Neurotoxic, biotransformation, oxidative stress and genotoxic effects in Astyanax altiparanae (Teleostei, Characiformes) males exposed to environmentally relevant concentrations of diclofenac and/or caffeine.

The present study evaluated neurotoxic, biotransformation, genotoxic and antioxidant responses to relevant environmental concentrations of diclofenac (0.4 µg L-1) and caffeine (27.5 µg L-1), separate and combined, in adult males of the freshwater fish Astyanax altiparanae after a subchronic exposure (14 days). Fish exposed to diclofenac and caffeine, both separate and combined, revealed a neurotoxic effect through the inhibition of acetylcholinesterase activity in the muscle, while diclofenac alone and in combination caused cyclooxygenase inhibition. Caffeine alone produces genotoxicity on this species but, when combined with diclofenac, it potentiates hepatic lipoperoxidation and the inhibition of oxidative stress enzymes, while diclofenac alone or in combination produces a general inhibition of important enzymes. This study suggests that aquatic contamination produced by these pharmaceuticals has the potential to affect homeostasis and locomotion in A. altiparanae and compromise their immune system and general health.

Global variation in freshwater physico-chemistry and its influence on chemical toxicity in aquatic wildlife.

Chemical pollution is one of the major threats to global freshwater biodiversity and will be exacerbated through changes in temperature and rainfall patterns, acid-base chemistry, and reduced freshwater availability due to climate change. In this review we show how physico-chemical features of natural fresh waters, including pH, temperature, oxygen, carbon dioxide, divalent cations, anions, carbonate alkalinity, salinity and dissolved organic matter, can affect the environmental risk to aquatic wildlife of pollutant chemicals. We evidence how these features of freshwater physico-chemistry directly and/or indirectly affect the solubility, speciation, bioavailability and uptake of chemicals [including via alterations in the trans-epithelial electric potential (TEP) across the gills or skin] as well as the internal physiology/biochemistry of the organisms, and hence ultimately toxicity. We also show how toxicity can vary with species and ontogeny. We use a new database of global freshwater chemistry (GLORICH) to demonstrate the huge variability (often >1000-fold) for these physico-chemical variables in natural fresh waters, and hence their importance to ecotoxicology. We emphasise that a better understanding of chemical toxicity and more accurate environmental risk assessment requires greater consideration of the natural water physico-chemistry in which the organisms we seek to protect live.

Aluminum at environmental concentrations affects the sperm quality of the freshwater teleost Astyanax altiparanae: An in vitro study.

Aluminum (Al) is present in rivers and reservoirs in concentrations above that is allowed by regulatory agencies (e.g. 0.5 mg L-1 Al), which can impair fish reproduction. The present study evaluated the in vitro effects on the sperm of Astyanax altiparanae upon Al exposure at different concentrations (0, 0.05, 0.1, 0.3, and 0.5 mg L-1) with various exposure periods (50 s, 10 min, and 30 min). The following biomarkers were evaluated: membrane vitality, DNA fragmentation, morphology, kinetics (10 s and 30 s after sperm activation), and sperm mitochondrial activity. Al damages the membrane vitality of gametes at 0.3 and 0.5 mg L-1 after 50 s of exposure. After 30 min of exposure, there was a decrease in membrane vitality at 0.1 and 0.5 mg L-1, and the membrane vitality decreased with increased exposure time. Within 30 s after sperm activation, Al (0.3 and 0.5 mg L-1) reduced sperm motility by more than 50% at the longest exposure time, while at 0.1 and 0.5 mg L-1, Al exposure reduced motility over time. The average path speed (VAP; 10 s post-sperm activation) was reduced at longer exposure times at 0.05 and 0.5 mg L-1 of Al. Increased exposure time had deleterious effects on mitochondrial activity at the highest concentrations tested. Al did not damage DNA and sperm morphology. In conclusion, Al negatively influences the sperm quality of A. altiparanae with a potential effect of exposure time and increasing concentrations.

Diclofenac and caffeine inhibit hepatic antioxidant enzymes in the freshwater fish Astyanax altiparanae (Teleostei: Characiformes).

