and immunoglobulins, as well as group-specific correlation matrices of immunoglobulins and RFs (P < 0.05) of seropositive and seronegative patients, were found. Higher IgM-RF and IgA-RF levels in seropositives and IgG-RF in seronegatives were evident compared with controls (P < 0.05). Immunoglobulin and RF profiles did not reflect gender disparity of RA (P > 0.05). Proportions of seropositives with nodules and erosions were significantly higher than seronegatives (P < 0.05). While IgM-RF and erosions positively correlated in the seropositives (P < 0.05), the seronegatives showed an inverse correlation between IgG-RF and erosions (P < 0.01). Familial clustering imposed a relative risk of 4.7 for developing seropositive RA. Conclusions: This model study provides baseline information on pathogenetic aspects of RA in Sri Lanka, which may have implications for further research on management of the disease. [Copyright &y& Elsevier]

An 11-month-old infant presented with a subcutaneous nodule in the right cheek which was found by ultrasonography to be a worm. Following treatment with di-ethylcarbamazine, a worm emerged from the left upper eyelid which was confirmed to be Dirofilaria repens. Dirofilariasis usually manifests as a single lesion and is rare in infants. [ABSTRACT FROM AUTHOR]

Inflammation-derived oxidative stress is postulated to contribute to neuronal damage leading to poor clinical outcomes in Acute Ischemic Stroke (AIS). We aimed to investigate the association between serum levels of selected cytokines (IL-1β, IFN-γ, IL-4), and vitamin D in ischemic stroke progression, and their accuracy in predicting AIS prognosis, among Sri Lankans. We compared 60 AIS patients admitted in 4 phases post-stroke onset (<6 h; 6-24 h; 24-48 h; 48-96 h; n = 15/phase), with 15 age- and sex-matched controls. The 30-day functional outcome (FO) was assessed using the modified Rankin Scale (mRS). Serum cytokine and vitamin D levels were quantified using sandwich ELISAs, and competitive ELISA, respectively. The CombiROC web tool established optimal prognostic biomarker combinations. Serum IL-1β and IFN-γ were elevated in all four phases following stroke onset while IL-4 was elevated exclusively in the recovery phase (48-96 h) (p<0.05). Th1 bias polarization of the Th1:Th2 cytokine (IFN-γ:IL-4) ratio occurred with AIS progression while a Th2 bias occurred during AIS recovery (p<0.05). Lower serum IL-1β and higher IL-4 levels were associated with a good FO (p<0.05), while lower Vitamin D levels were related to a poor FO (p = 0.001). The triple-biomarker panel, IL-4- IFN-γ -Vit D, accurately predicted AIS prognosis (sensitivity = 100%, specificity = 91.9%, area under the curve = 0.98). Serum immunologic mediators IFN-γ, IL-4, and vitamin D may be useful biomarkers of AIS prognosis and may serve as therapeutic targets in improving stroke outcomes. Vitamin D supplementation may improve the prognosis of AIS patients. Furthermore, binary logistic model fitted for FO indicated Th1:Th2 cytokine ratio (IFN-γ:IL-4), vitamin D status, history of stroke, and ischemic heart disease as significant predictors of AIS prognosis., Competing Interests: The authors declare that no competing interests exist., (Copyright: © 2024 Samarakoon et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Ethnopharmacological Relevance: Sri Lankan traditional medicine uses Vernonia zeylanica and Mallotus repandus broadly for the treatment of a multitude of disease conditions, including wound healing., Aim of the Study: We aimed to scientifically validate the safety and efficacy of wound healing of an aqueous distillate of Vernonia zeylanica and Mallotus repandus (ADVM) mature leaves, tested on primary human dermal fibroblasts., Materials and Methods: Human dermal fibroblasts isolated from clinical waste from circumcision surgery were characterized by flowcytometry and trilineage differentiation. The MTT dye reduction assay, and the ex vivo wound healing scratch assay established wound healing properties of ADVM using the primary human dermal fibroblast cell line. Upregulation of genes associated with wound healing (MMP3, COL3A1, TGFB1, FGF2) were confirmed by RT qPCR. GC-MS chromatography evaluated the phytochemical composition of ADVM., Results: Compared to the synthetic stimulant, β fibroblast growth factor, ADVM at 0.25% concentration on the primary dermal fibroblast cell line exhibited significant ex vivo, (i) 1.7-fold % cell viability (178.7% vs 304.3 %, p < 0.001), (ii) twofold greater % wound closure (%WC) potential (47.74% vs 80.11%, p < 0.001), and (iii) higher rate of % WC (3.251 vs 3.456 % WC/h, p < 0.05), sans cyto-genotoxicity. Up regulated expression of FGF2, TGFB1, COL3A1 and MMP3, genes associated with wound healing, confirmed effective stimulation of pathways of the three overlapping phases of wound healing (P < 0.05). GC-MS profile of ADVM characterized four methyl esters, which may be posited as wound healing phytochemicals., Conclusions: Exceeding traditional medicine claims, the exvivo demonstration of rapid skin regeneration, reiterated by upregulated expression of genes related to wound healing pathways, sans cytotoxicity, propounds ADVM, cued from traditional medicine, as a potential safe and effective natural stimulant for rapid wound-healing. Additionally, it may serve as an effective proliferative stimulant of dermal fibroblasts for cell therapy, with potential in reparative and regenerative therapy of skin disorders., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Understanding variations in host-parasite relationships with urbanization is vital for both, public health management and conservation of endemic animals with high anthropogenic interactions. Toque macaques ( Macaca sinica ) are such endemic old-world monkeys in Sri Lanka. Three macaque sub species inhabit the main climatic zones of the island; M . s. sinica , M . s. aurifrons and M . s. opisthomelas inhabit the dry zone, wet zone, and montane regions of the island, respectively. This study aimed to examine parasite prevalence in this host in association with urbanization. A total of 180 fecal samples were collected from the three sub species of toque macaques inhabiting the main climatic zones (dry, wet, and montane) in Sri Lanka; 20 samples each were collected from urban, suburban, and wild populations representing each climatic zone. Twenty gastrointestinal (GI) parasite genera types i.e . five types of protozoan cysts, two types of trematode ova, four types of cestode ova, eight types of nematode ova, and a single type of acanthocephalan ova were identified. The overall prevalence of parasites was 62% (112/180) with the highest prevalence of Strongyloides infection. In all three sub species, toque macaque populations with proximity to human settlements, including urban and suburban populations, manifested a greater GI parasitic prevalence, mean ova/cyst counts and species richness, compared to their wild counterparts. Importantly, records of five parasite types in toques in Sri Lanka are reported for the first time, while Moniliformis type was reported as a first record in free ranging macaques, globally. This study clearly demonstrated that human contact and habitat modification may influence patterns of parasitic infections in macaques. As most of the parasite types identified manifest zoonotic potential, with public health implications, close associations of macaques may cause a threat to human well-being., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Authors. Published by Elsevier Ltd on behalf of Australian Society for Parasitology.)

