Leishmania amazonensis exhibits phosphatidylserine-dependent procoagulant activity, a process that is counteracted by sandfly saliva

Leishmania parasites expose phosphatidylserine (PS) on their surface, a process that has been associated with regulation of host's immune responses. In this study we demonstrate that PS exposure by metacyclic promastigotes of Leishmania amazonensis favours blood coagulation. L. amazonensis accelerates in vitro coagulation of human plasma. In addition, L. amazonensis supports the assembly of the prothrombinase complex, thus promoting thrombin formation. This process was reversed by annexin V which blocks PS binding sites. During blood meal, Lutzomyia longipalpis sandfly inject saliva in the bite site, which has a series of pharmacologically active compounds that inhibit blood coagulation. Since saliva and parasites are co-injected in the host during natural transmission, we evaluated the anticoagulant properties of sandfly saliva in counteracting the procoagulant activity of L. amazonensis . Lu. longipalpis saliva reverses plasma clotting promoted by promastigotes. It also inhibits thrombin formation by the prothrombinase complex assembled either in phosphatidylcholine (PC)/PS vesicles or in L. amazonensis . Sandfly saliva inhibits factor X activation by the intrinsic tenase complex assembled on PC/PS vesicles and blocks factor Xa catalytic activity. Altogether our results show that metacyclic promastigotes of L. amazonensis are procoagulant due to PS exposure. Notably, this effect is efficiently counteracted by sandfly saliva.

Depressão, cortisol urinário e perfil sócio-demográfico de portadores de diabetes mellitus tipo 2

O objetivo deste estudo foi investigar a relação entre indicadores de depressão e perfil sócio-demográfico de portadores de Diabetes Mellitus tipo 2 (DM2).A avaliação sócio-demográfica foi conduzida em amostra composta por 40 pacientes na Liga de Diabetes (HC-FMUSP).Os indicadores de depressão foram investigados a partir do Inventário de Depressão de Beck (IBD) em associação com cortisol urinário (CORT).Os resultados mostraram que indivíduos portadores de DM2 com alta escolaridade, baixo poder aquisitivo individual e familiar e com história de rompimento de relação conjugal estável estão mais propensos a sintomas de depressão.

Stress among nurses: an examination of salivary cortisol levels on work and day off

The present study evaluates the use of salivary cortisol concentration as a physiological index of the stress level among nurses on their work day and day off and correlates it with the questionnaire used to measure occupational stress in nurses (Inventário de Estresse em Enfermeiros - IEE). This is a comparative, cross-sectional descriptive study in which sociodemographic data, IEE results and salivary cortisol levels were used. Fifty-seven nurses participated in the study (80.7% females and a mean age of 37.1 years old). The IEE average score was 124.5. The average cortisol level was 564.1 ng/m on work day and 354.1 ng/mL on day off. Nurses who had double workdays presented high values of salivary cortisol during the work day (638.1 ng/mL). In conclusion, salivary cortisol identified the nurses’ stress level, and differences were found between a work day and day off. On the nurses’ day off, their salivary cortisol levels and stress scores were lower.

Vitamin C and endogenous cortisol in foreign-body inflammatory response in pacus

The objective of this work was to evaluate the effect of food supplementation with vitamin C on macrophage and multinucleated giant cell (MGC) activities of pacus at two stocking densities. The experiment was carried out in a 2x2x3 split-plot factorial arrangement with: 0 and 500 mg kg-1 vitamin C; 5 and 20 kg m-3 stocking densities; and evaluation times at 3, 6, and 12 days after the subcutaneous implantation of glass coverslips (DPI). The number of macrophages and MGC, as well as cortisol and glucose plasma levels were determined. The number of macrophages and MGC with two to five nuclei was significantly greater in fish supplemented with vitamin C at 5 kg m-3 stocking density at 3 DPI in comparison to nonsupplemented ones. The macrophage and MGC counts were lower in fish with high-plasma cortisol concentration. Supplementation with 500 mg vitamin C benefits macrophage activity on foreign-body inflammation, and high-cortisol concentration has suppressive effects on this response.

