Cortisol plays a role in the high environmental ammonia associated suppression of the immune response in zebrafish.

Exposure to high environmental ammonia (HEA) levels increases the vulnerability of fishes to parasitic, viral and bacterial diseases. We tested the hypothesis that elevated plasma cortisol levels play a role in the HEA-mediated immunosuppression in fishes. To this end, we tested the effect of exogenous cortisol treatment on the lipopolysaccharide (LPS)-induced immune response in zebrafish (Danio rerio). Also, to test whether glucocorticoid receptor (GR) signaling is involved in HEA-mediated immunosuppression, zebrafish were treated with mifepristone, a GR antagonist, and the LPS-induced immune response assessed after HEA exposure. We evaluated a panel of important immunity-related genes including interleukin 1ß (il1b) and suppressor of cytokine signaling (socs-1a, 2, 3) and acute phase response genes [serum amyloid A (saa), transferrin (tfa), leukocyte cell-derived chemotaxin 2-like (lect2l), haptoglobin (hp), hepcidin (=hepatic anti-microbial peptide hamp), and complement component 3b (c3b)] by real-time quantitative PCR. Our results demonstrate that exogenous cortisol administration as well as elevated cortisol levels in response to HEA exposure modulate mRNA transcript levels of key mediators of the innate immune response in zebrafish. Mifepristone treatment reduced whole body cortisol levels and eliminated the HEA-mediated changes in transcript abundance of socs1a, il1b, as well as APR genes. Together, these results suggest that the HEA effect on the innate immune response is in part mediated by cortisol signaling, while the mode of action, including the receptors involved remains to be elucidated.

Frações solúveis e insolúveis do hidrolisado proteico de resíduo de sardinha na alimentação do jundiá: consumo de ração e excreção de amônia / Soluble and insoluble fractions of the sardine waste protein hydrolysate in silver catfish feeding: feed intake and excretion of ammonia

O hidrolisado proteico de pescado é produzido por meio da hidrólise enzimática de resíduos da indústria de pescado, resultando em um ingrediente de excelente qualidade para ser utilizado na alimentação de peixes. O objetivo deste estudo foi determinar o efeito das frações do hidrolisado proteico de resíduo de sardinha sobre o consumo de ração e excreção de amônia de juvenis de jundiá. Foram testadas as frações solúveis e insolúveis do hidrolisado de músculo de sardinha, avaliadas individualmente e combinadas entre si. Juvenis de jundiás (9,76±0,55g) foram divididos em 12 aquários de 20L, na densidade de quatro peixes por aquário, resultando em uma biomassa média de 1,90±0,17g/L. O hidrolisado proteico foi produzido com carcaças limpas de sardinhas mediante o uso da enzima protease bacteriana Protamex(r) (Novozymes A/S) e dele derivaram as frações solúveis e insolúveis. As dietas foram isoproteicas (39% PB) e isoenergéticas (4450kcal EB/kg), e as frações do hidrolisado foram incluídas de forma a fornecerem 50% da proteína das rações. A fração solúvel apresentou grau de hidrólise mais elevado (20,1%) em relação à fração insolúvel (9,97%). As duas dietas contendo a fração solúvel do hidrolisado foram as mais consumidas. Com cinco horas de avaliação, a dieta contendo a fração solúvel do hidrolisado aumentou a excreção de amônia em relação à dieta contendo a fração insolúvel. Após 25 horas, a fração solúvel também aumentou a excreção de amônia, mas, desta vez, em relação à dieta contendo a combinação das duas frações. Conclui-se que a inclusão da fração solúvel do hidrolisado de músculo de sardinha estimula o consumo de ração, todavia, quando a inclusão é elevada, pode haver aumento da excreção de amônia de juvenis de jundiá.(AU)