Although concentrations of pharmaceutical compounds in aquatic ecosystems are low, they can cause toxic effects on organisms. The aim of this study was to evaluate the effects of diclofenac (DCF), a non-steroidal anti-inflammatory drug, and caffeine (CAF), a central nervous system stimulant, both alone or combined, in Astyanax altiparanae males under acute exposure (96 h), measuring neurotoxicity biomarkers, antioxidant response and damage at biochemical and cellular levels. DCF concentration in water, separated and combined, was 3.08 mg L-1 and that of CAF was 9.59 mg L-1. To assess neurotoxicity, brain and muscle acetylcholinesterase (AChE) activities were measured. To evaluate oxidative stress, the enzymatic activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and glutathione S-transferase (GST), as well as lipoperoxidation (LPO), were analyzed in liver and gills. Activity of hepatic cyclooxygenase (COX) was also evaluated. Genotoxicity was assessed in blood using comet assay and micronucleus test, as well as nuclear abnormalities. DCF and CAF, alone or combined, had neither effect on AChE activity, nor in the activity of SOD, CAT, GPx and GST in gills. In liver, DCF inhibited SOD and GPx activity, CAF inhibited CAT activity, the mixture inhibited SOD and GST activity; although only fish exposed to CAF showed increased hepatic LPO. Under these experimental conditions, no effect on COX activity was observed, nor cytotoxic and genotoxic damage. The most pronounced effects were caused by the drugs separately, since both compounds altered the enzymes, but only CAF triggered LPO, showing more harmful effects.

Paternal exposure to aluminum, acidity, and temperature affect fatty acid seminal profile, embryonic and larval development of Astyanax altiparanae.

We investigated the effects of water acidity, temperature, and aluminum (Al) on the fatty acid (FA) seminal profile, reproductive parameters (fertilization and hatching) and embryonic development of Astyanax altiparanae. We treated males with different experimental treatments, corresponding to the combination of water temperature (20 °C; 25 °C), pH (neutral - 7.0; acidic - 5.5), and the absence or presence of Al (0.5 mg L-1). After 96 h, we analyzed the FA profile of semen and performed artificial fertilization in activating medium with neutral pH or activating medium in the same experimental conditions of the males (neutral pH, acidic pH, and Al) to evaluate fertilization and hatching rates and to monitor embryonic development. Polyunsaturated FA percentage decreased in semen of fish from the neutral group, while monounsaturated FA increased in all groups maintained at 20 °C compared to 25 °C. Aluminum exposure decreased the percentage of C20:4n6 and increased the percentage of C22:5n3 at 20 °C. Males exposed to acidic pH and Al showed lower fertilization and hatching rates, as well as increased mortality of embryos and larvae. Moreover, Al favoured a higher percentage of abnormal larvae. Fertilization in Al activating medium harmed the embryos and larvae since fertilization and hatching rates decreased. Finally, temperature influenced fertilization time, hatching rate, and the morphology of embryos and larvae. Males exposed to Al had lower fertilizing capacity, which negatively affected the embryonic development of the species. Furthermore, Al activating medium reduced the number of fertilized oocytes, hatched embryos, and normal larvae. All events were temperature dependent.

[Association of empathy and occupational stress with burnout among primary health care professionals]. / Associação da empatia e do estresse ocupacional com o burnout em profissionais da atenção primária à saúde.

There are indications that burnout syndrome affects primary health care (PHC) professionals. The scope of this study was to investigate the association of empathy and occupational stress with the burnout of PHC professionals. A total of 348 workers filled out an an online assessment including occupational stress, empathy, and burnout scales. The empathetic-concern and perspective-taking empathy dimensions were negatively correlated with burnout (p<.01), while personal anguish was positively related to it (p<.01). The reward and effort occupational stress dimensions revealed greater weight in explaining the job disillusion and psychic exhaustion burnout subscales (p<.001), respectively, while empathetic concern and personal anguish revealed the greatest weight for the explanation of indolence and guilt (p<.001). Reward was the significant predictor of Profile 1 (p=.008), the less severe form of the syndrome, while personal anguish (p=.028) and effort (p=.012) revealed the same weight in the model for Profile 2, namely the most severe level of burnout. The results suggest interventions that focus on both work stressors and the empathy of the professionals involved.

