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Shearing during the latter half of pregnancy is a common practice to improve flock health and productivity. Previous studies have demonstrated that shearing pregnant ewes at mid or late pregnancy is associated with an increase in lamb birth weight. In the present study, we used singleton Polypay × Dorset pregnant sheep, to investigate the potential roles of placental function and changes in maternal metabolism in underlying this increased birth weight response. Two groups were randomly established and blocked at enrollment by animal BW, body condition score and subcutaneous adipose tissue depth. The groups were shorn (SH; n = 18) or not (C; n = 20) at gestational day (GD) 107 ± 1 (mean ± SEM). Weekly maternal plasma samples were collected between shearing and birth, but only six samples were assayed for progesterone, pregnancy-associated glycoproteins (PAG1), glucose and non-esterified fatty acids (NEFAs). At birth, sex, birth weight, and newborn body mass index (BMI) were recorded. Maternal BW during mid- to late-pregnancy was similar between groups. Shearing resulted in increased lamb birth weight and BMI (P < 0.05) regardless of fetal sex but did not affect the maternal concentration of PAG1 or progesterone from GDs 100 to 142. After shearing (GD100) and up to lambing, shorn females had higher circulating glucose concentrations (P < 0.05), but not NEFA, compared to the control group. Maternal circulating PAG1, progesterone, glucose or NEFA concentration across pregnancy did not differ according to lamb sex. Across pregnancy, birth weight was positively associated with PAG1 (P < 0.001), but not with progesterone concentrations. In conclusion, weight and BMI at birth were higher in both sexes upon shearing in singleton pregnancies. Despite PAG1 being associated with birth weight, late-pregnancy shearing did not alter the placental endocrine response. Whether other placental factors are altered upon shearing and may influence the increase in birth weight and BMI remain to be investigated.
In 2005, the Portuguese government launched a primary care (PC) reform. After a promising start, the reform is still incomplete and has been compromised by low investment. The incomplete nature of the reforms has resulted in the coexistence of different models of care delivery and heterogeneity in resource allocation and performance. PC has been extensively evaluated, but little is known about the patients’ views and preferences regarding PC and the ongoing reform.
This study aims to examine patients’ experiences of and preferences for PC in Portugal and to explore their experience of the recent reforms.
A qualitative study was undertaken which collected data from eight focus groups in the city of Braga, Portugal. Participants were recruited with the collaboration of eight local institutions. Focus groups’ discussions focused on patients’ experiences of and preferences for PC as well as their views on the reforms. Audio recordings were transcribed and analysed using an inductive thematic content analysis.
The majority of participants perceived that the reform was positive. However, the improvements achieved by the reform were insufficient to lead to most participants having a positive experience of PC delivery in Portugal. Participants’ satisfaction/dissatisfaction with primary care was strongly associated with interpersonal relations and communication with doctors. Participants valued continuity of care, but felt the levels of responsiveness, flexibility and coordination in the current system were still unsatisfactory. Access and waiting times were seen as challenging and led participants to seek PC from emergency departments and private doctors.
The perception of increased inequity and the lack of effective choice undermined the social acceptability of the reform. Policies aimed at improving doctor–patient communication and continuity of care, as well as choice, may therefore lead to better satisfaction and more efficient use of health care settings.
We studied the variability of germination, dormancy and viability loss of Hirschfeldia incana seeds in relation to seed size. Seeds were stored at 35°C under humid [75% relative humidity (RH)] or dry (33% RH) conditions. Seed germination and electrolyte leakage were evaluated periodically. Small seeds had lower longevity at humid or dry storage conditions (5 or 407 days, respectively) than large or intermediate seeds (7–9 or 536–727 days, respectively). Moreover, H. incana shows variability in seed dormancy related to seed size within a population, with small seeds having lower dormancy (13%) than intermediate (50%) or large seeds (72%). Dormancy was partially released after a short storage at 35°C and humid conditions. Under dry storage conditions, endogenous dormancy cycles were observed for over a year, and longer times of storage had a dormancy-breaking effect through dry after-ripening. Results suggest a dual strategy producing non-dormant seeds with low longevity that will germinate immediately after dispersal, and seeds with greater longevity that will delay germination. Membrane permeability increased linearly with ageing at both humid and dry storage (R2 = 0.60). Small seeds showed greater conductivity than intermediate or large seeds (0.7, 0.4 or 0.3 mS g–1 dry weight, respectively, at the 80% germination). The conductivity test could be used to evaluate the quality of H. incana seeds and would allow us to identify dormant (non-germinating) seed lots as viable. However, the influence of storage conditions and variability within a seed population on seed longevity should be taken into account when evaluating seed quality.