Inflammation contributes to knee osteoarthritis (KOA) where many immunological mediators participate in its initiation and progression. Most clinicians manage primary (pKOA) and secondary osteoarthritis (sKOA) alike. Whether immunological profiles of pKOA and sKOA differ remains obscure. Hence, we aimed to differentially identify potential serum immunologic diagnostic markers of pKOA and of sKOA. This case control study used 46 KOA patients (pKOA, n = 30; sKOA, n = 16), and 60 age, gender matched controls (normal healthy, n = 30; systemic lupus erythematosus [SLE] disease controls, n = 30) where serum was assayed for cytokines (TNF-α, IL-1β, IL-6, IL-10) and nitric oxide derivatives (NOx). Sandwich ELISA assessed cytokine levels, while the 'Griess assay' quantified NOx levels. The diagnostic accuracy of optimal marker combinations was evaluated by the CombiROC web tool. Compared with pKOA, sKOA serum displayed significantly elevated levels of pro inflammatory cytokines (TNF-α, IL-1β, IL-6) with a concurrent decrease in the anti-inflammatory cytokine, IL-10 (P<0.05). This was reiterated by significantly higher Th1:Th2 (TNF-α: IL-10) serum cytokine ratio observed in sKOA compared to that of pKOA. The CombiROC curves identified TNF-α, IL-1β, IL-6 and NOx as the best performing panel of potential diagnostic markers to discriminate pKOA from control groups (~97% accuracy, 90% Sensitivity [SE] and 98% specificity [SP]), while TNF-α, IL-1β and IL-6 discriminated sKOA from control groups (~100% accuracy, 100% SE, and 98% SP). The study identified discrete serum immune biomarker panels to differentiate between pKOA (TNF-α, IL-1β, IL-6 and NOx) and sKOA (TNF-α, IL-1β and IL-6). These findings may assist in developing distinct therapeutic agents for the two types of KOA., Competing Interests: The authors have declared that no competing interests exist.