The fish protein hydrolysate is produced by the enzymatic hydrolysis of seafood industry wastes, resulting in a high quality ingredient to be used in fish feed. The aim of this study was to determine the effect of the fractions of the sardine protein waste hydrolysate fractions on feed intake and ammonia excretion of silver catfish. The soluble and insoluble fractions of the sardine muscle hydrolysate, evaluated individually and in combination were tested. Juvenile silver catfish (9.76±0.55g) were divided into twelve 20L aquaria, in the density of four fish per tank, resulting in an average biomass of 1.90±0.17g/L. The protein hydrolysate was produced with fresh sardine carcasses using the enzyme Protamex(r) bacterial protease (Novozymes A/S) and soluble and insoluble fractions were derived. The diets were isonitrogenous (39% CP) and isocaloric (4450kcal EB/kg) and the hydrolysate fractions were included in order to provide 50% of the protein of the diet. The soluble fraction has higher degree of hydrolysis (20.1%) compared to the insoluble fraction (9.97%). The two diets containing the hydrolysate soluble fraction were the most consumed. With 5 hours of evaluation, the diet containing the hydrolysate soluble fraction increased the excretion of ammonia in relation to diet containing the insoluble fraction. After 25 hours, the soluble fraction also increases the excretion of ammonia but this time in relation to the diet containing a combination of two fractions. It was concluded that the inclusion of the sardine protein hydrolysate soluble fraction in the diet stimulates the silver catfish feed intake, but when inclusion is high it can increase ammonia excretion of juvenile silvercatfish.(AU)

Tuberculosis screening in patients receiving biological therapy.

AIM: Biological therapies are a risk factor for tuberculosis (TB). Portuguese recommendations recommend universal baseline screening for TB before starting biologics (2006) and annually thereafter if screened negative (2012 update). The gain with re-screening remains unknown. We aimed to i)identify the risk of latent TB infection at baseline screening among patients candidates to initiate biologics ii)present follow-up results for patients receiving different biological therapies and analyse intolerance or toxicity related to preventive therapy, conversions of immunodiagnostic tests under biological therapy and development of active TB. METHODS: Patients screened for TB at a reference centre before starting biological therapy between 2008-2012 were identified. Medical files were retrospectively reviewed. Demographic data, screening and follow-up results and information on biological therapy were collected. EXCLUSION CRITERIA: unavailable information on initiation of biological therapy. RESULTS: 183 patients were included in the study, with 115 starting biological therapy. The baseline screening was positive in 52(45,2%) patients - 50(96,2%) were proposed for preventive treatment (2 had abnormal liver enzymes). Mild hepatotoxicity occurred in 4(8%) patients without need to interrupt TB prophylaxis. No cases of active TB occurred during follow-up in patients with positive baseline screening. Among the 63(54,8%) patients who screened negative, 2(3,2%) developed active TB (under infliximab and adalimumab) more than one year after initiation of biologics. 26(41,3%) patients were re-screened at the TB centre. 5(19,2%) had tuberculin skin test (TST) conversion and one concomitantly undetermined IGRA. No IGRA conversions were observed. The follow-up period was 4,0 years. TB baseline screening's negative predictive value (NPV) was 96,8% (95%CI: 89,0% to 99.5%). A low rate of re-screening was observed. CONCLUSION: The rate of latent TB at baseline screening was higher than expected. Preventive treatment was well tolerated. No patients with positive baseline screening developed active TB. Efforts should be made to raise awareness concerning the risk of TB exposure, specially considering that the active TB cases were compatible with new infection. The rate of re-screening suggests a low awareness regarding current recommendations Nation-wide studies are necessary to evaluate the efficacy of the re-screening strategy and to clarify what risk groups most benefit from it.

Development of a powdered activated carbon in bar adsorptive micro-extraction for the analysis of morphine and codeine in human urine.

In the present work, bar adsorptive microextraction using an activated carbon (AC) adsorbent phase followed by liquid desorption and high-performance liquid chromatography with diode array detection was developed to monitor morphine (MOR) and codeine (COD) in human urine. Under optimized experimental conditions, assays performed in aqueous media spiked at the 30.0 µg/L level yielded recoveries of 41.3 ± 1.3% for MOR and 38.4 ± 1.7% for COD, respectively. The textural and surface chemistry properties of the AC phase were also correlated with the analytical data for a better understanding of the overall enrichment process. The analytical performance showed good precision (relative standard deviation < 8.0%), suitable detection limits (0.90 and 0.06 µg/L for MOR and COD, respectively) and convenient linear dynamic ranges (r(2) > 0.991) from 10.0 to 330.0 µg/L. By using the standard addition methodology, the applications of this analytical approach to water and urine matrices allowed remarkable performance to monitor MOR and COD at the trace level. This new confirmatory method proved to be a suitable alternative to other sorptive micro-extraction methodologies in monitoring trace levels of opiate-related compounds, because it was easy to implement, reliable, sensitive and required a low sample volume.

