Innovative vs classical methods for drying heterotrophic Chlorella vulgaris: Impact on protein quality and sensory properties.

Drying is a necessary step in the microalgae production chain to reduce microbial load and oxidative degradation of the end product. Depending on the differences in applied temperature and treatment time, the process of drying can have a substantial impact on protein quality and aroma, important characteristics determining the incorporation potential in food products. In this study, we compared the drying of heterotrophic Chorella vulgaris with both innovative (agitated thin film drying (ATFD), pulse combustion drying (PCD) and solar drying (SolD)) and commonly used drying techniques (spray drying (SprD) and freeze drying (FD)). To evaluate the impact on protein quality, we evaluated techno-functional properties, in vitro digestibility (INFOGEST) as well as protein denaturation using differential scanning calorimetry (DSC). A sensory analysis was performed by a trained expert panel, combined with headspace solid-phase microextraction (HS-SPME) - gas chromatography-mass spectrometry (GC-MS) to determine volatile organic compounds (VOCs). ATFD was found to increase techno-functional properties such as gelling, water holding and solubility as well as in vitro protein digestibility. These observations could be related to induced cell disruption and protein denaturation by ATFD. Sensory analysis indicated an increased earthy off-flavor after ATFD. Interestingly, the high-temperature PCD led to an increase in cacao odor while low-temperature FD resulted in lower flavor, odors and VOCs. These results demonstrate that protein quality and sensorial properties of C. vulgaris can be steered through the type of drying, which could help in the selection of application-specific drying methods. Overall, this work could promote the incorporation of microalgal single cell proteins in different innovative food products.

Polar Lipids of Marine Microalgae Nannochloropsis oceanica and Chlorococcum amblystomatis Mitigate the LPS-Induced Pro-Inflammatory Response in Macrophages.

Microalgae are recognized as a relevant source of bioactive compounds. Among these bioactive products, lipids, mainly glycolipids, have been shown to present immunomodulatory properties with the potential to mitigate chronic inflammation. This study aimed to evaluate the anti-inflammatory effect of polar lipids isolated from Nannochloropsis oceanica and Chlorococcum amblystomatis. Three fractions enriched in (1) digalactosyldiacylglycerol (DGDG) and sulfoquinovosyldiacylglycerol (SQDG), (2) monogalactosyldiacylglycerol (MGDG), and (3) diacylglyceryl-trimethylhomoserine (DGTS) and phospholipids (PL) were obtained from the total lipid extracts (TE) of N. oceanica and C. amblystomatis, and their anti-inflammatory effect was assessed by analyzing their capacity to counteract nitric oxide (NO) production and transcription of pro-inflammatory genes Nos2, Ptgs2, Tnfa, and Il1b in lipopolysaccharide (LPS)-activated macrophages. For both microalgae, TE and Fractions 1 and 3 strongly inhibited NO production, although to different extents. A strong reduction in the LPS-induced transcription of Nos2, Ptgs2, Tnfa, and Il1b was observed for N. oceanica and C. amblystomatis lipids. The most active fractions were the DGTS-and-PL-enriched fraction from N. oceanica and the DGDG-and-SQDG-enriched fraction from C. amblystomatis. Our results reveal that microalgae lipids have strong anti-inflammatory capacity and may be explored as functional ingredients or nutraceuticals, offering a natural solution to tackle chronic inflammation-associated diseases.

Scenedesmus rubescens Heterotrophic Production Strategies for Added Value Biomass.

Microalgae attract interest worldwide due to their potential for several applications. Scenedesmus is one of the first in vitro cultured algae due to their rapid growth and handling easiness. Within this genus, cells exhibit a highly resistant wall and propagate both auto- and heterotrophically. The main goal of the present work is to find scalable ways to produce a highly concentrated biomass of Scenedesmus rubescens in heterotrophic conditions. Scenedesmus rubescens growth was improved at the lab-scale by 3.2-fold (from 4.1 to 13 g/L of dry weight) through medium optimization by response surface methodology. Afterwards, scale-up was evaluated in 7 L stirred-tank reactor under fed-batch operation. Then, the optimized medium resulted in an overall productivity of 8.63 g/L/day and a maximum biomass concentration of 69.5 g/L. S. rubescens protein content achieved approximately 31% of dry weight, similar to the protein content of Chlorella vulgaris in heterotrophy.

Differences and Similarities in Lipid Composition, Nutritional Value, and Bioactive Potential of Four Edible Chlorella vulgaris Strains.

The microalga Chlorella vulgaris is a popular food ingredient widely used in the industry, with an increasing market size and value. Currently, several edible strains of C. vulgaris with different organoleptic characteristics are commercialized to meet consumer needs. This study aimed to compare the fatty acid (FA) and lipid profile of four commercialized strains of C. vulgaris (C-Auto, C-Hetero, C-Honey, and C-White) using gas- and liquid-chromatography coupled to mass-spectrometry approaches, and to evaluate their antioxidant and anti-inflammatory properties. Results showed that C-Auto had a higher lipid content compared to the other strains and higher levels of omega-3 polyunsaturated FAs (PUFAs). However, the C-Hetero, C-Honey, and C-White strains had higher levels of omega-6 PUFAs. The lipidome signature was also different between strains, as C-Auto had a higher content of polar lipids esterified to omega-3 PUFAs, while C-White had a higher content of phospholipids with omega-6 PUFAs. C-Hetero and C-Honey showed a higher content of triacylglycerols. All extracts showed antioxidant and anti-inflammatory activity, highlighting C-Auto with greater potential. Overall, the four strains of C. vulgaris can be selectively chosen as a source of added-value lipids to be used as ingredients in food and nutraceutical applications for different market needs and nutritional requirements.

