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The COVID-19 pandemic affected many aspects of human mobility and resulted in unprecedented changes in population dynamics, including lifestyle and mobility. Recognizing the effects of the pandemic is crucial to understand changes and mitigate negative impacts. Spatial data on human activity, including mobile phone data, has the potential to provide movement patterns and identify regularly visited locations. Moreover, crowdsourced geospatial information can explain and characterize the regularly visited locations. The analysis of both mobility and routine locations in the same study has seldom been carried out using mobile phone data and linked to the effects of the pandemic. Therefore, in this article we study human mobility patterns within Portugal, using mobile phone and crowdsourced data to compare the population's mobility and routine locations after the pandemic's peak. We use clustering algorithms to identify citizens' stops and routine locations, at an antenna level, during the following months after the pandemic's first wave and the same period of the following year. Results based on two mobile phone datasets showed a significant difference in mobility in the two periods. Nevertheless, routine locations slightly differ. [ABSTRACT FROM AUTHOR]

• 13 rare LDLR missense variants were functionally characterised. • Our results identified 11 additional functionally abnormal LDLR variants. • 22 patients received a definite diagnosis with the new functional data. Familial Hypercholesterolemia (FH) is a semidominant disorder of the lipid metabolism associated with premature atherosclerosis and coronary heart disease. So far, about 3,000 unique LDLR variants have been described, most of which lack functional evidence proving their effect on LDLR function, despite the important role that functional studies play in variant classification. In this work, we aimed to functionally characterize 13 rare missense variants, identified worldwide and in Portugal, in clinical FH patients. LDLR -deficient CHO-ldlA7 cells were transfected with plasmids carrying different LDLR variants generated by site-directed mutagenesis. LDLR activity and expression were assessed by FACS. 11/13 variants affect LDLR function (p.Cys109Phe; p.Cys143Arg; p.Glu267Lys; p.Cys352Ser; p.Ile451Thr; p.His485Gln; p.Asp492Asn; p.Val500Ala; p.Gly529Arg; p.Phe614Ile; p.Glu626Lys) and 2/13 are inconclusive (p.Arg81Cys; p.Gly98Arg;). Of the 13 variants studied, 8 were classified as VUS by ACMG criteria, but for 7 of these 8, our functional studies were able to reassign them as Likely pathogenic or Pathogenic. For an accurate diagnosis, an effort must be made to improve functional characterization of putative disease-causing variants. [ABSTRACT FROM AUTHOR]

Familial hypercholesterolaemia (FH) is an autosomal disorder of lipid metabolism presenting with increased cardiovascular risk. LDLR mutations are the cause of disease in 90% of the cases but functional studies have only been performed for about 15% of all LDLR variants. In the Portuguese Familial Hypercholesterolemia Study (PFHS), 142 unique LDLR alterations were identified and 44 (30%) lack functional characterization. The aim of the present work is to increase evidence for variant classification by performing functional characterization of 13 LDLR missense alterations found in Portugal and in 20 other countries. Different LDLR mutants were generated by site-directed mutagenesis and expressed in CHO–ldlA7 cells lacking endogenous expression of LDLR. To determine the effects of alterations on LDLR function, cell surface expression, binding and uptake of FITC-LDL were assessed by flow cytometry and Western blot. Of the 13 variants studied 7 were shown to affect LDLR function - expression, binding or uptake, with rates lower than 60%: p.(Cys184Tyr), p.(Gly207_Ser213del); p.(His211Asp); p.(Asp221Tyr); p.(Glu288Lys); p.(Gly592Glu) and p.(Asp601Val)). The remaining 6 variants do not alter the LDLR function. These studies contributed to an update of these variants classification: from the 9 variants classified as variants of unknown significance, 7 have reached now a final classification and 3 variants have improved classification from likely pathogenic to pathogenic. In Portugal, an additional 55 patients received an FH definite diagnosis thanks to these studies. Since only likely pathogenic and pathogenic variants are clinically actionable, this work shows the importance of functional studies for variant classification. [Display omitted] • This work shows the importance of functional studies for variant classification, since only likely pathogenic and pathogenic variants are clinically actionable. • This work contributed to an update of 10 variants classification. • Variants presented in this work were also found in 20 different countries and many other patients worldwide will benefit from these. • Functional studies are very important to add evidence to reach a final classification with the ACMG algorithm. [ABSTRACT FROM AUTHOR]

