Hypoxia alters gene expression in the gonads of zebrafish (Danio rerio)

The objectives of this study were to characterize gene expression responses to hypoxia in gonads of mature zebrafish (Danio rerio), and to start characterizing modes of action by which hypoxia could potentially alter reproduction. Adult male and female zebrafish were maintained under normoxia (7mgO(2)/L), moderate hypoxia (3mgO(2)/L), and severe hypoxia (1mgO(2)/L) for 4 and 14 days and changes in gene expression in gonadal tissues (n=5 per sex per treatment) were evaluated using a commercial 21,000 gene zebrafish oligonucleotide microarray. Differentially expressed genes were determined using ANOVA (p<0.05), and enriched gene ontology (GO) categories (p<0.01) identified using GeneSpring GX software. Short-term (4d) exposure to hypoxia affected expression of genes associated with the initial adaptive responses such as: metabolism of carbohydrates and proteins, nucleotide metabolism, haemoglobin synthesis, reactive oxygen species metabolism, and locomotion. Prolonged (14d) hypoxia affected a suite of genes belonging to different GO categories: lipid metabolism, reproduction (e.g., steroid hormone synthesis), and immune responses. Results of the present study demonstrate that reproduction likely would be affected by hypoxia via multiple modes of action. These include previously hypothesized mechanisms such as modulation of expression of steroidogenic genes, and downregulation of serotonergic pathway. In addition, we propose that there are multiple other points of disruption of reproductive system function linked, for example, to reorganization of lipid transport and other mechanisms involved in responding to hypoxia (e.g., hydroxysteroid dehydrogenase alterations, downregulation of contractile elements, etc.).     

Scanning Electron Microscopy Observations of Failures of Implant Overdenture Bars: A Case Series Report

Background: Soldered or cast bars are used as a standard of care in attachment systems supporting maxillary and mandibular implant overdentures. When failures of these bars occur, currently there is a lack of evidence in relation to their specific etiology, location, or nature. Purpose: To investigate the failure process of a case series of six failed soldered bars, four intact soldered bars, and one intact cast milled bar, which had been supporting implant overdentures. Materials and Methods: A total of 11 different overdenture bars were removed from patients with different configuration of opposing arches. A failed bar (FB) group (n = 6) had failed soldered overdenture bars, which were recovered from patients following up to 2 years of wear before requiring prosthodontic maintenance and repair. An intact bar (IB) group (n = 5) had both soldered bars and a single cast milled bar, which had been worn by patients for 2 to 5 years prior to receiving other aspects of prosthodontic maintenance. All bars were examined using scanning electron microscopy to establish the possible mode of failure (FB) or to identify evidence of potential failure in the future (IB). Results: Evidence of a progressive failure mode of corrosion fatigue and creep were observed on all the FB and IB usually around the solder areas and nonoxidizing gold cylinder. Fatigue and creep were also observed in all the IB. Where the level of corrosion was substantial, there was no evidence of wear from the matrices of the attachment system. Evidence of an instantaneous failure mode, ductile and brittle overload, was observed on the fracture surfaces of all the FB, within the solder and the nonoxidizing gold cylinders, at the solder/cylinder interface. Conclusion: Corrosion, followed by corrosion fatigue, appears to be a key factor in the onset of the failure process for overdenture bars in this case series of both maxillary and mandibular overdentures. Limited sample size and lack of standardization identify trends only but prevent broad interpretation of the findings.     

SELECTIVE ENDOSCOPY IN MANAGEMENT OF INGESTED FOREIGN BODIES OF THE UPPER GASTROINTESTINAL TRACT: IS IT SAFE?

During a four-year period, 308 patients presented following ingestion of foreign bodies. Ingestion was accidental in 272 cases (88.3%) and deliberate in the remainder. Symptoms at presentation included dysphagia, odynophagia, nausea and vomiting, chest pain and pharyngeal discomfort. Sixty-eight patients were asymptomatic. A policy of expectant management and selective endoscopy was employed. Following initial assessment 202 patients (65.6%) were discharged without treatment, 30 (9.7%) of whom were later reviewed as outpatients and did not require admission. Forty-nine patients (16%) were admitted for treatment; 27 had oesophagoscopy, five bronchoscopy and two had foreign body extraction with direct laryngoscopy. In nine patients who were endoscoped, no foreign body was identified. Twenty-seven others were referred to the otorhinolaryngology service in another hospital. There were no deaths in the group and morbidity was 1.2%. We conclude that a policy of selective endoscopy is safe and effective in the management of patients following ingestion of foreign bodies.     

