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The objectives of this study were to determine selected Hofmeister anions and cations that are important for kidney health, in raw rice samples from selected Chronic Kidney Disease of unknown etiology (CKDu) endemic and non-endemic areas in Sri Lanka and their intake. The anions and cations were analyzed by Ion Chromatography and Microwave Plasma Atomic Emission Spectrometry (MP-AES), respectively, after alkaline and acid digestion in thirty raw rice samples each from CKDu endemic and non-endemic areas, and the dietary intake was estimated. The mean concentrations of fluoride (F-), chloride (Cl-), phosphate (PO43-), sulfate (SO42-), sodium (Na+), magnesium (Mg2+), potassium (K+), and calcium (Ca2+) in raw rice in CKDu endemic areas were 53.317, 1515.3, 2799.6, 2704.9, 30.603, 300.76, 1001.3, and 90.075 mg/kg, respectively. The mean concentration of the anions and cations in raw rice from CKDu non-endemic areas were 22.850, 947.52, 4418.7, 6080.2, 23.862, 364.45, 955.78, and 96.780 mg/kg, respectively. Significantly higher differences (p < 0.05) were reported in the mean concentration of F-, Cl-, and Na+ in raw rice from CKDu endemic areas in comparison with the samples from non-endemic areas. The aggregated estimated daily intake (EDI) and cumulative EDI of F- via consumption of cooked non-traditional samba rice from CKDu endemic areas for adults were the highest (0.155 and 0.172 mg/kg bw/d, respectively), which were higher than the recommended tolerable upper intake value (0.15-0.2 mg/kg bw/d). In contrast, the traditional rice from CKDu non-endemic areas for adolescents, reported the lowest values (0.0210 and 0.0470 mg/kg bw/d, respectively). Adults who consume non-traditional samba rice from CKDu endemic areas were at health risk, while children were the most vulnerable group due to their low body weight. These results indicate that the consumption of rice rich in Hofmeister ions may contribute to the total intake and act as risk factors to negatively affect weak kidneys in CKDu endemic areas. Further research to analyze Hofmeister ions in cooked rice and rice from different countries is recommended.
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Oryza , Insuficiência Renal Crônica , Adulto , Criança , Humanos , Adolescente , Oryza/química , Sri Lanka/epidemiologia , Insuficiência Renal Crônica/induzido quimicamente , Insuficiência Renal Crônica/epidemiologia , Doenças Renais Crônicas Idiopáticas , Fluoretos , Ânions , CátionsRESUMO
BACKGROUND: Tuberculosis (TB), caused by the bacterium Mycobacterium tuberculosis is one of the top thirteen causes of death worldwide. The major challenge to control TB is the emergence of drug-resistant tuberculosis (DR-TB); specifically, multi-drug resistant TB which are resistant to the most potent drugs; rifampin and isoniazid. Owing to the inconsistencies of the current diagnostic methods, a single test cannot identify the whole spectrum of DR-TB associated mutations. Recently, host blood transcriptomics has gained attention as a promising technique that develops disease-specific RNA signatures/biomarkers. However, studies on host transcriptomics infected with DR-TB is limited. Herein, we intended to identify genes/pathways that are differentially expressed in multi-drug/rifampin resistant TB (MDR/RR-TB) in contrast to drug susceptible TB. METHOD AND RESULTS: We conducted blood RNA sequencing of 10 pulmonary TB patients (4; drug susceptible and 6; DR-TB) and 55 genes that were differentially expressed in MDR/RR-TB from drug-susceptible/mono-resistant TB were identified. CD300LD, MYL9, VAMP5, CARD17, CLEC2B, GBP6, BATF2, ETV7, IFI27 and FCGR1CP were found to be upregulated in MDR/RR-TB in all comparisons, among which CLEC2B and CD300LD were not previously linked to TB. In comparison pathway analysis, interferon alpha/gamma response was upregulated while Wnt/beta catenin signaling, lysosome, microtubule nucleation and notch signaling were downregulated. CONCLUSION: Up/down-regulation of immunity related genes/pathways speculate the collective effect of hosts' attempt to fight against continuously multiplying DR-TB bacteria and the bacterial factors to fight against the host defense. The identified genes/pathways could act as MDR/RR-TB biomarkers, hence, further research on their clinical use should be encouraged.
