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We are increasingly confronted with external factors that affect or promote sustainable tourism development. The pandemic triggered by COVID-19 has shown that to effectively manage uncertainty, it is essential to include perceived risk as a predictor variable. This study analyses residents′ support for sustainable tourism development in low-density territories and proposes a model that takes into account several known antecedents for this support (community attachment, community involvement, economic dependence, local government management of tourism, tourism impact) and gives relevance to the perception of risk, largely ignored in academic studies and from the residents′ perspective. A structured questionnaire was used for data collection, applied to a sample of 250 residents from Lamego-Portugal, a low-density territory. The data was analysed using the Structural Equation Modelling (SEM). The results show that perceived risk (associated with COVID) significantly affects the perceived costs of tourism, but has no direct influence on residents′ support for sustainable tourism development. This support is positively affected by community involvement, economic dependence, local government management of tourism and perceived costs. The results also show that community involvement and local government management of tourism have a positive impact on perceived benefits. The results can help tourism professionals strengthen sustainable tourism development programmes. • Community support is key for sustainable development in sparse areas. • Residents' support for STD is crucial for all stakeholders. • A social exchange theory model explores factors in residents' STD support. • Studying risk perception offers insights for sustainable tourism destinations. [ABSTRACT FROM AUTHOR]

We aimed to study whether the percentwise age distribution of RSV cases changes over time during annual epidemics. We used surveillance data (2008–2019) from the Netherlands, Lyon (France), Portugal, Singapore, Ecuador, South Africa, and New Zealand. In each country, every season was divided into "epidemic quarters", i.e. periods corresponding to each quartile of RSV cases. Multinomial logistic regression models were fitted to evaluate whether the likelihood of RSV cases being aged <1 or ≥5 years (vs. 1 to <5) changed over time within a season. In all countries, RSV cases were significantly more likely to be aged <1 year in the 4th vs. 1st epidemic quarter; the relative risk ratio [RRR] ranged between 1.35 and 2.56. Likewise, RSV cases were significantly more likely to be aged ≥5 years in the 4th vs. 1st epidemic quarter (except in Singapore); the RRR ranged from 1.75 to 6.70. The results did not change when stratifying by level of care or moving the lower cut-off to 6 months. The age profile of RSV cases shifts within a season, with infants and adolescents, adults, and the elderly constituting a higher proportion of cases in the later phases of annual epidemics. These findings may have implications for RSV prevention policies with newly approved vaccines. • We studied whether the age of RSV cases changes over time during annual epidemics. • We used surveillance data (2008–2019) from seven countries around the world. • RSV cases become more likely to be aged <1 or >5 years as the epidemics unfolds. • The trend is largely consistent across countries and robust to sensitivity analyses. [ABSTRACT FROM AUTHOR]

Background and Aims: Little is known about differences of cardiometabolic risk factors (CMRF) and the function of Framingham Risk Score (FRS) within severe obesity, thus we aimed to study not only CMRF and FRS, but to determine significant differences between BMI ranges within severe obesity.Methods and Results: In this baseline analysis of the Traditional Brazilian Diet (DieTBra) Trial, several CMRF were assessed in 150 adult patients in two BMI ranges: 35.0-44.9 kg/m2 (n = 76) and ≥45 kg/m2 (n = 74). Body composition was evaluated by multifrequency bioelectrical impedance analysis to measure the percent of body fat, visceral fat area and waist circumference. Pearson's Chi-squared, Fisher's Exact, Student's t-test, and Mann-Whitney's test were used in the statistical analysis with a 5% significance level. Hypertension, C-reactive protein, systolic and diastolic blood pressure and positive family history for heart diseases were more prevalent in BMI ≥45.0 kg/m2 (p < 0.05). Mean values of waist circumference, body fat %, visceral fat area, and systolic blood pressure were significantly higher in patients with BMI ≥45.0 kg/m2. Regarding the function of FRS, 40.0% of the patients were at high risk. No differences were found for diabetes, lifestyle, lipid parameters, and FRS within different BMI ranges, except for dyslipidemia, significantly higher among participants with BMI 35.0-44.9 kg/m2.Conclusion: BMI >45 kg/m2 was associated with higher prevalence of hypertension, systolic and diastolic blood pressure, C-reactive protein, waist circumference, body fat % and family history of heart diseases, enhancing the risk for the occurrence of cardiovascular diseases. [ABSTRACT FROM AUTHOR]

The present study evaluated the ecological risk of metal contamination in sediments of Guanabara Bay (GB) by combining multiple lines of evidence (LOEs). Chemical analysis and a set of whole-sediment toxicity assays were conducted with Tiburonella viscana , Kalliapseudes schubartii , Anomalocardia flexuosa , and Nitocra sp. Results were integrated by multivariate analysis and qualitative methods. Additionally, a whole-sediment Toxicity Identification Evaluation technique (TIE) was applied to identify the chemical groups responsible for the effects. Sediments from harbor and industrial areas exhibited toxicity linked to moderate to high concentrations of Zn, Pb, Cu and Cr. The TIE technique confirmed such effects, but it also indicates the contribution of ammonia and organic compounds to the observed toxicity. Our results demonstrate that the combination of multiple LOEs improves the effectiveness of environmental risk assessment of chemical stressors and management of coastal ecosystems in tropical regions. Unlabelled Image • Ecological risk assessment of sediment contamination in a tropical bay • Multiple endpoints evaluated in whole-sediment toxicity testing • Bioavailable fraction of metals associated with toxic effects • The whole-sediment toxicity identification evaluation (TIE) was assessed. • A qualitative analysis integrating geochemical and ecotoxicological outcomes is provided. [ABSTRACT FROM AUTHOR]

