Morphological and molecular data on Phyllodistomum (Digenea: Gorgoderidae) from Brazil, with the description of a new species parasitizing Hoplias malabaricus (Bloch, 1794) (Osteichthyes, Erythrinidae).

Phyllodistomum pepirense n. sp. is described from the urinary bladder of Hoplias malabaricus (Bloch, 1794), sampled in the Jacaré-Pepira River in São Paulo state, Brazil. The isolates of the new species were recovered as a monophyletic group in the phylogenetic analysis of the 28S rRNA gene, which showed the new species as the sister taxa of Phyllodistomum virmantasi Pinacho-Pinacho, Sereno-Uribe, Hernández-Orts, García-Varela & Pérez-Ponce de León, 2021, a species sampled from an eleotrid fish in Southeastern Mexico. The new species differs morphologically from P. virmantasi by having a larger body size, slightly lobed testes and ovary, a mostly intercaecal uterus, slightly diverticulated caeca, and vitelline masses irregularly shaped. The new species is also readily distinguished from other species of Phyllodistomum Braun, 1899 reported from freshwater fishes in Brazil - namely, P. rhamdiae Amato & Amato, 1993 and P. spatula Odhner, 1902. The new species is herein described based on morphological characteristics, molecular data from D1-D3 domains of the 28S rRNA gene, host association, and geographical distribution.

Wildfires in Europe: Burned soils require attention.

Annually, millions of hectares of land are affected by wildfires worldwide, disrupting ecosystems functioning by affecting on-site vegetation, soil, and above- and belowground biodiversity, but also triggering erosive off-site impacts such as water-bodies contamination or mudflows. Here, we present a soil erosion assessment following the 2017's wildfires at the European scale, including an analysis of vegetation recovery and soil erosion mitigation potential. Results indicate a sharp increase in soil losses with 19.4 million Mg additional erosion in the first post-fire year when compared to unburned conditions. Over five years, 44 million Mg additional soil losses were estimated, and 46% of the burned area presented no signs of full recovery. Post-fire mitigation could attenuate these impacts by 63-77%, reducing soil erosion to background levels by the 4th post-fire year. Our insights may help identifying target policies to reduce land degradation, as identified in the European Union Soil, Forest, and Biodiversity strategies.

Detecting the PEX Like Domain of Matrix Metalloproteinase-14 (MMP-14) with Therapeutic Conjugated CNTs.

Matrix metalloproteinases (MMPs) are essential proteins acting directly in the breakdown of the extra cellular matrix and so in cancer invasion and metastasis. Given its impact on tumor angiogenesis, monitoring MMP-14 provides strategic insights on cancer severity and treatment. In this work, we report a new approach to improve the electrochemical interaction of the MMP-14 with the electrode surface while preserving high specificity. This is based on the detection of the hemopexin (PEX) domain of MMP-14, which has a greater availability with a stable and low-cost commercial molecule, as a recognition element. This molecule, called NSC-405020, is specific of the PEX domain of MMP-14 within the binding pocket. Through the covalent grafting of the NSC-405020 molecule on carbon nanotubes (CNTs), we were able to detect and quantify MMP-14 using electrochemical impedance spectroscopy with a linear range of detection of 10 ng⋅mL-1 to 100 ng⋅mL-1, and LOD of 7.5 ng⋅mL-1. The specificity of the inhibitory small molecule was validated against the PEX domain of MMP-1. The inhibitor loaded CNTs system showed as a desirable candidate to become an alternative to the conventional recognition bioelements for the detection of MMP-14.

Crude extract of Jatobá leaves promotes canine osteosarcoma cell D17 proliferation.

