Earlier biomarkers in fish evidencing stress responses to metal and organic pollution along the Doce River Basin.

The Doce River Basin (DRB) represents a well-described watershed in terms of contamination by metals, especially after a major environmental disaster of a mining tailing dam failure. Despite the massive mortality of the ichthyofauna, very few studies addressed the risks to the health of wild fish exposed to complex mixtures of pollutants from multiple sources. The present study proposed to investigate earlier responses of fish for assessing the impacts of multiple sources of pollution, considering: different niches of fish and target organs; and the influence of seasonality, associated with their probable sources of pollution. To achieve that, fish were collected along the DRB, and biomarkers responses were assessed in target organs and correlated with the levels of inorganic and organic contaminants. As one of the most prominent responses, fishes from the Upper DRB showed the highest expression of the metallothionein and oxidative stress parameters which were related to the higher levels of metals in this region due to the proximity of mining activities. On the other hand, higher levels of DNA damage and increased AChE activity from fish sampled in the Mid and Lower DRB were more associated with organic contaminants, from other sources of pollution than mining residues. The integrated biomarker responses also revealed seasonal variations, with higher values in fishes from the dry season, and pelagic fish showing greater variation within the seasons. The multivariate analysis integrating suitable biomarkers with chemical data represented an adequate strategy for assessing the ecological risks in the DRB, allowing the identification of distinct spatio-temporal impacts from multiple sources of contaminants. The continued exposure of the ichthyofauna representing future risks reinforces the need for ecological restoration and the protection of the fauna from the Doce River.

Chemical data of contaminants in water and sediments from the Doce River four years after the mining dam collapse disaster.

Chemical datasets describing the occurrence of both inorganic and organic contaminants along the Doce River Basin (DRB) could provide a better understanding of the potential impacts of a major mining dam collapse disaster combined to additional chronic sources of contamination. This data article presents datasets of main contaminants detected in the water and sediments sampled four years after the mining dam collapse in the DRB. A summary table of data obtained in the literature is also provided to allow a comparison of the variation of chemicals before, right after in 2015/2016 and after the event (current data). In addition, there are also provided physical-chemical parameters of water and sediments of different sampling sites, which could support the investigation of chemicals distribution. For this purpose, triplicate samples of water and sediment were obtained in 8 sampling sites along the DRB during wet and dry seasons of 2019, totalizing 48 samples of each environmental matrix. The sampling sites were strategically selected according to their different main sources of pollution along the river. Concentrations of trace elements and organic contaminants (polycyclic aromatic hydrocarbons, and pyrethroids) were determined in samples of water and sediments by inductively coupled plasma mass spectrometry (ICP-MS) and gas chromatography - mass spectrometry GC-MS, respectively. Main data obtained in the literature consisted in published reports from environmental agencies (IGAM) and private companies (RENOVA) as well as journal articles. The datasets provided may be useful to the stakeholders, which include scientific community, authorities and public agencies, and private companies interested to understand the impacts of the contaminants introduced along the River Basin four years after the environmental disaster.

GnRH analog is ineffective in increasing adult height in girls with puberty onset after 7 years of age: a systematic review and meta-analysis.

Objective We assessed the effectiveness of puberty blockade with a gonadotropin-releasing hormone (GnRH) analog in increasing adult height (AH) in girls with puberty onset between 7 and 10 years of age. Methods We performed a systematic review and included controlled studies in which girls with early puberty (EP) were assigned to the GnRH analog or no treatment groups. The primary outcome analyzed was AH. Search strategies were applied to the MEDLINE, EMBASE, LILACS and CENTRAL databases. Results We identified 1514 references, and six studies fulfilled our eligibility criteria. Two studies were randomized and four were not randomized. At the baseline of each trial, height, chronological age, bone age, predicted AH (PAH) and target height (TH) were equal between the groups. All studies used intramuscular triptorelin every 28 days in the intervention groups. The mean duration of the therapy was 2 years. Meta-analysis of AH among the six studies (comprising 332 girls) showed no significant difference between the groups (mean difference = 0.50 cm, 95% confidence interval = -0.72 to 1.73 cm, I 2 = 0%). In a sub-group analysis based on PAH (<155 cm and 155 cm and equal to TH), there was no difference in average AH between the groups. The quality of evidence according to the Grading of Recommendations Assessment, Development and Evaluation approach was low. Conclusion We found no evidence from controlled experimental and observational studies that compared with no treatment, the use of GnRH analogs improved AH in girls with EP.