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Mycobacterium tuberculosis , Tuberculose Resistente a Múltiplos Medicamentos , Tuberculose , Humanos , Rifampina/farmacologia , Projetos Piloto , Antituberculosos/farmacologia , Tuberculose/microbiologia , Tuberculose Resistente a Múltiplos Medicamentos/tratamento farmacológico , Tuberculose Resistente a Múltiplos Medicamentos/genética , Mycobacterium tuberculosis/genética , Isoniazida/farmacologia , Testes de Sensibilidade MicrobianaRESUMO
Contagious diseases are needed to be monitored to prevent spreading within communities. Timely advice and predictions are necessary to overcome the consequences of those epidemics. Currently, emphasis has been placed on computer modelling to achieve the needed forecasts, the best example being the COVID-19 pandemic. Scientists used various models to determine how diverse sociodemographic factors correlated and influenced COVID-19 Global transmission and demonstrated the utility of computer models as tools in disease management. However, as modelling is done with assumptions with set rules, calculating uncertainty quantification is essential in infectious modelling when reporting the results and trustfully describing the limitations. This article summarizes the infectious disease modelling strategies, challenges, and global applicability by focusing on the COVID-19 pandemic.
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Anthropogenic and natural sources influence trace metals (TMs) bound to different sized particulate matter (PM) in dry and wet atmospheric deposition, which can create ecosystem and human health issues in the long run. Limited reviews are available summarizing worldwide concentrations in TMs in atmospheric PMs, their sources and pathways. Simultaneously, quantitative assessment of the potential human and ecosystem health risks imposed by the atmospheric particulate matter has not been adequately reviewed. Addressing this gap, here we review, the concentration of TMs in dry deposition mainly varies with the responsible sources, whereas, in wet deposition, it depends on the solubility of TMs. Other than deposition on impervious surfaces, the TMs incorporated PM can be deposited on biological agents. Health risk assessments show that ingestion and dermal contact pathways are more likely to cause health issues, however, the probability of occurring ingestion and dermal contact pathways is limited. Attention must be paid to the contribution from non-exhaust and exhaust vehicular emissions for TMs in atmospheric deposition, understanding their impact on stormwater management and urban agriculture. Behaviors of TMs in the atmosphere depends on many complex factors including origin, wind patterns, and weather conditions. Therefore, future research needs to be carried to model and predict the fate and transfer of TMs once they are generated through natural and anthropogenic sources. We believe that such research would allow establishing pollution control policies and measures in urban environments which will be critical to reduce the levels of TMs associated with atmospheric deposition in the environment.
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Poluentes Atmosféricos , Metais Pesados , Oligoelementos , Poluentes Atmosféricos/análise , Ecossistema , Monitoramento Ambiental , Humanos , Metais Pesados/análise , Material Particulado/análise , Medição de Risco , Oligoelementos/análise , Emissões de VeículosRESUMO
The human urinary microbiome, also termed urobiome, has been overlooked due to the clinical dogma of sterile urine, as reported by routine culture. However, evolving sensitive tools such as expanded quantitative urine culture, 16S ribosomal RNA gene sequencing, and next-generation sequencing have discovered a vast number of microorganisms present in urine, even in healthy individuals. Microbiome dysbiosis and its links to disease is a heavily explored area in several microbial niches. Presently, urobiome dysbiosis and its correlation to urinary system-related diseases is at its infancy but rapidly emerging, as it provides potential therapeutic insights. This review outlines the changes in the human urinary microbiome concerning globally prevalent diseases affecting kidney function, such as chronic kidney disease (CKD), diabetes mellitus (DM), hypertension (HT), and urinary tract infection (UTI). Alterations to urine microbial diversity, including differences in the abundance and species richness of particular microbial genera, notably Lactobacillus, Prevotella, Streptococcus, Staphylococcus, Klebsiella, Enterococcus, between diseased and healthy samples are discussed utilising studies to date. Subsequent research needs to move beyond correlation to understand the roles of the urinary microbiota in diseases, thereby clarifying whether urinary dysbiosis has causal contributions that may provide important insight for diagnostics, pathophysiology, and therapy in renal pathologies.