The present study examined the association between high-quality diet (using the Mediterranean diet (MD) as an example) and well-being cross-sectionally and prospectively in Spanish children and adolescents. Participants included 533 children and 987 adolescents at baseline and 527 children and 798 adolescents at 2-year follow-up, included in the UP&DOWN study (follow-up in schoolchildren and adolescents with and without Down’s syndrome). The present study excluded participants with Down’s syndrome. Adherence to an MD was assessed using the KIDMED index. Well-being was measured using the Positive and Negative Affect Schedule and the KIDSCREEN-10 questionnaire. Associations between MD adherence and well-being were assessed using multi-level, mixed-effects linear regression. At baseline, MD adherence was positively related to health-related quality of life in secondary school girls and boys (β=0·41, se 0·10, P<0·001; β=0·46, se 0·10, P<0·001, respectively) and to positive affect in secondary school girls and boys (β=0·16, se 0·05, P=0·006; β=0·20, se 0·05, P<0·001, respectively) and in primary school boys (β=0·20, se 0·08, P=0·019). At 2-year follow-up, MD adherence was negatively related to negative affect in secondary school adolescent girls and boys (β=–0·15, se 0·07, P=0·047; β=–0·16, se 0·06, P=0·019, respectively), and MD adherence was associated with higher positive affect scores in secondary school girls (β=0·30, se 0·06, P<0·001) and in primary school boys (β=0·20, se 0·09, P=0·023). However, MD adherence at baseline did not predict well-being indicators at 2-year follow-up. In conclusion, higher MD adherence was found to behave as a protective factor for positive well-being in cross-sectional analysis.
This paper tests the factorial structure of a questionnaire comprising seven health care rationing criteria (waiting time, ‘rule of rescue’, parenthood of minors, health maximization, youngest first, positive and negative version of social merit) and explores the adherence to them of 254 Portuguese health care professionals, when considered individually and when confronted with two-in-two combinations. Data were collected through a self-administered questionnaire where respondents faced hypothetical rationing dilemmas comprising one rationing criterion and dichotomous options pairs with two rationing criteria. Confirmatory factor analysis and multinomial logistic regressions were used to validate the structure of the questionnaire and the data. The findings suggest that: (i) the hepta-factorial structure of the questionnaire presented a good fit of the data; and (ii) support for rationing criterion depends on whether they are individually considered or confronted in dichotomous options pairs. When only one criterion distinguishes the patients, healthcare professionals support six criteria (by descending order): waiting time, rule of rescue, health maximization, penalization of patients’ risky behaviors, youngest first and being parent of a young child. When two criteria were confronted, immediate threat of life/health and large expected benefits were the most preferred. Conversely, the positive version of social merit was an unappreciated rationing criterion.
Highlighting the dispersal ecology of parasites is important for understanding epidemiological, demographic and coevolutionary aspects of host–parasite interactions. Yet, critical aspects of the dispersal stage of parasites, such as longevity and the factors influencing it, are poorly known. Here we study the lifespan of the dispersal stage of an ectoparasitic dipteran, Carnus hemapterus, and the impact of gender, body size and food provisioning on longevity. We found that freshly emerged imagoes survive at most less than 4 days. Longevity increased with body size and, since this parasite exhibits sexual size dimorphism, the bigger females lived longer than males. However, controlling for body size suggests that males lived relatively longer than females. Furthermore, a humid environment and food provisioning (flowers) significantly increased individual life spans. We discuss the relative importance of spatial and temporal dispersal in relation to the infectious potential of this parasite.