Alterations of cytochrome P450 and the occurrence of persistent organic pollutants in tilapia caged in the reservoirs of the Iguaçu River.

Environmental chemicals originating from human activities, such as persistent organic pollutants (POPs), may interfere with the endocrine system of aquatic organisms. The effect of these chemicals on biota and human populations is of high public concern but remains poorly understood, especially in aquatic environments of South America. The aim of this study was to investigate the bioavailability of POPs and the related effects in caged male tilapia (Oreochromis niloticus) in four cascading reservoirs of the Iguaçu River, Southern Brazil. POPs including organochlorine pesticides (OCPs), polychlorinated biphenyl (PCBs), and polybrominated diphenyl ethers (PBDEs) were determined in the reservoir water and tissue samples of tilapia after two months of exposure. The PCB levels in water (14.7 ng L-1) were 14 times higher than the limits permitted by the Brazilian legislation in the Salto Santiago (SS) reservoir. Similarly, concentrations of aldrin and its metabolites (6.05 ng L-1) detected in the water sample of the Salto Osório (SO) reservoir were also above the permitted limits. RT-qPCR analysis revealed different transcript levels of cytochrome P450 enzymes (CYP1A and CYP3A) in the liver among the four groups, with induced activity in tilapia from the SS reservoir. Quantification of the CYP3A mRNA expression and catalytic activity showed higher values for fish caged at the SS reservoir. The fish from this site also had a higher number of eosinophils observed in the testes. Although overt measurements of endocrine disruption were not observed in caged fish, alteration of CYP enzymes with co-occurrence of organochlorine contaminants in water may suggest bioavailability of contaminants from agricultural sources to biota. Additional studies with feral or caged animals for a longer duration may be necessary to evaluate the risks of the waterways to humans and wildlife.

Vitellogenin levels and others biomarkers show evidences of endocrine disruption in fish species from Iguaçu River - Southern Brazil.

The adverse effects of endocrine disrupting chemicals (EDCs) on aquatic wildlife and human health represent a current issue of high public concern. Even so, they are still poorly studied in aquatic environments of South America. The aim of the present study was to investigate the impact of EDCs in five cascading reservoirs from the Iguaçu River, evaluating reproductive endpoints in three native fish species (Astyanax bifasciatus, Chrenicicla iguassuensis and Geophagus brasiliensis). Additionally, a polyclonal antiserum anti-vitellogenin from G. brasiliensis and a capture ELISA assay were developed for detection of estrogenic or anti-estrogenic activities in male and female fish, respectively. Vitellogenin (VTG) levels in male fish from the Iguacu River was observed, as well as decreased levels of vitellogenin and estradiol in the plasma of female fish. These findings were associated with immature gonads and lower gonadosomatic index in G. brasiliensis adult females from the Foz do Areia (FA) Reservoir. Additionally, both endemic species (Astyanax bifasciatus and Chrenicicla iguassuensis) displayed immature gonads and histological changes, such as degeneration of germ cells, in other studied reservoirs. The current results suggest that these reproductive responses may be associated with the bioavailability of EDCs in the Iguaçu River. These impacts are likely related to chemicals released by human activities, especially from sewage and industrial sources and agricultural production, detected in previous studies. Overall, the FA reservoir was potentially the most affected by chemicals with endocrine properties, and further studies are necessary to identify and quantify these chemicals.