Determination of free urinary cortisol in cushing's syndrome using reversed-phase high performance liquid chromatography

Determination of free urinary cortisol is a test of choice in the diagnosis of Cushing's syndrome. In this study, cortisol was quantified using reversed-phase high-performance liquid chromatography (RP-HPLC) in urine samples previously extracted with ether and using triamcinolone acetonide as internal standard (IS). A BDS-Hypersil-C18® column, water-acetonitrile (72:28; v/v), with a flow rate of 1.0 mL/min and detection at 243 nm were used. This method showed to be both effective and efficient, with sensitivity and linearity ranging from 2.50 to 150 μg/L, and can be used in substitution to the radioimmunoassay technique within this concentration range.

Avaliação dos parâmetros cardiorrespiratórios, hemogasométricos e da concentração plasmática de cortisol em bovinos submetidos à imobilização eletromagnética

As formas de imobilização em bovinos mais utilizadas levam a uma manipulação excessiva e consequentemente estressante para o animal. A dor é uma experiência emocional de sensação desagradável que vem associada a um prejuízo tecidual. Surge no mercado um método de contenção física para bovinos que propõe a utilização de um aparelho que funciona com ondas eletromagnéticas de baixa intensidade promovendo imobilização sem estresse e ou estímulo doloroso para o animal. O objetivo deste projeto foi avaliar os parâmetros cardiorrespiratórios e hemogasométricos bem como os níveis de cortisol de bovinos submetidos à imobilização eletromagnética, a fim de observar se tal método é eficaz na diminuição do estresse e/ou estímulo doloroso. Foram utilizados 6 bovinos, mestiços, peso médio de 300,3 kg (±85,76). Os animais foram alocados aleatoriamente em dois grupos (controle dele mesmo): grupo controle (GC), introdução da sonda via retal (desligada); grupo Imobilizador (GI) introdução da sonda e permanência desta ligada por 5 minutos. Análise estatística utilizou o Teste T pareado entre os grupos (P<0,05) e a avaliação entre tempos dentro de cada grupo análise de variância de uma única via com repetições múltiplas (ANOVA - RM). Momentos avaliados M0: basal; M1: imediatamente após a retirada da sonda; M2 e M3 (30 e 60 minutos após M1 respectivamente). Houve um aumento significativo nas variáveis: frequências cardíaca e respiratória em (M1), na Pressão arterial média (M1, M2, M3) e níveis de cortisol nos momentos M1 e M2 no grupo imobilizador quando comparado ao grupo controle. Com relação à resposta ao estímulo, o GI teve resposta negativa no momento da estimulação, comparado ao GC (resposta máxima). De acordo com os resultados obtidos, podemos concluir que o imobilizador eletromagnético é um método, exclusivamente, de contenção física, não oferecendo nenhum suporte anestésico, para o conforto ou bem estar ao animal.

Avaliação física e dos níveis séricos de cortisol de bezerros neonatos da raça Nelore, nascidos de partos normais e auxiliados

O presente trabalho visou avaliar a vitalidade e os níveis séricos de cortisol de bezerros neonatos, nascidos de partos eutócicos e auxiliados, bem como determinar os índices de partos distócicos em vacas receptoras de embriões nelores. Cerca de 310 bezerros foram avaliados. Observou-se a ocorrência de 38 partos distócicos (38/310) e de oito abortos (8/310). Os bezerros nascidos de partos distócicos mais duradouros, entre quatro e seis horas, apresentaram os maiores valores médios de frequência respiratória e dos níveis séricos de cortisol. A temperatura retal foi mais elevada em animais nascidos de partos laboriosos do que naqueles nascidos de partos normais. As taxas de morbidade e mortalidade em animais neonatos nelores foram de 77,93% e 3,44%, respectivamente. O emprego do escore APGAR, ao nascimento, demonstrou que 90% dos bezerros apresentavam-se deprimidos, independente do tipo de nascimento.