Background and Aim: New substances for neoplasm treatment have to be carefully studied to minimize adverse effects and prevent disease progression stimulation. Jatobá is a typical tree of the Cerrado and Caatinga biome, with antifungal, antimicrobial, larvicide, antioxidant, and antiproliferative properties. This study aimed to investigate the action of the crude extract of Jatobá leaves (EBFJ) on canine osteosarcoma (CO) cells and analyze the expression of biomarkers in neoplasm progression. Materials and Methods: D17 cells were cultured and subjected to treatment with EBFJ at different concentrations (10 µg/mL; 100 µg/mL; 1000 µg/mL; 2000 µg/mL; and 5000 µg/mL) and exposure times (24 h, 48 h, and 72 h). The tetrazolium reduction assay and the immunocytochemistry technique, with anti-Bcl2, anti-p53, and anti-Ki-67 antibodies, were used to observe the effect of the extract on cell proliferation. Results: Doses of 2000 µg and 5000 µg had cell viability of 300.80% and 361.84%, respectively. The extract did not show significant cytotoxicity of samples with the control group. The confluence of cells, the number of labeled cells, and the expression of Bcl2, Ki-67, and p53 were higher in the groups treated with EBFJ, with a statistical difference from the group without treatment. Conclusion: EBFJ was not cytotoxic and had a proliferative effect on CO D17 cells. The confluence of cells, the number of labeled cells, and the expression of Bcl2, Ki-67, and p53 were higher in the groups treated with the extract.

Prediction, validation, and uncertainties of a nation-wide post-fire soil erosion risk assessment in Portugal.

Wildfires are a recurrent and increasing threat in mainland Portugal, where over 4.5 million hectares of forests and scrublands have burned over the last 38 years. These fire-affected landscapes have suffered an intensification of soil erosion processes, which can negatively affect soil carbon storage, reduce fertility and forest productivity, and can become a source of pollutants. The main objective of the present study is to produce a post-fire soil erosion risk map for the forest and shrubland areas in mainland Portugal and assess its reliability. To this end, the semi-empirical Morgan-Morgan-Finney erosion model was used to assess the potential post-fire soil erosion according to distinct burn severity and climate scenarios, and the accuracy of the predictions was verified by an uncertainty analysis and validated against independent field datasets. The proposed approach successfully allowed mapping post-fire soil erosion in Portugal and identified the areas with higher post-fire erosion risk for past and future climate extremes. The outcomes of this study comprise a set of tools to help forest managers in their decision-making for post-fire emergency stabilization, ensuring the adequate selection of areas for mitigation to minimize the economic and environmental losses caused by fire-enhanced soil erosion.

European Council of Legal Medicine (ECLM) on-site inspection forms for forensic pathology, anthropology, odontology, genetics, entomology and toxicology for forensic and medico-legal scene and corpse investigation: the Parma form.

Further to a previous publication by the European Council of Legal Medicine (ECLM) concerning on-site forensic and medico-legal scene and corpse investigation, this publication provides guidance for forensic medical specialists, pathologists and, where present, coroners' activity at a scene of death inspection and to harmonize the procedures for a correct search, detection, collection, sampling and storage of all elements which may be useful as evidence, and ensure documentation of all these steps. This ECLM's inspection form provides a checklist to be used on-site for the investigation of a corpse present at a crime or suspicious death scene. It permits the collection of all relevant data not only for the pathologist, but also for forensic anthropologists, odontologists, geneticists, entomologists and toxicologists, thus supporting a collaborative work approach. Detailed instructions for the completion of forms are provided.

Boat noise impacts early life stages in the Lusitanian toadfish: A field experiment.

Marine traffic is the most common and chronic source of ocean noise pollution. Despite the evidence of detrimental effects of noise exposure on fish, knowledge about the effects on the critical early life stages - embryos and larvae - is still scarce. Here, we take a natural habitat-based approach to examine potential impacts of boat noise exposure in early life stages in a wild fish population of the Lusitanian toadfish (Halobatrachus didactylus). In-situ experiments were carried out in the Tagus estuary, an estuary with significant commercial and recreational boat traffic. Nests with eggs were exposed to either ambient (control) or boat noise (treatment), for 1 fortnight. Eggs were photographed before being assigned to each treatment, and after exposure, to count number of eggs and/or larvae to assess survival, and sampled to study development and oxidative stress and energy metabolism-related biomarkers. Data concerns 4 sampling periods (fortnights) from 2 years. Results indicate that offspring survival did not differ between treatments, but boat noise induced a detrimental effect on embryos and larvae stress response, and on larvae development. Embryos showed reduced levels of electron transport system (ETS), an energy metabolism-related biomarker, while larvae showed higher overall stress responses, with increased levels of superoxide dismutase (SOD) and DNA damage (oxidative stress related responses), ETS, and reduced growth. With this study, we provided the first evidence of detrimental effects of boat noise exposure on fish development in the field and on stress biomarker responses. If these critical early stages are not able to compensate and/or acclimate to the noise stress later in the ontogeny, then anthropogenic noise has the potential to severely affect this and likely other marine fishes, with further consequences for populations resilience and dynamics.