Toxicity reduction evaluation at a municipal wastewater treatment plant using mutagenicity as an endpoint

Previous work revealed substantial levels of mutagenicity in effluents from certain municipal wastewater treatment plants. One of these treatment plants was selected for further study to track the effluent mutagenicity to its sources, to chemically characterize the mutagenicity, and to assess the treatability of the mutagens. Mutagenicity testing using the Salmonella/microsome assay was performed on methylene chloride extracts of influent and effluent samples from the municipal wastewater treatment plant, as well as on four selected industrial effluents entering the plant. The mutagenicity of the influent samples was detected only in the presence of a microsomal metabolic activation system and was highest in Salmonella strain TA98. About two-thirds of the mutagenicity passed through the treatment plant, suggesting that the mutagenic compounds were refractory to conventional biological treatment. No significant mutagenic activity was detected in three of the industrial waste streams, all paper products plant discharges. However, a high level of mutagenicity (1.2 million TA98 revertants/liter) was detected in the effluent from a coke oven plant. This source could account for all of the mutagenicity entering the wastewater treatment plant. After fractionation of the coke oven effluent by sequential extraction at neutral, acidic and basic pH with methylene chloride, 93% of the TA98 (+S9) mutagenicity was found in the neutral fraction. A C₁₈ column fractionation scheme using a methanol/water elution gradient revealed that 92% of the mutagenicity eluted with the 75% and the 80% methanol in water fractions. The C₁₈ fractionation also provided good separation of mutagenicity from toxicity to fathead minnows. This study has demonstrated the potential of toxicity reduction evaluation (TRE) methodology for tracing effluent toxicity to its source, using genotoxicity as an endpoint.Disclaimer. This article has been reviewed by the U.S. Environmental Protection Agency and approved for publication. Mention of trade names or commercial products does not constitute endorsement or recommendation for use.     

Persistent diarrhea associated with AIDS

Chronic diarrhea and wasting are very common manifestations of AIDS in adults in developing countries. Etiologic studies show that protozoa (including Cryptosporidium parvum, Isospora belli , and Enterocytozoon bieniusi ) and Mycobacterium avium-intracellulara are the most frequently identified pathogens. Limited data in children suggest that common enteric pathogens are equally as likely in HIV⁺ and HIV^- babies. Preliminary analysis of an ongoing longitudinal study of 469 babies born to mothers with known HIV serostatus in Kinshasa, Zaire, reveals progression of acute to persistent diarrhea is six times greater in HIV⁺ compared to HIV^- babies, and 3.5 times greater in HIV^- babies born of HIV⁺ mothers in comparison to HIV^- babies with HIV^- mothers. HIV⁺ babies were also at greater risk than HIV^- babies to have recurrent episodes of diarrhea (RR = 2.3). Fifty percent of the deaths were due to acute or persistent diarrhea, and were strongly associated with HIV infection. Efforts to improve child survival in AIDS infected populations will need to address HIV infections in both mothers and infants.     

阿克拉霉素A聚氰基丙烯酸异丁酯毫微粒冻干针剂体内外抗肝癌活性

阿克拉霉素A聚氰基丙烯酸异丁酯毫微粒的冻干针剂,能明显抑制体外培养人肝癌细胞株7703的生长,IC50为0.28μg·ml^-1。在0.8μg·ml^-1浓度时,克隆形成抑制率为90%,抑制作用有明显剂量依赖关系而未见明显时间依赖关系。静脉给药后,对常位移植人肝癌模型裸小鼠的抑瘤率为86.84%,肿瘤细胞增殖活性阳性率为20.83%。体内外均显示明显的抗肝癌活性,且体内抗肝癌活性比阿克拉霉素A冻干针剂强。     