The Rodrigo de Freitas Lagoon (RFL - Rio de Janeiro, RJ, Brazil) is a highly polluted and eutrophic lacustrine system, which has been often used for the practice of aquatic sports, including during the 2016 Summer Olympic Games. This study proposes the evaluation of metal concentrations in surface sediments from the RFL before and after urban interventions performed for the 2016 Olympics, as well as their toxicity to the benthic amphipod Tiburonella viscana and embryos of the sea-urchin Echinometra lucunter. Metal concentrations determined in 2017 were significantly higher than those obtained in 2015 (especially Cu, Cd and Ni), suggesting that the interventions performed to fulfill the requirements of the Olympics increased metal contents in sediments. The sediments from the northern sector of the RFL were muddier, more organically enriched, exhibited higher metal concentrations and were more toxic to T. viscana when compared to the sediments collected in the southern sector. This fact is particularly important since the practice of sports, including during the 2016 Olympics, has been preferably performed in the northern sector. Metal distribution was strongly correlated with organic matter and mud contents. The toxicity to E. lucunter embryos was high for both northern and southern sediments; most of the samples led to 100% lack or abnormal embryonic development. The integration of physical, chemical and ecotoxicological data indicates that the mortality to T. viscana was correlated with metal contents, whereas the toxicity to E. lucunter was apparently related to the release of ammonia from the sediment to water column. Finally, high metal concentrations and the toxicity to aquatic organisms evidence the ecological risks to the biota from RFL. Image 1 • Sediments from Rodrigo de Freitas Lagoon are highly metal polluted. • Metal concentrations in sediments were higher after urban interventions for the 2016 Olympic Games. • Sediments were toxic to amphipods and embryos of sea urchin. High metal concentrations and the toxicity to aquatic organisms evidence the ecological risks to the biota from Rodrigo de Freitas Lagoon (Rio de Janeiro – Brazil). [ABSTRACT FROM AUTHOR]

Abstract Severe obesity and diabetes lead to a significant decrease in quality of life. Although controversial, population-wide studies have implicated obesity in the development of tuberculosis (TB). Non-classical monocytes have been described in obesity and TB, whereas in diabetes they have been associated with poorer clinical outcomes. The present study focuses on the functional significance of several monocyte populations of obese, obesity-related diabetic (OBDM), non-obese/diabetic tuberculosis and non-obese healthy control patients. Monocytes were evaluated by measuring expression of CD86, CD206, TLR-2 and TLR-4 as well as production of IL-6, IL-12, and by using a mycobacterial growth inhibition assay for both Mycobacterium tuberculosis and M. abscessus subsp. massiliense. Non-classical monocytes from OBDM and non-obese TB patients exhibited similar activation profiles (CD86/CD206/TLR-2 and TLR-4 expressions). Only monocytes from TB patients had a higher positivity for IL-12 and IL-6, whereas adiponectin serum levels increased similarly between TB and OBDM patients. Monocytes from active TB patients and OBDM were more permissive to Mtb growth than obese individuals, but this susceptibility was not observed for M. abscessus subsp. massiliense. From these findings, we conclude that diabetes and tuberculosis had similarities in the population of circulating non-classical monocytes, improving our understanding of the association of these diseases. [ABSTRACT FROM AUTHOR]

Abstract We present an alternative approach for establishing in situ bioaccumulation assessment of methyl-Hg (MeHg), total-Hg, Cd, Pb and Zn in bivalve Anomalocardia brasiliana from four bays of Rio de Janeiro presenting varying degrees of eutrophication, acid volatile sulfides (2–55 μmol g−1), simultaneously extracted metals (SEM) and total metals (TM) in sediments. Using metal concentrations of composite samples from three size classes of bivalve and their incorporation rates (IR = metal concentration / total length), which depend on exposure time, we calculated asymptotic IR and respective consequent metal concentrations. Both IR and the metal concentration presented inverse relationships with total length (excepting MeHg) and bay contamination. Lead and zinc concentrations were above Brazilian legal criteria in the most anoxic and contaminated bay, suggesting significant metal bioavailability (SEM/TM between 8% and 63%). Highlights • An alternative to ex situ bioassays in lab to bioaccumulation assessment of metals. • We estimated the metals incorporation rate using only three sizes of A. brasiliana. • Metal incorporation rates during asymptotic growth up decrease with size/age. • The results were consistent with geochemistry processes from four bays. • The approach is reliable to predict potential risk for human consumption. [ABSTRACT FROM AUTHOR]