Myxobolus spp. (Cnidaria: Myxobolidae) in the circulating blood of fishes from Goiás and Mato Grosso States, Brazil: case report / Myxobolus spp. (Cnidaria: Myxobolidae) no sangue circulante de peixes dos estados de Goiás e Mato Grosso, Brasil: relato de caso

Myxosporidiosis is an infectious disease caused by myxozoans of the Phylum Cnidaria, Class Myxosporea, and Order Bivalvulida, considered a common parasite in fresh and saltwater fishes that parasitize many organs, especially gills. In the present study, 49 specimens of fishes belonging to eight genera: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus, and Serrasalmus were collected and blood smears were made, fixed with absolute methanol, and stained with Giemsa 10% to survey hemoparasites. However, myxospores were found in the circulating blood of five (10.20%) fishes belonging to genus Tetragonopterus, Myleus, and Pygocentrus. Two morphological types of Myxobolus spp. were identified in all the five fish specimens analyzed. Usually, investigations on myxozoans in fish are carried out with the search for plasmodia or cysts in the fish organs and observation of the cavity of organs. Nevertheless, this study highlights the importance of also examining the blood of these animals, since these parasites can cause severe pathogenic diseases in fish. Thus, the blood analyses can proportionate preventive sanitary control for commercial fish avoiding economic loss.

A mixosporidiose é considerada uma doença infecciosa causada por mixozoários pertentences ao Filo Cnidaria, Classe Myxosporea e Ordem Bivalvulida, considerados parasitos comuns de peixes de água doce e salgada, parasitando vários órgãos, principalmente as brânquias. No presente estudo, 49 espécimes de peixes pertencentes a oito gêneros: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus and Serrasalmus foram coletados e extensões sanguíneas foram feitas com a finalidade de encontrar hemoparasitos. Entretanto, mixoesporos foram observados em cinco (10.20%) espécimes de peixes, pertencentes aos gêneros Tetragonopterus, Myleus e Pygocentrus. Dois morfotipos de Myxobolus spp. foram identificados parasitando esses peixes. Normalmente, as investigações sobre mixozoários em peixes é realizada com a procura de plasmódios ou cistos nos órgãos dos peixes e observação da cavidade de órgãos. Porém, esse trabalho ressalta a importância de se examinar também o sangue desses animais, já que esses parasitos podem ocasionar patogenias severas em peixes. Sendo assim, a análise sanguínea pode proporcionar controle sanatório preventivo para peixes comerciais e evitar perdas econômicas.

Myxobolus spp. (Cnidaria: Myxobolidae) in the circulating blood of fishes from Goiás and Mato Grosso States, Brazil: case report

Abstract Myxosporidiosis is an infectious disease caused by myxozoans of the Phylum Cnidaria, Class Myxosporea, and Order Bivalvulida, considered a common parasite in fresh and saltwater fishes that parasitize many organs, especially gills. In the present study, 49 specimens of fishes belonging to eight genera: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus, and Serrasalmus were collected and blood smears were made, fixed with absolute methanol, and stained with Giemsa 10% to survey hemoparasites. However, myxospores were found in the circulating blood of five (10.20%) fishes belonging to genus Tetragonopterus, Myleus, and Pygocentrus. Two morphological types of Myxobolus spp. were identified in all the five fish specimens analyzed. Usually, investigations on myxozoans in fish are carried out with the search for plasmodia or cysts in the fish organs and observation of the cavity of organs. Nevertheless, this study highlights the importance of also examining the blood of these animals, since these parasites can cause severe pathogenic diseases in fish. Thus, the blood analyses can proportionate preventive sanitary control for commercial fish avoiding economic loss.