Development and validation of a 2,000-gene microarray for the fathead minnow (Pimephales promelas)

Gene microarrays provide the field of ecotoxicology new tools to identify mechanisms of action of chemicals and chemical mixtures. Herein we describe the development and application of a 2,000-gene oligonucleotide microarray for the fathead minnow Pimephales promelas, a species commonly used in ecological risk assessments in North America. The microarrays were developed from various cDNA and subtraction libraries that we constructed. Consistency and reproducibility of the microarrays were documented by examining multiple technical replicates. To test application of the fathead minnow microarrays, gene expression profiles of fish exposed to 17beta-estradiol, a well-characterized estrogen receptor (ER) agonist, were examined. For these experiments, adult male fathead minnows were exposed for 24 h to waterborne 17beta-estradiol (40 or 100 ng/L) in a flow-through system, and gene expression in liver samples was characterized. Seventy-one genes were identified as differentially regulated by estradiol exposure. Examination of the gene ontology designations of these genes revealed patterns consistent with estradiol's expected mechanisms of action and also provided novel insights as to molecular effects of the estrogen. Our studies indicate the feasibility and utility of microarrays as a basis for understanding biological responses to chemical exposure in a model ecotoxicology test species.     

Effects of a glucocorticoid receptor agonist, dexamethasone, on fathead minnow reproduction, growth, and development

Synthetic glucocorticoids are pharmaceutical compounds prescribed in human and veterinary medicine as anti-inflammatory agents and have the potential to contaminate natural watersheds via inputs from wastewater treatment facilities and confined animal-feeding operations. Despite this, few studies have examined the effects of this class of chemicals on aquatic vertebrates. To generate data to assess potential risk to the aquatic environment, we used fathead minnow 21-d reproduction and 29-d embryo-larvae assays to determine reproductive toxicity and early-life-stage effects of dexamethasone. Exposure to 500?μg dexamethasone/L in the 21-d test caused reductions in fathead minnow fecundity and female plasma estradiol concentrations and increased the occurrence of abnormally hatched fry. Female fish exposed to 500?μg dexamethasone/L also displayed a significant increase in plasma vitellogenin protein levels, possibly because of decreased spawning. A decrease in vitellogenin messenger ribonucleic acid (mRNA) expression in liver tissue from females exposed to the high dexamethasone concentration lends support to this hypothesis. Histological results indicate that a 29-d embryo-larval exposure to 500?μg dexamethasone/L caused a significant increase in deformed gill opercula. Fry exposed to 500?μg dexamethasone/L for 29?d also exhibited a significant reduction in weight and length compared with control fry. Taken together, these results indicate that nonlethal concentrations of a model glucocorticoid receptor agonist can impair fish reproduction, growth, and development.     

A QSAR evaluation of Ah receptor binding of halogenated aromatic xenobiotics.

Because of their widespread occurrence and substantial biological activity, halogenated aromatic hydrocarbons such as polychlorinated biphenyls (PCBs), polychlorinated dibenzofurans (PCDFs), and polychlorinated dibenzo-p-dioxins (PCDDs) comprise one of the more important classes of contaminants in the environment. Some chemicals in this class cause adverse biological effects after binding to an intracellular cytosolic protein called the aryl hydrocarbon receptor (AhR). Toxic responses such as thymic atrophy, weight loss, immunotoxicity, and acute lethality, as well as induction of cytochrome P4501A1, have been correlated with the relative affinity of PCBs, PCDFs, and PCDDs for the AhR. Therefore, an important step in predicting the effects of these chemicals is the estimation of their binding to the receptor. To date, however, the use of quantitative structure activity relationship (QSAR) models to estimate binding affinity across multiple chemical classes has shown only modest success possibly due, in part, to a focus on minimum energy chemical structures as the active molecules. In this study, we evaluated the use of structural conformations other than those of minimum energy for the purpose of developing a model for AhR binding affinity that encompasses more of the halogenated aromatic chemicals known to interact with the receptor. Resultant QSAR models were robust, showing good utility across multiple classes of halogenated aromatic compounds.