Development, implementation and evaluation of a multinational FLS mentorship programme in Latin America.

Realising the benefits of systematic secondary fracture prevention requires supporting local sites to get started and becoming effective. We here describe the development, implementation and impact of a regional fracture liaison service (FLS) mentorship programme in Latin America that led to 64 FLS getting started and coverage of 17,205 patients. INTRODUCTION: Despite treatments and service models to deliver effective secondary fracture prevention, most patients are left untreated after a fragility fracture. To improve the capability to get FLS started and more effective, we describe the development, implementation and evaluation of an international programme to develop national communities of FLS mentors as part of the Capture the Fracture Partnership in Latin America. METHODS: The IOF regional team and the University of Oxford developed the curriculum and associated resources for training mentors in setting up FLS, service improvement and mentorship. Mentors were selected during a preparatory meeting, trained using live online sessions followed by regular mentor-led post-training meetings. The programme was evaluated using a pre-training needs assessment and post-training evaluation based on Moore's outcomes. RESULTS: The mentorship programme was initiated in Mexico, Brazil, Colombia and Argentina. The mentors were multidisciplinary, including orthopaedic surgery, rehabilitation, rheumatology, endocrinology, geriatrics, gynaecology and internal medicine. There was 100% participation in training sessions and reported satisfaction with the training. Since the initiation of the training programme, 22 FLS have been set up in Mexico, 30 in Brazil, 3 in Colombia and 9 in Argentina, in comparison with two in Chile and none in any other LATAM countries that were not involved in the mentorship programme. This equates to approximately 17,025 additional patients identified from 2019 to 2021 after initiation of mentorship. The mentors have engaged with 58 FLS for service development. Post-training activities include two published national best practice guidelines and other country-specific resources for FLS in the local language. CONCLUSION: Despite the COVID pandemic, the mentorship pillar of the Capture the Fracture Partnership has developed a community of FLS mentors with measurable improvement in national FLS provision. The programme is a potentially scalable platform to develop communities of mentors in other countries.

Evidence of Antineutrinos from Distant Reactors Using Pure Water at SNO.

The SNO+ Collaboration reports the first evidence of reactor antineutrinos in a Cherenkov detector. The nearest nuclear reactors are located 240 km away in Ontario, Canada. This analysis uses events with energies lower than in any previous analysis with a large water Cherenkov detector. Two analytical methods are used to distinguish reactor antineutrinos from background events in 190 days of data and yield consistent evidence for antineutrinos with a combined significance of 3.5σ.

Aberrant hippocampal transmission and behavior in mice with a stargazin mutation linked to intellectual disability.

Mutations linked to neurodevelopmental disorders, such as intellectual disability (ID), are frequently found in genes that encode for proteins of the excitatory synapse. Transmembrane AMPA receptor regulatory proteins (TARPs) are AMPA receptor auxiliary proteins that regulate crucial aspects of receptor function. Here, we investigate a mutant form of the TARP family member stargazin, described in an ID patient. Molecular dynamics analyses predicted that the ID-associated stargazin variant, V143L, weakens the overall interface of the AMPAR:stargazin complex and impairs the stability of the complex. Knock-in mice harboring the V143L stargazin mutation manifest cognitive and social deficits and hippocampal synaptic transmission defects, resembling phenotypes displayed by ID patients. In the hippocampus of stargazin V143L mice, CA1 neurons show impaired spine maturation, abnormal synaptic transmission and long-term potentiation specifically in basal dendrites, and synaptic ultrastructural alterations. These data suggest a causal role for mutated stargazin in the pathogenesis of ID and unveil a new role for stargazin in the development and function of hippocampal synapses.

Influences of environmental variables on the weight-length relationship of the shrimp Xiphopenaeus kroyeri: Do variations occur along time?