Key message: The wood ofPinus nigrapopulations planted in Portugal, comparatively toPinus pinaster, has higher total extractive content, lower Klason lignin and H/G ratio, and similar mechanical properties, presenting advantages for industrial purposes. Context: P. nigra was used in the reforestation of mountainous areas in Portugal, but its wood chemical and mechanical properties were never studied. Aims: This work intends to evaluate the chemical and mechanical wood properties of the P. nigra populations planted in Portugal, to relate these properties with previously characterised physical features and to compare these data with other European P. nigra stands and species, namely, P. pinaster. Methods: Wood chemical and mechanical properties were analysed in 90 trees from six Portuguese sites, using near-infrared (NIR) spectrometry and the three-point bending test. Results: The wood of the P. nigra populations planted in Portugal presented average values of total extractive content = 9.4%, Klason lignin = 26.69%, MORRad = 14.93 MPa and MOERad = 1200.98 MPa. Ring density showed no significant correlation with ring width. Conclusion: The P. nigra populations planted in Portugal presented qualitative and quantitative properties similar to P. pinaster wood, the main resinous species in Portugal. Facing the lack of raw material for wood industry due to frequent forest fires in the Mediterranean region, P. nigra could be used to reforest mountainous areas of those regions. [ABSTRACT FROM AUTHOR]

Background: Prevalence of fatty liver (FL) and nonalcoholic fatty liver disease (NAFLD) depends mainly on obesity, diabetes and genetic factors. FL and NAFLD prevalence was evaluated in Portuguese adult population and correlated with several risk factors and related mortality data, within the same period. Materials and methods: A cross‐sectional, population‐based multicenter study, voluntary and randomly selected in 834 Portuguese adults (18‐79 years). Participants were evaluated after 12‐hour fasting. Anthropometric data, past history including alcohol consumption, and associated diseases were registered. Blood samples were collected for biochemical testing. Dietary intake was evaluated using a semi‐quantitative food frequency questionnaire. Presence of FL was evaluated using ultrasound, and NAFLD was diagnosed after exclusion of other causes for liver disease. Results: Adjusted prevalence of FL and NAFLD was 37.8% and 17.0%, respectively. FL individuals were older, more frequently males, with increased probability of having obesity, diabetes or harmful alcohol consumption (HAC). NAFLD individuals were also older, but had a similar sex distribution and an increased probability of obesity and diabetes. In both groups, no differences were found regarding dietary pattern or physical activity. During the same time period, nonalcoholic steatohepatitis (NASH) liver‐related deaths in Portugal were 0.105/100 000, while alcohol‐related liver disease mortality was 6.790/100 000. Conclusion: The large spectrum of FL was present in more than one third of the population, although only less than half could be classified as NAFLD. Other significant risk factors, such as HAC, are probably implicated in FL, explaining the low NASH‐related mortality compared with the high alcohol‐related mortality during the same time period. [ABSTRACT FROM AUTHOR]

Background There is little information about familial hypercholesterolemia (FH) epidemiology and care in Ibero-American countries. The Ibero-American FH network aims at reducing the gap on diagnosis and treatment of this disease in the region. Objective To describe clinical, molecular, and organizational characteristics of FH diagnosis in Argentina, Brazil, Chile, Colombia, Mexico, Portugal, Spain, and Uruguay. Methods Descriptive analysis of country data related to FH cascade screening, molecular diagnosis, clinical practice guidelines, and patient organization presence in Ibero-America. Results From a conservative estimation of an FH prevalence of 1 of 500 individuals, there should be 1.2 million heterozygous FH individuals in Ibero-America and roughly 27,400 were diagnosed so far. Only Spain, Brazil, Portugal, and Uruguay have active cascade screening programs. The prevalence of cardiovascular disease ranged from 10% to 42% in member countries, and the highest molecular identification rates are seen in Spain, 8.3%, followed by Portugal, 3.8%, and Uruguay with 2.5%. In the 3 countries with more FH patients identified (Spain, Portugal, and Brazil) between 10 and 15 mutations are responsible for 30% to 47% of all FH cases. Spain and Portugal share 5 of the 10 most common mutations (4 in low density lipoprotein receptor [ LDLR ] and the APOB3527). Spain and Spanish-speaking Latin American countries share 6 of the most common LDLR mutations and the APOB3527. LDL apheresis is available only in Spain and Portugal and not all countries have specific FH diagnostic and treatment guidelines as well as patient organizations. Conclusions Ibero-American countries share similar mutations and gaps in FH care. [ABSTRACT FROM AUTHOR]

The aim of this study was to analyze the interference of strength and aerobic training order over an 8-week period on explosive skills and maximal oxygen uptake (...O2max) in prepubes-cent children. One hundred twenty-eight prepubescent children aged 10-11 years (10.9 ± 0.5 years) were randomly selected and assigned to 1 of the 3 groups: intrasession concurrent aerobic before (GAS: n = 39) or after strength training (GSA: n = 45) or control group (GC: n = 44; no training program). The GC maintained their baseline level performance, and training-induced differences were found in the experimental groups. Increases were found in the 1-kg and 3-kg medicine ball throws: GAS: +3%, +5.5%, p ≤ 0.05, p < 0.001 ; GSA: +5.7%, +8.7%, p < 0.001, respectively; in the counter movement jump height and standing long jump length: GAS: +6.5%, +3.4%, p ≤ 0.05; GSA: +7%, +4.5%, p < 0.001, respectively; in the 20-m shuttle-run time: GAS: +2.3%; GSA: +4.6%, p < 0.001 ; and, in the ...O2max: GAS: +7.3%, p < 0.001 ; GSA: +3.8%, p < 0.001 from pretraining to post-training. All programs were effective, but GSA produced better results than GAS for muscle strength variables, and GAS produced better results than GSA for aerobic capacity variables. The present study explored an unknown issue and added useful information to the literature in this area. These training methods should be taken into consideration to optimize explosive strength and cardiorespiratory fitness training in school-based programs and sports club programs. [ABSTRACT FROM AUTHOR]