Cortisol influence on testicular testosterone secretion in domestic cat: An in vitro study

The aim of the present experiment was to investigate the effect of corticosteroids (exogen) on in vitro testosterone secretion after stress by transportation (40 minutes). Feline testes (Felis silvestris catus) were incubated in the following media: TCM 199; TCM 199 + hCG 10_7M; TCM 199 + hydrocortisone 10_7M, or TCM 199 + hCG + hydrocortisone. The animals (n=21) were allocated into three groups: (S) that arrived at 3 h prior to surgery, (A) that remained in the laboratory for 36 h before being submitted to surgical procedure, and (C) that were also allowed to remain for 36 hours in the laboratory before the surgical procedure, but whose testes had been incubated with hydrocortisone prior to incubation in the referred media. The results showed that group S secreted higher levels of testosterone, regardless of the culture media. It is noteworthy that the suppressing action of hydrocortisone sodium succinate led to a reduction in the testosterone concentration, despite the presence of hCG.

Glicemia e concentrações séricas de insulina, triglicérides e cortisol em equinos da raça Mangalarga Marchador após exercício físico

O presente estudo teve por objetivo avaliar a influência do exercício físico de intensidade submáxima (provas de marcha) sobre as variáveis bioquímicas sanguíneas usadas na avaliação do metabolismo energético em equinos da raça Mangalarga Marchador criados no Estado do Espírito Santo. Para tal foram obtidas amostras de soro e plasma de 15 equinos, da raça Mangalarga Marchador, em quatro momentos assim definidos: antes (T0) e com 5 minutos (T1), 30 minutos (T2) e 2 horas (T3) após o término do exercício. A análise dos resultados demonstrou a não influência do exercício físico imposto sobre a glicose plasmática, com valores médios de 117,1±35,8mg/dL, 122,6±59,6mg/dL, 124,8± 48,6mg/dL e 112,9±49,1mg/dL, e sobre a insulina sérica, com valores de mediana de 6,50mUI/mL, 2,00mUI/mL, 5,85mUI/mL e 11,60mUI/mL, respectivamente, nos tempos T0, T1, T2 e T3. De forma oposta, foi possível observar uma influência significativa sobre triglicérides séricos, com valores médios de 25,4±14,9mg/dL, 42,3±17,8mg/dL, 31,4±17,7mg/dL e 25,1±15,1mg/dL, e sobre o cortisol sérico, com valores médios de 7,46±4,37mg/dL, 12,45±3,08mg/dL, 11,40±2,52mg/dL e 6,89±1,78mg/dL, respectivamente nos tempos T0, T1, T2 e T3. A interpretação destes resultados permitiu concluir que a marcha elevou as concentrações séricas de triglicérides e cortisol. Também foi possível destacar que, por tais valores encontrarem-se dentro de intervalos fisiológicos, os equinos usados estavam aptos ao nível de exercício físico imposto na ocasião.

Profile of cortisol, glycaemia, and blood parameters of American Bullfrog tadpoles Lithobates catesbeianus exposed to density and hypoxia stressors

The aim of this study was to evaluate alterations to the physiological profile (cortisol, glycaemia, and blood parameters) of Lithobates catesbeianus caused by the stressors density and hypoxia. The organisms were in the prometamorphosis stage and exposed to different tadpole densities: 1 tadpole/L (T1), 5 tadpoles/L (T2), and 10 tadpoles/L (T3) for 12 days. The blood was collected through the rupture of the caudal blood vessel and collected under normoxia (immediate collection) and hypoxia (after 15 minutes of air exposure) conditions. Cortisol levels rose on the fourth and eighth days of treatment and returned to basal levels by the end of the experiment. The stressor mechanisms tested did not affect glycaemia. White blood cells (total number of lymphocytes, neutrophils, and eosinophils) showed a significant difference at the twelfth day of the experiment when compared with the start of the experiment. We concluded that, under controlled conditions, a density of up to 10 tadpoles/L and air exposure for 15 minutes did not cause harmful physiological alterations during the experimental period. The answer to these stressors maybe was in another hormonal level (corticosterone).

