Polymorphisms in ARMS2/HTRA1 and complement genes and age-related macular degeneration in India: findings from the INDEYE study.

PURPOSE: Association between genetic variants in complement factor H (CFH), factor B (CFB), component 2 (C2), and in the ARMS2/HTRA1 region with age-related macular degeneration (AMD) comes mainly from studies of European ancestry and case-control studies of late-stage disease. We investigated associations of both early and late AMD with these variants in a population-based study of people aged 60 years and older in India. METHODS: Fundus images were graded using the Wisconsin Age-Related Maculopathy Grading System and participants assigned to one of four mutually exclusive stages based on the worse affected eye (0 = no AMD, 1-3 = early AMD, 4 = late AMD). Multinomial logistic regression was used to derive risk ratios (RR) accounting for sampling method and adjusting for age, sex, and study center. RESULTS: Of 3569 participants, 53.2% had no signs of amd, 45.6% had features of early amd, and 1.2% had late amd. CFH (RS1061170), C2 (RS547154), OR CFB (RS438999) was not associated with early or late AMD. In the ARMS2 locus, RS10490924 was associated with both early (adjusted RR 1.22, 95% confidence interval [CI]: 1.13-1.33, P < 0.0001) and late AMD (adjusted RR 1.81, 95% CI: 1.15-2.86; P = 0.01); rs2672598 was associated only with early AMD (adjusted RR 1.12, 95% CI: 1.02-1.23; P = 0.02); rs10490923 was not associated with early or late AMD. CONCLUSIONS: Two variants in ARMS2/HTRA1 were associated with increased risk of early AMD, and for one of these, the increased risk was also evident for late AMD. The study provides new insights into the role of these variants in early stages of AMD in India

Genetic variants in a sodium-dependent vitamin C transporter gene and age-related cataract.

BACKGROUND: Cataract is a major health burden in many countries and a significant problem in India. While observational studies show lower cataract risk with increasing dietary or plasma vitamin C, randomised controlled trials of supplements have been negative. Genetic variants in vitamin C transporter proteins (SLC23A1), especially rs33972313, may provide evidence on a causal association of vitamin C with cataract. METHODS: We used data from a randomly selected population-based study in people aged 60 years and above in north and south India. Of 7518 sampled, 5428 (72%) were interviewed for socioeconomic and lifestyle factors, attended hospital for lens imaging and blood collection and were subsequently genotyped for rs33972313 and rs6596473. Mixed or pure types of cataract were graded by the Lens Opacity Classification System III as nuclear (2404), cortical (494) or posterior subcapsular cataract (PSC) (1026); 1462 had no significant cataract and no history of cataract surgery and 775 had bilateral aphakia/pseudophakia. RESULTS: rs33972313 was associated with cortical (OR 2.16; 95% CI 1.34 to 3.49, p=0.002) and PSC (OR 1.68; 95% CI 1.06 to 2.65, p=0.03) but not with nuclear cataract. In analyses of pure cataracts, associations were found only between rs33972313 and pure cortical cataracts (OR 2.29; 95% CI 1.12 to 4.65, p=0.03) and with a standardised cortical opacity score. There was no association with rs6596473 and any cataract outcomes. CONCLUSIONS: Using an established genetic variant as a proxy for lifetime ascorbate concentrations, our results support a causal association of vitamin C with cataract

Use of Cooking Fuels and Cataract in a Population-Based Study: The India Eye Disease Study.

BACKGROUND: Biomass cooking fuels are commonly used in Indian households, especially by the poorest socioeconomic groups. Cataract is highly prevalent in India and the major cause of vision loss. The evidence on biomass fuels and cataract is limited. OBJECTIVES: To examine the association of biomass cooking fuels with cataract and type of cataract. METHODS: We conducted a population-based study in north and south India using randomly sampled clusters to identify people ≥ 60 years old. Participants were interviewed and asked about cooking fuel use, socioeconomic and lifestyle factors and attended hospital for digital lens imaging (graded using the Lens Opacity Classification System III), anthropometry, and blood collection. Years of use of biomass fuels were estimated and transformed to a standardized normal distribution. RESULTS: Of the 7,518 people sampled, 94% were interviewed and 83% of these attended the hospital. Sex modified the association between years of biomass fuel use and cataract; the adjusted odds ratio (OR) for a 1-SD increase in years of biomass fuel use and nuclear cataract was 1.04 (95% CI: 0.88, 1.23) for men and 1.28 (95% CI: 1.10, 1.48) for women, p interaction = 0.07. Kerosene use was low (10%). Among women, kerosene use was associated with nuclear (OR = 1.76, 95% CI: 1.04, 2.97) and posterior subcapsular cataract (OR = 1.71, 95% CI: 1.10, 2.64). There was no association among men. CONCLUSIONS: Our results provide robust evidence for the association of biomass fuels with cataract for women but not for men. Our finding for kerosene and cataract among women is novel and requires confirmation in other studies. Citation: Ravilla TD, Gupta S, Ravindran RD, Vashist P, Krishnan T, Maraini G, Chakravarthy U, Fletcher AE. 2016. Use of cooking fuels and cataract in a population-based study: the India Eye Disease Study. Environ Health Perspect 124:1857-1862; http://dx.doi.org/10.1289/EHP193