High prevalence of cardiovascular disease and risk factors among type 2 diabetes patients followed in a hospital setting in Portugal: The PICT2RE observational study.

INTRODUCTION AND OBJECTIVES: Portugal is a country with a high prevalence of type 2 diabetes (T2D) and cardiovascular disease (CVD). The prevalence of CVD and cardiovascular (CV) risk factors among T2D patients followed in hospitals in Portugal is not known. The primary objective of this study was to assess the prevalence of CVD and CV risk factors among T2D patients in a hospital setting in Portugal. The clinical management of CVD in the hospital setting was also assessed. METHODS: We performed a non-interventional, multicenter, cross-sectional study with a retrospective phase. T2D patients were consecutively invited to participate. Data were collected retrospectively. RESULTS: A total of 715 patients were included in the study. Mean age and diabetes duration were 66.6 and 17.4 years, respectively. Of these, 286 patients (40.0%) had been diagnosed with CVD, mostly ischemic heart disease (50.4%). All patients had at least one CV risk factor. CVD was significantly associated with hypertension, hypercholesterolemia, low high-density lipoprotein cholesterol, hypertriglyceridemia and smoking. During the three years prior to study inclusion, the incidence of hyperglycemic episodes and T2D complications increased among patients with established CVD, but the numbers of hospitalization episodes and specialist appointments remained stable. An improvement was observed in key cardiometabolic risk factors. CONCLUSIONS: Our study revealed a high prevalence of CVD and CV risk factors among a sample of T2D patients in a hospital setting. Patients with established CVD seem to be adequately managed but further efforts are needed at the prevention stage for better control of risk factors and morbidity.

Evaluation of the fertilizer potential of Chlorella vulgaris and Scenedesmus obliquus grown in agricultural drainage water from maize fields.

Producing microalgae with agricultural drainage water (ADW) allows recycling water and nutrients, with the production of a biofertilizer, avoiding receiving waters' contamination. Chlorella vulgaris and Scenedesmus obliquus were cultivated using ADW and standard media supplementation and presented higher productivities, relatively to the control industrial growth medium (using freshwater). Selected strains were grown outdoors in pilot flat panel photobioreactors, reaching 2.20 g L-1 for S. obliquus and 1.15 g L-1 for C. vulgaris, and degrading herbicides in the ADW to non-quantifiable concentrations. The potential of the C. vulgaris and S. obliquus suspensions to replace 50% of nitrogen (N) mineral fertilization of lettuce (0.5 g pot-1) was evaluated through a pot trial, also using a 2-times (1.0 g pot-1) and 5-times (2.5 g pot-1) higher dose, applied 31 days before lettuce transplanting. Even the lower dose of N, applied via C. vulgaris or S. obliquus suspensions, was able to provide significantly higher lettuce fresh matter yield, relatively to the mineral fertilized control. Soil enzymatic activities were improved, with significantly higher dehydrogenase, ß-glucosidase, and acid phosphatase activities for the 2.5 g pot-1 dose, more marked for S. obliquus, which was also able to increase soil organic matter content. Both the non-fertilized control and microalgae fertilized pots led to similar soil electrical conductivities, 3-fold lower than in the N-mineral fertilized pots, evidencing the capacity of microalgae fertilizers to avoid soil secondary salinization. Results suggest benefits from using ADW from maize cultivation to produce C. vulgaris or S. obliquus suspensions, that can be further used as liquid organic slow-release fertilizer.

Graves' disease with spontaneous resolution following ocrelizumab in primary progressive multiple sclerosis.

Objectives. Immune reconstitution therapies (IRT), which include antibody-based cell-depleting therapies targeting CD52+ (alemtuzumab) or CD20+ (rituximab, ocrelizumab) leukocytes, are approved for the treatment of multiple sclerosis. Thyroid autoimmunity is a common adverse effect of alemtuzumab treatment, Graves' disease (GD) being the most prevalent manifestation. To date, thyroid autoimmunity events have not been reported with CD20-targeting monoclonal antibodies. Case Report. A 59-year-old woman with primary progressive multiple sclerosis with no prior personal history of thyroid disease or autoimmunity, was diagnosed with GD 6 months following the first ocrelizumab infusion. She was asymptomatic and had no signs of ophthalmopathy. Due to the temporal association of GD diagnosis with ocrelizumab infusion, absence of symptoms and our experience with alemtuzumab-induced GD, we decided for an active surveillance strategy and antithyroid drugs were not started. She underwent spontaneous resolution of hyperthyroidism with thyroid-stimulating hormone (TSH) receptor antibodies (TRAb) negativity and a mild and transitory period of subclinical hypothyroidism, while she continued the biannually ocrelizumab administration schedule. To present date, she has maintained close clinical and biochemical surveillance with normal TSH, free thyroxine (fT4) and free triiodothyronine (fT3) levels and undetectable TRAb. Conclusions. This is the first case of GD reported after ocrelizumab administration. The timing, onset and course of this case is similar to alemtuzumab-induced GD, usually interpreted as an "immune reconstitution syndrome"; however, ocrelizumab cell count depletion is inferior in severity, cell population affected and duration of depletion. This case highlights the importance of pre-screening and follow-up with thyroid function tests in patients treated with ocrelizumab. As a novel therapeutic antibody, further investigation is required to unravel the causes of thyroid autoimmunity.