Resumo A mixosporidiose é considerada uma doença infecciosa causada por mixozoários pertentences ao Filo Cnidaria, Classe Myxosporea e Ordem Bivalvulida, considerados parasitos comuns de peixes de água doce e salgada, parasitando vários órgãos, principalmente as brânquias. No presente estudo, 49 espécimes de peixes pertencentes a oito gêneros: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus and Serrasalmus foram coletados e extensões sanguíneas foram feitas com a finalidade de encontrar hemoparasitos. Entretanto, mixoesporos foram observados em cinco (10.20%) espécimes de peixes, pertencentes aos gêneros Tetragonopterus, Myleus e Pygocentrus. Dois morfotipos de Myxobolus spp. foram identificados parasitando esses peixes. Normalmente, as investigações sobre mixozoários em peixes é realizada com a procura de plasmódios ou cistos nos órgãos dos peixes e observação da cavidade de órgãos. Porém, esse trabalho ressalta a importância de se examinar também o sangue desses animais, já que esses parasitos podem ocasionar patogenias severas em peixes. Sendo assim, a análise sanguínea pode proporcionar controle sanatório preventivo para peixes comerciais e evitar perdas econômicas.

Myxobolus spp. (Cnidaria: Myxobolidae) in the circulating blood of fishes from Goiás and Mato Grosso States, Brazil: case report / Myxobolus spp. (Cnidaria: Myxobolidae) no sangue circulante de peixes dos estados de Goiás e Mato Grosso, Brasil: relato de caso

Myxosporidiosis is an infectious disease caused by myxozoans of the Phylum Cnidaria, Class Myxosporea, and Order Bivalvulida, considered a common parasite in fresh and saltwater fishes that parasitize many organs, especially gills. In the present study, 49 specimens of fishes belonging to eight genera: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus, and Serrasalmus were collected and blood smears were made, fixed with absolute methanol, and stained with Giemsa 10% to survey hemoparasites. However, myxospores were found in the circulating blood of five (10.20%) fishes belonging to genus Tetragonopterus, Myleus, and Pygocentrus. Two morphological types of Myxobolus spp. were identified in all the five fish specimens analyzed. Usually, investigations on myxozoans in fish are carried out with the search for plasmodia or cysts in the fish organs and observation of the cavity of organs. Nevertheless, this study highlights the importance of also examining the blood of these animals, since these parasites can cause severe pathogenic diseases in fish. Thus, the blood analyses can proportionate preventive sanitary control for commercial fish avoiding economic loss.

A mixosporidiose é considerada uma doença infecciosa causada por mixozoários pertentences ao Filo Cnidaria, Classe Myxosporea e Ordem Bivalvulida, considerados parasitos comuns de peixes de água doce e salgada, parasitando vários órgãos, principalmente as brânquias. No presente estudo, 49 espécimes de peixes pertencentes a oito gêneros: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus and Serrasalmus foram coletados e extensões sanguíneas foram feitas com a finalidade de encontrar hemoparasitos. Entretanto, mixoesporos foram observados em cinco (10.20%) espécimes de peixes, pertencentes aos gêneros Tetragonopterus, Myleus e Pygocentrus. Dois morfotipos de Myxobolus spp. foram identificados parasitando esses peixes. Normalmente, as investigações sobre mixozoários em peixes é realizada com a procura de plasmódios ou cistos nos órgãos dos peixes e observação da cavidade de órgãos. Porém, esse trabalho ressalta a importância de se examinar também o sangue desses animais, já que esses parasitos podem ocasionar patogenias severas em peixes. Sendo assim, a análise sanguínea pode proporcionar controle sanatório preventivo para peixes comerciais e evitar perdas econômicas.