Genetic risk factors for late age-related macular degeneration in India.

BACKGROUND/AIMS: There are limited data from India on genetic variants influencing late age-related macular degeneration (AMD). We have previously reported associations from a population-based study in India (the India age-related eye disease study (INDEYE)) of early AMD and single nucleotide polymorphisms (SNPs) in ARMS2/HTRA1 and no association with CFH, C2 or CFB. Late AMD cases were too few for meaningful analyses. We aimed to investigate SNPs for late AMD through case enrichment and extend the loci for early AMD. METHODS: Fundus images of late AMD hospital cases were independently graded by the modified Wisconsin AMD grading scheme. In total 510 cases with late AMD (14 geographic atrophy and 496 neovascular AMD (nvAMD)), 1876 with early AMD and 1176 with no signs of AMD underwent genotyping for selected SNPs. We investigated genotype and per-allele additive associations (OR and 95% CIs) with nvAMD or early AMD. Bonferroni adjusted P values are presented. RESULTS: We found associations with nvAMD for CFHY402H variant (rs1061170) (OR=1.99, 95% CI 1.67 to 2.37, P=10-6), ARMS2 (rs10490924) (OR=2.94, 95% CI 2.45 to 3.52, P=10-9), C2 (rs547154) (OR=0.67, 95% CI 0.53 to 0.85, P=0.01), ABCA1 (rs1883025) (OR=0.77, 95% CI 0.65 to 0.92, P=0.04) and an SNP near VEGFA (rs4711751) (OR=0.64, 95% CI 0.54 to 0.77, P=10-3). We found no associations of TLR3 (rs3775291), CFD (rs3826945), FRK (rs1999930) or LIPC (rs10468017) or APOE ε4 alleles with nvAMD or early AMD, nor between early AMD and rs1883025 or rs4711751. CONCLUSIONS: The major genetic determinants of nvAMD risk in India are similar to those in other ancestries, while findings for early AMD suggest potential differences in the pathophysiology of AMD development

EPHA2 Polymorphisms and Age-Related Cataract in India

Objective: We investigated whether previously reported single nucleotide polymorphisms (SNPs) of EPHA2 in European studies are associated with cataract in India. Methods: We carried out a population-based genetic association study. We enumerated randomly sampled villages in two areas of north and south India to identify people aged 40 and over. Participants attended a clinical examination including lens photography and provided a blood sample for genotyping. Lens images were graded by the Lens Opacification Classification System (LOCS III). Cataract was defined as a LOCS III grade of nuclear >= 4, cortical >= 3, posterior sub-capsular (PSC) >= 2, or dense opacities or aphakia/pseudophakia in either eye. We genotyped SNPs rs3754334, rs7543472 and rs11260867 on genomic DNA extracted from peripheral blood leukocytes using TaqMan assays in an ABI 7900 real-time PCR. We used logistic regression with robust standard errors to examine the association between cataract and the EPHA2 SNPs, adjusting for age, sex and location. Results: 7418 participants had data on at least one of the SNPs investigated. Genotype frequencies of controls were in Hardy-Weinberg Equilibrium (p > 0.05). There was no association of rs3754334 with cataract or type of cataract. Minor allele homozygous genotypes of rs7543472 and rs11260867 compared to the major homozygote genotype were associated with cortical cataract, Odds ratio (OR) = 1.8, 95% Confidence Interval (CI) (1.1, 3.1) p = 0.03 and 2.9 (1.2, 7.1) p = 0.01 respectively, and with PSC cataract, OR = 1.5 (1.1, 2.2) p = 0.02 and 1.8 (0.9, 3.6) p = 0.07 respectively. There was no consistent association of SNPs with nuclear cataract or a combined variable of any type of cataract including operated cataract. Conclusions: Our results in the Indian population agree with previous studies of the association of EPHA2 variants with cortical cataracts. We report new findings for the association with PSC which is particularly prevalent in Indians