Diagnostic accuracy of Bilateral Inferior Petrosal Sinus Sampling: The Experience of a Tertiary Centre.

PURPOSE: To assess the accuracy of bilateral inferior petrosal sinus sampling both in the differential diagnosis of adrenocorticotrophic hormone dependent Cushing's syndrome and predicting the localization on the pituitary. METHODS: The authors evaluated all patients that undergone bilateral inferior petrosal sinus sampling in a tertiary centre, between January 1995 and March 2018. The probable diagnosis of Cushing's disease was made when the basal central/peripheral gradient was>2 and/or>3 after stimulation with corticotrophin releasing hormone. The localization was suggested when the inter-sinus gradient was>1.4. The results obtained were compared with the post operatory results: compatible histology and positive immunohistochemistry to adrenocorticotrophic hormone and/or the presence of criteria of cure. Sensitivity, specificity and predictive positive value were calculated. RESULTS: A total of 49 patients were evaluated (75.5% female; mean age 45.4±16.3 years old). Bilateral inferior petrosal sinus sampling was compatible with Cushing's disease in 27 out of 28 confirmed cases in histology or by criteria of cure, and was compatible with ectopic secretion in the 2 cases confirmed as ectopic secretion of adrenocorticotrophic hormone (sensitivity 96.4%; specificity 100%). The lateralization calculated was concordant with the results after surgery in 17 out of 27 patients with Cushing's disease - predictive positive value of 63%. Magnetic resonance had a higher predictive value to lateralization - 70.0%. CONCLUSIONS: Bilateral inferior petrosal sinus sampling is a safe and reliable procedure to diagnose Cushing's disease, with great sensitivity and specificity. Nevertheless, the capacity of this procedure to lateralize the lesion inside the pituitary is limited.

Severe symptomatic hyperparathyroidism-Is it carcinoma?-Case report and literature review.

Parathyroid carcinoma is a rare disease, difficult to diagnose and associated with a poor prognosis. It must be suspected preoperatively, based on clinical and imaging grounds, in order to perform the best surgical option and avoid compromising patient's prognosis.

Diagnosis, treatment, and survival analysis of adrenocortical carcinomas: a multicentric study.

INTRODUCTION: Current guidelines specify controversial areas in adrenocortical carcinomas (ACC), such as optimal follow-up time after remission and identification of prognostic markers. We aim to address these topics by analyzing four reference centers in our country. METHODS: Cross-sectional multicentric study of 69 patients (mean age: 51.7 ± 16.7 years-old; women, 72.5%). Kaplan-Meier survival curves and Cox regression analysis were used to calculate overall survival and its predictors. RESULTS: Thirty-eight individuals (55.0%) had hormonal autonomous production, and 40.6% of the patients presented with metastasis. Surgery was performed in 84.1% of them. Most of these patients (72.4%) were then assigned to adjuvant therapy, while 27.6% were actively surveilled. Among patients undergoing surgery, those who achieved transient remission presented a longer survival time (66 months) than those who never reached the disease-free status (21 months) (p = 0.021). One patient presented with recurrence more than 7 years after complete tumor resection. The lowest overall survival was observed in patients (n = 11) assigned to palliative care since diagnosis (9 months). Tumor stage was identified as the only independent predictor of survival in our cohort (p = 0.006). Five-year survival was 67% for tumors confined to the adrenal space (stage I/II), 56% for locally advanced disease (stage III), and 0% for metastatic disease (stage IV). CONCLUSION: This study reinforces the dismal prognosis of ACC, the need for long-term follow-up, and tumor stage as the most important survival predictor. Reviewing medical records in such rare conditions is an opportunity to identify insufficiencies and to improve medical care.

Oxidation of benzoic acid from biomass burning in atmospheric waters.

This work evaluates the degradation of benzoic acid, a tracer from biomass burning, by different oxidation agents (Fe (III); H2O2; sunlight; and combinations of the previous ones) in model solutions and in real atmospheric waters. The extent of reactions was assessed by Ultraviolet-Visible and molecular fluorescence spectroscopies. The oxidation of benzoic acid occurred with the chemical oxidants Fe (III), H2O2, Fe (III) and H2O2 simultaneously in the presence of sunlight, and with Fe (III) and H2O2 simultaneously in the absence of light. The decrease of the pH value from neutral to acid for atmospheric waters generally increased the extent of oxidation. Sunlight was an important oxidation agent, and its combination with chemical oxidants increased the oxidation rate of benzoic acid, possibly due to the photogeneration of hydroxyl radicals. The results also suggested the occurrence of direct and indirect photolysis of benzoic acid in atmospheric waters. Moreover, the oxidation of benzoic acid produced new and more complex chromophoric compounds, which were then degraded. In addition, the nocturnal period is not sufficient for the full degradation of benzoic acid and of the intermediates formed by Fenton-like oxidation. The diurnal period may be enough for their full degradation through photo-Fenton-like oxidation, but this depends on the composition of the atmospheric waters, namely of the chromophoric content. Thus, this study highlights that benzoic acid from biomass burning, and its derivatives, may persist in atmospheric waters for periods of longer than one day, becoming available for other reactions, and may also affect the terrestrial and aquatic ecosystems through the wet depositions.

Lipoprotein apheresis in the management of severe hypercholesterolemia and hyperlipoproteinemia(a)-The Portuguese experience.