Bioavailability of pollutants sets risk of exposure to biota and human population in reservoirs from Iguaçu River (Southern Brazil).

The Iguaçu River, located at the Southern part of Brazil, has a great socioeconomic and environmental importance due to its high endemic fish fauna and its potential to generate hydroelectric power. However, Iguaçu River suffers intense discharge of pollutants in the origin of the river. In a previous report, the local environmental agency described water quality to improve along the river course. However, no study with integrated evaluation of chemical analysis and biological responses has been reported so far for the Iguaçu River. In the current study, three different Brazilian fish species (Astyanax bifasciatus, Chrenicicla iguassuensis, and Geophagus brasiliensis) were captured in the five cascading reservoirs of Iguaçu River for a multi-biomarker study. Chemical analysis in water, sediment, and muscle indicated high levels of bioavailable metals in all reservoirs. Polycyclic aromatic hydrocarbons (PAHs) were detected in the bile of the three fish species. Integration of the data through a FA/PCA analysis demonstrated the poorest environmental quality of the reservoir farthest from river's source, which is the opposite of what has been reported by the environmental agency. The presence of hazardous chemicals in the five reservoirs of Iguaçu River, their bioaccumulation in the muscle of fish, and the biological responses showed the impacts of human activities to this area and did not confirm a gradient of pollution between the five reservoirs, from the source toward Iguaçu River's mouth. Therefore, diffuse source of pollutants present along the river course are increasing the risk of exposure to biota and human populations.

Very high resolution single pass HLA genotyping using amplicon sequencing on the 454 next generation DNA sequencers: Comparison with Sanger sequencing.

Compared to Sanger sequencing, next-generation sequencing offers advantages for high resolution HLA genotyping including increased throughput, lower cost, and reduced genotype ambiguity. Here we describe an enhancement of the Roche 454 GS GType HLA genotyping assay to provide very high resolution (VHR) typing, by the addition of 8 primer pairs to the original 14, to genotype 11 HLA loci. These additional amplicons help resolve common and well-documented alleles and exclude commonly found null alleles in genotype ambiguity strings. Simplification of workflow to reduce the initial preparation effort using early pooling of amplicons or the Fluidigm Access Array™ is also described. Performance of the VHR assay was evaluated on 28 well characterized cell lines using Conexio Assign MPS software which uses genomic, rather than cDNA, reference sequence. Concordance was 98.4%; 1.6% had no genotype assignment. Of concordant calls, 53% were unambiguous. To further assess the assay, 59 clinical samples were genotyped and results compared to unambiguous allele assignments obtained by prior sequence-based typing supplemented with SSO and/or SSP. Concordance was 98.7% with 58.2% as unambiguous calls; 1.3% could not be assigned. Our results show that the amplicon-based VHR assay is robust and can replace current Sanger methodology. Together with software enhancements, it has the potential to provide even higher resolution HLA typing.

Altered expression of dermokine in skin disorders.

BACKGROUND: Although dermokine-ß, a glycoprotein expressed in epithelial cells, does not have significant homology to other proteins, its carboxyl-terminal domain shares a high pI value with many cytokines, suggesting similar functions. OBJECTIVE: To better understand the biology of dermokine, we here determined its localization under pathological conditions and examined factors that regulate its expression. METHODS: We generated an anti-human dermokine-ß/γ monoclonal antibody cross-reacting with the mouse protein. Using this antibody, immunohistological staining and Western blotting of dermokine-ß/γ were performed with various tissue samples. RESULTS: Although human dermokine-ß/γ was expressed in almost all granular layers, upper spinous layers of the skin were also stained with anti-dermokine-ß/γ antibody in inflammatory skin disorders. Dermokine-ß/γ was expressed in keratoacanthoma and a part of well-differentiated squamous cell carcinoma (SCC). However, dermokine-ß/γ was not detected in poorly differentiated SCC or tumours derived from non-keratinocytes. In mice, dermokine-ß/γ-expressed keratinocytes were increased in models of contact hypersensitivity, ultraviolet-irradiated skin injury and wound healing. Consistent with expanded distribution in inflammatory skin diseases, proinflammatory cytokines such as interleukin-1ß, interleukin-12, and tumour necrosis factor-α augmented dermokine-ß/γ expression in cultured human keratinocytes. In contrast, growth factors including epidermal growth factor, insulin-like growth factor-I, keratinocyte growth factor and transforming growth factor-α significantly reduced dermokine expression. CONCLUSION: These results provide novel insights into the physiological and pathological significance of dermokine in the epidermis.

