Cyanotoxin Contamination in Human Urine in CKDu High Prevalence Padaviya and Low Prevalence Rajanganaya and Galnewa in Anuradhapura District, Sri Lanka

The naturally derived cyanotoxin, Cylindrospermopsin (CYN) and Microcystin-LR (MC-LR), has been recorded in freshwater systems worldwide and this toxin poses a threat on human health. The extraction and detection of CYN and MC-LR in source water was well documented and information on contamination of CYN and MC-LR in biological samples are limited. CYN and MC-LR are the most toxic cyanotoxins that affects the function of kidney, liver and listed as one of the hypothesis for CKDu in Sri Lanka. Thus, the present study was carried out to determine contamination of CYN and MC-LR in randomly selected CKDu patients’ and non-patients’ (as control) urine. According to the statistics published by the Ministry of Health, Padaviya was selected as high prevalence of CKDu area for the study where Rajanganaya and Galnewa were selected as low prevalence of CKDu in Anuradhapura District. Angunakolapallassa in Hambanthota district was selected as control area considering the occupational exposure, climate and other demographic factors. Eight medical clinics were conducted in two districts (Anuradhapura and Hambanthota) and 225 people including CKDu patients and non- patients were recruited to collect urine samples. Along with the medical clinics, well water sample from each recruitment was collected for CYN and MC-LR analysis using the ELISA method. The collected, urine samples were subjected to CYN and MC-LR analysis, following centrifugation, pH manipulation and Solid Phase Extraction (SPE). CYN and MC-LR quantification was done using the High Performance Liquid Chromatography (HPLC). CYN and MC-LR concentrations in well water of Padaviya-Bogaswewa, Padaviya-Parakramapura, Galnewa and Rajanganaya were ranged between; MC-LR=1.0±0.01 μgl-1-2.6±0.16 μgl-1, CYN=1.0±0.01 μgl-1–3.6±0.18 μgl-1, MC-LR=1.0±0.01 μgl-1-2.4±0.12μ gl-1, CYN=1.0±0.01 μgl-1-7.6±0.34 μgl-1, MC-LR=1.0±0.01 μgl-1-1.6±0.02 μgl-1, CYN=1.0±0.01 μgl-1-2.9±0.06 μgl-1, MC-LR=1.0±0.01 μgl-1 -1.9±0.07 μgl-1, CYN=1.0±0.01 μgl-1-2.2±0.03 μgl-1 respectively. Excitingly in Angunakolapellessa samples, neither MC-LR nor CYN detected in well water. Mean concentrations of CYN and MC-LR in urine samples were ranged between 1.0±0.01 μgml-1-8.6±0.26 μgml-1 and 0.2±0.01 μgml-1-2.4±0.12 μgml-1 in Padaviya-Bogaswewa where concentration between 0.8±0.23 μgml-1-4.1 ±0.16 μgml-1 and 0.6±0.01 μgml-1-1.2±0.12 μgml-1 recorded in Padaviya-Parakramapura. In Galnewa and Rajanganaya, mean concentrations of CYN and MC-LR were vary between 0.1± 0.01 μgml-1-1.6±0.09 μgml-1 and 0.1±0.01 μgml-1-0.7±0.04 μgml-1 respectively. However, none of urine samples were positive for either MC-LR or CYN in Angunakolapallessa. Thus, the results of the present study showed a positive relationship between MC-LR, CYN contaminations in well water and urine in CKDu patients of the study area. However, to clear out the hypothesis, further comprehensive studies are being investigated.Keywords: Cylindrospermopsin (CYN), Microcystin-LR (MC-LR), Human Urine, Chronic Kidney Disease (CKD), HPL