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The invasive nature of cyanotoxin-producing cyanobacteria and the adverse effects concerning their toxic impacts have gained heightened scientific attention of late. The persistence of cyanotoxins in irrigation water leads to bioaccumulation in plants, the development of phytotoxic effects, and the threat of groundwater contamination. The accumulation of cyanotoxins in plants is caused by several factors leading to severe toxic effects, including reduced plant growth and seed germination, enhanced oxidative stress, lowered rate of mineral uptake, decreased photosynthetic efficiency, and loss of chlorophyll content. The uptake and accumulation of cyanotoxins in plants can be concentration-dependent, as reported in a myriad of studies. Even though several studies have reported phytotoxic effects of cyanotoxin contamination, field-related studies reporting phytotoxic effects are particularly inadequate. Paradoxically, at realistic conditions, some plants are reported to be tolerant of cyanotoxins. Furthermore, the breadth of adverse impacts of cyanotoxins on human health is significant. Cyanotoxins cause major health effects including cancer, oxidative stress, organelle dysfunction, DNA damage, and enzyme inhibition. This review intends to present compelling arguments on microcystins (MCs), cylindrospermopsins (CYN), ß-N-methylamino-L-alanine (BMAA), and anatoxin-a (ANTX-a), their uptake and accumulation in crop plants, phytotoxic effects on plants, and potential health implications to humans. The accumulation of cyanotoxins implants cultivated as food crops, resulting in phytotoxic effects and adverse impacts on human health are serious issues that require scientific inputs to be addressed.
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Toxinas de Cianobactérias , Cianobactérias , Clorofila , Humanos , Microcistinas/toxicidadeRESUMO
Particulate matter (PM) is a complex mixture of solid particles and liquid droplets suspended in the air with varying size, shape, and chemical composition which intensifies significant concern due to severe health effects. Based on the well-established human health effects of outdoor PM, health-based standards for outdoor air have been promoted (e.g., the National Ambient Air Quality Standards formulated by the U.S.). Due to the exchange of indoor and outdoor air, the chemical composition of indoor particulate matter is related to the sources and components of outdoor PM. However, PM in the indoor environment has the potential to exceed outdoor PM levels. Indoor PM includes particles of outdoor origin that drift indoors and particles that originate from indoor activities, which include cooking, fireplaces, smoking, fuel combustion for heating, human activities, and burning incense. Indoor PM can be enriched with inorganic and organic contaminants, including toxic heavy metals and carcinogenic volatile organic compounds. As a potential health hazard, indoor exposure to PM has received increased attention in recent years because people spend most of their time indoors. In addition, as the quantity, quality, and scope of the research have expanded, it is necessary to conduct a systematic review of indoor PM. This review discusses the sources, pathways, characteristics, health effects, and exposure mitigation of indoor PM. Practical solutions and steps to reduce exposure to indoor PM are also discussed.
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Poluentes Atmosféricos , Poluição do Ar em Ambientes Fechados , Poluição do Ar , Poluentes Atmosféricos/análise , Poluição do Ar em Ambientes Fechados/análise , Culinária , Monitoramento Ambiental , Humanos , Tamanho da Partícula , Material Particulado/análiseRESUMO
Hot springs harbour diverse and interesting groups of microorganisms adapted to extreme conditions. However, due to limitations in the culture-dependent approach, most of such thermophiles remain uncultured and unexplored. Hence, this study was conducted to gain a comprehensive understanding of the bacterial diversity of Mahapelessa hot spring, Sri Lanka using both culture-dependent and culture-independent approaches. The in situ temperature of the water sample was 44.5 °C and the pH was 8.14. 16S rRNA Sanger sequencing of DNA extracted from the 18 bacterial isolates revealed the presence of eight genera belonging to two phyla: Proteobacteria (84%) and Firmicutes (16%) and the most abundant genus being Klebsiella. A total of 23 bacterial phyla representing 80 classes, 43 orders, 123 families, 205 genera and 83 species were detected by 16S rRNA V3-V4 region by amplicon metagenome sequencing of DNA extracted from water samples, where the most abundant phylum was the Proteobacteria (57.39%), followed by Firmicutes (23.7%) and Chloroflexi (4.14%). The three phyla Actinobacteria, Planctomycetes and Bacteroidetes were also detected less than 3% in abundance while 4.48% of bacteria could not be fit into any known phylum. The most abundant genera were Burkholderia (14.87%), Desulfotomaculum (7.23%) and Stenotrophomonas (6.1%). Four strictly anaerobic bacteria, Anaerosolibacter carboniphilus (0.71%), Bellilinea caldifistulae (0.04%), Salimesophilobacter vulgaris (0.1%), Anaerobacterium chartisolvens (0.12%); two potential plant growth-promoting bacteria, Azospirillum halopraeferens (0.04%) and Bradyrhizobium liaoningense (0.16%) and one potential alkali tolerant and sulphate-reducing bacterium, Desulfovibrio alkalitolerans (0.45%) were recorded. Pigmentiphaga sp. was isolated from Mahapelessa hot spring and to the best of our knowledge, this is the first record of this genus from a hot spring. This study gives insight into the vast bacterial diversity present in the Mahapelessa hot spring from the culture-independent approach which could not be identified using standard culturing techniques.