Amending reduced fish-meal feeds with marine lecithin, but not soy lecithin, improves the growth of juvenile cobia and may attenuate heightened responses to stress challenge.

Sparing of marine resources in aquafeeds can be environmentally and economically advantageous; however, fish meal (FM) replacement can affect the production performance and physiological competence. Phospholipids are increasingly understood to be involved in maintaining growth and vigour in fish and may be deficient in reduced FM formulations. Accordingly, we evaluated the growth and stress tolerance of juvenile cobia fed typical (50% FM) or reduced FM feeds (12% FM) with or without phospholipid amendment [1% marine lecithin (12% FM + Marine PL) or soy lecithin (12% FM + Soy PL)] for 6 weeks in triplicate tanks (N = 3) in a recirculation aquaculture system. The 50% FM feed yielded significantly superior growth and growth efficiency in comparison with the 12% FM and 12% FM+ Soy PL feeds, but the 12% FM+ Marine PL feed yielded comparable results to 50% FM feed. A low-water stress challenge induced elevated plasma glucose, cortisol and lactate levels in all treatments. However, a significant interaction (diet × stress) effect suggested a lesser cortisol response among fish fed the 12% FM+ Marine PL and 50% FM diets. These findings demonstrate that growth performance and, perhaps, resilience of cobia raised on reduced FM feeds may be improved by the addition of marine-origin phospholipid to the diet.

Cadmium effects on early development of chick embryos.

The toxic potential of cadmium (Cd) is well-documented for young and adult vertebrates, but it is still poorly understood in the early stages of development. In this study, cadmium effects were investigated on Gallus gallus embryos after injection of CdCl(2) (5µM and 50µM) within the egg air chamber, and incubation for 48 and 72h. After exposure, morphological and enzymatic analyses for glucose-6-phosphate dehydrogenase (G6PDH) and glutathione S-transferase (GST) were performed. Critical morphological abnormalities occurred after exposure to the highest concentration of cadmium, mainly in the cephalic region, indicating the powerful teratogenic effect of Cd to chick embryos. Cd exposure did not alter enzymatic activities when compared to the control group, but the levels of G6PDH activity were highest in older embryos at stage 19, indicating that antioxidant defenses are not so robust in the earliest embryo stages.

Blood cardioplegia serves as a bicarbonate donor to the myocardium during ischemia: effects of anoxia and hypercapnia on acid-base characteristics of blood cardioplegic solution.

OBJECTIVES: We investigated the alterations of acid-base characteristics of the blood cardioplegia (BCP) solution during aortic cross-clamping in hearts arrested with BCP and during in vitro-simulated ischemia. METHODS: Following aortic cross-clamping, the hearts of 40 patients undergoing cardiac surgery were intermittently infused with an 18°C BCP solution and finally with a 34°C BCP solution prior to aortic cross-clamp release. We measured the pH, partial CO(2) pressure (pCO(2)), [HCO(3)(-)], and [Cl(-)] of the coronary sinus effluent in the final BCP solution. The BCP solution was assessed under in vitro gassing at 34°C with 95% N(2) + 5% CO(2) (n = 6), 50% N(2) + 50% CO(2) (n = 3), or 100% CO(2) (n = 6). RESULTS: The coronary sinus effluent, compared with the preinfusion BCP solution, exhibited a significantly lower pH and a greater pCO(2) with no change in the [HCO(3)(-)] level. In vitro, the 95% N(2) + 5% CO(2) gassing (simulated hypoxia) group exhibited a slight increase in [HCO(3)(-)] with no change in pCO(2) or pH whereas the 50% N(2) + 50% CO(2) gassing and the 100% CO(2) gassing (simulated hypoxia and hypercapnia) groups exhibited a significant increase in [HCO(3)(-)] under high pCO(2)-induced acidification. CONCLUSIONS: Under anoxia and CO(2) retention during aortic cross-clamping, the BCP solution can be a bicarbonate donor to the myocardium.