Aluminum, at an environmental concentration, associated with acidic pH and high water temperature, causes impairment of sperm quality in the freshwater teleost Astyanax altiparanae (Teleostei: Characidae).

Given the toxicity of metals, including aluminum (Al), and the effects of water temperature on ectotherms, we investigated the individual or association effect of these variables (Al + acidic pH + temperature changes) on sperm quality of Astyanax altiparanae. Mature males were divided into nine experimental groups based on the combination of each of three water temperatures (20, 25, and 30 °C) with neutral and acidic pH values (7.0 and 5.5, respectively) with or without 0.5 mg L-1 Al. The fish were subjected to subacute, semi-static exposure and at 24 and 96 h were evaluated for seminal parameters: (1) pH; (2) osmolality; (3) sperm concentration; (4) sperm morphology; (5) sperm kinetics; and (6) sperm ultrastructure. At 30 °C, Al caused a reduction in osmolality (24 and 96 h) and sperm concentration (24 h). When analysing sperm kinetics (30 s post-activation), Al caused a reduction in total motility at all temperatures (24 h), and when this exposure time was longer (96 h), both acidic pH and Al addition to the water caused sperm motility reduction. By analysing curvilinear velocity (VCL) 30 s after sperm activation (24 and 96 h), the acidic pH caused a reduction in sperm movement at 20 and 30 °C, but at 25 °C Al triggered this reduction. Finally, Al in the water caused ultrastructural changes in the sperm head, midpiece, and flagella regardless of water temperature. Also, it was found that the combination of Al at 30 °C caused a reduction in sperm head area while at 20 °C, Al triggered a reduction in the midpiece area. Therefore, acidity influenced some A. altiparanae sperm parameters but Al in the water accentuated these effects on seminal quality, especially seminal osmolality and sperm concentration, kinetics, and ultrastructure. This toxicity was also influenced by changes in water temperature.

Associação da empatia e do estresse ocupacional com o burnout em profissionais da atenção primária à saúde / Association of empathy and occupational stress with burnout among primary health care professionals

Resumo Há evidências de que a Síndrome de Burnout afeta profissionais da atenção primária à saúde (APS). Objetivou-se investigar a associação da empatia e do estresse ocupacional com o Burnout de profissionais da APS. 348 trabalhadores responderam a versões online de escalas de estresse ocupacional, empatia e Burnout. As dimensões de empatia preocupação empática e tomada de perspectiva correlacionaram-se negativamente com a síndrome (p<.01), enquanto angústia pessoal demonstrou relação positiva (p<.01). As dimensões de estresse ocupacional recompensa e esforço apresentaram maior peso nos modelos das subescalas de Burnout ilusão pelo trabalho e desgaste psíquico (p<.001), respectivamente, enquanto a preocupação empática e angústia pessoal demonstraram os maiores pesos para a explicação de indolência e culpa (p<.001). Recompensa foi o preditor significativo de Perfil 1 (p=.008), forma menos grave da síndrome, enquanto angústia pessoal (p=.028) e esforço (p=.012) demonstraram o mesmo peso no modelo para Perfil 2, nível mais severo de Burnout. Os resultados sugerem intervenções que enfoquem tanto estressores do trabalho quanto a empatia dos profissionais.

Abstract There are indications that burnout syndrome affects primary health care (PHC) professionals. The scope of this study was to investigate the association of empathy and occupational stress with the burnout of PHC professionals. A total of 348 workers filled out an an online assessment including occupational stress, empathy, and burnout scales. The empathetic-concern and perspective-taking empathy dimensions were negatively correlated with burnout (p<.01), while personal anguish was positively related to it (p<.01). The reward and effort occupational stress dimensions revealed greater weight in explaining the job disillusion and psychic exhaustion burnout subscales (p<.001), respectively, while empathetic concern and personal anguish revealed the greatest weight for the explanation of indolence and guilt (p<.001). Reward was the significant predictor of Profile 1 (p=.008), the less severe form of the syndrome, while personal anguish (p=.028) and effort (p=.012) revealed the same weight in the model for Profile 2, namely the most severe level of burnout. The results suggest interventions that focus on both work stressors and the empathy of the professionals involved.