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Fontes Termais , Azospirillum , Bactérias/genética , Bradyrhizobium , Chloroflexi , Clostridiaceae , Clostridiales , Desulfovibrionaceae , Humanos , Filogenia , RNA Ribossômico 16S/genéticaRESUMO
The prevalence of chronic kidney disease of unknown etiology (CKDu) is receiving considerable attention due to the serious threat to human health throughout the world. However, the roles of geo-socio-environmental factors in the prevalence of the CKDu endemic areas are still unknown. Sri Lanka is one of the countries most seriously affected by CKDu, where 10 out of 25 districts have been identified as the areas with the high prevalence of CKDu (10-20%). This review summarizes the geographical distribution of CKDu and its probable geochemical, behavioral, sociological, and environmental risk factors based on research related to hydrogeochemical influences on CKDu in Sri Lanka. More than 98% of CKDu patients have consumed groundwater as their primary water source in daily life, indicating the interactions of geogenic contaminants (such as F-, total dissolved solids, Hofmeister ions) in groundwater is responsible for the disease. Apart from the hydrogeochemical factors, mycotoxins, cyanotoxins, use of some herbal medicines, dehydration, and exposure to agrochemicals were alleged as risk factors. Sociological factors, including poverty, living habits and anthropogenic activities, may also provoke the emergence of CKDu. Therefore, the interaction of geo-socio environmental risk factors should be sociologically and scientifically considered to prevent the prevalence of CKDu. Future in-depth studies are required to reveal the individual role of each of the postulated etiological factors, possibly using machine learning and advanced statistics.
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Insuficiência Renal Crônica , Água , Humanos , Insuficiência Renal Crônica/epidemiologia , Insuficiência Renal Crônica/etiologia , Fatores de Risco , Sri Lanka/epidemiologia , Abastecimento de ÁguaRESUMO
Mycobacterium tuberculosis belonging to Beijing sublineage (BL) is associated with high tuberculosis (TB) transmission, multidrug resistance, and adverse treatment outcomes. Sri Lanka experiences an increase in the number of travellers/workers to and from high TB-burden countries, and there is risk of getting BL strains imported into the country. In this context, a cohort study was conducted to assess the prevalence of BL strains among pulmonary tuberculosis (PTB) patients in the Kandy district of Sri Lanka (a popular tourist destination) and its association with patients' sociodemographic and clinical characteristics. The study population included sputum smear-positive PTB patients diagnosed from February 2018-July 2019. Fresh sputum samples were collected for culturing and conducted polymerase chain reaction using BL-specific primers. Among the 101 patients recruited, presence of BL strains could be ascertained in 94 patients of which 24 (26%; 95% CI: 18%-35%) had BL strains. Prevalence of BL strains was higher among those with high sputum smear grades (2+ and 3+) (P < 0.05) and those who had travelled abroad (P < 0.05). The prevalence was also higher among young people (aged <35 years). Treatment success rates were similar in patients with (83%) and without BL strains (83% vs. 81%; P value = 0.8375). The prevalence of BL strains in Kandy, Sri Lanka, was high compared to previously reported figures in Sri Lanka, and the percentage drives closer to the countries in South East Asia. International travel raises itself as an emerging issue in BL transmission urging the need of policies and practices in immigration/emigration strategies. The study findings have the potential to alter the TB epidemiology in the country and might represent the situation in other underexplored countries as well. Therefore, it is important to monitor the trends and factors related to the prevalence of Beijing strains globally and make decisions as a whole.