[Surgical repair of extracardiac unruptured aneurysm of the sinus of Valsalva].

An isolated extracardiac unruptured aneurysm of the non-coronary sinus of Valsalva was detected incidentally in a 49-year-old woman with chronic obstructive pulmonary disease. The orifice of the aneurysm was closed with XAG-400 patch. The aortic valve was normal. 20 cases of successful operation have been reported in Japan with extracardiac unruptured aneurysm. In the review of the literature, patch closure of the orifice of the aneurysm was the standard procedure in most cases. However, aortic root replacement was required in some cases with concomitant cardiac and/or aortic disease.

[Management of mediastinitis and preventions of perioperative nosocomial infection after cardiovascular surgery].

This article reviews risk factors and treatment of perioperative nosocomial infections. The primary prophylactic antibiotic is recommended to be a cefazolin or penicillin with sulbactam, because the most frequent organism cultured in cardiac surgical site infection (SSI) is Staphyloccocus. Antibiotic prophylaxis of 48 hours' duration after cardiovascular surgery is clinically effective in minimizing infectious complication. In patients considered at high risk for a staphylococcal infection, vancomycin may be recommended. In the treatment of postoperative infections, Gram-stain-based antibiotic selection is necessary and the initial empirical therapy to ensure adequate coverage of potentially infective organisms should be accompanied by de-escalation until microbiological data become available. Mediastinitis, which is one of the important infectious complications after cardiovascular surgery, requires surgical drainage and debridement, as well as antimicrobial therapy. Vacuum-assisted closure (VAC) is an effective therapy compared with the conventional technique of open packing. Continuous clinician's efforts and prolonged infection control programs are very important for prevention of perioperative nosocomial infection after cardiac surgery.

A DNA microarray-based methylation-sensitive (MS)-AFLP hybridization method for genetic and epigenetic analyses.

We previously developed a PCR-based DNA fingerprinting technique named the Methylation Sensitive (MS)-AFLP method, which permits comparative genome-wide scanning of methylation status with a manageable number of fingerprinting experiments. The technique uses the methylation sensitive restriction enzyme NotI in the context of the existing Amplified Fragment Length Polymorphism (AFLP) method. Here we report the successful conversion of this gel electrophoresis-based DNA fingerprinting technique into a DNA microarray hybridization technique (DNA Microarray MS-AFLP). By performing a total of 30 (15 x 2 reciprocal labeling) DNA Microarray MS-AFLP hybridization experiments on genomic DNA from two breast and three prostate cancer cell lines in all pairwise combinations, and Southern hybridization experiments using more than 100 different probes, we have demonstrated that the DNA Microarray MS-AFLP is a reliable method for genetic and epigenetic analyses. No statistically significant differences were observed in the number of differences between the breast-prostate hybridization experiments and the breast-breast or prostate-prostate comparisons.

Concentration of clarithromycin and 14-R-hydroxy-clarithromycin in plasma of patients with Mycobacterium avium complex infection, before and after the addition of rifampicin.