Water temperature and acid pH influence the cytotoxic and genotoxic effects of aluminum in the freshwater teleost Astyanax altiparanae (Teleostei: Characidae).

The toxicity of metals, including aluminum (Al), can be potentiated by temperature and acid pH, a concern in view of the current global warming scenario. The aim of this study was to evaluate the bioconcentration of Al in the testes and semen of Astyanax altiparanae and the potential of this metal, at different environmental temperatures and acid pH, to cause cytotoxicity and genotocixity in erythrocytes and spermatozoa. A. altiparanae males were divided into nine experimental groups: at each of three different water temperatures (20, 25 and 30 °C), the fish were exposed to a neutral pH, an acid pH and acidic water containing Al (0.5 mg.L-1). The fish were subjected to subacute, semi-static exposure and sampled at 24 and 96 h. After each exposure period the comet assay (blood and semen) and micronucleus test (blood) were performed. Bioconcentration of Al was evaluated in the testes and semen. Exposure time and temperature influenced the Al bioconcentration pattern in the testes. Al concentration in the semen was higher in fish exposed at 20 and 25 °C (24 h). The DNA fragmentation score for the semen and blood was higher in fish exposed to Al at 20 (24 h) and 30 °C (96 h). The frequency of nuclear abnormalities in erythrocytes was higher in the group exposed to Al at 30 °C (96 h). It was concluded that Al bioconcentrates in the testes and semen of A. altiparanae at different temperatures and is potentially cytotoxic and genotoxic to erythrocytes and spermatozoa in this species.

Killer-cell immunoglobulin-like receptors genotyping of 127 individuals from Terceira Island, Azores, Portugal.

One hundred and twenty-seven unrelated Azorean individuals were randomly selected to study the gene frequencies of Killer-cell immunoglobulin-like receptors (KIR) in the Azorean (Terceira) population. KIR genotyping was performed by polymerase chain reaction using commercial sequence-specific oligonucleotide probe kits. All loci were in HWE, showing no locus-level deviations. The genotype data is available in the Allele Frequencies Net Database under the population name "Azores Terceira Island KIR".

Os outros também têm problemas: a percepção dos usuários acerca de uma prática de acolhimento em saúde mental na atenção básica em saúde

A atenção básica em saúde tem demonstrado grande potencial para o cuidado em saúde mental. Contudo, ainda existem problemas a serem superados, entre eles, o desenvolvimento e qualificação de dispositivos de cuidado alternativos à medicalização do sofrimento psíquico. O presente estudo investigou a percepção dos usuários acerca do Acolhimento Coletivo em Saúde Mental de uma unidade de atenção básica. Trata-se de uma pesquisa qualitativa e foram sujeitos de pesquisa os usuários de uma unidade de atenção básica em saúde que participaram de um dispositivo de acolhimento coletivo em saúde mental de julho de 2012 a fevereiro de 2013. Foram realizadas entrevistas semi-estruturadas individuais, e as informações obtidas foram trabalhadas a partir da análise de conteúdo. Foi possível averiguar que entre os entrevistados há o predomínio de concepções centradas na individualização, na patologização e medicalização do sofrimento, que são reforçadas pela forma como a equipe acolhe as demandas e oferece o cuidado. Ainda assim, dispositivo de acolhimento em questão mostrou-se potente em gerar demanda de grupalidade e coletivização das questões em saúde mental. Apesar de ter sido considerado insuficiente em seus benefícios pelos usuários, entendeu-se que essa avaliação do dispositivo de acolhimento precisa ser tomada em um contexto mais amplo, do funcionamento da equipe e da cultura técnica em que esta está inserida. Concluiu-se que o dispositivo em questão precisa potencializar sua capacidade de corresponsabilizar equipe, usuários, comunidade e rede de saúde e intersetorial, promovendo um cuidado voltado à promoção de autonomia e cidadania, assim como à consolidação da Reforma Psiquiátrica e Sanitária.

Randomized comparison of safety and pharmacokinetics of caspofungin, liposomal amphotericin B, and the combination of both in allogeneic hematopoietic stem cell recipients.