Background: Parasitic diseases of companion animals comprise a group of globally distributed and rapidly spreading illnesses that are caused by a wide range of arthropods, helminths and protozoa. In addition to their veterinary importance, many of these parasites can also affect the human population, due to their zoonotic potential. The aim of the present work was to evaluate the knowledge of Portuguese pet owners regarding the zoonotic potential of parasites that dogs and cats can harbour, most common drugs, frequency of use and reasons for endo- and ectoparasite control. Methods: Seventy hundred and fifty multiple-choice questionnaires designed to obtain data knowledge about the meaning of zoonosis, knowledge about parasitic diseases and perception regarding their zoonotic potential, as well as the drugs, frequency and reason for deworming their animals were delivered to dog and/or cat owners from non-rural (i.e. urban or semi-urban) and rural parishes who attended veterinary medical centres from continental Portugal. Results: A total of 536 (71.5 %) questionnaires were retrieved. Two hundred and ninety five (56.5 %) responders had heard of zoonosis/zoonoses, but only 184 (35.2 %) knew their meaning. Tick fever, mange, leishmaniosis and ascaridiosis/roundworms were the parasitic diseases from pets most frequently identified. The number of owners who recognized the different parasitoses, who stated to have heard about zoonoses and who were aware of the potential transmission of parasites from animals to humans was significantly higher in those with intermediate (i.e. =9 and= 12 years of schooling) and/or higher academic degree (i.e. licentiate, master's and/or doctorate degrees). The combinations of febantel-pyrantel-praziquantel (23.5 %) and milbemycin-praziquantel (34.5 %) were the most widely endoparasitic drugs used in dogs and in cats, respectively. The most common ectoparasiticide used in dogs was a combination of imidacloprid-permethrin (33.4 %), while in cats it was imidacloprid (26.3 %) followed by fipronil (25.4 %). The most used treatment schedule against internal and external parasites in dogs and cats was an administration every three months and the main reason to do it was as a prophylactic purpose. Conclusions: The majority of Portuguese owners that attended veterinarian clinics use endoparasiticides and ectoparasiticides in/on their pets as a prophylactic measure, although in many cases not in the correct schedule of treatment. In addition, most of them are not aware of the possible transmission of parasites from their dogs and cats to themselves, a fact which highlights the important role of veterinarians in the continuous implementation of effective control measures to reduce the risk of parasitic infections in both humans and companion animals. [ABSTRACT FROM AUTHOR]

Characterization of water quality from indoor swimming pools, using chorine-based disinfection techniques, was performed during a 6-mo period to study the occurrence, distribution, and concentration factors of trihalomethanes (THM). Several parameters such as levels of water THM, water and air chloroform, water bromodichloromethane (BDCM), water dibromochloromethane (DBCM), water bromoform (BF), free residual chlorine (FrCl), pH, water and air temperature, and permanganate water oxidizability (PWO) were determined in each pool during that period. Chloroform (CFW) was the THM detected at higher concentrations in all pools, followed by BDCM, DBCM, and BF detected at 99, 34, and 6% of the samples, respectively. Water THM concentrations ranged from 10.1 to 155 μg/L, with 6.5% of the samples presenting values above 100 μg/L (parametric value established in Portuguese law DL 306/2007). In this study, air chloroform (CFAir) concentrations ranged from 45 to 373 μg/m3 with 24% of the samples presenting values above 136 μg/m3 (considered high exposure value). Several significant correlations were observed between total THM and other parameters, namely, CFW, CFAir, FrCl, water temperature (TW), and PWO. These correlations indicate that FrCl, TW and PWO are parameters that influence THM formation. The exposure criterion established for water THM enabled the inclusion of 67% of Lisbon pools in the high exposure group, which reinforces the need for an improvement in pool water quality. [ABSTRACT FROM PUBLISHER]