Monocytes are mononuclear phagocytes in peripheral blood that can differentiate into macrophages and dendritic cells. Macrophages play a specific role in the inflammatory process and are essential for the innate response. Given the important role of monocytes/macrophages in the immune response, this study aimed to evaluate the activity of kojic acid (KA), a natural product of certain fungal species, on human peripheral blood monocytes in vitro . Purified monocytes isolated from human blood were incubated with KA (50 μg/mL for 48 h) and analyzed by light microscopy, scanning electron microscopy, transmission electron microscopy and flow cytometry. Host cell cytotoxicity was measured by the colorimetric MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. KA treatment induced morphological alterations in monocytes, such as increased cell size, as well as numerous cellular projections. Furthermore, flow cytometry revealed increased labeling of cell surface EMR1-F4/80 but decreased labeling of CD11b and CD14. KA also promoted increased IL-6 cytokine production but did not cause cytotoxic effects in monocytes. In conclusion, our results show that KA promotes the differentiation of monocytes into macrophages and can act as an immunomodulatory agent. [ABSTRACT FROM AUTHOR]

Summary Obesity is recognised as a serious public health issue, due to its associated morbidity and mortality. This study aimed at estimating the prevalence of overweight and obesity in the Portuguese population through direct measurements obtained by the 1st National Health Examination Survey (INSEF 2015) and to identify its associated sociodemographic factors. INSEF was a nationally representative cross-sectional prevalence study conducted on 4911 Portuguese adults aged 25–74 years, in 2015. Height and weight were measured according to the European Health Examination Survey procedures. Poisson regression was used to estimate the adjusted prevalence ratios of overweight and obesity according to age, marital status, occupational activity, education, urbanization of living area and smoking status. Overall prevalences of overweight and obesity were 39.1% and 28.6%, respectively. The prevalence of overweight was higher among men (45.5% vs 33.2%) while the prevalence of obesity was higher among women (32% vs 25%). The sociodemographic factors associated with overweight and obesity were age, marital status and education. Smoking status was associated with overweight and obesity but only in women. INSEF suggests that a high prevalences of overweight and obesity are found in older individuals, married, with lower education levels and non-smoking women. Public health interventions are urgently required for obesity prevention, namely throughout health literacy strategies. [ABSTRACT FROM AUTHOR]

A sediment quality assessment was performed near to the main industrial source of metal contamination in Sepetiba Bay, Brazil, which represents one of the worst cases of trace metal contamination reported for coastal areas. Acute and chronic toxicity tests, benthic fauna community analysis and metal bioavailability evaluations were applied to identify risks to the benthic community. Significant amphipod mortality was observed close to the major pollution source and lower copepod fertility was observed for all stations. Equilibrium-partitioning and biotic-ligand models to predict pore water metal toxicity, which were based on acid-volatile sulfide (AVS) and organic carbon fraction ( f OC ) normalization approaches, suggested that metals are not likely to be available in sediment pore water. However, Cd, Pb and Zn concentrations were mainly (>50%) weakly bound to sediments, suggesting high potential bioavailability. Linking the chemical results with ecotoxicological responses, we observed that sediment-feeding organisms presented acute and chronic toxicities that were positively correlated to the metal concentrations in the sediments. Additionally, benthic fauna composition was dominated by tolerant species, revealing a trophic structure response to environmental contamination. These results reinforce the necessity of a multiple lines of evidence approach to establish sediment quality and to support environmental management decisions that are based on observed effects and potential extrapolation scenarios into the future. [ABSTRACT FROM AUTHOR]

Omnivorous fish utilize dietary carbohydrates better than carnivorous ones due to striking morphological and physiological differences in their digestive tracts. However, digestive tract morphology and physiology also varies among omnivorous fishes, which can lead to different dietary utilization of plant sources as well. Starch, energy, and dry matter apparent digestibility coefficient (ADC) of wheat bran, cassava residue, ground corn, and broken rice were compared between the omnivorous freshwater jundiá catfish (93.9 ± 34.0 g; mean ± standard deviation) and Nile tilapia (93.7 ± 51.6 g) by using 5 g kg − 1 chromic oxide as a diet marker. Starch ADCs were significantly higher in tilapia for all plant sources (92.02% to 99.74% versus 55.87% to 90.61%), except for wheat bran, which was similar to that found in jundiá. Starch-richer plant sources (ground corn and broken rice) showed the lowest starch digestibility for jundiá. However, ground corn provided significantly higher energy digestibility in jundiá (55.35%), as opposed to broken rice (86.59%) and ground corn (71.68%), in tilapia. Dry matter ADCs ranged from 22.89% for cassava residue in jundiá to 89.17% for broken rice in tilapia. Higher dry matter ADCs were registered in tilapia for all plant sources, except for wheat bran. Despite presenting lower starch and energy digestibilities, jundiá catfish showed an adaptive capacity to utilize starch sources by presenting significantly higher specific activity of amylase (58 U mg protein − 1 versus 29 U mg protein − 1 ) and maltase than Nile tilapia for all plant sources, except for wheat bran. Maltase activities in the gut of jundiá varied from 2.5 to 3.6 U mg protein − 1 , depending on the plant source, while in tilapia it averaged 1.7 U mg protein − 1 , regardless of the plant source. The scanning electron microscopy approach used to study the influence of fish digestion on the structure of starch granules from different plant sources revealed the presence of spherulites in the fish feces. Starch spherulite formation in jundiá intestines could be related to lower starch digestibility. Our findings demonstrate different carbohydrate digestion abilities among omnivores. [ABSTRACT FROM AUTHOR]