Acute toxicity assay using mysid as an alternative test organism in the assessment of the aqueous fraction of sediment.

The evaluation of sediment quality through biological testing of the aqueous fraction can be applied in dredging situations and is associated with a small number of confounding factors. The use of test organisms that are relatively resistant to contaminants is recommended when working with complex mixtures such as sediments. In this study, the sensitivity of the mysid Mysidopsis juniae to ammonia (NH3) and metals was investigated in acute toxicity laboratory tests and the results were compared to those obtained in the traditional test with sea urchin embryos and larvae. The ability of the toxicity identification and evaluation (TIE) technique, with the use of EDTA and sodium thiosulfate, to indicate the factors responsible for adverse effects was determined. The results indicate that mysids are almost 5 times more resistant than the sea urchin to ammonia (NH3), more sensitive to Cd2+ and Ni2+ and more resistant to Zn2+ and Pb2+. With the TIE technique and with the use of EDTA as a complexing agent, a greater resolution was observed in the diagnosis of the presence of the metals Cd2+ and Ni2+, which could be applied to the aqueous fraction of the sediment.

Application of proteomic to investigate the different degrees of meat tenderness in Nellore breed.

This study describes the association between meat tenderness and abundance of soluble muscle proteins in Nellore bulls (Bos indicus) using a proteomic approach. We evaluated shear force (SF) of Longissimus thoracis muscle 24 h after slaughter and selected three experimental groups of animals with moderately tender (TE; SF = 3.9 ± 0.7 kg), moderately tough (TO; SF = 5.6 ± 0.7 kg) and very tough meat (TO+; SF = 7.9 ± 1.4 kg). Proteome was investigated by two-dimensional electrophoresis (2D-PAGE) in combination with electrospray ionization-tandem mass spectrometry (ESI-MS/MS). The metabolic proteins triosephosphate isomerase (TPI1) and phosphoglucomutase 1 (PGM1), the structural protein profilin 1 (PFN1), and cytosol aminopeptidase (LAP3) were up-regulated (P < 0.05) in the TE meat group when compared to the TO and TO+ groups. Actin structural proteins (ACTA1, ACTB, and ACTG1), the oxidative stress protein peroxiredoxin (PRDX6, PRDX2, PRDX1, and PARK7), heat shock protein isoforms, and co-chaperones (CDC37 and STIP1) were up-regulated (P < 0.05) in the TO and TO+ meat groups. In addition, we also identified proteins PFN1, LAP3, PRDX1, PRDX2, HSPD1, and ARHGDIA to be associated with beef tenderness. The results reported herein demonstrated that meat tenderness in Nellore cattle depends on the modulation and expression of a set of proteins involved in different biological pathways. SIGNIFICANCE: The manuscript entitled "Application of proteomic to investigate the different degrees of meat tenderness in Nellore breed" describes a classical proteomics work using two-dimensional gel electrophoresis (2D-PAGE), followed by mass spectrometry coupled to electrospray ionization ion trap (ESI-MS/MS) in order to understand the biochemical engineering involved in the process of meat tenderness. We evaluated shear force (SF) of Longissimus thoracis muscle samples of Nellore cattle (n = 90) and select three experimental groups of animals with moderately tender (TE; SF = 3.9 ± 0.7), moderately tough (TO; SF = 5.6 ± 0.7) and very tough meat (TO+; SF = 7.9 ± 1.4). The proteomic approach allowed observing that meat tenderness is influenced by structural proteins (ACTA1, ACTG1, ACTB, MYL1 and PFN1), co-chaperones (CDC37 and STIP1), heat shock proteins (HSP90AA1, HSP90AB1, HSPD1, HSPA1L, HSPA1A and HSPB1), regulatory protein (ARHGDIA), metabolic proteins (TPI1 and PGM1) and oxidative stress proteins (PRDX1, PRDX2, PRDX6, PARK7). Our results suggest that meat tenderness in Nellore depends on the modulation and expression of a set of proteins involved in different biological pathways.

Myxobolus spp. (Cnidaria: Myxobolidae) in the circulating blood of fishes from Goiás and Mato Grosso States, Brazil: case report.