Polychaete annelids are some of the most heavily harvested invertebrates collected in coastal areas and estuaries for their economic value as live fishing bait or as food supplement in aquaculture activities. However, information on bait digging in Portugal is scarce or incomplete. Thus, the present study aimed at investigating the bait digging activity in the four most relevant brackish water systems in Portugal: Ria de Aveiro and Ria Formosa coastal lagoons and Tagus and Sado estuaries. Direct observations were performed in each brackish water system 1 h and a half before and after the diurnal low spring tide (<0.8 m height), on working days. Additionally, logbooks were delivered to bait diggers and two types of surveys were conducted in every system: one to all the intertidal users and the other only to bait diggers. The estimated daily weighted number of bait diggers varied between 31 in the Tagus estuary and 69 in the Ria Formosa coastal lagoon. Diopatra neapolitana (Delle Chiaje, 1841) was the main target species in Ria de Aveiro (more than 83%), Tagus estuary (more than 60%) and Ria Formosa (more than 90%), while in Sado estuary most of the bait diggers targeted Marphysa sp. (92.6%). The highest and lowest amount of polychaete annelid catches were estimated for Ria de Aveiro coastal lagoon (almost 50 tons per year) and Tagus estuary (11 tons per year), respectively. A total annual bait catch value in these four systems was estimated as €3.84 millions. The most important brackish water systems concerning bait diggers' number and bait captures were Sado estuary and Ria de Aveiro coastal lagoon where management measures should be considered, although managing polychaete harvesting activity is still difficult due to limited information on harvesting and on direct and indirect impacts on population dynamics. • Information regarding the harvest of polychaetes in Portugal is scarce. • Surveys and bait diggers' observations were performed in four brackish water systems. • Preferential areas and target species (D. neapolitana & Marphysa sp.) were determined. • A total annual bait catch value in the four systems was estimated as €3.84 millions. • The establishment of rotational closed areas was proposed as a management measure. [ABSTRACT FROM AUTHOR]

Ultramafic areas are critical for nickel (Ni) phytomining due to the high concentration of this element in their soils and the number of hyperaccumulators they harbor. The aim of the present study was to evaluate the potential of the Morais massif, an ultramafic area in Portugal, for phytomining using the hyperaccumulator species Alyssum serpyllifolium subsp. lusitanicum. Soil samples and A. serpyllifolium specimens were collected in four locations of the Morais massif. After determination of Ni concentrations in the samples, the results show that soil pseudo-total Ni concentrations in sites number 1 and 2 are significantly higher than in the soil samples collected in the other two locations, with 1918 and 2092 mg kg−1, respectively. Nickel accumulation is significantly greater in the aerial parts of plants collected at sites 1, 2, and 4, presenting Ni harvestable amount means of 88.36, 93.80, and 95.56 mg per plant, respectively. These results suggest that the sites with highest potential for phytomining are sites 1, 2, and 4. A nickel agromining system in these locations could represent an additional source of income to local farmers, since ultramafic soils have low productivity for agriculture and crop production. [ABSTRACT FROM AUTHOR]

The early diagnosis of familial hypercholesterolaemia is associated with a significant reduction in cardiovascular disease (CVD) risk. While the recent use of statistical and machine learning algorithms has shown promising results in comparison with traditional clinical criteria, when applied to screening of potential FH cases in large cohorts, most studies in this field are developed using a single cohort of patients, which may hamper the application of such algorithms to other populations. In the current study, a logistic regression (LR) based algorithm was developed combining observations from three different national FH cohorts, from Portugal, Brazil and Sweden. Independent samples from these cohorts were then used to test the model, as well as an external dataset from Italy. The area under the receiver operating characteristics (AUROC) and precision-recall (AUPRC) curves was used to assess the discriminatory ability among the different samples. Comparisons between the LR model and Dutch Lipid Clinic Network (DLCN) clinical criteria were performed by means of McNemar tests, and by the calculation of several operating characteristics. AUROC and AUPRC values were generally higher for all testing sets when compared to the training set. Compared with DLCN criteria, a significantly higher number of correctly classified observations were identified for the Brazilian (p < 0.01), Swedish (p < 0.01), and Italian testing sets (p < 0.01). Higher accuracy (Acc), G mean and F 1 score values were also observed for all testing sets. Compared to DLCN criteria, the LR model revealed improved ability to correctly classify observations, and was able to retain a similar number of FH cases, with less false positive retention. Generalization of the LR model was very good across all testing samples, suggesting it can be an effective screening tool if applied to different populations. [Display omitted] • Early diagnosis of familial hypercholesterolemia is associated with a significant reduction in cardiovascular disease risk. • The development of a multi-cohort classification model can allow for better generalization of results. • Compared to traditional clinical criteria, accuracy was higher with the developed classification model. • Furthermore, sensitivity is not compromised with this model. [ABSTRACT FROM AUTHOR]

Infectious diseases with epizootic consequences have not been fully studied in marine mammals. Presently, the unprecedented depth of sequencing, made available by high-throughput approaches, allows detailed comparisons of the microbiome in health and disease. This is the first report of the striped dolphin microbiome in different body sites. Samples from one striped female edematous dolphin were acquired from a variety of body niches, including the blowhole, oral cavity, oral mucosa, tongue, stomach, intestines and genital mucosa. Detailed 16S rRNA analysis of over half a million sequences identified 235 OTUs. Beta diversity analyses indicated that microbial communities vary in structure and cluster by sample origin. Pathogenic, Gram-negative, facultative and obligate anaerobic taxa were significantly detected, including Cetobacterium, Fusobacterium and Ureaplasma . Phocoenobacter and Arcobacter dominated the oral-type samples, while Cardiobacteriaceae and Vibrio were associated with the blowhole and Photobacterium were abundant in the gut. We report for the first time the association of Epulopiscium with a marine mammal gut. The striped dolphin microbiota shows variation in structure and diversity according to the organ type. The high dominance of Gram-negative anaerobic pathogens evidences a cetacean microbiome affected by human-related bacteria. [ABSTRACT FROM AUTHOR]

