RESUMO
The lack of standardized diet for laboratory animals can have profound effects on animal health and lead to less reproducible research outcomes. Live diets are commonly used in zebrafish culture and, although they are a more natural feed than flake or pellet food, are also a potential source of pathogens and toxic compounds. Heavy metals are a group of such compounds, which can accumulate in fish leading to developmental abnormalities, reduced growth, and increased rates of mortality. Two to three weeks after feeding adult zebrafish a new lot of nonhatching decapsulated brine shrimp cysts (Decaps), embryos at the University of Minnesota Zebrafish Core Facility (ZCF) and the University of Utah Centralized Zebrafish Animal Resource (CZAR) began to exhibit an orange color in the yolk, and larval health began to decline. The concentration of chromium in the Decaps (69.6 mg/kg) was more than 30 times that of other zebrafish diets tested (up to 2.1 mg/kg) and is thought to be the cause of the observed symptoms. Within 3 weeks of removing the Decaps from the feeding regimen, the orange coloration in the yolks began to diminish, the morphological abnormalities began to subside, and larval survival rates began to increase. Thus, implementation of standardized zebrafish diets and regular feed-quality testing may help to prevent the introduction of contaminants to zebrafish research facilities.
Assuntos
Fenômenos Fisiológicos da Nutrição Animal , Cromo/toxicidade , Dieta/veterinária , Peixe-Zebra/crescimento & desenvolvimento , Ração Animal , Animais , Larva/crescimento & desenvolvimentoRESUMO
BACKGROUND: Severe malaria (SM) syndromes caused by Plasmodium falciparum infection result in major morbidity and mortality each year. However, only a fraction of P. falciparum infections develop into SM, implicating host genetic factors as important determinants of disease outcome. Previous studies indicate that tumour necrosis factor (TNF) and lymphotoxin alpha (LTα) may be important for the development of cerebral malaria (CM) and other SM syndromes. METHODS: An extensive analysis was conducted of single nucleotide polymorphisms (SNPs) in the TNF, LTA and LTB genes in highland Papuan children and adults, a population historically unexposed to malaria that has migrated to a malaria endemic region. Generated P-values for SNPs spanning the LTA/TNF/LTB locus were corrected for multiple testing of all the SNPs and haplotype blocks within the region tested through 10,000 permutations. A global P-value of < 0.05 was considered statistically significant. RESULTS: No associations between SNPs in the TNF/LTA/LTB locus and susceptibility to SM in highland Papuan children and adults were found. CONCLUSIONS: These results support the notion that unique selective pressure on the TNF/LTA/LTB locus in different populations has influenced the contribution of the gene products from this region to SM susceptibility.
Assuntos
Predisposição Genética para Doença , Linfotoxina-alfa/genética , Linfotoxina-beta/genética , Malária Falciparum/genética , Malária Falciparum/patologia , Polimorfismo de Nucleotídeo Único , Fator de Necrose Tumoral alfa/genética , Adulto , Criança , Pré-Escolar , Humanos , Linfotoxina-alfa/imunologia , Linfotoxina-beta/imunologia , Malária Falciparum/complicações , Malária Falciparum/imunologia , Papua Nova Guiné , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Nitric oxide (NO) mediates host resistance to severe malaria and other infectious diseases. NO production and mononuclear cell expression of the NO producing enzyme-inducible nitric oxide synthase (NOS2) have been associated with protection from severe falciparum malaria. The purpose of this study was to identify single nucleotide polymorphisms (SNPs) and haplotypes in the NOS2 promoter, to identify associations of these haplotypes with malaria severity and to test the effects of these polymorphisms on promoter activity. We identified 34 SNPs in the proximal 7.3 kb region of the NOS2 promoter and inferred NOS2 promoter haplotypes based on genotyping 24 of these SNPs in a population of Tanzanian children with and without cerebral malaria. We identified 71 haplotypes; 24 of these haplotypes comprised 82% of the alleles. We determined whether NOS2 promoter haplotypes were associated with malaria severity in two groups of subjects from Dar es Salaam (N = 185 and N = 250) and in an inception cohort of children from Muheza-Tanga, Tanzania (N = 883). We did not find consistent associations of NOS2 promoter haplotypes with malaria severity or malarial anemia, although interpretation of these results was potentially limited by the sample size of each group. Furthermore, cytokine-induced NOS2 promoter activity determined using luciferase reporter constructs containing the proximal 7.3 kb region of the NOS2 promoter and the G-954C or C-1173T SNPs did not differ from NOS2 promoter constructs that lacked these polymorphisms. Taken together, these studies suggest that the relationship between NOS2 promoter polymorphisms and malaria severity is more complex than previously described.
Assuntos
Haplótipos/genética , Malária/genética , Óxido Nítrico Sintase Tipo II/genética , Polimorfismo de Nucleotídeo Único , Regiões Promotoras Genéticas/genética , Alelos , Sequência de Bases , Linhagem Celular Tumoral , Criança , Pré-Escolar , Frequência do Gene , Heterogeneidade Genética , Genótipo , Humanos , Lactente , Quênia , Desequilíbrio de Ligação , Malária/patologia , Dados de Sequência Molecular , Mutação , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Índice de Gravidade de Doença , TanzâniaRESUMO
Ethanol (EtOH), isopropyl alcohol (IPA), and propylene glycol (PG) increase topical drug delivery, but are sometimes associated with erythema. A potential genetic basis for alcohol-associated erythema was investigated as the function of polymorphisms in coding and non-coding regions of class IB alcohol dehydrogenase (ADHIB) and evaluated for altered gene expression in vitro and metabolic activity in vivo via altered skin blood flow (Doppler velocimeter) and erythema (reflectance colorimeter a*) following topical challenge to 5 M EtOH, IPA, PG, and butanol (ButOH). Promoter polymorphisms G-887A and C-739T and exon G143A form eight ADHIB haplotypes with different frequencies in Caucasians vs Asians and exhibit variable gene expression and metabolic activity. Polymorphisms C-739T and G-887A independently alter gene expression, which is further increased by IPA and PG, but not EtOH or ButOH. EtOH and ButOH increase erythema as a function of skin blood flow. IPA increases skin blood flow without erythema and PG increased erythema with decreased skin blood flow, all as a function of ADHIB haplotype. PG-induced erythema was uniquely associated with tumor necrosis factor-alpha expression. Thus, erythema following alcohol exposure is alcohol type specific, has a pharmacogenetic basis related to ADHIB haplotype and can be functionally evaluated via Doppler velocimetry and reflectance colorimetry in vivo.