Natural secondary metabolites of sponges of the genus Haliclona are associated with an array of biological activity with therapeutic usage. We investigated the immunopharmacological properties of a presumably novel marine sponge species from Sri Lanka, Haliclona ( Soestella ) sp. Sponge material was collected from southern Sri Lanka by scuba diving. Sponge identification was based on spicule and skeleton morphology using light microscopy. Selected in vivo and ex vivo tests investigated nonfunctional and functional immunomodulatory activity of the Haliclona ( Soestella ) sp. crude extract (HSCE) in the Wistar rat model. Compared to the controls, rats orally gavaged daily for 14 consecutive days with 15 mg/kg dose of the HSCE manifested a significant reduction of immune cell counts of total WBCs (by 17%; p < 0.01), lymphocytes (38%), platelets (52%), splenocytes (20%), and bone marrow cells (BMC; 60%) ( p < 0.001), with a concurrent increase in the neutrophil : lymphocyte ratio ( p < 0.05); RBC counts abated by 53% ( p < 0.001). A significant reduction of the splenosomatic index was evident with the 10 and 15 mg/kg doses ( p < 0.001). Rat plasma TNF- α cytokine level was augmented by tenfold ( p < 0.001), IL-6 level by twofold ( p < 0.01) with the 15 mg/kg HSCE treatment, while IL-10 was detectable in rat plasma only with this treatment; the corresponding Th 1  : Th 2 cytokine ratio (TNF- α  : IL-10) was indicative of an unequivocal Th1-skewed cytokine response ( p < 0.01). Ex vivo bone marrow cell and splenocyte proliferation were significantly and dose dependently impaired by HSCE (IC 50 0.719 and 0.931  μ g/mL, respectively; p < 0.05). Subacute toxicity testing established that HSCE was devoid of general toxic, hepatotoxic, and nephrotoxic effects. In conclusion, HSCE was orally active, nontoxic, and effectively suppressed nonfunctional and functional immunological parameters of Wistar rats, suggestive of the potential use of the HSCE as an immunosuppressant drug lead., Competing Interests: The authors do not have any conflicts of interest regarding this document., (Copyright © 2020 Varuni Gunathilake et al.)

Silencing of tumor-suppressor genes (TSGs) by DNA promoter hypermethylation is an early event in carcinogenesis; hence, TSGs may serve as early tumor biomarkers. We determined the promoter methylation levels of p16 INK4a , RASSF1A , TIMP3 , and PCQAP/MED15 TSGs in salivary DNA from oral cancer (OC) and oropharyngeal cancer (OPC) patients, using methylation-specific PCR coupled with densitometry analysis. We assessed the association between DNA methylation of individual TSGs with OC and OPC risk factors. The performance and the clinical validity of this quadruple-methylation marker panel were evaluated in discriminating OC and OPC patients from healthy controls using the CombiROC web tool. Our study reports that RASSF1A , TIMP3 , and PCQAP/MED15 TSGs were significantly hypermethylated in OC and OPC cases compared to healthy controls. DNA methylation levels of TSGs were significantly augmented by smoking, alcohol use, and betel quid chewing, indicating the fact that frequent exposure to risk factors may drive oral and oropharyngeal carcinogenesis through TSG promoter hypermethylation. Also, this quadruple-methylation marker panel of p16 INK4a , RASSF1A , TIMP3 , and PCQAP/MED15 TSGs demonstrated excellent diagnostic accuracy in the early detection of OC at 91.7% sensitivity and 92.3% specificity and of OPC at 99.8% sensitivity and 92.1% specificity from healthy controls., Competing Interests: The authors declare no conflicts of interest.

Weligama coconut leaf wilt disease (WCLWD) causes heavy losses in the coconut cultivations of southern Sri Lanka. The in-house developed and validated indirect ELISA was based on specific polyclonal antibodies raised in female New Zealand White rabbits, against partially purified WCLWD associated phytoplasma. This ELISA has the potential to distinguish secA PCR confirmed, WCLWD associated phytoplasma positive palms from phytoplasma free palms at high accuracy (93%) and sensitivity (92.7%), but with marginal specificity (79%). The calculated ELISA cross reactivity index (CRI) values were low for sugarcane white leaf (7%) and sugarcane grassy shoot (8%) infected leaves, but with marked highCRIfor both Bermuda grass white leaf (69%) and areca nut yellow leaf (70%) infected leaves. SecA gene based phylogenetic relationships of the WCLWD associated phytoplasma with these other locally prevalent phytoplasma strains elucidated this immunological cross reactivity, which was further reiterated by virtual restriction fragment length polymorphism analysis. Based on scanning electron microscopy, this study provides additional visual evidence, for the presence of phytoplasmas in WCLWD infected tissues., Competing Interests: The authors have declared that no competing interests exist.