Abstract: Do nematode and macrofauna assemblages provide similar ecological assessment information? To answer this question, in the summer of 2006, subtidal soft-bottom assemblages were sampled and environmental parameters were measured at seven stations covering the entire salinity gradient of the Mondego estuary. Principal components analysis (PCA) was performed on the environmental parameters, thus establishing different estuarine stretches. The ecological status of each community was determined by applying the Maturity Index and the Index of Trophic Diversity to the nematode data and the Benthic Assessment Tool to the macrofaunal data. Overall, the results indicated that the answer to the initial question is not straightforward. The fact that nematode and macrofauna have provided different responses regarding environmental status may be partially explained by local differentiation in microhabitat conditions, given by distinct sampling locations within each estuarine stretch and by different response-to-stress times of each benthic community. Therefore, our study suggests that both assemblages should be used in marine pollution monitoring programs. [Copyright &y& Elsevier]

The objective of this analytical and interventional prospective quantitative study was to assess the effect of an educational intervention performed by nurses for mental adjustment to chronic disease in patients with hypertension. A convenience sample was studied, composed of 329 participants with chronic hypertension, followed in a primary healthcare unit in the Central Region of Portugal. Data collection was carried out by applying the Mental Adjustment to Disease Scale (MADS) before and 1 month after the educational nursing intervention between September 2017 and February 2018. Prior to the application of the educational intervention, 43.5% of the participants were classified as "unadjusted" in at least one of the subscales of MADS. After the educational intervention, 21.3% of the participants classified as "unadjusted" became "adjusted" in all MADS subscales. The success rate of the intervention varied from 26.9% (in the fatalism subscale) to 44.6% (for the anxious concern subscale). Participants were more likely to be mentally "unadjusted" to hypertension if they lived with other family members, had an active professional situation before the diagnosis of hypertension, still had an active professional situation now, were under 65 years old, had a shorter time to diagnosis (1-2 years), and measured blood pressure less regularly. The educational intervention performed by nurses is relevant for the mental adjustment of hypertensive patients, contributing to increased knowledge, as well as improvement in preventive and self-care practices, facilitating the experience of the health/disease transition process.

Background: The current standard of treatment in primary biliary cholangitis (PBC) is ursodeoxycholic acid (UDCA), although a considerable proportion of patients show incomplete response resulting in disease progression., Objective: This study aimed to assess the prevalence of incomplete response to UDCA and determine associated patients' characteristics., Methods: Patients with PBC as main diagnosis were included from a national multicentric patient registry-Liver.pt. Main endpoints included incomplete response to UDCA treatment according to Barcelona, Paris I and Paris II criteria, Globe and UK PBC scores and the association between baseline characteristics and incomplete response according to Paris II criteria., Results: A total of 434 PBC patients were identified, with a mean age of 55 years and 89.2% females. Nearly half of patients were asymptomatic at diagnosis and 93.2% had positive anti-mitochondrial antibodies. Almost all patients (95.6%) had been prescribed at least one drug for PBC treatment. At the last follow-up visit, 93.3% were under treatment of which 99.8% received UDCA. Incomplete response to UDCA was observed in 30.7%, 35.3%, 53.7% and 36.4% of patients according to Barcelona, Paris I, Paris II criteria and Globe score, respectively. After adjusting for age and sex, and accordingly to Paris II criteria, the risk for incomplete biochemical response was 25% higher for patients with cirrhosis at diagnosis (odds ratio [OR] = 1.25; 95% confidence interval [95%CI]: 1.02-1.54; p = 0.033) and 35% (95%CI:1.06-1.72; p = 0.016) and 5% (OR = 1.05; 95%CI:1.01-1.10; p = 0.013) for those with elevated gamma-glutamyl transferase (GGT) and alkaline phosphatase (ALP)., Conclusion: A considerable proportion of patients showed incomplete biochemical response to UDCA treatment according to Paris II criteria. Cirrhosis, elevated GGT and ALP at diagnosis were identified as associated risk factors for incomplete response. Early identification of patients at risk of incomplete response could improve treatment care and guide clinical decision to a more careful patient monitorization., (© 2021 The Authors. United European Gastroenterology Journal published by Wiley Periodicals LLC. on behalf of United European Gastroenterology.)