The combination of liposomal amphotericin B (LAMB) and caspofungin (CAS) holds promise to improve the outcome of opportunistic invasive mycoses with poor prognosis. Little is known, however, about the safety and pharmacokinetics of the combination in patients at high risk for these infections. The safety and pharmacokinetics of the combination of LAMB and CAS were investigated in a risk-stratified, randomized, multicenter phase II clinical trial in 55 adult allogeneic hematopoietic stem cell recipients (aHSCT) with granulocytopenia and refractory fever. The patients received either CAS (50 mg/day; day 1, 70 mg), LAMB (3 mg/kg of body weight/day), or the combination of both (CASLAMB) until defervescence and granulocyte recovery. Safety, development of invasive fungal infections, and survival were assessed through day 14 after the end of therapy. Pharmacokinetic sampling and analysis were performed on days 1 and 4. All three regimens were well tolerated. Premature study drug discontinuations due to grade III/IV adverse events occurred in 1/18, 2/20, and 0/17 patients randomized to CAS, LAMB, and CASLAMB, respectively. Adverse events not leading to study drug discontinuation were frequent but similar across cohorts, except for a higher frequency of hypokalemia with CASLAMB (P < 0.05). Drug exposures were similar for patients receiving combination therapy and those randomized to monotherapy. There was no apparent difference in the occurrence of proven/probable invasive fungal infections and survival through day 14 after the end of therapy. CASLAMB combination therapy in immunocompromised aHSCT patients was as safe as monotherapy with CAS or LAMB and had similar plasma pharmacokinetics, lending support to further investigations of the combination in the management of patients with invasive opportunistic mycoses.

Role of human leukocyte antigen, killer-cell immunoglobulin-like receptors, and cytokine gene polymorphisms in leptospirosis.

Leptospirosis is an emerging zoonotic disease caused by pathogenic species of the genus Leptospira. It has a broad range of clinical presentations in humans. Although progress has been made in the characterization of the host immune system factors that may affect disease progression and outcome, to date few reports have addressed the role of genetic polymorphisms in the susceptibility to leptospirosis. In this work a group of patients with a history of leptospiral infection and a control group were compared for polymorphisms in the human leukocyte antigen (HLA), in killer-cell immunoglobulin-like receptors (KIR), and in cytokine genes. Alleles in the HLA-A and -B loci were associated with susceptibility, as were the class I haplotype A*01-B*08-Cw*07 and the 8.1 ancestral haplotype (A*01-B*08-Cw*07-DRB1*03-DQB1*02). Single nucleotide polymorphisms in the interleukin (IL)-4 and IL-4Ralpha genes also had significantly higher frequencies in the patient group. No association was reported between KIR gene profile and leptospirosis. This work highlights the importance of using genetic polymorphisms to better understand the mechanisms involved in the immune response to leptospirosis.

Analytical validation of a microplate reader-based method for the therapeutic drug monitoring of L-asparaginase in human serum.

The enzyme L-asparaginase (ASNASE), which hydrolyzes L-asparagine (L-Asn) to ammonia and L-aspartic acid (L-Asp), is commonly used for remission induction in acute lymphoblastic leukemia. To correlate ASNASE activity with L-Asn reduction in human serum, sensitive methods for the determination of ASNASE activity are required. Using L-aspartic beta-hydroxamate (AHA) as substrate we developed a sensitive plate reader-based method for the quantification of ASNASE derived from Escherichia coli and Erwinia chrysanthemi and of pegylated E. coli ASNASE in human serum. ASNASE hydrolyzed AHA to L-Asp and hydroxylamine, which was determined at 710 nm after condensation with 8-hydroxyquinoline and oxidation to indooxine. Measuring the indooxine formation allowed the detection of 2 x 10(-5)U ASNASE in 20 microl serum. Linearity was observed within 2.5-75 and 75-1,250 U/L with coefficients of correlation of r(2)>0.99. The coefficients of variation for intra- and interday variability for the three different ASNASE enzymes were 1.98 to 8.77 and 1.73 to 11.0%. The overall recovery was 101+/-9.92%. The coefficient of correlation for dilution linearity was determined as r(2)=0.986 for dilutions up to 1:20. This method combined with sensitive methods for the quantification of L-Asn will allow bioequivalence studies and individualized therapeutic drug monitoring of different ASNASE preparations.