Perfil hematológico, bioquímico sérico, proteína C reativa e cortisol de ararajubas (Guaroba guarouba) mantidas em cativeiro

Objetivou-se determinar os valores hematológicos, bioquímicos séricos, proteína C reativa e cortisol de 14 ararajubas (Guaroba guarouba) clinicamente saudáveis mantidas em cativeiro no zoológico do Parque Estadual de Dois Irmãos Recife/PE. Amostras de sangue foram obtidas da veia jugular com volume médio de 0,8ml, fracionando em duas porções, a primeira depositada em tubo MiniCollect®CE contendo EDTA e a outra tubo com gel separador, para a separação do soro sanguíneo. Os indicadores bioquímicos e PCR foram determinados através do analisador ARCHITECT c8000. O cortisol foi analisado no Cobas E411 da Roche. Para a determinação dos números de eritrócitos e leucócitos, foi utilizada a metodologia da contagem em câmara de Neubauer. O hematócrito foi avaliado pelo método do microhematócrito e a hemoglobina pelo método da cianometahemoglobulina. Para a contagem diferencial de leucócitos, foi utilizada a técnica de Shilling. Os índices hematimétricos (VCM, HCM E CHCM) foram determinados com os valores encontrados na série eritrocítica. Os dados foram caracterizados por dispersão de freqüências, utilizando-se as seguintes medidas de tendência central: Média, desvio-padrão, mediana e percentil de 25 e percentil 75. A determinação de alguns parâmetros como ferro, triglicerídeos, PCR e cortisol foram identificados como sendo pioneiramente referenciados nesta espécie de ave. Os dados são apresentados como sendo de referência para a ararajuba (Guaroba guarouba) criada em cativeiro em condições similares de manejo e higidez e ainda pode contribuir para os trabalhos de conservação ex situ desta espécie.

Fecal cortisol metabolites as indicators of stress in crab-eating-fox (Cerdocyoun thous) in captivity

Abstract: Blood samples collection is a common method in biological research using domestic animals. However, most blood sampling techniques are complicated and highly invasive and may therefore not be appropriate for wildlife animals in research concerning stress. Thus, a non-invasive method to measure steroid hormones is critically needed. The first goal of this study was to determine how glucocorticoids concentrations are impacted by translocation and reproductive activity in crab-eating-fox (Cerdocyoun thous) in captivity. The physiological relevance of fecal glucocorticoid metabolites was further validated by demonstrating: (1) The translocation of a male to a females enclosure resulted in a 3.5-fold increase compared to baseline concentrations, (2) changes in adrenocortical activity, as reflected in concentrations of fecal cortisol metabolites during reproduction, gestation and lactation in females foxes, indicating that social interactions resulted in large increases of fecal glucocorticoids metabolites during the reproductive season. From these findings we conclude that fecal samples can be used for the non-invasive assessment of adrenocortical status in crab-eating-fox.

