Solation and Characterization of Putative Nitrogen-Fixing Endophytes from Three Distinct Wild Rice Species in Sri Lanka

There are 22 recorded wild rice species on earth, of which 5 are found in Sri Lanka. Knowledge on the endophytic diazotrophs of wild rice species, which can be adopted for paddy cultivation, is still at a fledgling stage of discovery. The identification of stably maintained endophytic diazotrophic bacteria in the genus Oryza requires further attention, especially due to skyrocketing global prices of chemical fertilizers. The main objective of our study was the isolation and characterization of endophytic and rootsurface colonizing nitrogen fixers from three distinct wild rice species found in Sri Lanka, namely Oryza eichingeri, Oryza granulata and Oryza nivara. Endophytic nitrogen fixers were isolated from surface– sterilized stems, leaves, and roots, while root surface colonizing bacteria were isolated from washed roots. There were eleven endophytic bacteria and four root surface colonizing bacteria isolated by using nitrogenfree yeast mannitol/malate agar plates; six isolates from O. granulata, five isolates from O. eichingeri and four isolates from O. nivara. Out of the 15 isolates, 13 were Gram negative and 2 stained Gram positive. Six of the eleven isolates secreted three enzymes tested, primarily pectinases, proteases, and cellulases, suggesting their prospective involvement in endophytic lifestyles. Most isolates showed either bulls eyes, dendritic or featureless patterns of swarming chemotaxis for the chemoattractant proline, a common promoter of bacterial motility. We also tested the genomes of the isolates for the presence of the nifH gene. DNA from seven isolates gave a PCR amplicon of the expected size (360 bp) using universal nifH primers, which proves that the genetic foundation for the production of nitrogenase reductase subunit was found in these seven isolates. We also tested each of the bacterial isolates against a devastating fungal pathogen of rice (Rhizoctonia solani). We found 4 potential candidates that show partial inhibitory activity against the growth of this rice sheath blight causative agent

An integrated 3D approach for effective mine risk education in post war-zone areas

International Mine Action Standards define Mine Risk Education (MRE) as activities that seek to reduce the risk of death and injury from mines and explosive remnants of war, by raising awareness and promoting safe behaviour. MRE systems and practices in post clearance areas in conflict affected regions worldwide promote two main types of activities, identification of explosive remnants of war and making the correct response ensuring individual safety as well as the safety of the community. This can be viewed as a two dimensional approach spanned by Identification and Response dimensions. This paper based on the findings from a recent empirical study conducted specifically in the northern Sri Lanka introducing Reconciliation as a third dimension to further improve the present two dimensional approach of mine risk education. The authors acknowledge that MRE has had a positive impact on war affected communities in global context. However, it needs considerable integration of a learning module which should reflect the different causes of life catastrophe risks are similar to mine risks in extreme scenarios- they constitute non-diversifiable risks, so that MRE helps maintain a significant effect and long- lasting impact on attitudes towards landmines and other explosive remnants of war at the community level.Les normes internationales d'action contre les mines définissent l'éducation au risque des mines (MRE) comme une activité visant à réduire les risques de mort et de blessures dues aux mines et aux restes explosifs de guerre, en sensibilisant l'opinion publique et en encourageant les comportements sans danger. Les systèmes et les pratiques d’EDM dans les zones de dépollution dans les régions touchées par les conflits dans le monde encouragent deux types d’activités: l’identification des restes explosifs de guerre et la réaction appropriée assurant la sécurité individuelle ainsi que la sécurité de la communauté. Cela peut être considéré comme une approche bidimensionnelle englobée par les dimensions Identification et Réponse. Ce document s'appuie sur les conclusions d'une étude empirique récente menée spécifiquement dans le nord de Sri Lanka, introduisant la réconciliation en tant que troisième dimension pour améliorer encore l'approche en deux dimensions actuelle de l'éducation au risque des mines. Les auteurs reconnaissent que l'ERM a eu un impact positif sur les communautés touchées par la guerre dans le contexte mondial. Cependant, il faut intégrer un module d’apprentissage qui devrait refléter les différentes causes des risques de catastrophe dans la vie. Ces derniers sont similaires aux risques des mines dans les scénarios extrêmes. Ils constituent des risques non diversifiables, de sorte que les MRE aident à maintenir un effet significatif et durable sur les projets

Wearable Bluetooth Triage Healthcare Monitoring System.