Introduction: Nonalcoholic fatty liver disease (NAFLD) is the liver manifestation of adiposopathy. Recently, a new score was developed to estimate body fat percentage (relative fat mass, RFM). We aimed to evaluate the value of RFM in predicting the presence and severity of NAFLD, compared with other anthropometric measurements., Methods: RFM, body mass index (BMI), and other anthropometric measurements were evaluated in two cohorts of subjects: a cohort from a Portuguese prospective epidemiological study (e&#95;Cor) and morbidly obese patients with biopsy-proven NAFLD. We evaluated if RFM and BMI were related with the presence and severity of liver disease, which was assessed by noninvasive tools in the first cohort and by liver histology in the morbidly obese cohort. The independence of relations found in univariate analysis was assessed with multivariable logistic regression analysis., Results: In the general population cohort, 744 subjects (48% male) were enrolled. BMI-defined obesity was present in 23% and RFM-defined obesity in 86%. Insulin resistance (IR) related with BMI-defined obesity (OR 4.37 [2.16-8.84]) and weight (OR 1.05 [1.02-1.08]) in men, and waist circumference (WC) (OR 1.07 [1.03-1.11]) in women. Dyslipidemia and hypertension related with RFM-defined obesity in men (OR 2.96 [1.36-6.47] and OR 5.37 [1.31-22.06], respectively). Ultrasound-diagnosed NAFLD in 33% related with weight in men (OR 1.03 [1.003-1.06] and WC in women (OR 1.06 [1.02-1.10]). In men, ALT elevation related with weight (OR 1.04 [1.02-1.07]). In women, advanced fibrosis (estimated by NAFLD Fibrosis Score) associated with BMI-defined obesity (OR 42.43 [3.61-498.13]). In the morbidly obese cohort, 152 subjects were enrolled, of whom 84% were female, 37% had steatohepatitis, and 9.4% had advanced fibrosis. Adiponectin associated inversely and leptin positively with RFM in men. The severity of steatosis increased linearly with BMI and WC in women. Higher BMI associated with steatohepatitis in women and advanced fibrosis in men., Conclusion: RFM-defined obesity better predicted dyslipidemia and hypertension (though not IR) and adipokine imbalance; however, it did not add value to BMI-defined obesity in predicting NAFLD or liver injury.

Introduction and aims: Nonalcoholic fatty liver disease (NAFLD) has become highly prevalent, paralleling the pandemic of obesity and diabetes, and represents an important burden. Nutrition knowledge is fundamental, in prevention, evolution and treatment of NAFLD. Association of low serum levels of vitamin D (VD) with several diseases, including NAFLD, has been emphasized in the last decade. We evaluated how serum levels of VD correlate with the presence of hepatic steatosis, and VD intake, in a random sample of the Portuguese adult population. Methods: Participants underwent a dietary intake inquiry, using a semi-quantitative food frequency questionnaire representative of the usual intake over the previous year. Anthropometric measures, blood tests and ultrasound were done. Hepatic steatosis was quantified according to Hamaguchi's ultrasonographic score (steatosis defined by a score ≥ 2). Results: We recruited 789 adult individuals, 416 males (52.7%), mean age of 49.9 ± 17.0 years (18-79). Prevalence of hepatic steatosis was 35.5%, and after exclusion of excessive alcohol consumption, 28.0%. Mean VD serum levels were 26.0 ± 9.8 ng/ml and 68.4% participants had serum VD levels below 30 ng/ml. Mean serum levels of VD were not significantly different between participants with steatosis vs . no steatosis: 25.2±8.7 vs . 26.4±10.3 ng/ml, respectively (p=0.071). There was no correlation between VD serum levels and VD intake, measured by the FFQ, r=0.075 (p= 0.383). Conclusions: In spite of a high prevalence rate, there was no evidence that decreased VD serum levels were associated with hepatic steatosis. No significant correlation was found between VD dietary ingestion and VD serum levels., Competing Interests: Competing Interests: A Carvalho, received fees on consultancy, from Intercept and H Cortez-Pinto received fees on consultancy from Genfit, Intercept and Gilead. The other authors declare that they have no conflict of interest.

Background Amd Aims: APOB mutations are a rare cause of familial hypercholesterolaemia (FH) and, until recently, routine genetic diagnosis only included the study of two small APOB fragments. In previous years, 5 novel functional mutations have been described in APOB fragments not routinely studied, our group having functionally characterized 2 of them. The main aim of this work was to identify and characterize novel alterations in APOB to assess the genetic cause of hypercholesterolemia in patients with a clinical diagnosis of FH., Methods: We performed next generation sequencing of 48 Portuguese clinical FH patients, who were apparently mutation negative. All variants found in APOB were annotated. For functional studies, LDL from index patients and relatives was separated and marked with FITC-LDL for flow cytometry assays in lymphocytes and U937 growth assays., Results: A total of 11 potential pathogenic variants were identified. Variants p.(Pro994Leu) and p.(Thr3826Met) in exons 19 and 26 were found in 4 patients, and in vitro analysis was performed for these variants. An exon 26 alteration (p.(Thr3826Met)) showed a decrease in binding and internalization of LDL, and in U937 growth assays that was similar to the effect with p.(Arg3527Gln). An alteration in exon 19 had a neutral effect., Conclusions: The spectrum of functional alterations in APOB outside the fragments routinely screened is slowly growing. Screening of all 29 exons of APOB is advised for FH routine diagnosis, but functional characterization is necessary for pathogenicity assessment. It is expected that the number of patients with functional APOB mutations will increase in the near future., (Copyright © 2018. Published by Elsevier B.V.)