Abstract: Horseradish peroxidase (HRP) is a plant enzyme widely used in biotechnology, including antibody-directed enzyme prodrug therapy (ADEPT). Here, we showed that HRP is able to catalyze the autoxidation of acetylacetone in the absence of hydrogen peroxide. This autoxidation led to generation of methylglyoxal and reactive oxygen species. The production of superoxide anion was evidenced by the effect of superoxide dismutase and by the generation of oxyperoxidase during the enzyme turnover. The HRP has a high specificity for acetylacetone, since the similar β-dicarbonyls dimedon and acetoacetate were not oxidized. As this enzyme prodrug combination was highly cytotoxic for neutrophils and only requires the presence of a non-human peroxidase and acetylacetone, it might immediately be applied to research on the ADEPT techniques. The acetylacetone could be a starting point for the design of new drugs applied in HRP-related ADEPT techniques. [Copyright &y& Elsevier]

: ObjectiveTo verify the viability of isolated primordial follicles after exposure to different concentrations of dimethyl sulfoxide (DMSO) and after cryopreservation.: DesignRandomized control trial.: SettingLaboratorio Renzo Giuliani, University of Florence, Italy.: Animal(s)Thirty- to 40-day-old lambs.: Main outcome measure(s)Isolated primordial follicles were stained with trypan blue to evaluate the effect of different DMSO concentrations before and after the cryopreservation. Histological structure and follicular mortality were evaluated.: Result(s)After the isolation procedure (control), a mean (±SE) of 800 ± 203.86 live primordial follicles/mL were obtained. The number of live follicles in the toxicity test using the DMSO at 0, 0.5, 1.0, 1.5, 2.0, and 2.5 M was 782 ± 193.96, 754 ± 172.03, 778 ± 191.58, 736 ± 191.92, 476 ± 122.9, and 316 ± 83.52, respectively. The number of live follicles at 2.5 M was lower than that in the control procedure. After cryopreservation, the numbers decreased to 0 ± 0, 232 ± 44.20, 636 ± 161.82, 628 ± 181.28, 208 ± 11.57, and 184 ± 47.07, respectively at 0, 0.5, 1.0, 1.5, 2.0, and 2.5 M. The number of live follicles at 0, 0.5, 2.0, and 2.5 M were lower than that in the control procedure.: Conclusion(s)After cryopreservation, only DMSO concentrations of 1.0 and 1.5 M showed a number of live follicles similar to that of the control procedure. [Copyright &y& Elsevier]

This study proposes a pro-active approach for evaluations of methylmercury (MeHg), total mercury (THg), arsenic (As), cadmium (Cd) and lead (Pb) in situ bioaccumulation in fish (Atherinella brasiliensis) muscles, using specimens from the external sector of Guanabara Bay as a study case. This approach included an hierarchical sequence: analysis of the pollutants concentrations and their comparison to safety criteria; correlations between specimens concentrations vs length (as a proxy of exposure time); projections of concentrations in key lengths (sexual maturation, asymptotic, length limits for fishing and median of fish population) through polynomial regressions, dose-response analysis (Probit), decreasing curves and incorporation rates (using only three length intervals). The incorporation rates were ascending for MeHg and THg (continued bioaccumulation) and descending for As, Pb and Cd (possible biological dilution). The projections were satisfactory, evidencing their use for an improvement on the risks monitoring of fishing and fish consumption by humans in coastal environments. • We reveal a Probit dose-response relationship for metal bioaccumulation in fish; • We predicted the metal concentrations in fish muscles at sexual maturity, legal and asymptotic lengths; • We estimated the metals incorporation rate using only three sizes and this decreasing rate towards the asymptotic size; • The tested approaches could be useful tools for in situ monitoring of coastal ecosystems having fish species as bioindicators; • The approach is reliable to predict potential risk for human consumption. [ABSTRACT FROM AUTHOR]

In autumn 2023, European vaccination campaigns predominantly administered XBB.1.5 vaccine. In a European multicentre study, we estimated 2023 COVID-19 vaccine effectiveness (VE) against laboratory-confirmed symptomatic infection at primary care level between September 2023 and January 2024. Using a test-negative case–control design, we estimated VE in the target group for COVID-19 vaccination overall and by time since vaccination. We included 1057 cases and 4397 controls. Vaccine effectiveness was 40 % (95 % CI: 26–53 %) overall, 48 % (95 % CI: 31–61 %) among those vaccinated < 6 weeks of onset and 29 % (95 % CI: 3–49 %) at 6–14 weeks. Our results suggest that COVID-19 vaccines administered to target groups during the autumn 2023 campaigns showed clinically significant effectiveness against laboratory-confirmed, medically attended symptomatic SARS-CoV-2 infection in the 3 months following vaccination. A longer study period will allow for further variant-specific COVID-19 VE estimates, better understanding decline in VE and informing booster administration policies. [ABSTRACT FROM AUTHOR]

In vitro studies have demonstrated that GM-CSF in combination with other stimulatory factors induces a microbicidal response that control T. gondii infection. We assessed whether GM-CSF alone can control T. gondii replication in murine microglial cultures. Microglia were collected and cultured with or without GM-CSF and the half of each group was infected with T. gondii . We determined the T. gondii infectivity, cytokines levels, NO and superoxide detection. GM-CSF alone primes microglia, which after infection induces the production of TNF-α and IL-6, leading to NO and superoxide production, without any stimulus from IL-12p70 and IFN-γ. [ABSTRACT FROM AUTHOR]