Plasma cortisol levels in captive wild felines after chemical restraint

Eight Panthera onca (Po), 13 Felis concolor (Fc), 7 Felis yagouaroundi (Fy), 7 Felis tigrina (Ft) and 5 Felis pardalis (Fp) specimens from São Paulo State zoos were used. All animals were restrained with darts containing 10 mg/kg ketamine and 1 mg/kg xylazine. Venous blood samples were collected as soon as possible (within 15-20 min) and serum was frozen until the time for cortisol quantification. Cortisol was determined using a solid phase radioimmunoassay with an intra-assay coefficient of 8.51%. Data were analyzed statistically by the Kruskal-Wallis test, followed by Dunn's multiple comparisons test, and the one-sample t-test, with the level of significance set at P<0.05. Data are reported as means ± SEM. Cortisol levels differed among the captive felines: Po = 166 ± 33a, Fc = 670 ± 118b, Fy = 480 ± 83b, Ft = 237 ± 42ab, Fp = 97 ± 12a nmol/l (values followed by different superscript letters were significantly different (P<0.001)). Since most of the veterinary procedures on these species involve chemical restraint, these results show the necessity of preventive measures in order to minimize the effect of restraint stress on more susceptible species

Salivary cortisol as a tool for physiological studies and diagnostic strategies

Salivary cortisol is an index of plasma free cortisol and is obtained by a noninvasive procedure. We have been using salivary cortisol as a tool for physiological and diagnostic studies, among them the emergence of circadian rhythm in preterm and term infants. The salivary cortisol circadian rhythm in term and premature infants was established between 8 and 12 postnatal weeks. In the preterm infants the emergence of circadian rhythm was parallel to the onset of sleep rhythm. We also studied the use of salivary cortisol for screening for Cushing's syndrome (CS) in control and obese outpatients based on circadian rhythm and the overnight 1 mg dexamethasone (DEX) suppression test. Salivary cortisol was suppressed to less than 100 ng/dl after 1 mg DEX in control and obese patients. A single salivary cortisol measurement at 23:00 h and again after 1 mg DEX above the 90th percentile of the obese group values had sensitivity and specificity of 93 and 93% (23:00 h), and 91 and 94% (after DEX), respectively. The sensitivity improved to 100% when we combined both parameters. We also studied 11 CS children and 21 age-matched primary obese children for whom salivary cortisol sensitivity and specificity were 100/95% (23:00 h), and 100/95% (1 mg DEX), respectively. Similar to adults, sensitivity and specificity of 100% were obtained by combining 23:00 h and 1 mg DEX. The measurement of salivary cortisol is a useful tool for physiological studies and for the diagnosis of CS in children and adults on an outpatient basis.

Effect of the establishment of dominance relationships on cortisol and other metabolic parameters in Nile tilapia (Oreochromis niloticus)

The objective of the present study was to investigate the influence of the establishment of dominance relationships and social stress on plasma cortisol and metabolite levels in Nile tilapia (Oreochromis niloticus). During the 30-day experiment, the fish weighing 236 ± 29 g were kept in individual aquaria, except for two pairings lasting 6 h each. Blood samples were taken from the animals before and after pairing. Display, approach, attack, rebuff, chase flight, and coloration were carried out on days 16 and 30. Activities and behaviors characteristic of the establishment of dominance relationships were described. It was possible to classify all experimental fish (N = 30) as dominant or subordinate. No differences were detected between dominant (N = 15) and subordinate (N = 15) fish during isolation or after pairing in cortisol (isolated: 5.76 ± 0.98 vs 5.42 ± 0.63; paired: 10.94 ± 1.62 vs 11.21 ± 2.45 µg/dl), glucose (isolated: 60.02 ± 4.9 vs 67.85 ± 16.16; paired: 110.44 ± 15.72 vs 136.26 ± 22.46 mg/dl), triglyceride (isolated: 167.87 ± 5.06 vs 185.68 ± 7.24; paired: 210.85 ± 13.40 vs 221.82 ± 12.70 mg/dl) or total protein levels (isolated: 7.01 ± 0.42 vs 6.69 ± 0.59; paired: 9.21 ± 0.62 vs 9.51 ± 0.66 g/dl). However, when isolated (N = 30) and paired (N = 30) tilapia were compared, there were significant differences in cortisol and metabolite levels. The similar response presented by dominant and subordinate tilapia indicates that establishment of dominance relationships was a stressor for both groups.