Host range and parasite specificity determine key epidemiological, ecological and evolutionary aspects of host–parasite interactions. Parasites are usually classified as generalists or specialists based on the number of hosts they feed on. Yet, the requirements of the various stages of a parasite may influence the suitability of a given host species. Here, we investigate the generalist nature of three common ectoparasites (the dipteran Carnus hemapterus and two species of louse flies, Pseudolynchia canariensis and Ornithophila metallica), exploiting two avian host species (the European roller Coracias garrulus and the Rock pigeon Columba livia), that frequently occupy the same breeding sites. We explore the prevalence and abundance of both the infective and the puparial stages of the ectoparasites in both host species. Strong preferences of Pseudolynchia canariensis for pigeons and of Carnus hemapterus for rollers were found. Moderate prevalence of Ornithophila metallica was found in rollers but this louse fly avoided pigeons. In some cases, the infestation patterns observed for imagoes and puparia were consistent whereas in other cases host preferences inferred from imagoes differed from the ones suggested by puparia. We propose that the adult stages of these ectoparasites are more specialist than reported and that the requirements of non-infective stages can restrict the effective host range of some parasites.
The aim of this study was to optimize protocols for electroporation (EP) and polyfection (PLF) using polyethyleneimine (PEI) for pig sperm transfection and to determine which method was the most efficient. For EP standardization, different voltages, amounts and times of electric pulses were tested using propidium iodide (PI) as reporter. For PLF standardization, different concentrations of fluorescein isothiocyanate (FITC)-labelled PEI (PEI/FITC) were incubated with sperm for different periods of time. Flow cytometry was performed to evaluate the best protocol in terms of cell viability, including cytoplasmic membrane, acrosome, chromatin integrities and mitochondrial potential using the FITC probe, PI, acridine orange (AO) and JC1. Transfections with the plasmid pmhyGENIE-5 were carried out under optimum conditions for each procedure (EP: 500 volts, 500 μs and two pulses; PLF: PEI 0.5 mg/ml and incubation time 10 min). Transfection efficacy was assessed by fluorescence in situ hybridization (FISH). A lower transfection rate was observed for sperm in the control group (17.8%) compared with EP (36.7%), with PLF (76.8%) being the most efficient. These results suggest that the EP and PEI could be an efficient and low cost transfection method for swine sperm. Notably, treated cells showed higher plasmatic the membrane damage (PMD) and/or acrosome damage (AD) indexes, therefore the combination of this procedure with biotechniques that facilitate fecundation (i.e. in vitro fertilization or intracytoplasmic sperm injection) or even inclusion of antioxidant or anti-apoptotic drugs to improve spermatozoa viability would be important.
Shortly after the implementation of comprehensive chromosome screening (CCS) techniques for preimplantation genetic testing for aneuploidies (PGT-A), the discussion about the transition from day 3 to blastocyst stage biopsy was initiated. Trophectoderm biopsy with CCS is meant to overcome the limitations of cleavage-stage biopsy and single-cell analysis. The aim of this study was to assess the results obtained in our PGT-A programme after the implementation of this new strategy. Comparisons between the results obtained in 179 PGT-A cycles with day 3 biopsy (D+3) and fresh embryo transfer, and 204 cycles with trophectoderm biopsy and deferred (frozen–thawed) embryo transfer were established. Fewer embryos were biopsied and a higher euploidy rate was observed in the trophectoderm biopsy group. No differences in implantation (50.3% vs. 61.4%) and clinical pregnancy rate per transfer (56.1% vs. 65.3%) were found. Although the mean number of euploid embryos per cycle did not differ between groups (1.5 ± 1.7 vs. 1.7 ± 1.8), the final number of euploid blastocysts available for transfer per cycle was significantly higher in the trophectoderm biopsy group (1.1 ± 1.3 vs. 1.7 ± 1.8). This factor led to an increased cumulative live birth rate in this last group (34.1% vs. 44.6%). Although both strategies can offer good results, trophectoderm biopsy offers a more robust diagnosis and the intervention is less harmful for the embryos so more euploid blastocysts are finally available for transfer and/or vitrification.