Background: The standard dose of rituximab used in rheumatoid arthritis (RA) is 1000 mg but recent studies have shown that low dose (500 mg) is also effective. Efficacy of low dose rituximab in rheumatoid arthritis (RA) refractory to first-line non-biologic Disease Modifying Anti Rheumatic Drugs (DMARDs), compared to leflunomide is unknown. In a tertiary care referral setting, we conducted a randomized, double blind controlled clinical trial comparing the efficacy and safety of low-dose rituximab-methotrexate combination with leflunomide-methotrexate combination., Methods: Patients on methotrexate (10-20 mg/week) with a Disease Activity Score (DAS) > 3.2 were randomly assigned to rituximab (500 mg on days 1 and 15) or leflunomide (10-20 mg/day). The primary end-point was ACR20 at 24 weeks. Sample of 40 had 70% power to detect a 30% difference. ACR50, ACR70, DAS, EULAR good response, CD3 + (T cell), CD19 + (B cell) and CD19 + CD27+ (memory B cell) counts, tetanus and pneumococcal antibody levels were secondary end points., Results: Baseline characteristics were comparable in the two groups. At week 24, ACR20 was 85% vs 84% (p = 0.93), ACR50 was 60% vs. 64% (p = 0.79) and ACR70 was 35% vs 32% (P = 0.84), in rituximab and in leflunomide groups respectively. Serious adverse events were similar. With rituximab there was significant reduction in B cells (p < 0.001), memory B cells (p < 0.001) and pneumococcal antibody levels (P < 0.05) without significant changes in T cells (p = 0.835) and tetanus antibody levels (p = 0.424) at 24 weeks. With leflunomide, significant reduction in memory B cells (p < 0.01) and pneumococcal antibody levels (p < 0.01) occurred without significant changes in B cells (P > 0.05), T cells (P > 0.05) or tetanus antibody levels (P > 0.05)., Conclusions: Leflunomide-methotrexate combination is as efficacious as low-dose rituximab-methotrexate combination at 24 weeks, in RA patient's refractory to initial DMARDs. The high responses seen in both groups have favorable cost implications for patients in developing countries. Changes in immune parameters with leflunomide are novel and need further characterization., Trial Registration: The trial was registered with the Sri Lanka Clinical Trials Registry (SLCTR), a publicly accessible primary registry linked to the registry network of the International Clinical Trials Registry Platform of the WHO (WHO-ICTRP) (registration number: SLCTR/2008/008 dated 16th May 2008).

Phytoplasmas that infect gramineous plants, including Napier grass stunt, sugarcane whiteleaf, sugarcane grassy shoot, and Bermuda grass whiteleaf, have been classified into two closely related groups, 16SrXI and 16SrXIV, based on the 16S ribosomal RNA (rRNA) gene. Subsequently, phytoplasmas associated with coconut and Areca palm in southern India and Sri Lanka have been added into the 16SrXI group. However, the 16S rRNA gene gives relatively poor resolution between these phytoplasmas. In this study, a new set of universal phytoplasma primers that amplify approximately 1 kb of the leucyl transfer RNA synthetase (leuS) gene have been validated on a broad range of phytoplasma taxonomic groups. These have been used along with partial sequences of the secA gene to clarify the taxonomic classification of 16SrXI and 16SrXIV phytoplasmas. Based on this data, the sugarcane whiteleaf and grassy shoot phytoplasmas appear to be the same phytoplasma. The Napier grass stunt phytoplasma forms a distinct group from the Bermuda grass whiteleaf and sugarcane phytoplasmas, suggesting that Napier grass stunt should be in its own 'Candidatus Phytoplasma sp.'. The phytoplasmas associated with coconut and arecanut in southern India and Sri Lanka, which are in the same 16SrXI group, appear in different groups based on secA analysis.

One approach towards the development of a vaccine against malaria is to immunize against the parasite sexual stages that mediate transmission of the parasite from man to mosquito. Antibodies against these stages, ingested with the blood meal, inhibit the parasite development in the mosquito vector, constituting "transmission blocking immunity." Most epitopes involved in transmission-blocking immunity depend on the tertiary conformational structure of surface antigens. However, one of the transmission-blocking monoclonal antibodies we have raised against Plasmodium vivax reacts with a linear epitope on both asexual stages and gametes. This monoclonal antibody (A12) is capable of totally blocking development of the parasite in the mosquito host when tested in membrane feeding assays with gametocytes from P. vivax-infected patients. Immune screening of a P. vivax lambda gt11 genomic expression library with A12 led to the isolation of a clone to which was mapped the six-amino acid epitope recognized by A12. Antisera raised in mice against a 12-mer synthetic peptide containing this epitope coupled to bovine serum albumin not only had high titers of antipeptide antibodies as measured by enzyme-linked immunosorbent assay, but in addition recognized the same 24- and 57-kD parasite components as A12 on Western blots and reacted with the parasite by immunofluorescence. When tested in membrane feeding assays, these antibodies have significant suppressive effects on parasite development in the mosquito.