In-vitro Toxicity Study of Cylindrospermopsin on Wistar Rats

Naturally derived cyanotoxin, Cylindrospermopsin (CYN) found in fresh water systems poses a threat to human health worldwide. Cylindrospermopsin (CYN) is most potent cyanotoxin which affects the functions of kidney and liver. The present study was carried out to determine the possible effects of Cylindrospermopsin on mammals using male Wistar rats as an animal model. All experimental protocols were approved by the ethics committee of Faculty of Medical Science, University of Sri Jayewardenepura (No. 17/18). Thirty-five Wistar rats were divided into five groups (n=7) and rats were orally treated with different concentrations of cyanotoxin; Cylindrospermopsin (2.5 μg/L, 2 μg/L, 1.5 μg/L) and control group was treated with distilled water for 90 days. Fifth group received environmental water sample contaminated with Cylindrospermopsin (2.3 μg/L) obtained from randomly selected well in Padaviya. Blood and urine samples from each individual were collected at 0, 7, 14, 28, 42, 60, 90 days intervals and the collected samples were subjected to serum creatinine and urine creatinine analysis using creatinine assay kits. Cylindrospermopsin in urine samples were quantified by the ELISA methods. Aspartate aminotransferase (AST), Aspartate alaninetransferase (ALT) and Full Blood Count (FBC) were analysed. The mean body weight of treated (200 to 310 g) and control groups (200-335 g) of the experiment gradually increased until fourteenth week. There was no significant difference of body weights between treated and control groups (p=0.08). The absolute and relative (% body weight) weights of liver and kidneys of the treated groups were less than control group. Cylindrospermopsin dose 2.5 μg/L, 2 μg/L , 1.5 μg/L, 2.3 μg/L and control showed increased serum creatinine levels from 0.62 to 0.87 mg/dL, 0.64 to 0.86 mg/dL, 0.64 to 0.86 mg/dL, 0.61 to 0.83 mg/dL and 0.6 to 0.79 mg/dL respectively after 90 days treatment. The control group did not show a significant cause. Rat treated with different concentrations of Cylindrospermopsin, 2.5 μg/L, 2 μg/L, 1.5 μg/L, 2.3 μg/L and control showed gradually decreased of urine creatinine level from 34 to 55 mg/dL, 40 to 54 mg/dL, 38 to 54 mg/dL, 32 to 53 mg/dL and 43 to 54 mg/dL respectively after 90 days treatment and statistically significant (p<0.05) difference was found between treated and control groups. The highest Aspartate Aminotransferase (AST) and Aspartate Alaninetransferase (ALT) levels were obtained from Cylindrospermopsin dose 2.5 μg/L at 90 days exposure. Cylindrospermopsin concentration in urine gradually increased from 1.5 to 2.32 μg/L, 0.9 to 1.7 μg/L, 0.8 to 1.1 μg/L and 0.4 to 1.5 μg/L when animal exposed to 2.5 μg/L, 2 μg/L, 1.5 μg/L and 2.3 μg/L concentrations of Cylindrospermopsin where control did not show. Thus, the result of present study show consumption of Cylindrospermopsin contaminated water may lead to liver and kidney injuries.Keywords: Cylindrospermopsin (CYN), Wistar rats, ELISA, AST, AL

Chronic kidney disease of unknown aetiology in Sri Lanka: is cadmium a likely cause?

<p>Abstract</p> <p>Background</p> <p>The rising prevalence of chronic kidney disease (CKD) and subsequent end stage renal failure necessitating renal replacement therapy has profound consequences for affected individuals and health care resources. This community based study was conducted to identify potential predictors of microalbuminuria in a randomly selected sample of adults from the North Central Province (NCP) of Sri Lanka, where the burden of CKD is pronounced and the underlying cause still unknown.</p> <p>Methods</p> <p>Exposures to possible risk factors were determined in randomly recruited subjects (425 females and 461 males) from selected areas of the NCP of Sri Lanka using an interviewer administered questionnaire. Sulphosalicylic acid and the Light Dependent Resister microalbumin gel filtration method was used for initial screening for microalbuminuria and reconfirmed by the <it>Micral </it>strip test.</p> <p>Results</p> <p>Microalbumnuria was detected in 6.1% of the females and 8.5% of the males. Smoking (p < 0.001), alcohol use (p = 0.003), hypertension (p < 0.001), diabetes (p < 0.001), urinary tract infection (UTI) (p = 0.034) and consumption of water from wells in the fields (p = 0.025) were associated with microalbuminuria. In the binary logistic regression analysis, hypertension, diabetes mellitus, UTI, drinking well water in the fields, smoking and pesticide spraying were found to be significant predictors of microalbuminuria.</p> <p>Conclusions</p> <p>Hypertension, diabetes mellitus, UTI, and smoking are known risk factors for microalbuminuria. The association between microalbuminuria and consumption of well water suggests an environmental aetiology to CKD in NCP. The causative agent is yet to be identified. Investigations for cadmium as a potential causative agent needs to be initiated.</p

Knowledge and awareness of preventive measures in influenza H1N1 infection among Students of University of Sri Jayewardenepura

Objective: To assess the knowledge and awareness of preventive measures in influenza H1N1 infection among students of University of Sri Jayewardenepura Methods: In a descriptive cross sectional study, 384 of randomly selected students from four faculties of University of Sri Jayewardenepura were given standard interview administered questionnaires (Balkhy et al ,2010 ) to ascertain knowledge and awareness of preventive measures in  H1N1 infection.  Sample characteristics were analyzed using the SPSS statistical package (16 Version). Results: Response rate was 100% and 53.9% of participants were males. Knowledge of H1N1 infection was significantly different among students in four faculties. 43.5% of the participants had average knowledge and 81.2% of them knew correct mode of transmission of the H1N1 infection.  Among the participant who had high awareness of preventive measures from medical faculty (61.5%). Only 12.3% of the participants used face masks during the pandemic episode and 63.8% of them did hands washing. Among the study group 48.2% had not used face makes as they did not like that practice. Almost half of the students who do not use handkerchief were used elbow joint to prevent droplet infections when they sneeze or cough during endemic season. 65% students had not practice hand washing and use elbow joint to prevent droplet infections due to lack of awareness. Conclusion: Student’s knowledge on H1N1 infection was average. Awareness of preventive measures was high level among Faculty of Medical Sciences. Preventive practices were at low percentage in all the faculties.