BACKGROUND: Low-density lipoprotein cholesterol (LDL-C) and lipoprotein(a) (Lp(a)) are established causal risk factors for cardiovascular disease (CVD). Lipoprotein apheresis is often required for treatment of patients with a high risk for CVD due to hypercholesterolemia and/or hyperlipoproteinemia(a). AIM: To describe our experience with lipoprotein apheresis in patients with severe hypercholesterolemia or with hyperlipoproteinemia(a). METHODS: We retrospectively investigated patients treated with Lipoprotein apheresis using direct adsorption of lipoproteins (DALI) technique, between December 2008 and March 2018, in our center. Adverse events, acute and long term reductions in lipid parameters were analyzed. RESULTS: Between December 2008 and March 2018, a total of 950 treatments were performed in five patients, four with heterozygous familial hypercholesterolemia (HeFH), all on maximally tolerated cholesterol-lowering drug therapy and in one patient with hyperlipoproteinemia(a) and progressive CVD. In the four patients with HeFH we obtained mean acute reductions in LDL-C and non-high-density lipoprotein cholesterol (non-HDL-C) of 62.0 ± 7.8% and 60.4 ± 6.8%, respectively. Regarding long-term efficacy we achieved a mean reduction of 43.1% in LDL-C and of 41.2% in non-HDL-C. In the patient with hyperlipoproteinemia(a) we attained mean acute reductions of 60.4 ± 6.4% in Lp(a) and of 75.4 ± 7.3% in LDL-C per session and long term reductions in Lp(a) and LDL-C of 67.4% and 40.5%, respectively. Adverse events were recorded in only 1.2% of treatments. CONCLUSION: Lipoprotein apheresis is an efficient and safe treatment in severely hypercholesterolemic patients who are refractory to conservative lipid-lowering therapy or with hyperlipoproteinemia(a) and progressive CVD.

Visceral Obesity and Metabolic Syndrome Are Associated with Well-Differentiated Gastroenteropancreatic Neuroendocrine Tumors.

The determinants for gastroenteropancreatic neuroendocrine tumors (GEP-NET) recent burden are matters of debate. Obesity and metabolic syndrome (MetS) are well established risks for several cancers even though no link with GEP-NETs was yet established. Our aim in this study was to investigate whether well-differentiated GEP-NETs were associated with obesity and MetS. Patients with well-differentiated GEP-NETs (n = 96) were cross-matched for age, gender, and district of residence with a control group (n = 96) derived from the general population in a case-control study. Patients presented gastro-intestinal (75.0%) or pancreatic (22.9%) tumors, grade G1 (66.7%) or G2 (27.1%) with localized disease (31.3%), regional metastasis (16.7%) or distant metastasis (43.8%) at diagnosis, and 45.8% had clinical hormonal syndromes. MetS was defined according to Joint Interim Statement (JIS) criteria. Well-differentiated GEP-NETs were associated with MetS criteria as well as the individual components' waist circumference, fasting triglycerides, and fasting plasma glucose (p = 0.003, p = 0.002, p = 0.011 and p < 0.001, respectively). The likelihood of the association was higher when the number of individual MetS components was greater than four. MetS and some individual MetS components including visceral obesity, dyslipidemia, and increased fasting glucose are associated with well-differentiated GEP-NET. This data provides a novel insight in unraveling the mechanisms leading to GEP-NET disease.

Dreno de sucção fechado em mastectomias unilaterais totais em cadelas / Closed suction drain in total unilateral mastectomy in bitches

O objetivo deste estudo foi avaliar a utilização de dreno de sucção fechado em feridas cirúrgicas de mastectomias unilaterais totais em cadelas. Foram utilizadas 18 cadelas com peso médio de 14,87±9,29kg, idade média 9,34±2,68 anos, provenientes da rotina hospitalar, as quais foram selecionadas por meio de avaliação clínica e oncológica. Os animais foram alocados em dois grupos: Convencional (GC, n=9) que foram submetidas à cirurgia de mastectomia unilateral total, sem colocação de dreno e grupo Dreno (GD, n=9) que foram submetidas à mastectomia unilateral total com colocação do dreno de sucção fechado (Biovácuo?), sendo o diâmetro do dreno determinado conforme peso do animal. Foram realizadas cinco avaliações durante o período pós-operatório: avaliação clínica diária dos animais através da frequência cardíaca (FC), frequência respiratória (f), temperatura retal (TR), coloração de mucosas, tempo de preenchimento capilar (TPC), grau de desidratação, hemograma, perfil bioquímico (ureia, creatinina, colesterol, triglicérides e glicose); avaliação qualitativa do dreno; avaliação qualitativa da ferida cirúrgica, sendo esta também realizada em 14 e 30 dias quanto à presença ou não de exsudato, edema, integridade da pele, deiscência, necrose e fibrose cicatricial; avaliação quantitativa do fluido drenado e avaliação qualitativa do fluido drenado, quanto à celularidade, tipo e morfologia celular. Houve aumento dos valores médios de densidade, neutrófilos, linfócitos, e aumento do valor de mediana de fibrinogênio do fluido drenado em 72 horas em relação às 24 horas no grupo GD. Ocorreu aumento do exsudato em 24, 48 e 72 horas no GD quando comparado ao GC. Houve aumento do volume de fluido drenado em 60 e 72 horas em relação às 12 horas no grupo GD. Houve redução do valor médio de eritrócitos em 72 horas em relação ao basal no grupo GD. Observou-se aumento nos valores médios de hemoglobina e hematócrito em 24 horas no grupo GC quando comparado ao GD. Houve aumento no valor médio de neutrófilos segmentados no basal no GD quando comparado ao GC. Conclui-se que a técnica de colocação de dreno de sucção fechado apresenta maior teor de exsudato. Recomenda-se que o vácuo do dreno seja refeito de 12 em 12 horas, sendo inicialmentrealizado 6 horas após o término da cirurgia. O dreno de sucção fechado não gera desconforto adicional ao paciente; recomenda-se que o mesmo seja removido 72 horas após o término da cirurgia, quando a produção de fluido diminui, porém mesmo assim alguns indivíduos podem apresentar formação de seroma no pós-operatório tardio.(AU)