Clarithromycin (CAM) and rifampicin (RFP) have both been recognized to be effective antibiotic agents against Mycobacterium avium complex (MAC) infection. Rifamycin derivatives including RFP and rifabutin modulate the CAM metabolism by inducing the hepatic cytochrome p-450 3A4. To clarify the effect of RFP on the CAM metabolism, we measured the plasma concentration of CAM and 14-R-hydroxyclarithromycin (M-5), the major metabolite of CAM, in 9 patients suffering from MAC infection before and after the addition of RFP. After the addition of RFP, the mean plasma concentration of CAM significantly decreased, while that of M-5 did not. In addition, the amount of CAM + M-5 concentration also significantly decreased. As M-5 is less effective against MAC infection than CAM, more attention should thus be paid to the plasma CAM concentration in patients administered CAM and RFP concomitantly.

Tei index determined by tissue Doppler imaging in patients with pulmonary regurgitation after repair of tetralogy of Fallot.

The myocardial performance index (Tei index) determined by the pulsed Doppler method is a simple and noninvasive measurement for assessing global right ventricular (RV) function. This index can also be obtained by tissue Doppler imaging (TDI). The effects of significant pulmonary regurgitation (PR) on the determination of the Tei index by these two methods have not been investigated. We examined 15 patients (6.3 +/- 2.2 years) with significant PR after repair of tetralogy of Fallot (TOF) and 24 age-matched healthy children. Myocardial wall motion velocities at the tricuspid annulus were assessed during systole (Sa), early diastole (Ea), and late diastole (Aa) from a four-chamber view. Pulsed Doppler-Tei index and TDI-Tei index were measured as reported previously. The Tei index obtained by the pulsed Doppler method in TOF patients did not differ from that in normal children (0.30 +/- 0. 12 vs 0.32 +/- 0.07, p = not significant). TDI showed that TOF patients had significantly decreased Ea, Aa, and Sa velocities compared to those of normal children. Both isovolumic contraction time and isovolumic relaxation time in TOF patients were significantly longer than those in normal children (88 +/- 18 vs 62 +/- 23 msec and 46 +/- 11 vs 21 +/- 12 msec, respectively; p < 0.0001), although the duration of Sa did not differ between the two groups. Consequently, the Tei index as measured by TDI was significantly greater in TOF patients than in normal children (0.48 +/- 0.07 vs 0.30 +/- 0.07, p < 0.0001). The Tei index measured by the pulsed Doppler method is limited in its ability to assess RV function in patients with significant PR. However, the Tei index determined by TDI is a sensitive indicator of RV function in these patients, and it appears to be a promising new means of assessing global RV function in patients with significant PR.

A base substitution in the interleukin-10 (IL-10) promoter between Sp1 and ets-1 binding sites is not associated with variation of IL-10 levels.

Interleukin 10 (IL-10) may play an important anti-inflammatory and immunoregulatory role in asthma. In this study, we investigated the role of a C to A substitution at position -627 of the IL-10 promoter, located in a necessary transcriptional region, which contains a number of putative transcriptional binding sites. The -627 nucleotide position is itself flanked by Sp-1 and ets-1 binding sites. We studied the allele frequency in 53 unrelated subjects from an admixed Caucasian, Asian and Pacific Islander group with personal or family histories of asthma. The frequency of homozygous C/C, heterozygous C/A, and homozygous A/A alleles at position -627 was 0.28, 0.44 and 0.28, respectively. In vitro assays indicated no differences between the C/C and A/A forms in binding transcriptional factors, especially Sp-1 factor, or in promoter activity. Moreover, in this selected population, there was no association between the C to A substitution and serum IL-10 levels. The mean level of IL-10 serum was determined to be 3.87 +/- 1.23 pg/ml in subjects carrying the A/A genotype, 3.47 +/- 0.57 for C/C genotype and 3.13 +/- 0.41 for the heterozygous (C/A genotype). This requires confirmation by comparing to non-asthmatic subjects. We conclude that although the -627 A allele occurs frequently (50% of alleles) in this selected group, in vitro assays and serum IL-10 levels suggest that the -627C-->A substitution represents a silent or neutral variant in the IL-10 promoter.