Aspectos biológicos da inoculação experimental e atividade malaricida da 4-(6-mercaptopurina)-7-cloroquinolina em Gallus gallus Linnaeus, 1758 experimentalmente infectados por Plasmodium (Novyella) juxtanucleare Versiani & Gomes, 1941 (Apicomplexa, Plasmodiidae)

Plasmodium juxtanucleare is the agent of the avian malaria that occurs in some states of Brazil. This malaria is related to several clinical signs and it can cause damages in the poultry section. The aviary model was already used for the investigation of drugs in the combat to the malaria and nowadays the model more used is the Plasmodium berghei in rodents. The search for anti-malarial drugs is of extreme importance. The aim of this study was to accomplish experimental infections of Plasmodium juxtanucleare in Gallus gallus and to test a substance recently synthesized, the 4-(9H-purin-6-ylthio)-7-cloroquinoline, derived of the cloroquine, to verify their effects on the avian malaria and to improve the aviary model for these types of tests. For a bird donor's encounter, two chicken farms were visited in the Juiz de Fora city and blood smears made in 30 hens. The prevalence was 100%. Among the 30 examined hens, six with the largest parasitemia values had been acquired and taken to the laboratory. To verify which the best day to retreat the blood to infected hens, five of the six hens received only dose of the imunossupressor substance (metilprednisolon acetate) and one served as control. The parasitaemia of the hens was accompanied by 26 days after the day of the imunossupression, through blood smears prepared each two days. The weight and corporal temperature were checked and made blood hematocrits. It was verified that to the 10th day powder-imunossupression it happened parasitaemia pick. There were fall of body weight and correlation with the parasitaemia. There was a little variation in the body temperature and hematocrite and there was not correlation of these with the parasitaemia. In other eight acquired hens in commercial house with 15 days old, experimental infections were accomplished with blood inoculated through intramuscle and intraperitoneally in the 0,3mL and 0,5mL for the two routes. During one month the hens had the value of parasites accompanied to compare which the most effective route and difference among the doses tested in the establishment of the infection. Significant difference was not observed among the xvii doses but it was possible to verify that the infection through intraperitonial reaches the parasitemia pick more quickly, with higher averages of parasites, however to the end of the experiment the total number of parasites differed hardly between the doses and routes. To test the effect a derived drug of the cloroquine, 45 chicks were infected, white Leghorn, through intramuscle route . The hens were separate in four experimental groups, 15 chicks for group (Group1 - no infected, Group 2 - infected and without treatment, Group 3 - infected and treated with the cloroquine and Group 4 - infected and treated with derived of the cloroquine) The drug was administered four consecutive days in the 100mg/Kg of alive weight dose. For the evaluation of the antimalarial effect of the drug, the hens had the number of parasites accompanied by blood smears done each two days after the tenth day of the inoculation. Also the weight and corporal temperature were checked each two days and hematocrit four days. Derived of the cloroquine had antimalarial activity, reducing the number of parasites and maintaining the lowest parasitaemia in relation to the group not treated and to the group treated with cloroquine.Plasmodium juxtanucleare é o agente causador da malária aviária que ocorre em alguns estados do Brasil. Esta malária está relacionada a diversos sinais clínicos e pode causar danos em criações rústicas de aves. O modelo aviário já foi utilizado para a investigação de drogas no combate à malária e hoje em dia o modelo mais utilizado é o Plasmodium berghei em roedores. A busca por anti-maláricos e malaricidas é de extrema relevância. O objetivo deste trabalho foi verificar o efeito da 4-(6-MERCAPTOPURINA)-7-CLOROQUINOLINA, uma substância recém sintetizada a partir da cloroquina, sobre a malária aviária em Gallus gallus e aprimorar o modelo aviário para testes de potenciais malaricidas. Para o encontro de uma ave doadora, foram visitadas no município de Juiz de Fora duas granjas e feitos esfregaços sangüíneos de 30 aves. A prevalência foi de 100%. Dentre as 30 aves examinadas, as seis com os maiores valores de parasitemia foram adquiridas e levadas para o laboratório. Para verificar qual o melhor dia para a retirada de sangue de aves infectadas e imunossuprimidas pelo acetato de metilprednisolona, cinco das seis aves receberam em dose única este imunossupressor e uma serviu como controle. A parasitemia das aves foi acompanhada por 26 dias após o dia da imunossupressão, por meio de esfregaços sangüíneos preparados a cada dois dias. Também foram aferidos o peso e temperatura corporal e feitos microhematócritos sangüíneos. Verificou-se que ao 10º dia pós-imunossupressão ocorreu pico de parasitemia. Houve queda de peso corporal e correlação com a parasitemia. Ocorreu pouca variação na temperatura corporal e hematócrito e não houve correlação destes com a parasitemia. Em outras oito aves adquiridas em casa comercial com 15 dias de idade, foram realizadas infecções experimentais com sangue inoculado via intramuscular e via intraperitonial nas doses de 0,3mL e 0,5mL para as duas vias. Durante um mês as aves tiveram o valor médio de parasitos acompanhado para comparar qual a via mais efetiva e se havia diferença entre as doses testadas no estabelecimento da infecção. Não foi observada diferença entre as xv dosagens, mas foi possível verificar que a infecção via intraperitonial atinge mais rapidamente o pico de parasitemia, com médias de parasitos mais altas, entretanto ao fim do experimento o número total de parasitos quase não diferiu entre as doses e vias. Para testar o efeito malaricida da 4-(6-MERCAPTOPURINA)-7-CLOROQUINOLINA, uma droga derivada da cloroquina, recém sintetizada, foram infectados 45 pintos, Leghorn branco, via intramuscular. As aves foram separadas em quatro grupos experimentais com 15 aves por grupo (Grupo1- não infectado, Grupo 2- infectado e sem tratamento, Grupo 3- infectado e tratado com a cloroquina e grupo 4- infectado e tratado com o derivado da cloroquina). A droga foi administrada via gavagem por 4 dias consecutivos na dose de 100mg/Kg de peso vivo. Para a avaliação do efeito malaricida da droga, as aves tiveram o número médio de parasitos encontrados acompanhados por esfregaços sangüíneos feitos a cada dois dias após o décimo dia da inoculação. Também foram aferidos o peso e temperatura corporal a cada dois dias e hematócrito a cada quatro dias. O derivado da cloroquina teve atividade malaricida, mantendo a parasitemia mais baixa em relação ao grupo controle não tratado e ao grupo controle tratado com a cloroquina. Entretanto em todos os grupos a parasitemia se manteve baixa. Sugere-se a investigação da ação malaricida desta droga em modelos com P. berghei ou culturas com P. falciparum