This study analyzes the influences of environmental variables on the weight-length relationship of the penaeid shrimp Xiphopenaeus kroyeri during nine consecutive years (2005 to 2013) in northern Rio de Janeiro State. The main questions raised are: i) Does species relative growth vary along time? and ii) How are the environmental variables sea surface temperature, salinity, chlorophyll-a and/or organic matter related to the relative growth? For males, the long-term mean of the allometric coefficient stayed <3 (immature: 2.49; mature: 2.91), whereas for females they were >3 (immature: 3.08; mature: 3.10). The sine-consine models highlighted the wave T period in which the allometric coefficient values complete one cycle of increase-decrease: 4.72 years and 14.72 years for immatures female and male, respectively; and 9.08 years and 9.98 years for matures male and females, respectively. The relative growth behavior varies in a predictable time scale; however, none of the environmental variables strongly supported the variation. Changes in the relative growth behavior are probably drive by intrinsic mechanisms to maintain the population locally.

External contamination of broilers by Campylobacter spp. increases from the farm to the slaughterhouse.

1. In this study, classical and molecular microbiological methods for detection and quantification of Campylobacter spp. were used to estimate their prevalence in faecal samples and skin swabs collected from 31 broiler flocks (20 farms) in Portugal, and measure the impact of transport-related factors on the expected rising excretion rates from the farm to the slaughterhouse. 2. Data on husbandry practices and transport conditions were gathered, including time in transit, distance travelled or ante-mortem plant-holding time. 3. A generalised linear mixed model was used to evaluate the significance of a potential post-transport rise in Campylobacter spp. counts and to assess risk determinants. 4. At least one flock tested positive for Campylobacter spp. in 80% of the sampled farms. At the slaughterhouse, Campylobacter spp. were detected in all faecal samples, C. jejuni being the most commonly isolated. 5. A post-transport rise of Campylobacter spp. counts from skin swabs was observed using classical microbiological methods (from a mean of 1.43 to 2.40 log10 CFU/cm2) and molecular techniques (from a mean of 2.64 to 3.31 log10 genome copies/cm2). 6. None of the husbandry practices or transport-related factors were found to be associated with Campylobacter spp. counts. 7. This study highlights the need for more research to better understand the multi-factorial nature of Campylobacter spp., a public health threat that was found to be highly prevalent in a sample of Portuguese poultry farms.

Bioconcentration and translocation of Cd and Hg in a tomato (Solanum lycopersicum) from cultivated soils in southeastern Brazil.

Food is the main source of toxic metals like mercury (Hg) and cadmium (Cd) to humans. This study evaluated the accumulation and translocation of Cd and Hg in a soil-plant system in four tomato plantations and assessed the risk associated with ingestion of these metals. Ten soil samples (rhizosphere) and 10 samples of plant tissues (leaf, ripe fruit, green fruit, and roots) were collected in each plantation. Spatial variation in Cd and Hg concentrations was negligible. The Cd level in rhizosphere was lower in comparison with that of plant tissues. Hg levels in rhizosphere were similar to roots and higher than the value observed in aerial parts of plants. The Cd bioconcentration factor was approximately five times higher compared to that of Hg. Approximately 93% and 48.6% of Cd and Hg accumulated in roots reached aerial parts, respectively. Our results indicate that tomato readily absorbs Cd accumulated in soil, translocating it to aerial parts. Comparatively, the absorption of Hg is not efficient. Levels of Cd in tomatoes were over 17 times higher than the maximum residual levels in 57.5% of ripe fruits and in 27.5% of green tomatoes.

Total Hg and methylmercury dynamics in a river-floodplain system in the Western Amazon: Influence of seasonality, organic matter and physical and chemical parameters.