The inhibitory effect of environmental ammonia on Danio rerio LPS induced acute phase response.

Ammonia is a toxic by-product of amino acid catabolism and a common environmental pollutant that has been associated with increased disease susceptibility in fish although the mechanism is not well understood. We addressed the hypothesis that elevated environmental ammonia acts by impairing the acute phase response (APR). Specifically, we determined the impact of sub-lethal acute (24 h) and chronic (14 d) ammonia exposure on acute phase protein gene expression in zebrafish (Danio rerio) in response to a challenge with bacterial lipopolysaccharide (LPS: i.p. 10 µg/g after 24h). A panel of LPS-responsive genes (SAA, HAMP, LECT2, Hp and IL1ß) were identified and evaluated by real-time quantitative PCR. Ammonia was found to impair induction of SAA, HAMP and LECT2 by 50-90%. Both short (15 min, 1h and 24h) and long-term (14 days) exposure to high environmental ammonia concentrations significantly elevated whole-body cortisol levels compared with control fish. Our results reveal for the first time that exposure to high environmental levels of ammonia suppresses the innate immune response in fish. We hypothesize that high environmental ammonia-mediated elevation of cortisol levels in zebrafish may be playing a key role in this immunosuppression, while the mechanisms involved remains to be elucidated.

Sex-related macrostructural organization of the deer's brachial plexus.

We describe the morphological organization of the deer brachial plexus in order to supply data to veterinary neuroclinics and anaesthesiology. The deer (Mazama gouazoubira) brachial plexus is composed of four roots: three cervical (C6, C7 and C8) and one thoracic (T1). Within each sex group, no variations are observed between the left and the right brachial plexus, though sex-related differences are seen especially in its origin. The origin of axillary and radial nerves was: C6, C7, C8 and T1 in males and C8-T1 (radial nerve) and C7, C8 and T1 (axillary nerve) in females; musculocutaneous nerve was: C6-C7 (males) and C8-T1 (females); median and ulnar nerves was: C8-T1 (males) and T1 (females); long thoracic nerve was: C7 (males) and T1 (females); lateral thoracic nerve was: C6, C7, C8 and T1 (males) and T1 (females); thoracodorsal nerve was: C6, C7, C8 and T1 (males) and C8-T1 (females); suprascapular nerve was: C6-C7 (males) and C6 (females) and subscapular nerve was: C6-C7 (males) and C7 (females). This study suggests that in male deer the origin of the brachial plexus is more cranial than in females and the origin of the brachial plexus is slightly more complex in males, i.e. there is an additional number of roots (from one to three). This sexual dimorphism may be related to specific biomechanical functions of the thoracic limb and electrophysiological studies may be needed to shed light on this morphological feature.

[The prevalence of cerebrovascular stroke in Coimbra]. / Prevalência dos acidentes vasculares cerebrais em Coimbra.

The purpose of this study was to assess the prevalence of strokes in a community-based on a sample of the population of the county of Coimbra in 1992 (139,000 inhabitants). The study was made with the collaboration of local general practitioners who randomly selected a sample of 950 individuals over 50 years of age. These individuals were screened for the presence of vascular risk factors and strokes in the past, and submitted to clinical and laboratory examinations. There were 86 persons with strokes (crude prevalence 8%: 10.2% male, 6.6% female) the diagnosis being confirmed by neurological examination or analysis of hospital registers. Hypertension was significantly more frequent in patients with strokes (56.8%). Other risk factors had no significant differences. The magnitude of disability associated with stroke was assessed according to the modified Rankin scale. About fifty percent had no significant disability or a slight disability and 20% were severely disabled.