The present study investigated the effect of adding allopathic doxorubicin (DOX 0.3 µg/mL), the vehicle of ultradiluted/dynamized doxorubicin (0.2 % ethanol), different dynamizations of ultradiluted/dynamized doxorubicin (DOX 6CH, DOX 12CH and DOX 30CH), both in the absence or presence of chemical stress induced by doxorubicin at 0.3 µg/mL on follicular survival and activation, antioxidant capacity of the medium, Catalase activity (CAT), production of reactive protein thiol, maintenance of type I and III collagen fibers and accumulation of lipofuscin in porcine ovarian tissue cultured in vitro for 48 hours. To do this, part of the ovarian tissue fragments was fixed for the uncultured control and the rest were cultured in: MEM (cultured control), DOX 0.3 µg/mL, Ethanol, DOX 6CH, DOX 12CH, DOX 30CH, DOX (0.3 µg/mL) + DOX 6CH, DOX (0.3 µg/mL) + DOX 12CH, DOX (0.3 µg/mL) + DOX 30CH treatments. The results showed that, in general, ultradiluted/dynamized doxorubicin (DOX 6CH, DOX 12CH and DOX 30CH) mitigated the toxic effect of allopathic doxorubicin (0.3 µg/mL) on the morphology of preantral follicles, the content of type I and III collagen fibers, and the production of lipofuscin in the tissue. However, only DOX (0.3 µg/mL) + DOX 6CH attenuated the oxidative stress induced by DOX (0.3 µg/mL), maintaining adequate CAT activity that was similar to the uncultured control. Additionally, when the three isolated ultradiluted/dynamized doxorubicin were considered, only DOX 12CH increased the reduced thiol levels compared to the uncultured control and MEM. In conclusion, supplementing the culture medium with ultradiluted/dynamized DOX (DOX 6CH, DOX 12CH and DOX 30CH) attenuated the toxicity induced by allopathic doxorubicin during the in vitro culture of pig preantral follicles enclosed in ovarian tissue. • DOX ultradiluted/dynamized attenuated the toxicity induced by allopathic doxorubicin. • After 48 h, DOX ultradiluted/dynamized improved the survival of porcine preantral follicles. • The accumulation of lipofuscin was significantly reduced. • Proportions of type I and III collagen fibers and Catalase activity were maintained adequate. [ABSTRACT FROM AUTHOR]

[Display omitted] Takotsubo Syndrome (TS) is a heart disease caused by extreme exposure of the body to physical or psychological stress. In the context of COVID-19, the virus can be a significant source of stress, with particular attention being paid to the cytokine storm as a cause of damage to the body. New research shows that the production of specific cytokines is linked to the activation of immune checkpoint proteins such as PD-1, PD-L1, and CTLA-4 on T cells. Although initially beneficial in combating infections, it can suppress defense and aid in disease progression. Therefore, checkpoint inhibitor therapy has been highlighted beyond oncological therapies, given its effectiveness in strengthening the immune system. However, this treatment can lead to excessive immune responses, inflammation, and stress on the heart, which can cause Takotsubo Syndrome in patients. Several studies investigate the direct link between this therapy and cardiac injuries in these patients, which can trigger TS. From this perspective, we must delve deeper into this treatment and consider its effects on the prognosis against SARS-CoV-2 infection. [ABSTRACT FROM AUTHOR]

Abstract: This study investigates the effects on fish and assesses human health hazards from mercury released in two gold mining areas in Indonesia: Tatelu (North Sulawesi Province) and Galangan (Katingan District, Central Kalimatan Province). In Tatelu, 154 fish specimens of 10 freshwater species were collected, as well as five marine species from the fish market. The mean concentration of total mercury in muscles of freshwater fish from this area was 0.58±0.44μg/g, with more than 45% of fish having Hg levels above the WHO guideline for human consumption of 0.5μg/g. In Galangan, where 263 fish specimens of 25 species were collected, the total mercury in muscles averaged 0.25±0.69μg/g. Excluding data from flooded open pits in sub-area P4, mean Hg levels in fish from Galangan were 2 to 4 times lower than 0.5μg/g, while fewer than 10% of fish from Galangan exceeded WHO guidelines. The Hazard Quotient (HQ) was applied to both areas to determine the threat of MeHg exposure for communities in both areas. The HQ is a risk assessment indicator which defines the ratio of exposure level to a single substance in relation to a reference dose. Samples from Tatelu (excluding marine species) had an HQ above one, while those from Galangan resulted in values of 2.4 for the whole area and 9.9 for sub-area P4, pointing to potentially harmful fish consumption for the local population. By using the single-compartment model to estimate mercury levels in blood and hair from daily intake dose, sub-area P4 showed the highest levels, higher than the upper limit guideline for pregnant women, but still lower than threshold levels associated with observed clinical effects. [Copyright &y& Elsevier]