Differentiation of niche by means of resource partitioning facilitates coexistence of species with similar requirements. Here we analyse the association between different habitats (i.e. nest types) and two Diptera species of the poorly known Family Carnidae that coexist during their larval and pupal stage in the nests of troglodytic bird species. We also describe for the first time the puparium of Hemeromyia anthracina and Hemeromyia longirostris and offer morphometric data of the puparia of these two species and of Carnus hemapterus. Both the smaller size and the occurrence of well-developed spiracles allow easy discrimination of the puparium of C. hemapterus. The puparia of both Hemeromyia species is very similar and only differ in the distance between the small spiracles. Hemeromyia anthracina and C. hemapterus coexisted in nest boxes but the former species did not occur in natural sandy cavities where, in turn, C. hemapterus was highly prevalent. Carnus hemapterus prevalence did not differ between nest boxes and natural cavities but its abundance was higher in the first type of nest. This study shows clear associations of the two dipteran species with specific types of nests. Yet, some conditions are seemingly acceptable for both species.
The Unified Health System (SUS) is the Brazilian set of public health services that offers global access to health care and disease treatments for all citizens. These services have been evaluated by means of a national survey assessing the users’ perceptions.
To explore and characterize the SUS users’ perceptions regarding primary dental team practices in the five Brazilian geographical regions.
Descriptive study. The sample consisted of 37 262 subjects. Data were collected by means of the Ministry of Health survey, conducted between 2012 and 2014. Variables used in the present study are associated with SUS users’ perspectives of satisfaction, access, and use of services. The study utilized bivariate data analysis, and dichotomous variables were derived for analysis following 95% reliability.
This study observed similarities and proportionality of perceptions in the Brazilian territory. In most macro-regions, dental teams did not develop an active search for dental treatment absentees. However, the SUS users reported very good and good perceptions, which were homogeneously distributed across five Brazilian regions, thereby showing an overall positive perception of primary dental treatment.
This research study aimed to evaluate the use of the milk leukocyte differential (MLD) to: (a) identify quarter milks that are culture-positive; and (b) characterize the milk leukocyte responses to specific groups of pathogens causing subclinical mastitis. The MLD measures the absolute number and relative percentage of inflammatory cells in milk samples. Using the MLD in two dairy herds (170 and 172 lactating cows, respectively), we studied all lactating cows with a most recent monthly Dairy Herd Improvement Association somatic cell count (SCC) >200 × 103 cells/ml. Quarter milk samples from 78 cows meeting study criteria were analysed by MLD and aseptically collected milk samples were subjected to microbiological culture (MC). Based upon automated instrument evaluation of the number and percentage of inflammatory cells in milk, samples were designated as either MLD-positive or – negative for subclinicial mastitis. Positive MC were obtained from 102/156 (65·4%) of MLD-positive milk samples, and 28/135 (20·7%) of MLD-negative milk samples were MC-positive. When MC was considered the gold standard for mastitis diagnosis, the calculated diagnostic Se of the MLD was 65·4% (IC95% = 57·4 to 72·8%) and the Sp was 79·3% (IC95% = 71·4 to 85·7%). Quarter milks positive on MC had higher absolute numbers of neutrophils, lymphocytes and macrophages, with higher neutrophils% and lymphocytes% but lower macrophages%. The Log10 (N/L) ratios were the most useful ratio to differentiate specific subclinical mastitis quarters from healthy quarters. Use of the MLD on cows with monthly composite SCC > 200 × 103 cells/ml for screening at quarter level identified quarters more likely to be culture-positive.
In conclusion, the MLD can provide an analysis of mammary quarter status more detailed than provided by SCC alone; however, the MLD response to subclinical mastitis was not found useful to specifically identify the causative pathogen.