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Respiratory bacterial microbiota plays a key role in human health. Lung cancer microbiome is a significant yet an understudied area while bronchiectasis microbiome is often studied. We assessed the bacterial microbiota in the upper and lower respiratory tract of the patients with lung cancer and bronchiectasis against a healthy group and their variations in individuality. 16S rRNA gene based metagenomic sequencing was used to detect entire bacterial community along with conventional aerobic bacterial culturing. In comparison to healthy, increased bacterial diversity was observed in diseased population. Abundance of more than 1% was considered and bacteria were identified in 97% similarity. Only lung cancer patients exhibited bacteria specific to the disease: Corynebacterium tuberculostearicum and Keratinibaculum paraultunense. However, Enterococcus faecalis and Delftia tsuruhatensis were also observed limited to lung cancer and bronchiectasis respectively, in less than 1% but supported with bacterial culturing. In conclusion the disease condition and intra-group variability should be considered in future with larger cohorts to understand individual patient variability highlighting the social habits and gender of the individual.
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BACKGROUND: The transmission dynamics of Mycobacterium tuberculosis (Mtb) using various genotyping tools has been studied globally and a particular tool for genotyping Mtb is the mycobacterial interspersed repetitive units-variable number tandem repeats (MIRU-VNTR). Tuberculosis (TB) remains an important public health problem worldwide and Sri Lanka being a country of tourist destination; because of major development projects undergoing, it has a high proportion of tourists and immigrants from Asia and Europe that are characterized with highest TB incidences and drug-resistant clinical isolates. Hence, in order to address the question of Mtb genetic diversity, we investigated the discriminatory power of both MIRU-VNTR typing of 15 and 24 loci with spoligotyping to differentiate Mtb isolates. METHOD: Acid-fast bacilli positive sputum samples (n = 150) from first visit patients were collected. Decontamination of sputum and extraction of genomic DNA were carried out using standard techniques. The isolates were characterized by MIRU-VNTR for both the 15 and 24 loci and spoligotyping. RESULTS: In our study population, MIRU-VNTR 15 and 24 loci did not show a significant difference among the identified M. tuberculosis strains. However, MIRU 24 loci yielded an additional strain LAM, which is of T1 origin. 15 loci strain grouping had more clusters of strains grouped together while 24 loci differentiated the same cluster of strains into distinct strain types. CONCLUSION: We conclude that the use of 15-locus MIRU-VNTR typing is sufficient for a first-line epidemiological study to genotype M. tuberculosis, but the additional discriminatory power of 24 loci MIRU-VNTR has been able to differentiate samples within highly homologous groups.
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Técnicas de Genotipagem/métodos , Sequências Repetitivas Dispersas/genética , Repetições Minissatélites/genética , Mycobacterium tuberculosis/genética , Reação em Cadeia da Polimerase/métodos , Tuberculose Pulmonar/microbiologia , Feminino , Genes Bacterianos , Variação Genética , Técnicas de Genotipagem/instrumentação , Humanos , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Tipagem Molecular/métodos , Mycobacterium tuberculosis/classificação , FilogeniaRESUMO
PURPOSE: Chronic kidney disease of unknown etiology (CKDu), having epidemic characteristics, is being diagnosed increasingly in certain tropical regions of the world, mainly Latin America and Sri Lanka. They have been observed primarily in farming communities and current hypotheses point toward many environmental and occupational triggers. CKDu does not have common etiologies of chronic kidney disease (CKD) such as hypertension, diabetes, or autoimmune disease. We aimed to understand the molecular processes underlying CKDu in Sri Lanka using transcriptome analysis. METHODS: RNA extracted from whole blood was reverse transcribed and used for microarray analysis using the Human HT-12 v.4 array (Illumina). Pathway analysis was carried out using ingenuity pathway analysis (IPA-Qiagen). Microarray results were validated using real-time PCR of five selected genes. RESULTS: Pathways related to innate immune response, including interferon signaling, inflammasome signaling and TREM1 signaling had the most significant positive activation z scores, where as EIF2 signaling and mTOR signaling had the most significant negative activation z scores. Pathways previously linked to fluoride toxicity; G-protein activation, Cdc42 signaling, Rac signaling and RhoA signaling were activated in CKDu patients. The most significantly activated biological functions were cell death, cell movement and antimicrobial response. Significant toxicological functions were mitochondrial dysfunction, oxidative stress and apoptosis. CONCLUSIONS: Based on the molecular pathway analysis in CKDu patients and review of literature, viral infections and fluoride toxicity appear to be contributing to the molecular mechanisms underlying CKDu.