The aim of this study was to evaluate the use of closed suction drain in surgical wounds of total unilateral mastectomy in dogs. Eighteen dogs were used with an average weight of 14.87±9.29kg, average age 9.34±2.68 years, from the hospital routine, which were selected through clinical and oncological evaluation. The animals were divided into two groups: Conventional (CG, n=9) which underwent total unilateral mastectomy surgery without drain and Drain group (DG, n=9) which underwent complete unilateral mastectomy with placement closed suction drain (Biovácuo?). The diameter of the drain was determined according to the animal weight. Five evaluations were performed during the postoperative period: clinical evaluation of the animals daily through the heart rate (HR), respiratory rate (RR), rectal temperature (RT), color of mucous membranes, capillary refill time (CRT), degree of dehydration, blood count, chemistry profile (urea, creatinine, cholesterol, triglycerides and glucose); qualitative assessment of the drain; qualitative assessment of the surgical wound, is also being held on 14 and 30 days for the presence or absence of exudate, edema, skin integrity, dehiscence, necrosis and scarring; quantitative evaluation of the drained fluid; qualitative assessment of fluid drained, as the cellularity type and cell morphology. Surgical time was recorded in both groups. There were an increase in average values of density, neutrophils, lymphocytes, and increased fibrinogen median value of drained fluid in 72 hours compared to 24 hours in DG group. There was an increase of exudate within 24, 48 and 72 hours in DG when compared to GC. There was an increase in the volume of drained fluid in 60 and 72 hours compared to 12 hours in DG. There was a reduction in the average value of red blood cells within 72 hours from baseline in DG group. There was an increase in average hemoglobin and hematocrit in 24 hours in the control group compared to the DG. There was an increase in the average value of neutrophils targeted at baseline in DG when compared to CG. It follows that, the closed suction drain placement technique presents more exudate. It is recommended that the drain vacuum is redone 12 in 12 hours, initially being performed 6 hours after surgery. The closed suction drain does not generate additional discomfort to the patient; it is recommended that it be removed 72 hours after the surgery, when the fluid production decreases, but even so some individuals may present seroma formation in the late postoperative.(AU)

Echocardiographic evaluation of dogs subjected to different protocols of anesthetic induction / Avaliação ecocardiográfica de cães submetidos a diferentes protocolos de indução anestésica

ABSTRACT: The objective of this study was to investigate the echocardiographic changes during anesthesia induction in dogs sedated with acepromazine (0.05mg/kg) and butorphanol (0.3mg/kg) (AB). Twenty-four male dogs, with a mean weight of 12.40kg±3.1kg, were randomly assigned to 4 groups (n=6). Fifteen minutes after administering pre-anesthetic medication, anesthesia with diazepam (0.5mg/kg) and etomidate (1mg/kg) (group DE); diazepam (0.5mg/kg) and ketamine (3mg/kg) (group CD); propofol (4mg/kg) (group P); or ketamine (1mg/kg) and propofol (3mg/kg) (group CP) was administered to the 6 dogs in each group. Systolic blood pressure (SBP) was measured and echocardiography was performed immediately prior to the application of the sedation protocol (baseline), 15 minutes after sedation (M1), and immediately after anesthesia induction (M2). No significant differences were observed in SBP and in hemodynamic variables such as cardiac index, shortening fraction, and ejection fraction, between groups at all time points (M0, M1, and M2) evaluated. The SBP was significantly reduced after anesthetic induction in the dogs of the DE and CP groups. It can be concluded that the protocols DE and CP reduce similarly to SPB in dogs medicated with CD and P to SBP remain stable after anesthetic induction. All anesthetic induction protocols maintained a stable IC in premedicated dogs. None of the protocols evaluated promoted significant echocardiographic changes. Furthermore, the ketamine and diazepam combination had a negative impact on myocardial relaxation.

RESUMO: O aumento crescente da expectativa de vida dos cães, faz com que muitos animais cardiopatas necessitem de um procedimento anestésico-cirúrgico. Os objetivos do estudo foram investigar as alterações ecocardiográficas de protocolos de indução anestésica, em cães sedados com acepromazina (0,05mg/kg) e butorfanol (0,3mg/kg) (AB). Foram utilizados 24 cães, machos, SRD, com peso médio de 12,40±3,1kg, os quais foram alocados aleatoriamente em quatro grupos (n=6). Após 15 minutos da medicação pré-anestésica, foi realizada indução anestésica com diazepam (0,5mg/kg)/etomidato (1mg/kg) (DE), ou diazepam (0,5mg/kg)/cetamina (3mg/kg) (CD), oupropofol (4mg/kg) (P) ou cetamina (1mg/kg)/propofol (3mg/kg) (CP). Aferiu-se a PAS e ecocardiografia imediatamente antes da aplicação do protocolo de sedação (basal), 15 minutos após a sedação (M1) e imediatamente após a indução anestésica (M2). Não foram observadas diferenças significativas na PAS e nas variáveis hemodinâmicas como índice cardíaco, fração de encurtamento, fração de ejeção, entre os grupos, em todos os momentos avaliados. A PAS reduziu significativamente, após indução anestésica, nos cães do grupo DE e CP. A FC reduziu, após indução, em relação aos valores pós sedação, somente no grupo CD, mantendo-se estável nos demais grupos estudados. Conclui-se que os protocolos DE e CP reduzem de maneira semelhante a PAS, nos cães medicados com CD e P a PAS mantêm-se estável, após indução anestésica. Todos os protocolos de indução anestésica mantém estáveis o IC em cães pré-medicados. Nenhum dos protocolos avaliados promove alterações ecocardiográficas significativas. Aassociação cetamina/diazepam tem um impacto negativo no relaxamento miocárdico.