BackgroundObesity is often associated with iron deficiency in children and adolescents. We aimed to study the effect of an 8-month physical exercise (PE) intervention on hepcidin and other markers of inflammation and on iron status in overweight/obese children and adolescents.MethodsSeventy-three overweight/obese children and adolescents participated in the 8-month-long longitudinal study. They were divided into two groups according to their participation in an after-school PE program: the PE group (n=44) and the control group (n=29). Hepcidin, interleukin (IL)-6, C-reactive protein (CRP), iron, ferritin, transferrin, and soluble transferrin receptor (sTfR) were evaluated.ResultsAt baseline, IL-6 correlated positively with hepcidin and negatively with iron and transferrin saturation, suggesting that increasing adiposity associates with increasing IL-6 and hepcidin synthesis, reducing iron availability. After 8 months, the PE group showed a decrease in BMI z-score (P=0.003), body fat mass (P=0.012), CRP (P=0.002), IL-6 (P=0.048), ferritin (P=0.013), hepcidin (P=0.040), and sTfR (P=0.010), and an increase in iron concentration (P=0.002). Moreover, the PE group, when compared with the control group, showed lower weight (P=0.026), BMI (P=0.040), waist circumference (P=0.010), and waist-to-height ratio (P=0.046).ConclusionWe showed that an 8-month-long intervention at school allowed a reduction in BMI z-score and an improvement in inflammation, reducing hepcidin levels and the disturbances in iron status.

Purpose: Familial hypercholesterolemia (FH) is one of the most common monogenic disorders, and the high concentrations of low-density lipoprotein (LDL) cholesterol presented since birth confers on these patients an increased cardiovascular risk. More than 1,600 alterations have been described in the LDL receptor gene (LDLR), but a large number need to be validated as mutations causing disease to establish a diagnosis of FH. This study aims to characterize, both at the phenotypic and genotypic levels, families with a clinical diagnosis of FH and present evidence for the importance of the integration of clinical, molecular, and functional data for the correct diagnosis of patients with FH., Methods: A detailed analysis of the phenotype and genotype presented by 55 families with 13 different alterations in the LDLR was conducted. For eight of these, an extensive functional characterization was performed by flow cytometry, confocal microscopy, and reverse transcriptase polymerase chain reaction., Results: Carriers of neutral alterations presented a significantly lower incidence of premature cardiovascular disease, lower levels of atherogenic lipoproteins and a large number of these individuals had LDL-cholesterol values below the 75(th) percentile. presented a significantly lower incidence of premature cardiovascular disease, lower levels of atherogenic lipoproteins and a large number of these individuals had LDL-cholesterol values below the 75th percentile However, the functional study was essential to determine the pathogenicity of variants., Conclusion: The data collected illustrate the importance of this integrated analysis for the correct assessment of patients with FH who can otherwise be misdiagnosed.