Myxosporidiosis is an infectious disease caused by myxozoans of the Phylum Cnidaria, Class Myxosporea, and Order Bivalvulida, considered a common parasite in fresh and saltwater fishes that parasitize many organs, especially gills. In the present study, 49 specimens of fishes belonging to eight genera: Tetragonopterus, Leporinus, Myleus, Pirinampus, Rhapiodon, Pygocentrus, Ageneiosus, and Serrasalmus were collected and blood smears were made, fixed with absolute methanol, and stained with Giemsa 10% to survey hemoparasites. However, myxospores were found in the circulating blood of five (10.20%) fishes belonging to genus Tetragonopterus, Myleus, and Pygocentrus. Two morphological types of Myxobolus spp. were identified in all the five fish specimens analyzed. Usually, investigations on myxozoans in fish are carried out with the search for plasmodia or cysts in the fish organs and observation of the cavity of organs. Nevertheless, this study highlights the importance of also examining the blood of these animals, since these parasites can cause severe pathogenic diseases in fish. Thus, the blood analyses can proportionate preventive sanitary control for commercial fish avoiding economic loss.

Risk and limits in dental practice: a Portuguese approach to medical-legal evaluation and professional liability.

Patient safety and quality of healthcare delivery systems are an objective of WHO. This study aims to present and analyse Portuguese clinical data on risk and malpractice in dental practice. Data from the Forensic Dentistry Laboratory (Faculty of Medicine, University of Coimbra) was analysed, between the years of 2013 to 2018. One hundred and seven technical reports were selected, and seventy nine files were included in the iatrogenic sequelae group. Data included the analysis of the performance of dental professionals. Sequelae were divided in descending order of occurrence:1) mandibular dysfunction (53,2%)[(42)79]; 2) neurological deficit (39,2%)[(31)79]; 3) tooth loss (6,3%)[(5)79]; and 4) opening deficit (1,3%)[(1)79].Three major areas with significant expression in the field of expert evaluations were analysed: 1) orthodontic treatment (51,9%), implant rehabilitation (29,1%), and oral surgery. Given the prevalence of malpractice, the need to assess its causes and recognise standards for its prevention is necessary.

Microextraction by Packed Sorbent as a Novel Strategy for Sample Clean-Up in the Determination of Methadone and EDDP in Hair.

A microextraction by packed sorbent (MEPS) procedure for rapid concentration of methadone and its primary metabolite (EDDP) in hair samples was developed. The miniaturized approach coupled to gas chromatography with tandem mass spectrometry (GC-MS-MS) was successfully validated. Hair samples (50 mg) were incubated with 1 mL of 1 M sodium hydroxide for 45 min at 50°C, time after which the extract was neutralized by adding 100 µL of 20% formic acid. Subsequently, MEPS was applied using a M1 sorbent (4 mg; 80% C8 and 20% strong cation-exchange (SCX)), first conditioned with three 250-µL cycles of methanol and three 250-µL cycles of 2% formic acid. The extract load occurred with nine 150-µL cycles followed by a washing step involving three 50-µL cycles with 3.36% formic acid. For the elution of the analytes, six 100-µL cycles of 2.36% ammonium hydroxide in methanol were applied. The method was linear from 0.01 to 5 ng/mg, for both compounds, presenting determination coefficients greater than 0.99. Precision and accuracy were in accordance with the statements of international guidelines for method validation. This new miniaturized approach allowed obtaining recoveries ranging from 73 to 109% for methadone and 84 to 110% for EDDP, proving to be an excellent alternative to classic approaches, as well as other miniaturized procedures.

Evaluation of genotoxic potential of peptides used in nuclear medicine (PSMA -617 and -11, and ubiquicidine 29-41) using a flow-cytometric, semi-automated analysis of micronuclei frequency in cell cultures.