Total mercury (Hg) and methylmercury (MeHg) circulation in a connected river-floodplain system composed of two black water (a small forest river, igarapé, and Cuniã Lake) and a white water body (Madeira River), located in the Madeira River Basin were evaluated during the rising-water, early and late falling-water periods. We assessed organic matter (C and N composition, (C:N)a; and δ13C isotopic signature), and physical and chemical influences (pH, dissolved O2, electric conductivity) in relationship to Hg and MeHg concentrations. Hg and MeHg concentrations in a sediment profile and three aquatic macrophytes (E. crassipes, E. azuera and Oryza sp.) were measured. Igarapé and Cuniã Lake showed higher Hg and MeHg concentrations (115-709; 10-25 ng g-1) in the suspended particulate matter compared (SPM) compared to the Madeira River (Hg: 5-16; MeHg: 0.2-0.3 ng g-1), partially independent of seasonality (p = 0.06). Total Hg had higher affinity for the SPM (1.75 times) than for dissolved organic matter. Organic matter characteristics correlated with MeHg concentrations (δ13C and (C:N)a; r2 = 0.79; p < 0.0001), as well as physical and chemical parameters of the water column (dissolved O2 and pH; r2 = 0.80; p < 0.0001), demonstrating that physical and chemical changes between the river-floodplain system affect MeHg circulation and production. The inverse correlation of MeHg and SO42- concentrations (r2 = 0.73; p < 0.0001) suggests the action of sulfate-reducing bacteria. Total Hg and MeHg concentrations as well as %MeHg were detected in the sediment profile (Hg: 24-51; MeHg: 0.6-3.2 ng g-1; %MeHg: 1.8-6.2) and aquatic macrophytes (Hg: 1-30; MeHg: 0.3-7.5 ng g-1; %MeHg: 1.6-33.7). We conclude that the highest Hg and MeHg concentrations in Cuniã Lake, compared to the Madeira River, are due to the physical and chemical differences between these environments.

Methylmercury in environmental compartments of a hydroelectric reservoir in the Western Amazon, Brazil.

Damming rivers to generate electricity creates a lentic environment that favors methylmercury (MeHg) formation. Reservoirs in the Amazon are critical environments for MeHg formation, considering its old soils and the use of Hg in gold mining in the region. The objective of this study was to evaluate MeHg accumulation in three environmental compartments (aquatic macrophytes, suspended particulate matter (SPM) and sediment) of the Samuel reservoir (Western Amazon, Brazil), during the low water, ebb and high water hydrological periods, characterizing the dry season, the end of the rainy season and the rainy season, respectively. MeHg concentrations were determined through GC-AFS. The aquatic macrophytes presented higher %MeHg in their roots (up to 12%) compared to their other tissues. This ratio was 1.7 and 5.9 times higher than those observed for SPM and the sediment, but MeHg concentrations were the lowest (0.5-4.5 ng g-1) among the three environmental compartments. Contrary, the highest MeHg concentration was observed in SPM (104 ng g-1) during the low water period. The MeHg concentration in the sediment profile decreased with increasing depth (0.93-0.48 ng g-1) and with decreasing organic matter lability (increasing C:N ratio). In the SPM, on the other hand, MeHg concentration showed a positive association with increasing C:N ratio. We conclude that MeHg dynamics in the SPM are associated with the hydrological periods, with peaks during the low water period. The organic matter lability of the sediments is more limiting to the production of MeHg than the total Hg concentrations.

Isotopic profile and mercury concentration in fish of the lower portion of the rio Paraíba do Sul watershed, southeastern Brazil

The aim of this study was to evaluate the isotopic profile and mercury (Hg) concentrations in fish (carnivores, omnivores and detritivores) in the lower portion of the rio Paraíba do Sul watershed, southeastern Brazil. Carbon (δ13C) isotopic analyses revealed that pelagic and benthonic sources are part of the feeding of the fish from the different guilds (-14.0 to -24.8 ‰). The benthic sources are usually enriched in δ13C (-16.9‰, from watershed runoff) compared to pelagic sources because the phytoplankton, important primary producer that supports several pelagic chains, has δ13C signature lighter (-23.9‰). The nitrogen (δ15N) isotopic signatures indicated that most guilds were at the same trophic position (10.0 to 15.5 ‰), except for pelagic omnivorous fish, which had a lower trophic position. Niche overlap was observed among pelagic and demersal carnivorous fish, demersal omnivorous fish, and demersal detritivorous fish. The lower isotopic niche breadth of pelagic carnivorous fish reveals the specialized resource use by this guild. Hg concentrations (ng g-1 dry weight) differed significantly between demersal carnivorous fish (185.3 dry weight; 27.8 wet weight) and demersal omnivorous fish (277.9 dry weight; 41.7 wet weight) and between pelagic omnivorous fish (197.2 dry weight; 29.6 wet weight) and demersal omnivorous fish due to (1) differences in food sources: guilds that fed on bottom resources were more affected by contamination because the sediment is an important Hg accumulator in the study area, and (2) because of its trophic positions. Considering that the fish consumed prey of similar trophic positions, the guilds did not show a well-defined food hierarchy. Therefore, in this study, there was no clear relationship between Hg and δ15N.