Background & Aims: Liver steatosis measurement by controlled attenuation parameter (CAP) is a non-invasive method for diagnosing steatosis, based on transient elastography. Its usefulness as screening procedure for hepatic steatosis in general population has not been previously evaluated. The aim of this study was to evaluate the diagnostic accuracy of CAP and fatty liver index (FLI) for detection and quantification of steatosis in general population., Methods: Recruitment was done from a prospective epidemiological study of the general adult population. Steatosis was evaluated using CAP, FLI and ultrasound (US). Steatosis scored according to Hamaguchi's US scoring, from 0 (S0) to 6 (S6) points. Hepatic steatosis defined by score ≥2 (S≥2) and moderate/severe steatosis by score ≥4 (S≥4). Performance of CAP and FLI for diagnosing steatosis compared with US was assessed using areas under receiver operating characteristic curves (AUROC)., Results: From 219 consecutive individuals studied, 13 (5.9%) excluded because of failure/unreliable liver stiffness measurements. Steatosis prevalence: S≥2 38.4% and S≥4 12.1%. CAP significantly correlated with steatosis (ρ = 0.73, P < 0.0001), steatosis score (ρ = 0.76; P < 0.0001), FLI (ρ = 0.69), waist circumference (ρ = 0.62), body mass index (ρ = 0.55), triglyceride (ρ = 0.49), HOMA-IR (ρ = 0.26), alcohol consumption (ρ = 0.24) and cholesterol (ρ = 0.19), not with liver stiffness measurements. Using CAP and FLI, AUROC's were 0.94 (95% CI 0.91-0.97, P < 0.001) and 0.91 for S≥2; 0.95 (95% CI 0.90-0.99, P < 0.001) and 0.93 for S≥4 respectively. Optimal cut-off value of CAP and FLI were 243 dB/m and 48 for S≥2; 303.5 dB/m and 62 for S≥4 respectively., Conclusion: Controlled attenuation parameter and FLI seem promising tools for screening and steatosis quantification in the general population. Larger studies are needed for validation., (© 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Introduction: Occult hepatitis B virus (HBV) infection, defined as the presence of HBV DNA in the liver (with detectable or undetectable HBV DNA in the serum), has been reported in patients with chronic hepatitis C. Some data suggest its association with a more severe liver disease and a worse response to interferon therapy in this subgroup of patients. However, the clinical significance of this condition is still under debate., Aim: To determine the prevalence of occult HBV infection and its clinical significance in patients with chronic hepatitis C liver disease., Materials and Methods: A prospective analysis of consecutive outpatients with chronic hepatitis C who underwent a liver biopsy recruited between January 2008 and June 2011 was carried out. Data included patient's sex and age, source of hepatitis C virus (HCV) infection, HCV genotype and viral load, presence of serologic markers of previous HBV infection, HBV DNA presence in the liver, histologic findings, and response to interferon and ribavirin treatment. HBV DNA and HCV RNA detection were carried out using a sensitive commercially available PCR kit. HBV DNA was tested in liver samples using a nested PCR procedure., Results: One hundred patients were included, 73% men, mean age 49 ± 11.9 years. Most patients had a genotype 1, with a high viral load, HCV infection. Of the patients, 33% had HBV serologic markers of past infection. The presence of HBV DNA in liver samples was found in 57% of the patients. No statistically significant difference in the epidemiological, histological, or virological or response to therapy data was found in patients with occult HBV infection., Conclusion: Occult HBV infection occurred in a high percentage of patients but was not clinically significant.

Borrelia and Rickettsia bacteria are the most important tick-borne agents causing disease in Portugal. Identification and characterization of these circulating agents, mainly in recreational areas, is crucial for the development of preventive measures in response to the gradually increasing exposure of humans to tick vectors. A total of 677 questing ticks including Dermacentor marginatus, Rhipicephalus sanguineus, Ixodes ricinus, Hyalomma lusitanicum, H. marginatum, and Haemaphysalis punctata were collected in a Safari Park in Alentejo, Portugal, to investigate the prevalences of infection and characterize Borrelia and Rickettsia species. From a total of 371 ticks tested by PCR for Borrelia burgdorferi sensu lato (s.l.), of which 247 were tested for Rickettsia, an infection prevalence of 18.3% was found for B. lusitaniae and 55.1% for Rickettsia spp. Sequence analysis of positive amplicons identified the presence of B. lusitaniae (18.3%), R. monacensis strain IRS3 (51.7%), and R. helvetica (48.3%) in I. ricinus. R. slovaca (41.5%), R. raoultii (58.5%), and also B. lusitaniae (21%) were identified in D. marginatus ticks. One (5.9%) H. lusitanicum was infected with B. lusitaniae, and R. massiliae was found in one Rhipicephalus sanguineus. Coinfection was found in 7 (20%) I. ricinus and 34 (23.3%) D. marginatus ticks. We report, for the first time, simultaneous infection with R. helvetica and B. lusitaniae and also R. slovaca, the agent of TIBOLA/DEBONEL, with B. lusitaniae. Additionally, 6 isolates of B. lusitaniae were established, and isolates of Rickettsia were also obtained for the detected species using tick macerates cultured in mammalian and mosquito cell lines. This report describes the detection and isolation of tick-borne agents from a Portuguese Safari Park, highlighting the increased likelihood of infection with multiple agents to potential visitors or staff., (Copyright © 2010 Elsevier GmbH. All rights reserved.)

Abstract We conducted a four-year (2003-2006) retrospective study of yeasts recovered in a hospital laboratory in the centre of Portugal to evaluate the epidemiology of yeast infections. Clinical isolates and data were gathered from 751 patients corresponding to 906 episodes of yeast infection. The isolates were first identified using classical and commercial methods, routinely employed at the hospital laboratory. We then re-identified the same isolates using RFLP of the ITS 5.8S rRNA gene and sequence of the D1/D2 domain of the 26S rRNA gene. Candida parapsilosis isolates were re-identified using the Ban I digestion of the SADH gene. C. albicans was the most frequently isolated of the yeasts found in the analysed specimens, with an overall incidence of 69.6% and then in decreasing order, C. glabrata, C. tropicalis, C. parapsilosis and C. krusei. C. parapsilosis was most frequently recovered from younger patients, decreasing with age, while C. glabrata occurrence increased with age. We found an increased number of cases of fungemia per 100,000 people per year, reaching a maximum of 4.4 during 2006.