Descriptive Cross-Sectional Study on Knowledge, Awareness, and Adherence to Medication among Hypertensive Patients at a Tertiary Care Centre in Colombo District, Sri Lanka

Objective. This study was aimed to assess the patient’s knowledge and awareness about hypertension and adherence to antihypertensive medication among hypertensive patients. Methods. The descriptive cross-sectional study was conducted in three medical clinics of Colombo South Teaching hospital, Kalubowila, Sri Lanka, from April 2019 to September 2019. Total of 384 hypertensive patients were recruited by systematic randomized controlled sampling and interviewed with validated questionnaires to assess their knowledge about hypertension and adherence to antihypertensive medication. Data were analyzed using SPSS (version 21) analytical package, and the chi-squared test was performed. Results. The total sample consisted of 384 hypertensive patients with a mean age of 59.32 (±12.34SD). This included 180 (46.9%) males and 204 (53.1%) females. The male : female ratio was approximately 9 : 10. Most of patients were with primary and ordinary educational status (65.9%), normal body mass index (54.9%), mild elevation of LDL cholesterol (76.3%), and coexistent ischemic heart disease (39.6%). The knowledge about hypertension among majority of patients was reasonable. However, they were unaware about normal values of blood pressure (69%, 95% of CI 1.92–2.09) and diagnostic values of hypertension (90.1%, 95% of CI 2.26–2.40). Moreover, they were unaware of their blood pressure values at time of diagnosis (75.3%, 95% of CI 2.09–2.25), at recent clinic visit (71.3%, 95% of CI 2.0-2.17), and target level (81.8%, 95% of CI 2.25-2.41). Most patients had adequate knowledge about the risk factors and complications of hypertension and were aware of their target organ damage (70.3%). Most patients believed that medication alone is not sufficient to control blood pressure (41.7%, 95% of CI 1.40-1.51) and adequate control of their blood pressure reduces complications (68.2%, 95% of CI 1.37-1.51). Most of the patients (71.8%) had reasonable good drug compliance. The forgetfulness was commonly attributed for nonadherence (69%, 95% of CI 1.26-1.36). Conclusions. The knowledge about hypertension among majority of patients was reasonable. But, they were unaware about their disease status and their diagnosis, target, and recent blood pressure values. Most of patients had adequate knowledge about the risk factors and complications of hypertension. However, they were unaware about their target organ damage due to hypertension. The drug compliance was reasonable among them. The forgetfulness was common reason for nonadherence. Therefore, healthcare professionals should implement individualized educational programmes to increase the awareness of disease status, appropriate blood pressure levels, and adherence of treatment to improve the outcome of patients

Duration of Diabetes and eGFR -A Preliminary Study

Objective: The objective of this study was to assess whether there is a significant difference in duration of type 2 diabetes with estimated glomerular filtration rate (eGFR). Method: A descriptive study was carried out using 67 patients with type 2 diabetes between the ages 30- 60 years attending the diabetic clinic of Colombo South Teaching Hospital. The 4 variable Modification of Diet in Renal Disease (MDRD) Study Equation was used to calculate the eGFR using the latest available serum creatinine levels. Results: Total number of subjects in the study group was 67 with 11 males and 56 females. Among this diabetic population, 32 were having an eGFR in the range of 30 to 59 ml/min/1.732m2 while 35 had an eGFR in the range 60-89ml/min/1.732m2. Independent student t test values obtained by comparing the two groups showed no significant difference between the mean values and duration of diabetes mellitus (p>0.05). Conclusions: According to this study, there was no significant difference in the duration (from the date of diagnosis) of diabetes mellitus between the eGFR 30 to 59 ml/min/1.732m2 (CKD stage 3 ) group and the eGFR more than 60ml/min/1.732m2 group. This highlights the importance of assessing renal functions early in newly diagnosed type 2 diabetics since early or late diagnosis of diabetes and its control from the day of diagnosis may be factors affecting the their renal functions (eGFR)

Smchd1 is a maternal effect gene required for genomic imprinting

Genomic imprinting establishes parental allele-biased expression of a suite of mammalian genes based on parent-of-origin specific epigenetic marks. These marks are under the control of maternal effect proteins supplied in the oocyte. Here we report epigenetic repressor Smchd1 as a novel maternal effect gene that regulates the imprinted expression of ten genes in mice. We also found zygotic SMCHD1 had a dose-dependent effect on the imprinted expression of seven genes. Together, zygotic and maternal SMCHD1 regulate three classic imprinted clusters and eight other genes, including non-canonical imprinted genes. Interestingly, the loss of maternal SMCHD1 does not alter germline DNA methylation imprints pre-implantation or later in gestation. Instead, what appears to unite most imprinted genes sensitive to SMCHD1 is their reliance on polycomb-mediated methylation as germline or secondary imprints, therefore we propose that SMCHD1 acts downstream of polycomb imprints to mediate its function