Plasmodium spp. and Haemoproteus spp. infection in birds of the Brazilian Atlantic Forest detected by microscopy and polymerase chain reaction

In recent years haemosporidian infection by protozoa of the genus Plasmodium and Haemoproteus, has been considered one of the most important factors related to the extinction and/or population decline of several species of birds worldwide. In Brazil, despite the large avian biodiversity, few studies have been designed to detect this infection, especially among wild birds in captivity. Thus, the objective of this study was to analyze the prevalence of Plasmodium spp. and Haemoproteus spp. infection in wild birds in captivity in the Atlantic Forest of southeastern Brazil using microscopy and the polymerase chain reaction. Blood samples of 119 different species of birds kept in captivity at IBAMA during the period of July 2011 to July 2012 were collected. The parasite density was determined based only on readings of blood smears by light microscopy. The mean prevalence of Plasmodium spp. and Haemoproteus spp. infection obtained through the microscopic examination of blood smears and PCR were similar (83.19% and 81.3%, respectively), with Caracara plancus and Saltator similis being the most parasitized. The mean parasitemia determined by the microscopic counting of evolutionary forms of Plasmodium spp. and Haemoproteus spp. was 1.51%. The results obtained from this study reinforce the importance of the handling of captive birds, especially when they will be reintroduced into the wild

Blood samples from Liophis typhlus (Linnaeus, 1758) (Serpentes, Dipsadinae) by cardiac puncture without surgical procedures

Resumo. O desafio para os pesquisadores que trabalham com animais silvestres é a escolha apropriada do método de coleta sanguínea que não cause grande estresse ou danos aos animais. O método de coleta mais utilizado para serpentes é a punção da veia caudal, mas ele possui desvantagens, como o risco de colabamento da veia de animais de pequeno porte e o risco de traumatizar a glândula caudal de fêmeas e o hemipenis de machos, ocasionando o prolapso desses órgãos. Liophis typhlus é pobremente documentada com relação aos aspectos anatômicos, hematológicos e parasitológicos. O objetivo deste trabalho é apresentar um novo protocolo para coletar sangue de L. typhlus que é fácil de realizar, fornece uma quantidade suficiente de sangue e permite a sobrevivência do animal sem causar danos ou estresse desnecessários