: ObjectiveTo verify the viability of isolated primordial follicles to different propylene glycol (PROH) and glycerol (GLY) concentrations before and after cryopreservation.: DesignIsolated primordial follicles were stained with trypan blue to evaluate the effect of different PROH and GLY concentrations before and after cryopreservation.: SettingLaboratorio Renzo Giuliani, University of Florence, Italy.: Patient(s)Thirty- to forty-day-old lambs.: Intervention(s): Isolation of primordial follicles with subsequent exposure to cryoprotectant and freezing.: Main outcome measure(s)Histologic structure and follicular mortality.: Result(s)After the isolation procedure (control), the mean number of live primordial follicles/mL was 2,688 and 4,452 in the GLY and PROH groups, respectively. When GLY was used, the number of live follicles before cryopreservation was 820, 756, 640, 524, 564, and 460 follicles/mL with concentrations of 0, 0.5, 1.0, 1.5, 2.0, and 2.5 mol/L, respectively. After cryopreservation, this number decreased to 0, 12, 36, 100, 84, and 68 follicles/mL, respectively, with the same concentrations. When PROH was used, the number of live follicles before cryopreservation was 4,216, 3,880, 3,560, 1,812, 704, and 568 follicles/mL with concentrations of 0, 0.5, 1.0, 1.5, 2.0, and 2.5 mol/L, respectively. After cryopreservation, this number decreased to 0, 116, 336, 472, 360, and 244 follicles/mL, respectively, with the same concentrations.: Conclusion(s)Both cryoprotectants were shown to preserve isolated primordial follicles after cryopreservation. [Copyright &y& Elsevier]

Cryopreservation of primordial follicles represents an opportunity to preserve female gametes, and consequently to protect the reproductive capacity of humans and animals, as well as to safeguard genetic material from endangered animal species or rare breeds. The aim of this work was to assess the toxicity of different concentrations of ethylene glycol (EG) to primordial follicles, and verify the viability of these follicles after the freezing–thawing procedure. Primordial follicles were isolated from ovine ovaries and exposed to different EG concentrations to evaluate the cryoprotectant (CPA) toxicity before and after cryopreservation. After isolation of primordial follicle (control), the number (mean±S.E.M.) of viable primordial follicles/ml was 3764±795.21. The number of viable follicles in the toxicity test using EG at 0, 0.5, 1.0, 1.5, 2.0, and 2.5 M was 1096±447.9, 960±446.67, 948±366.14, 832±313.59, 856±280.67, and 700±255.02, respectively. The number of viable follicles at concentrations of 2.5 M was less than for controls. After cryopreservation, the numbers decreased to 0±0, 148±85.46, 764±246.69, 824±291.9, 844±296.27, and 588±200.65, respectively for 0, 0.5, 1.0, 1.5, 2.0, and 2.5 M EG. The number of viable follicles at 0, 0.5, and 2.5 M was less than for controls. In conclusion, after the freezing and thawing procedure, concentrations of 1.0, 1.5, and 2.0 M EG can be successfully used for the cryopreservation of isolated follicles in sheep. [Copyright &y& Elsevier]

• Melatonin did not provide protection and seems to have evidenced the toxic effect of withanolide D on primordial follicles. • Activated follicles were more resistant to toxic effects. • Withanolide D and/or melatonin could induce DNA DSBs on in situ preantral follicles or on isolated secondary follicles. Withanolide D (WD) has been investigated as an antineoplastic drug. This study aimed to evaluate whether melatonin (MT) could attenuate toxic effects on preantral follicles enclosed in the ovarian cortex (experiment 1 - E1) or on isolated secondary follicles (experiment 2 - E2) exposed to WD. For E1, ovarian cortex was incubated for 48 h to: (1) α-MEM+; (2) α-MEM+ plus 6 μM WD; (3) α-MEM+ plus 3 mmol/L MT or (4) α-MEM+ plus WD and MT. For E2, secondary follicles were exposed for until 96 h in. (1) only to basic medium (α-MEM++/α-MEM++); (2) α-MEM++ plus 3 mmol/L MT (MT/MT); (3) α-MEM++ until 48 h, followed by more 48 h in 6 μM WD (α-MEM++/WD) or (4) a pre-exposure to MT for until 48 h, followed by more 48 h of exposure to WD plus MT (MT/MT + WD). The main results obtained showed that exposure to drugs caused damage to follicular morphology (WD or WD + MT) and diameter (WD) in the ovarian cortex or in isolated follicles. In pre-antral follicles in situ , ATM expression increased in the presence of WD, MT or association. As for the secondary follicles, ATM and γH2AX were immunostained in the granulosa and theca cells and oocytes in all treatments. TAp63α was immunostained in follicles included in the ovarian cortex and in isolated follicles. We conclude that melatonin did not provide protection and could have enhanced the toxic effect of WD to follicles surrounded or not by the ovarian cortex. [ABSTRACT FROM AUTHOR]