The goals of this article are: (i) to understand how individual characteristics affect the likelihood of patients defaulting their pulmonary tuberculosis (PTB) treatment regimens; (ii) to quantify the predictive capacity of these risk factors; and (iii) to quantify and map spatial variation in the risk of defaulting. We used logistic regression models and generalized additive models with a spatial component to determine the odds of default across continental Portugal. We focused on new PTB cases, diagnosed between 2000 and 2013, and included some individual information (sex, age, residence area, alcohol abuse, intravenous drug use, homelessness, HIV, imprisonment status). We found that the global default rate was 4·88%, higher in individuals with well-known risk profiles (males, immigrants, HIV positive, homeless, prisoners, alcohol and drug users). Of specific epidemiological interest was that our geographical analysis found that Portugal's main urban areas (the two biggest cities) and one tourist region have higher default rates compared to the rest of the country, after adjusting for the previously mentioneded risk factors. The challenge of treatment defaulting, either due to other individual non-measured characteristics, healthcare system failure or patient recalcitrance requires further analysis in the spatio-temporal domain. Our findings suggest the presence of significant within-country variation in the risk of defaulting that cannot be explained by these classical individual risk factors alone. The methods we advocate are simple to implement and could easily be applied to other diseases.
In this paper the characterization of a gypsum plaster sample from the end of the 19th century simulating imperial red porphyry using a multi-analytical approach is presented and discussed. The results of X-ray diffraction (XRD), thermogravimetric and differential thermal analysis (TGA-DTA), physical and mechanical properties are summarized. In order to have further insight into the microstructure, polarized light microscopy (PLM), scanning electron microscopy coupled with energy dispersive X-ray spectrometer (SEM-EDS), and micro Raman spectroscopy analyzes were also made. They helped to clarify the main issues raised by the other complementary analytical techniques and allowed the establishment of interrelations between the different properties, providing important information about the materials, the skills, and the technological development involved in the art of imitating noble stones with gypsum pastes. This study also contributes to our knowledge concerning the preservation of these types of elements that are important in the context of European decorative arts and rarely reported in the literature.
Bisphenol-A (BPA) is a widely used endocrine-disrupting chemical. Prenatal exposure to BPA is known to affect birth weight, but its impact on the cardiovascular system has not been studied in detail. In this study, we investigated the effects of prenatal BPA treatment and its interaction with postnatal overfeeding on the cardiovascular system. Pregnant sheep were given daily subcutaneous injections of corn oil (control) or BPA (0.5 mg/kg/day in corn oil) from day 30 to day 90 of gestation. A subset of female offspring of these dams were overfed to increase body weight to ~30% over that of normal fed controls. Cardiovascular function was assessed using non-invasive echocardiography and cuff blood pressure (BP) monitoring at 21 months of age. Ventricular tissue was analyzed for gene expression of cardiac markers of hypertrophy and collagen at the end of the observation period. Prenatal BPA exposure had no significant effect on BP or morphometric measures. However, it increased atrial natriuretic peptide gene expression in the ventricles and reduced collagen expression in the right ventricle. Overfeeding produced a marked increase in body weight and BP. There were compensatory increases in left ventricular area and internal diameter. Prenatal BPA treatment produced a significant increase in interventricular septal thickness when animals were overfed. However, it appeared to block the increase in BP and left ventricular area caused by overfeeding. Taken together, these results suggest that prenatal BPA produces intrinsic changes in the heart that are capable of modulating morphological and functional parameters when animals become obese in later life.