Triage is the first interaction between a patient and a nurse/paramedic. This assessment, usually performed at Emergency departments, is a highly dynamic process and there are international grading systems that according to the patient condition initiate the patient journey. Triage requires an initial rapid assessment followed by routine checks of the patients' vitals, including respiratory rate, temperature, and pulse rate. Ideally, these checks should be performed continuously and remotely to reduce the workload on triage nurses; optimizing tools and monitoring systems can be introduced and include a wearable patient monitoring system that is not at the expense of the patient's comfort and can be remotely monitored through wireless connectivity. In this study, we assessed the suitability of a small ceramic piezoelectric disk submerged in a skin-safe silicone dome that enhances contact with skin, to detect wirelessly both respiration and cardiac events at several positions on the human body. For the purposes of this evaluation, we fitted the sensor with a respiratory belt as well as a single lead ECG, all acquired simultaneously. To complete Triage parameter collection, we also included a medical-grade contact thermometer. Performances of cardiac and respiratory events detection were assessed. The instantaneous heart and respiratory rates provided by the proposed sensor, the ECG and the respiratory belt were compared via statistical analyses. In all considered sensor positions, very high performances were achieved for the detection of both cardiac and respiratory events, except for the wrist, which provided lower performances for respiratory rates. These promising yet preliminary results suggest the proposed wireless sensor could be used as a wearable, hands-free monitoring device for triage assessment within emergency departments. Further tests are foreseen to assess sensor performances in real operating environments

Investigation of Optimum pH and Temperature for In-Vitro Crystallization of Urinary Cystine

Cystinuria contributes in formation of urinary stones. But, it has been reported that cystinuria is diagnosed when someone experiences with cystine stones. Therefore, early diagnosis of this condition is important. Thus, the objective of the study was to determine the optimum pH and temperature for crystallization of urine cystine in-vitro. Cystinuria solutions were prepared with the concentrations of 40, 60, 70, 75, 80, 90, 100 and 120 mg/dL. The pH of each solution was changed with the addition of acetic acid. Then solutions were exposed to temperature +4°C and 37°C, for 15, 30 and 45min. The sediments were observed microscopically for cystine crystals formation. Then acetone was added to cystinuria with the ratio of cystinuria:acetone, 8:1, 4:1, 2:1 and 1.1 and pH was altered with acetic acid and were subjected to +4 °C and 37 °C, for 15, 30 and 45 minutes and sediment was observed for cystine crystals under the microscope. Cystine crystallization had been occurred in the cystinuria of ≥100 mg/dL at pH 5 at 37 ° C and +4 °C, 30min after the addition of acetic acid whereas with the addition of acetone at cystinuria of ≥75mg/dL at pH 5 in both 37°C and at +4°C, 30min after the addition of acetic acid. The number of cystine crystals per High Power Field (HPF) was highest where cystinuria:acetone was 8:1.  The optimum conditions for cystine crystallization is at pH 5, 37 °C and +4 °C, 30min after acidifying with acetic acid at the minimum concentration of 100 mg/dL  of cystinuria. With the addition of acetone, at the ratio of cystinuria:acetone 8:1 with minimum concentration of cystinuria of 75 mg/dL.   KEYWORDS: Cystine, Crystallization, Acetic acid, Acetone, Temperature, p

Occurrence of Urinary Crystals among Urinary Tract Infections Suspected Paediatric Patients, Sri Lanka

Crystalluria has become one of the most vital biomarkers in urinalysis in detecting several disease conditions. It has been reported that urinary tract infections (UTI) may be the presenting sign of Urolithiasis in children. Therefore, the objective of this study was to identify and estimate the different types of crystals in the urine samples collected from UTI suspected children who admitted to the Lady Ridgeway Hospital for children, Sri Lanka. A descriptive cross-sectional study was conducted using 400 children belong to age<12 years suspected with UTI. The participants included 242 males and 158 females. The urine samples were collected prior to start antibiotics. Each sample was examined macroscopically and centrifuged at 2000 rpm for 5 minutes. The urine sediment was examined under the light microscope and different crystal types were identified and counted at x40 magnification. Out of 400 samples 82 samples (82/400) were positive for crystalluria. The crystal types present were uric acid, calcium oxalate, triple phosphate, ammonium biuate and ammonium urate. None of the samples showed abnormal crystal types. The distribution of each crystal type was as follow; uric acid 25/82, calcium oxalate 34/82, triple phosphate 12/82, ammonium biuate 7/82 and ammonium urate 4/82. The quantity of crystals per mL of urine was ranged as follow; uric acid 850-130,000, calcium oxalate 350- >250,000, triple phosphate 650-6,000, ammonium biurate and ammonium urate were presented in clumps. KEYWORDS: Crystalluria, Uric Acid, Calcium Oxalate, Triple Phosphate, Ammonium Biurate, Ammonium Urate, Urolithiasis, Urinary Tract Infections