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Fluoretos/efeitos adversos , Imunidade Inata/genética , Insuficiência Renal Crônica/etiologia , Transdução de Sinais/genética , Viroses/complicações , Adulto , Fator de Iniciação 2 em Eucariotos/genética , Proteínas de Ligação ao GTP/genética , Perfilação da Expressão Gênica , Humanos , Inflamassomos/genética , Interferons/genética , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos , RNA/análise , Sri Lanka , Serina-Treonina Quinases TOR/genética , Receptor Gatilho 1 Expresso em Células Mieloides/genética , Proteína cdc42 de Ligação ao GTP/genética , Proteínas rac de Ligação ao GTP/genética , Proteína rhoA de Ligação ao GTP/genéticaRESUMO
BACKGROUND: In Sri Lanka, there exists chronic kidney disease of both known (CKD) and unknown etiologies (CKDu). Identification of novel biomarkers that are customized to the specific causative factors would lead to early diagnosis and clearer prognosis of the diseases. This study aimed to find genetic biomarkers in blood to distinguish and identify CKDu from CKD as well as healthy populations from CKDu endemic and non-endemic areas of Sri Lanka. METHODS: The expression patterns of a selected panel of 12 potential genetic biomarkers were analyzed in blood using RT-qPCR. Fold changes of gene expressions in early and late stages of CKD and CKDu patients, and an apparently healthy population of a CKDu endemic area, Girandurukotte (GH) were calculated relative to apparently healthy volunteers from a CKDu non-endemic area, Kandy (KH) of Sri Lanka, using the comparative CT method. RESULTS: Significant differences were observed between KH and early stage CKDu for both the insulin-like growth factor binding protein 1 (IGFBP1; p = 0.012) and kidney injury molecule-1 (KIM1; p = 0.003) genes, and KH and late stage CKD and CKDu for the glutathione-S-transferase mu 1 (GSTM1; p < 0.05) gene. IGFBP1 and KIM1 genes showed significant difference between the early and late stage CKDu (p < 0.01). The glutamate cysteine ligase catalytic subunit (GCLC) gene had significantly different expression between KH and all the other study groups (p < 0.01). The GH group was significantly different from the KH group for the oxidative stress related genes, G6PD, GCLC and GSTM1 (p < 0.01), and also the KIM1 gene (p = 0.003). IGFBP1, insulin-like growth factor binding protein 3 (IGFBP3), fibronectin 1 (FN1) and KIM1 showed significant correlations with serum creatinine, and IGFBP1, KIM1 and kallikrein 1 (KLK1) with eGFR (p < 0.05). CONCLUSION: A panel consisting of IGFBP1, KIM1, GCLC and GSTM1 genes could be used in combination for early screening of CKDu, whereas these genes in addition with FN1, IGFBP3 and KLK1 could be used to monitor progression of CKDu. The regulation of these genes has to be studied on larger populations to validate their efficiency for further clinical use.
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Receptores ErbB/sangue , Glutamato-Cisteína Ligase/sangue , Glutationa Transferase/sangue , Receptor Celular 1 do Vírus da Hepatite A/sangue , Proteína 1 de Ligação a Fator de Crescimento Semelhante à Insulina/sangue , Insuficiência Renal Crônica/sangue , Insuficiência Renal Crônica/diagnóstico , Biomarcadores/sangue , Estudos de Viabilidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Prevalência , Insuficiência Renal Crônica/epidemiologia , Reprodutibilidade dos Testes , Fatores de Risco , Sensibilidade e Especificidade , Sri Lanka/epidemiologiaRESUMO
Objective. To infer the influence of internal and external oxidative stress in chronic kidney disease patients of unknown etiology (CKDu) in Sri Lanka, by analyzing expression of genes related directly or indirectly to oxidative stress: glutamate-cysteine ligase catalytic subunit (GCLC), glutathione S-transferase mu 1 (GSTM1), glucose-6-phosphate dehydrogenase (G6PD), fibroblast growth factor-23 (FGF23), and NLR family pyrin domain containing 3 (NLRP3). Methods. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was carried out for the selected populations: CKDu patients (n = 43), chronic kidney disease patients (CKD; n = 14), healthy individuals from a CKDu endemic area (GHI; n = 9), and nonendemic area (KHI; n = 16). Fold changes were quantified relative to KHI. Results. GCLC had greater than threefold upregulation in all three study groups, with a maximum of 7.27-fold upregulation in GHI (p = 0.000). GSTM1 was not expressed in 25.6% of CKDu and 42.9% of CKD patients, but CKDu patients expressing GSTM1 showed upregulation of 2.60-fold (p < 0.05). Upregulation of FGF23 and NLRP3 genes in CKD and CKDu was observed (p < 0.01), with greater fold changes in CKD. Conclusion. Results suggest higher influence of external sources of oxidative stress in CKDu, possibly owing to environmental conditions.