The aim of this study was to evaluate the effect of nerolidol free (N-F) and nerolidol-loaded in nanospheres (N-NS) on the hepatic antioxidant/oxidant status of mice experimentally infected by Trypanosoma evansi. In the liver it was measured: reactive oxygen species (ROS), thiobarbituric reactive acid substances (TBARS) and non-protein thiols (NPSH), catalase (CAT), superoxide dismutase (SOD) and glutathione-S-transferase (GST) and performed histopathological examination. In addition, seric levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured. Liver samples from mice infected by T. evansi showed increased (P < 0·05) ROS, TBARS, AST and ALT levels and SOD activity, and decreased NPSH levels and CAT activity (P < 0·05) compared with uninfected animals. N-NS treatment prevented (P < 0·05) ROS and TBARS increase, and increased NPSH levels, and ameliorate CAT and SOD activities on liver of infected mice. Moreover, N-NS treatment reduced (P < 0·05) AST and ALT levels, and prevented histopathological changes caused by the parasite. N-NS protected the liver from the oxidative stress caused by T. evansi, which might be due to its antioxidant properties. Nerolidol might be considered a promising therapeutic agent against oxidative stress, and nanotechnology is an encouraging approach to be explored.
While vitrification has become the method of choice for preservation of human oocytes and embryos, cryopreservation of complex tissues and of large yolk-containing cells, remains largely unsuccessful. One critical step in such instances is appropriate permeation while avoiding potentially toxic concentrations of cryoprotectants. Permeation of water and small non-charged solutes, such as those used as cryoprotectants, occurs largely through membrane channel proteins termed aquaporins (AQPs). Substitution of a Thr by an Ala residue in the pore-forming motif of the zebrafish (Dario rerio) Aqp3b paralog resulted in a mutant (DrAqp3b-T85A) that when expressed in Xenopus or porcine oocytes increased their permeability to ethylene glycol at pH 7.5 and 8.5. The main objective of this study was to test whether ectopic expression of DrAqp3b-T85A also conferred higher resistance to cryoinjury. For this, DrAqp3b-T85A + eGFP (reporter) cRNA, or eGFP cRNA alone, was microinjected into in vivo fertilized 1-cell mouse zygotes. Following culture to the 2-cell stage, appropriate membrane expression of DrAqp3b-T85A was confirmed by immunofluorescence microscopy using a primary specific antibody directed against the C-terminus of DrAqp3b. Microinjected 2-cell embryos were then cryopreserved using a fast-freezing rate and low concentration (1.5 M) of ethylene glycol in order to highlight any benefits from DrAqp3b-T85A expression. Notably, post-thaw survival rates were higher (P<0.05) for T85A–eGFP-injected than for -uninjected or eGFP-injected embryos (73±7.3 vs. 28±7.3 or 14±6.7, respectively). We propose that ectopic expression of mutant AQPs may provide an avenue to improve cryopreservation results of large cells and tissues in which current vitrification protocols yield low survival.
Within the COST action EMBOS (European Marine Biodiversity Observatory System) the degree and variation of the diversity and densities of soft-bottom communities from the lower intertidal or the shallow subtidal was measured at 28 marine sites along the European coastline (Baltic, Atlantic, Mediterranean) using jointly agreed and harmonized protocols, tools and indicators. The hypothesis tested was that the diversity for all taxonomic groups would decrease with increasing latitude. The EMBOS system delivered accurate and comparable data on the diversity and densities of the soft sediment macrozoobenthic community over a large-scale gradient along the European coastline. In contrast to general biogeographic theory, species diversity showed no linear relationship with latitude, yet a bell-shaped relation was found. The diversity and densities of benthos were mostly positively correlated with environmental factors such as temperature, salinity, mud and organic matter content in sediment, or wave height, and related with location characteristics such as system type (lagoons, estuaries, open coast) or stratum (intertidal, subtidal). For some relationships, a maximum (e.g. temperature from 15–20°C; mud content of sediment around 40%) or bimodal curve (e.g. salinity) was found. In lagoons the densities were twice higher than in other locations, and at open coasts the diversity was much lower than in other locations. We conclude that latitudinal trends and regional differences in diversity and densities are strongly influenced by, i.e. merely the result of, particular sets and ranges of environmental factors and location characteristics specific to certain areas, such as the Baltic, with typical salinity clines (favouring insects) and the Mediterranean, with higher temperatures (favouring crustaceans). Therefore, eventual trends with latitude are primarily indirect and so can be overcome by local variation of environmental factors.