Bioinformatic identification of proteins with tissue-specific expression for biomarker discovery

<p>Abstract</p> <p>Background</p> <p>There is an important need for the identification of novel serological biomarkers for the early detection of cancer. Current biomarkers suffer from a lack of tissue specificity, rendering them vulnerable to non-disease-specific increases. The present study details a strategy to rapidly identify tissue-specific proteins using bioinformatics.</p> <p>Methods</p> <p>Previous studies have focused on either gene or protein expression databases for the identification of candidates. We developed a strategy that mines six publicly available gene and protein databases for tissue-specific proteins, selects proteins likely to enter the circulation, and integrates proteomic datasets enriched for the cancer secretome to prioritize candidates for further verification and validation studies.</p> <p>Results</p> <p>Using colon, lung, pancreatic and prostate cancer as case examples, we identified 48 candidate tissue-specific biomarkers, of which 14 have been previously studied as biomarkers of cancer or benign disease. Twenty-six candidate biomarkers for these four cancer types are proposed.</p> <p>Conclusions</p> <p>We present a novel strategy using bioinformatics to identify tissue-specific proteins that are potential cancer serum biomarkers. Investigation of the 26 candidates in disease states of the organs is warranted.</p

A secretome profile indicative of oleate-induced proliferation of HepG2 hepatocellular carcinoma cells

Increased fatty acid (FA) is often observed in highly proliferative tumors. FAs have been shown to modulate the secretion of proteins from tumor cells, contributing to tumor survival. However, the secreted factors affected by FA have not been systematically explored. Here, we found that treatment of oleate, a monounsaturated omega-9 FA, promoted the proliferation of HepG2 cells. To examine the secreted factors associated with oleate-induced cell proliferation, we performed a comprehensive secretome profiling of oleate-treated and untreated HepG2 cells. A comparison of the secretomes identified 349 differentially secreted proteins (DSPs; 145 upregulated and 192 downregulated) in oleate-treated samples, compared to untreated samples. The functional enrichment and network analyses of the DSPs revealed that the 145 upregulated secreted proteins by oleate treatment were mainly associated with cell proliferation-related processes, such as lipid metabolism, inflammatory response, and ER stress. Based on the network models of the DSPs, we selected six DSPs (MIF, THBS1, PDIA3, APOA1, FASN, and EEF2) that can represent such processes related to cell proliferation. Thus, our results provided a secretome profile indicative of an oleate-induced proliferation of HepG2 cell

Low-Dose Cd Induces Hepatic Gene Hypermethylation, along with the Persistent Reduction of Cell Death and Increase of Cell Proliferation in Rats and Mice

Cadmium (Cd) is classified as a human carcinogen probably associated with epigenetic changes. DNA methylation is one of epigenetic mechanisms by which cells control gene expression. Therefore, the present study genome-widely screened the methylation-altered genes in the liver of rats previously exposed to low-dose Cd.Rats were exposed to Cd at 20 nmol/kg every other day for 4 weeks and gene methylation was analyzed at the 48(th) week with methylated DNA immunoprecipitation-CpG island microarray. Among the 1629 altered genes, there were 675 genes whose promoter CpG islands (CGIs) were hypermethylated, 899 genes whose promoter CGIs were hypomethylated, and 55 genes whose promoter CGIs were mixed with hyper- and hypo-methylation. Caspase-8 gene promoter CGIs and TNF gene promoter CGIs were hypermethylated and hypomethylated, respectively, along with a low apoptosis rate in Cd-treated rat livers. To link the aberrant methylation of caspase-8 and TNF genes to the low apoptosis induced by low-dose Cd, mice were given chronic exposure to low-dose Cd with and without methylation inhibitor (5-aza-2'-deoxyctidene, 5-aza). At the 48(th) week after Cd exposure, livers from Cd-treated mice displayed the increased caspase-8 CGI methylation and decreased caspase-8 protein expression, along with significant increases in cell proliferation and overexpression of TGF-β1 and cytokeratin 8/18 (the latter is a new marker of mouse liver preneoplastic lesions), all which were prevented by 5-aza treatment.These results suggest that Cd-induced global gene hypermethylation, most likely caspase-8 gene promoter hypermethylation that down-regulated its expression, leading to the decreased hepatic apoptosis and increased preneoplastic lesions