[Endocrinology in Portugal - Census 2016. Board of the Portuguese College of Endocrinology and Nutrition of the Portuguese Medical Association]. / A Endocrinologia em Portugal - Censo 2016. Direção do Colégio de Endocrinologia e Nutrição da Ordem dos Médicos.

INTRODUCTION: On September 2016, the Board of the College of Endocrinology and Nutrition of the Portuguese Medical Association carried out a national survey, about all Endocrinology, Diabetes and Metabolism Departments of the public hospitals included in the Portuguese National Health Service and a simplified version of this survey was sent to all endocrinologists working in Portugal and registered with the Portuguese Medical Association. MATERIAL AND METHODS: Data related to organizational and human resources were collected, reporting the situation by the end of year 2015. The census registered 107 individuals and 27 Departments. RESULTS: The ratio of endocrinologists-population was 1.4, much lower than in the other European countries (varies between 2 to 4), resulting in alarming shortages of services in some areas of Portugal and in worse quality indicators. DISCUSSION: These data suggest that actions should be taken to increase the number of endocrinologists and departments in the country. CONCLUSION: In recent years, the number of residents has significantly increased, which will make it possible to correct this situation.

Introdução: A Direção do Colégio de Endocrinologia e Nutrição da Ordem dos Médicos realizou um inquérito nacional em setembro de 2016, a todos os serviços de Endocrinologia, Diabetes e Metabolismo dos hospitais do Serviço Nacional de Saúde e uma versão simplificada do mesmo foi enviada a todos os endocrinologistas a trabalhar em Portugal e inscritos no colégio. Material e Métodos: O censo inclui dados organizacionais e de recursos humanos relativos ao fim do ano de 2015. Registou 107 respostas individuais e 27 serviços. Resultados: O ratio de endocrinologistas por 100 000 habitantes era de 1,4, muito inferior a outros países europeus (varia de 2 a 4), que resulta numa carência grave de serviços em algumas zonas do País e em piores indicadores de qualidade. Discussão: Estes dados indicam que devem ser implementadas medidas para aumentar o número de endocrinologistas e serviços em Portugal. Conclusão: Nos últimos anos, o número de internos tem vindo a aumentar, o que vai permitir melhorar esta situação.

Electrocardiographic, echocardiographic, and indirect blood pressure evaluation in dogs subjected to different sedation protocols / Avaliação eletrocardiográfica, ecocardiográfica e de pressão arterial não invasiva de cães submetidos a diferentes protocolos de sedação

ABSTRACT: The present study aimed to evaluate the effects of different sedation protocols on blood pressure and echocardiographic and electrocardiographic parameters in dogs. In total, 24 male mixed-breed dogs with a mean weight of 9.87±3.0kg were used.Animals were randomly divided into four groups (n=6), which were subjected to sedation using the following protocols: acepromazine (0.05mgkg-1) and butorphanol (0.3mgkg-1) (AB); acepromazine (0.05mgkg-1)and methadone (0.5mgkg-1) (AM); acepromazine (0.03mgkg-1), methadone (0.5mgkg-1), and midazolam (0.3mgkg-1)(MAM); and methadone only (0.5mgkg-1) (M). Indirect blood pressure (BP) measurements and computerized electrocardiography (ECG) and echocardiography (ECO) were performed immediately before the application of the sedation protocol (baseline), and the same evaluations were repeated after 15 minutes. BP decreased in groups AB, MAM, and AM compared to baseline values. Electrocardiographic measurements showed decreased heart rates (HRs) after sedation in all groups, and bradycardia was observed after sedation in two dogs from group M and one animal from group AM. The P-wave duration increased after sedation in groups AM and M. After sedation, no changes in cardiac dimensions were revealed byECO.Fractional shortening (FS) decreased after sedation in the AM group, and dogs from group AB exhibited a smaller decrease in FS compared with the other groups. The cardiac index (CI) was lower in groups AM and M than in the other groups. Animals from group AB were less resistant to examination and exhibited the most favorable sedation scores. It was concluded that the combination of acepromazine and butorphanol was the best sedation protocol for performing echocardiogram measurementsbecause dogs were less resistant to examinations and echocardiographic parameters of FS and CI remained stable.