Highlights • WD6.0 showed a toxic effect similar to PTX and higher than other treatments after 2 and 6 days. • WD6.0 reduced the cell proliferating compared to PTX or WD3.0. • Chemotherapeutic agents (PTX and WD) don't alter the expression of ABCB1. Abstract The Withanolide D is a chemotherapeutic potential against the human tumor cell. However, there is no report on the effect of this compound on ovarian function, especially on preantral folliculogenesis. The aim of this study was to evaluate the toxicity of a new candidate to anticancer drug, Withanolide D (WD) on morphologic integrity, development (activation and granulosa cell proliferation) and gene expression of ABCB1 protein of caprine preantral follicles. Ovarian fragments were cultured in vitro for 2 or 6 days in α-MEM or α-MEM added with paclitaxel (PTX −0.1 μg/mL; negative control) and different concentrations of WD (WD1.5, WD3.0 or WD6.0). The higher dose of WD showed a toxic effect similar to PTX and higher (P < 0.05) than other treatments after 2 and 6 days. In addition, WD6.0 reduced the cell proliferating compared to PTX or mild dose. The expression of ABCB1 remained unchanged in the presence of the chemotherapeutic agents (PTX and WD) throughout the culture period. In conclusion, WD exerted a toxic effect observed by decreasing follicular survival and cell proliferation, on the preantral caprine follicles similar to PTX, whose negative effect on folliculogenesis is already widely known. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the follicular morphology and development (follicular activation, cell proliferation, and hormone production), as well as the distribution pattern of Connexins 37 and 43 and SDF-1α after vitrification and in vitro culture of goat ovarian tissue. The study involved four experimental groups: fresh control , vitrified control , fresh culture and vitrified culture. The ovarian fragments were vitrified by a solid surface technique using the Ovarian Tissue Cryosystem and subsequently in vitro cultured for 7 days. The percentage of normal preantral follicles was similar between vitrified control and vitrified culture . However, both vitrified control and vitrified culture treatments showed a significant reduction of morphologically normal follicles in comparison to fresh control . A higher percentage of developing follicles (transition, primary and secondary) was observed in both fresh culture and vitrified culture treatments. Progesterone and estradiol production decreased (P < 0.05) during in vitro culture. SDF-1α and Cx37 proteins were detected in oocytes and granulosa cells from all the treatments. However, in vitrified cultured tissue, only granulosa cells were labeled with Cx37. Connexin 43 was detected in the granulosa, theca cells and zona pellucida in all the treatments. In conclusion, in vitro culture of vitrified goat ovarian cortex was able to promote follicle survival and did not alter the expression of SDF-1α and 43. However, the expression of Cx 37 was modified after in vitro culture of vitrified tissue. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate whether the addition of resveratrol to vitrification/thawing medium improves the cryotolerance of preantral follicles enclosed in bovine ovarian fragments. Ovarian fragments were obtained from bovine fetuses and distributed to the following groups: fresh ovarian fragments (control), vitrified (VIT), and vitrified with resveratrol (VIT + RESV). Overall, the mean percentage of normal follicles was greater (P < 0.05) in the VIT + RESV compared to the VIT group. Moreover, the probability of finding normal follicles was 2.5 greater (P < 0.05) in the VIT + RESV group. In class comparison, the primordial and transitional follicles have ∼3.0 times (P < 0.05) greater odds of being normal after vitrification compared to the secondary follicles. Additionally, a negative association (P < 0.05) was observed between the proportion of viable follicles and the stage of follicular development. ROS levels were similar (P > 0.05) between the VIT and VIT + RESV groups, and both were lower (P < 0.05) than the control group. The tissue viability in the VIT + RESV group was similar (P > 0.05) to the control group. In summary, the resveratrol provided greater ovarian tissue viability and has a positive effect against degeneration of preantral follicles enclosed in ovarian fragments. [ABSTRACT FROM AUTHOR]

Introduction Immune profile for influenza viruses is highly changeable over time. Serological studies can assess the prevalence of influenza, estimate the risk of infection, highlight asymptomatic infection rate and can also provide data on vaccine coverage. The aims of the study were to evaluate pre-existing cross-protection against influenza A(H3) drift viruses and to assess influenza immunity in the Portuguese population. Materials and methods We developed a cross-sectional study based on a convenience sample of 626 sera collected during June 2014, covering all age groups, both gender and all administrative health regions of Portugal. Sera antibody titers for seasonal and new A(H3) drift influenza virus were evaluated by hemagglutination inhibition assay (HI). Seroprevalence to each seasonal influenza vaccine strain virus and to the new A(H3) drift circulating strain was estimated by age group, gender and region and compared with seasonal influenza-like illness (ILI) incidence rates before and after the study period. Results Our findings suggest that seroprevalences of influenza A(H3) (39.9%; 95% CI: 36.2–43.8) and A(H1)pdm09 (29.7%; 95% CI: 26.3–33.4) antibodies were higher than for influenza B, in line with high ILI incidence rates for A(H3) followed by A(H1)pdm09, during 2013/2014 season. Low pre-existing cross-protection against new A(H3) drift viruses were observed in A(H3) seropositive individuals (46%). Both against influenza A(H1)pdm09 and A(H3) seroprotection was highest in younger than 14-years old. Protective antibodies against influenza B were highest in those older than 65 years old, especially for B/Yamagata lineage, 33.3% (95% CI: 25.7–41.9). Women showed a high seroprevalence to influenza, although without statistical significance, when compared to men. A significant decreasing trend in seroprotection from north to south regions of Portugal mainland was observed. Conclusions Our results emphasize that low seroprotection increases the risk of influenza infection in the following winter season. Seroepidemiological studies can inform policy makers on the need for vaccination and additional preventive measures. [ABSTRACT FROM AUTHOR]