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Estresse Oxidativo , Insuficiência Renal Crônica/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Regulação para Cima , Adulto , Idoso , Feminino , Fator de Crescimento de Fibroblastos 23 , Humanos , Masculino , Pessoa de Meia-Idade , Sri LankaRESUMO
Sri Lanka has rich cyanobacterial diversity, however, only few studies have been conducted to identify the potential toxin producers in water bodies used for human consumption. As the detection of cyanotoxin is vital in water quality management, a study was done by employing 16S rRNA gene to explore the genetic divergence, phylogenetic relationships and potential toxin producing cyanobacteria in reservoirs and well waters in the dry zone of Sri Lanka. Forty five, 16S rRNA gene sequences were assayed and phylogenetic tree was constructed. Among 45 isolates, 20 isolates were classified as unidentified cyanobacteria and considered as novel cyanobacterial genera. Of 25 identified isolates, seven isolates were identified up to species level. With 16S rRNA phylogeny, 20 unidentified cyanobacterial isolates were able to place on their taxonomic positions up to order level. Results revealed that water samples understudy had vast cyanobacterial diversity with potential microcystin (MC) and cylindrospermopsin (CYN) producers and eleven clusters clearly demonstrated five cyanobacterial orders with more than 90% similarity irrespective to their toxicity which showed the suitability of 16S rRNA gene for taxonomic differentiation. Sixteen isolates had the potential to produce MC and two isolates to produce CYN. Findings of the study confirm the rich cyanobacterial diversity and the divergence among the potential cyanotoxin producers in the dry zone water bodies of Sri Lanka.
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BACKGROUND: Mycobacteria have a spectrum of virulence and different susceptibilities to antibiotics. Distinguishing mycobacterial species is vital as patients with non-tuberculous mycobacterial (NTM) infections present clinical features that are similar to those of patients with tuberculosis. Thus, rapid differentiation of Mycobacterium tuberculosis complex from NTM is critical to administer appropriate treatment. Hence the aim of the study was to rapid identification of mycobacterial species present in bronchial washings using multiplex real time Polymerase Chain Reaction (PCR) and to determine the drug susceptibility in identified mycobacterial species. METHODS: Sputum smear negative bronchoscopy specimens (n = 150) were collected for a period of one year, from patients attending the General Hospital Kandy, Sri Lanka. The specimens were processed with modified Petroff's method and were cultured on Löwenstein- Jensen medium. DNA, extracted from the mycobacterial isolates were subjected to a SYBR green mediated real time multiplex, PCR assay with primers specific for the M. tuberculosis complex, M. avium complex, M. chelonae-M.abscessus group and M. fortuitum group. DNA sequencing was performed for the species confirmation, by targeting the 16S rRNA gene and the drug susceptibility testing was performed for the molecularly identified isolates of M. tuberculosis and NTM. RESULTS: The optimized SYBR Green mediated multiplex real-time PCR assay was able to identify the presence of genus Mycobacterium in 25 out of 26 AFB positive isolates, two M. tuberculosis complex, three M. avium complex and two isolates belonging to M. chelonae-M. abscessus group. DNA sequencing confirmed the presence of M. tuberculosis, M. chelonae-M. abscessus, M. intracellulare, M. avium, Rhodococcus sp. and M. celatum. Remaining isolates were identified as Mycobacterium sp. All the NTM isolates were sensitive to amikacin and seven were resistant to ciproflaxacin. Twenty two of the NTM isolates and the isolate Rhodococcus was resistant to clarithromycin. The two isolates of M. tuberculosis were sensitive to all first line anti tuberculosis drugs. CONCLUSION: The optimized SYBR Green mediated multiplex real time PCR assay could be an effective tool for the rapid differentiation of pathogenic M. tuberculosis complex from the opportunistic nontuberculous mycobacteria and also it confirmed the presence of NTM in 15.3 % of the study population.