RESUMO: O objetivo deste estudo foi avaliar os efeitos de diferentes protocolos de sedação sobre a pressão arterial, parâmetros ecocardiográficos e eletrocardiográficos em cães. Foram utilizados 24 cães, machos, SRD, com peso médio de 9,87±3,0kg, os quais foram alocados aleatoriamente em quatro grupos (n=6), que foram submetidos à sedação com os protocolos acepromazina (0,05mgkg-1) e butorfanol (0,3mgkg-1) (AB), acepromazina (0,05mgkg-1) e metadona (0,5mgkg-1) (AM), acepromazina (0,03mgkg-1), metadona (0,5mgkg-1) e midazolam (0,3mgkg-1) (MAM) e metadona isolada (0,5mgkg-1) (M). Foi realizada avaliação da pressão arterial sistólica (PAS) não invasiva, eletrocardiografia computadorizada e ecocardiografia imediatamente antes da aplicação do protocolo de sedação (basal) e repetindo-se as mesmas avaliações, 15 minutos após. Observou-se redução na PAS nos grupos AB, MAM, AM, em relação ao basal. Na eletrocardiografia, houve redução da FC após sedação em todos grupos, sendo observada bradicardia após sedação em dois cães do grupo M e um animal do grupo AM. A duração da onda P aumentou após sedação nos grupos AM e M. Não foram observadas alterações nas dimensões cardíacas, avaliadas pela ecocardiografia, após sedação. A fração de encurtamento (FS) reduziu após sedação no AM e os cães do AB apresentaram menor queda da FS, diferindo dos demais grupos. O índice cardíaco (IC) foi menor no AM e M em relação aos demais. Os animais do grupo AB foram menos resistentes à execução dos exames, apresentando melhores escores de sedação. Concluiu-se que a associação acepromazina e butorfanol foi o melhor protocolo de sedação para realização do ecocardiograma, sendo os cães menos resistentes à execução do exame, mantendo estáveis os parâmetros ecocardiográficos de fração de encurtamento e índice cardíaco.

Cardiac index by thermodilution or transthoracic echocardiography in dogs at different hemodynamic states / Índice cardíaco pelos métodos de termodiluição ou ecocardiografia transtorácica em cães sob diferentes situações hemodinâmicas

ABSTRACT: Proper monitoring of cardiac index (CI) in critically ill patients requires accurate and minimally invasive methods. The aim of this study was to compare the CI values obtained by thermodilution or echocardiography using different methods in dogs in different hemodynamic states. Nine dogs weighing 19.6±1.3kg were anesthetized with isoflurane at 1.4V% (Baseline) and subjected to mechanical ventilation (MV),a hypodynamic state (Hypo) with isoflurane at 3.5V% and hyperdynamic state (hyper) with dobutamine infusion at 5μgkg-1min-1. CI analysis was performed by thermodilution (TD) and using the modified Simpson's method, aortic velocity-time integral (A-VTI) method and pulmonary VTI (P-VTI) method. We performed Pearson's correlation and Bland-Altman analysis. The CI values (Lm-2min-1) of the animals in the Baseline, MV, Hypo and Hyper states were 4.3±1, 3.6±0.7, 2.9±0.66 and 6.1±2, for TD; 2.8±0.7, 2.4±0.3, 1.7±0.7 and 4.4±1.2, for Simpson's method; 3.4±0.9, 3.1±0.7, 2.6±3.4, 6.1±1.8 for A-VTI; and 3.6±0.8, 3.6±0.8, 2.7±0.6 and 6.2±1.5, for P-VTI. The CI values using Simpson's method were lower than those obtained by TD in all states, and it was observed a significant correlation in the Hypo (r=0.89) and Hyper (r=0.76) groups. In addition,the percent error in the Hypo group using Simpson's method was 26% relative to TD, which allowed for the identification of the different hemodynamic states. With respect to the other methods and states, there was no agreement or correlation between the methods and TD. We concluded that none of the tested echocardiography methods exhibited acceptable agreement with thermodilution at different hemodynamic states.

RESUMO: A adequada monitoração do índice cardíaco (IC) em pacientes críticos requer métodos acurados e minimamente invasivos. O objetivo deste estudo foi comparar o IC obtido por termodiluição ou ecocardiografia em cães sob alterações hemodinâmicas. Utilizaram-se nove cães pesando 19,6±1,3kg, os quais foram anestesiados com isofluoranoa1,4V% (Basal) e submetidos à ventilação mecânica (VM) e estados hipodinâmico (Hipo) com isofluoranoa3,5V% e hiperdinâmico (Hiper), com dobutaminaa5µgkg-1min-1. O IC foi obtido por termodiluição (TD) e pelos métodos ecocardiográficos de Simpson modificado, e pela velocidade em tempo integral (VTI) nas valvas aórtica (VTI-A) e pulmonar (VTI-P). Realizou-se a análise de correlação de Pearson e de concordância de Bland-Altman. O IC (Lm-2min-1) nas fases Basal, VM, Hipo e Hiper foi de 4,3±1, 3,6±0,7, 2,9±0,66 e 6,1±2 para TD; 2,8 ±0,7, 2,4±0,3, 1,7±0,7 e 4,4±1,2 para Simpson; 3,4±0,9, 3,1 ±0,7, 2,6±3,4, 6,1±1,8 para VTI-A e 3,6±0,8, 3,6±0,8, 2,7±0,6 e 6,2±1,5 para VTI-P. O método de Simpson foi menor que a TD em todas as fases, mas com correlação significativa nos estados Hipo (r=0,89) e Hiper (r=0,76) e percentagem de erro de 26% no Hipo em relação à TD, identificando os diferentes estados hemodinâmicos. Nos demais, não houve concordância ou correlação com a TD. Conclui-se que nenhum dos métodos testados apresentou concordância aceitável com a termodiluição nos diferentes estados hemodinâmicos.

[Congenital adrenal hyperplasia due to 21-hydroxylase deficiency: genotype-phenotype correlation]. / Hiperplasia Congénita da Suprarrenal por Deficiência de 21-Hidroxílase: Correlação Genótipo-Fenótipo.