In the last decade there was a significant increase dairy goat participation in the world and Brazilian agricultural scenario. Minas Gerais state is the third largest producer of goat milk in Brazil. Paratuberculosis is a chronic intestinal disease that mostly affects ruminants and can become a public health problem due to the possible association with Crohn's disease. It is transmitted by ingesting Mycobacterium avium subsp. paratuberculosis (MAP) contaminated food or water. In recent years, with the development of molecular techniques, the disease has been identified with greater precision and more quickly. MAP strains (S-Sheep, Cattle-C and B-Bison) have also been typed. The detection of a circulating MAP strain in a herd is important to develop epidemiological studies and thus establish better strategies to control paratuberculosis. This study aimed to identify and type MAP in dairy goat farms in the Zona da Mata, the main producing region of Minas Gerais state, Brazil. Feces and milk samples of 467 animals were collected, processed, inoculated in Herrold's Egg Yolk Agar (HEYM) and submitted to PCR and REA techniques. Eleven (2.36%) animals were positive for the presence of MAP in four properties and the isolates were characterized as type C strain. It was concluded that MAP is present in dairy goats from properties in Zona da Mata, MG and that strain type C circulates in the area. This is the first report of MAP typing isolated from dairy goat in Brazil. [ABSTRACT FROM AUTHOR]

Cryoinjuries caused by vitrification of tissues and organs lead to the loss of membrane proteins that mediate intercellular communications, such as connexins 37 (Cx37) and 43 (Cx43). Thus, the present study aimed to evaluate ovine Cx37 and Cx43 gene and protein expressions and developmental competence by in vitro –cultured secondary follicles retrieved from vitrified ovarian tissue. Ovarian fragments for the same ovary pair were distributed into six treatments: (1) fresh ovarian tissue (FOT); (2) vitrified ovarian tissue (VOT); (3) isolated follicles from fresh ovarian tissue (FIF); (4) isolated follicles from vitrified ovarian tissue; (5) isolated follicles from fresh ovarian tissue followed by in vitro culture (CFIF); (6) isolated follicles from vitrified ovarian tissue followed by in vitro culture (CVIF). In all treatments, Cx37 and Cx43 gene and protein expression patterns were evaluated by reverse transcription polymerase chain reaction and immunocytochemistry. In addition, secondary follicles were analyzed according to follicular integrity and growth, apoptosis, and cell proliferation. In vitro –cultured secondary follicles (CFIF and CVIF) were evaluated based on morphology (extruded follicles), antrum formation, and viability. The percentage of intact follicles was higher, whereas antrum formation, oocyte extrusion rate, and follicle viability were lower in CVIF than in CFIF treatment (P < 0.05). Terminal deoxynucleotidyl transferase–mediated biotinylated deoxyuridine triphosphates nick end-labeling assay demonstrated that apoptosis was absent in FIF, whereas follicles from all other treatments showed positive labeling. Cell proliferation index was higher in isolated follicles from vitrified ovarian tissue and CVIF treatments than in follicles from FIF. Expression of Cx43 messenger RNA was lower in CVIF treatment when compared with follicles from all other treatments (P < 0.05). Follicle Cx37 messenger RNA levels did not show alterations in any treatment (P > 0.05). Cx37 and Cx43 immunolabeling was localized mainly on granulosa cells and oocytes, respectively. In conclusion, isolation of ovine secondary follicles could be done successfully after vitrification of ovarian tissue, and the basement membrane integrity remained intact after in vitro culture. Although the gene and protein expression of Cx37 did not change after vitrification of ovarian tissue, Cx43 turned out to be altered in secondary follicles after vitrification and in vitro culture. [ABSTRACT FROM AUTHOR]

Cryopreservation of preantral follicles is a promising technique to preserve female fertility. The aim of this study was to evaluate the effect of vitrification on the development of secondary follicles included in ovarian tissue or isolated after microdissection. An important end point included is the capacity of grown oocytes to resume meiosis. Sheep ovarian cortexes were cut into fragments and split into three different groups: (1) fresh (control): secondary follicles isolated without any previous vitrification; (2) follicle-vitrification (follicle-vit): secondary follicles vitrified in isolated form; and (3) tissue-vitrification (tissue-vit): secondary follicles vitrified within fragments of ovarian tissue ( in situ former) and subsequently subjected to isolation. From the three groups, isolated secondary follicles were submitted to IVC for 18 days. After IVC, cumulus-oocyte complexes (COCs) were harvested from follicles. As an additional control group, in vivo grown, in vivo -grown COCs were collected from antral ovarian follicles. All, recovered COCs were matured and the chromatin configuration was evaluated. Data were analyzed by ANOVA, and the means were compared by Student-Newman-Keuls test, and by chi-square. Differences were considered to be significant when P < 0.05. Isolated preantral follicles from all treatments had normal morphology, antrum formation, and low follicle degeneration after IVC. The growth rate between control and follicle-vit did not differ (P > 0.05), and was higher (P < 0.05) than for tissue-vit. The percentage of follicles that decreased diameter during IVC was significantly higher in tissue-vit than the in follicle-vit. Recovery rate of oocytes from normal follicles was higher in follicle-vit than in tissue-vit. Furthermore, oocyte viability was lower in tissue-vit than other treatments, and follicle-vit did not differ from control and in vivo grown. The percentage of oocytes meiosis resuming was not different between treatments except for in vivo grown. After vitrification, only follicle-vit showed metaphase I oocyte. We conclude that secondary follicles vitrified after isolation displayed a better follicular growth rate, oocyte viability, percentage of oocytes reaching the metaphase I stage, and fewer follicles with decreased diameter after IVC. [ABSTRACT FROM AUTHOR]