Assays that rely on the assessment of frequency of micronuclei are important standard techniques currently used to quantify potential genotoxic damage after exposure to chemical or physical agents, such as ionizing radiation, or in pre-clinical studies, to assessment of the genotoxic potential of drugs or its components. The experiments are usually performed using conventional microscopy, but currently the protocols are being upgraded to automated approaches based on flow cytometry protocols based on the elimination of the plasma membrane by chemical agents, allowing quantification by flow cytometry. In this work, the genotoxic potential of peptides used as components of radiopharmaceuticals (PSMA-617 and 11 and Ubiquicidine) was evaluated exposing CHO-KI cells to a wide range of concentration (0.1X and 100X the maximum allowed concentration to human adults). Incubation with PSMA-11 or UBI29-41 did not induce genotoxicity. After 24 h of incubation, PSMA-617 induced genotoxicity only in non-practical concentration (100-fold). Results corroborate the safety of the pre-drugs and the wide detection range of technique.

Secondary dengue infection in immunocompetent murine model leads to heart tissue damage.

Dengue, considered the most important arthropod-borne viral disease affecting humans, is transmitted by the bite of mosquitoes of the genus Aedes and caused by one of the four distinct serotypes of dengue virus (DENV-1, -2, -3 and -4). Infection with one of the four serotypes provides lifelong homotypic immunity. However, immunity against the heterologous serotypes is transient. As a consequence, secondary infection may lead to severer manifestations due to cross-reactivity of antibodies and T-cells. Over 500,000 people are hospitalized every year and around 2,5 million, living in endemic areas, are at risk of infection. Given the background, the development of vaccines and anti-DENV drugs is of the utmost importance, as is the characterization of an animal model for testing them. The purpose of this study was to investigate ultrastructural alterations caused by DENV secondary infection in BALB/c mice heart. To achieve our goal, six BALB/c mice were infected with DENV-1 and, 4 months later, reinfected with DENV-2. Uninfected mice were used as negative controls. Heart samples were collected and processed for ultrastructural and histopathological analysis. Our results showed edema, endothelium activation characterized by the presence of transport vesicles, free platelets in interstitium, mitochondria presenting rarefied matrix and degenerated cristae, and disorganization of muscle fibers. These results point not only to BALB/c mice susceptibility to DENV infection, but also to the fact that, although it is not an often reported occurrence, dengue can lead to heart damage. Keywords: dengue; experimental model; reinfection; BALB/c mice.

European council of legal medicine (ECLM) guidelines for the examination of suspected elder abuse.

Article 25 of the Charter of Fundamental Rights of the European Union (adopted in Nice on 7 December 2000) recognizes and respects the rights of older people to lead a life of dignity and independence and to participate in social and cultural life. It also highlights the importance of prevention and recognition of elder abuse, especially since exposure to violence is likely as the population ages, either in familial or in institutional settings. Elder abuse has some issues in common with child abuse but in spite of this fact currently is less recognized. Health professionals have a major role to play in early detection and management of cases of elder abuse. This protocol summarizes some key concepts and approaches to assist in the timely detection and investigation of elder abuse cases by healthcare professionals and forensic practitioners.

Community-based native seed production for restoration in Brazil - the role of science and policy.

Large-scale restoration programmes in the tropics require large volumes of high quality, genetically diverse and locally adapted seeds from a large number of species. However, scarcity of native seeds is a critical restriction to achieve restoration targets. In this paper, we analyse three successful community-based networks that supply native seeds and seedlings for Brazilian Amazon and Cerrado restoration projects. In addition, we propose directions to promote local participation, legal, technical and commercialisation issues for up-scaling the market of native seeds for restoration with high quality and social justice. We argue that effective community-based restoration arrangements should follow some principles: (i) seed production must be based on real market demand; (ii) non-governmental and governmental organisations have a key role in supporting local organisation, legal requirements and selling processes; (iii) local ecological knowledge and labour should be valued, enabling local communities to promote large-scale seed production; (iv) applied research can help develop appropriate techniques and solve technical issues. The case studies from Brazil and principles presented here can be useful for the up-scaling restoration ecology efforts in many other parts of the world and especially in tropical countries where improving rural community income is a strategy for biodiversity conservation and restoration.