INTRODUCTION: Congenital adrenal hyperplasia due to 21-hydroxylase deficiency is one of the most frequent inborn conditions. It is caused by distinct mutations in the CYP21A2 gene and in the majority of cases the disease's severity correlates with CYP21A2 allelic variation Our aim was to describe the mutational spectrum of CYP21A2 and evaluate genotype-phenotype correlation in a cohort of portuguese patients with 21-hydroxylase deficiency. MATERIAL AND METHODS: Retrospective study of 22 patients with clinical diagnosis of 21-hydroxylase deficiency. Molecular analysis of CYP21A2 was performed and genotype-phenotype correlation was then established. RESULTS: Genotyping was performed in 22 unrelated patients: 5 with classic salt-wasting (average age of diagnosis 10,2 days; minimum 1, maximum 20 days), 7 with classic simple virilizing (average age of diagnosis 3,5 years; minimum 0 days, maximum 7 years) and 10 with nonclassical form (average age of diagnosis 5,7 years; minimum 4 years, maximum 8 years). The most frequent genetic defects in the classic forms were I2 splice (24%) and I172N (24%), followed by Q318X (16%) and gene deletions (16%) and in the nonclassical form, the V281L (80%). The overall concordance between genotype and phenotype was 81,8%. Genotype accurately predicted phenotype in 83,3%, 100% and 90% of patients with classic salt-wasting, classic simple virilizing and nonclassical mutations, respectively. DISCUSSION: The frequency of genetic defects in our patients was comparable to similar studies. In most cases there was a good correlation between genotype and phenotype. CONCLUSIONS: Molecular analysis of CYP21A2 provides useful information in terms of prediction of disease severity, genetic and prenatal counseling.

Introdução: A hiperplasia congénita da suprarrenal por deficiência de 21-hidroxílase constitui uma das doenças hereditárias mais comuns. Resulta de diferentes mutações no gene CYP21A2 e, na maioria dos casos, a gravidade da doença correlaciona-se com a variação alélica do CYP21A2. O objetivo deste estudo foi descrever o espectro mutacional do CYP21A2 e avaliar a correlação genótipo-fenótipo numa coorte de doentes portugueses com deficiência de 21-hidroxílase. Material e Métodos: Estudo retrospetivo de 22 doentes com diagnóstico clínico de deficiência de 21-hidroxílase. Foi feita análise molecular do CYP21A2 e estabelecida a correlação genótipo-fenótipo. Resultados: Foi realizada genotipagem em 22 doentes não relacionados: 5 com a forma clássica perdedora de sal (idade média ao diagnóstico de 10,2 dias; mínimo 1, máximo 20 dias), 7 com a forma clássica virilizante simples (idade média ao diagnóstico de 3,5 anos; mínimo 0 dias, máximo 7 anos) e 10 com a forma não clássica (idade média ao diagnóstico de 5,7 anos; mínimo 4 anos, máximo 8 anos). Os defeitos genéticos mais frequentes nas formas clássicas foram o I2 splice (24%) e I172N (24%), seguindo-se o Q318X (16%) e deleções de genes (16%) e, na forma não clássica, o V281L (80%). Verificou-se uma concordância genótipo-fenótipo global de 81,8%. O genótipo permitiu prever adequadamente o fenótipo em 83,3%, 100% e 90% dos doentes com mutações compatíveis com a forma clássica perdedora de sal, clássica virilizante simples e não clássica, respectivamente. Discussão: A frequência de defeitos genéticos observados nos nossos doentes é comparável a estudos semelhantes. Observou-se, na maioria dos casos, uma boa correlação genótipo-fenótipo. Conclusões: A análise molecular do CYP21A2 fornece informação importante relativamente à gravidade da doença e no aconselhamento genético e pré-natal.

Preditores da adesão à medicação na Epilepsia: Estudo Longitudinal / Medication adherence predictors in Epilepsy: Longitudinal Study

Introdução: A não adesão à medicação na epilepsia é prevalente, pelo que a compreensão dos fatores associados deve ser promovida. Objetivos: Analisar a capacidade preditiva das estratégias de coping e da espiritualidade em relação à adesão à medicação ao longo do tempo. Metodologia: Foram avaliados 60 indivíduos através de um Questionário Sociodemográfico e Clínico, a Medida de Adesão aos Tratamentos, o COPER e a Escala de Avaliação de Espiritualidade em Contextos de Saúde, em dois momentos. Resultados A espiritualidade Momento 1 não se relacionou com adesão à medicação no Momento 3, e as estratégias de coping: Desinvestimento Comportamental e Aceitação no Momento 1 predizem negativamente a adesão à medicação no Momento 2. Conclusão: A relação entre a adesão à medicação, estratégias de coping e espiritualidade varia emfunção do tempo, o que deve ser considerado ao nível da intervenção...

Introduction: The nonmedication adherence in epilepsy is prevalent by which the understanding of associated factors shouldbe promoted. Objectives: To analyze the predictive capacity of the coping strategies and spirituality in relationto medication adherence over time. Methodology: A sociodemographic and clinical questionnaire, medical adherence to treatment, COPER and spirituality evaluation scale in health contexts was undertaken to evaluate 60 individuals with epilepsy, in three moments. Results: Spirituality at Moment 1 is not related to medication adherence at Moment 3, and the coping strategies: behavioral disinvestment and acceptance at Moment 1 predict medication adherence at moment 2. Conclusion: The relation between medication adherence, coping strategies and spirituality change according to time, which should be considered on the level intervention...