O objetivo deste estudo foi avaliar o perfil isotópico e as concentrações de mercúrio (Hg) em peixes (carnívoros, onívoros e detritívoros) na bacia inferior do rio Paraíba do Sul, sudeste do Brasil. As análises isotópicas de carbono (δ13C) revelaram participação de fontes pelágica e bentônica na alimentação dos peixes das diferentes guildas (-14,0 a -24,8 ‰). As fontes bentônicas são usualmente mais enriquecidas em δ13C (-16,9‰, derivado do escoamento superficial da bacia de drenagem) comparadas às fontes pelágicas pois, o fitoplâncton, importante produtor primário que suporta inúmeras cadeias pelágicas, tem assinatura de δ13C mais leve (-23,9‰). As assinaturas isotópicas de nitrogênio (δ15N) indicaram que a maioria das guildas estava no mesmo nível trófico (10,0 a 15,5 ‰), exceto os peixes onívoros pelágicos cuja posição trófica foi inferior. Observou-se sobreposição de nicho entre peixes carnívoros pelágicos e demersais, onívoros demersais, e detritívoros demersais. A menor amplitude de nicho isotópico dos peixes carnívoros pelágicos revela o uso de recursos especializados por esta guilda. A concentração de Hg (ng g-1 peso seco) diferiu significativamente entre peixes carnívoros demersais (185,3 peso seco; 27,8 peso úmido) e onívoros demersais (277,9 peso seco; 41,7 peso úmido), e entre peixes onívoros pelágicos (197,2 peso seco; 29,6 peso úmido) e onívoros demersais devido a (1) diferenças nas fontes alimentares: guildas que utilizaram recursos de fundo foram mais afetadas pela contaminação pois o sedimento é um importante acumulador de Hg na área de estudo, e (2) por causa das suas posições tróficas. Tendo em vista que os peixes consumiram presas com níveis tróficos semelhantes, as guildas não apresentaram uma hierarquia alimentar definida. Desse modo, no presente estudo não se verificou uma relação clara entre Hg e δ15N.

Analysis of new isolates reveals new genome organization and a hypervariable region in infectious myonecrosis virus (IMNV).

Infectious myonecrosis virus (IMNV) has been the cause of many losses in shrimp farming since 2002, when the first myonecrosis outbreak was reported at Brazilian's northeast coast. Two additional genomes of Brazilian IMNV isolates collected in 2009 and 2013 were sequenced and analyzed in the present study. The sequencing revealed extra 643 bp and 22 bp, at 5' and 3' ends of IMNV genome respectively, confirming that its actual size is at least 8226 bp long. Considering these additional sequences in genome extremities, ORF1 can starts at nt 470, encoding a 1708 aa polyprotein. Computational predictions reveal two stem loops and two pseudoknots in the 5' end and a putative stem loop and a slippery motif located at 3' end, indicating that these regions can be involved in the start and termination of translation. Through a careful phylogenetic analysis, a higher genetic variability among Brazilian isolates could be observed, comparing with Indonesian IMNV isolates. It was also observed that the most variable region of IMNV genome is located in the first half of ORF1, coinciding with a region which probably encodes the capsid protrusions. The results presented here are a starting point to elucidate the viral's translational regulation and the mechanisms involved in virulence.

Brain ischemia downregulates the neuroprotective GDNF-Ret signaling by a calpain-dependent mechanism in cultured hippocampal neurons.

The glial cell line-derived neurotrophic factor (GDNF) has an important role in neuronal survival through binding to the GFRα1 (GDNF family receptor alpha-1) receptor and activation of the receptor tyrosine kinase Ret. Transient brain ischemia alters the expression of the GDNF signaling machinery but whether the GDNF receptor proteins are also affected, and the functional consequences, have not been investigated. We found that excitotoxic stimulation of cultured hippocampal neurons leads to a calpain-dependent downregulation of the long isoform of Ret (Ret51), but no changes were observed for Ret9 or GFRα1 under the same conditions. Cleavage of Ret51 by calpains was selectively mediated by activation of the extrasynaptic pool of N-methyl-d-aspartate receptors and leads to the formation of a stable cleavage product. Calpain-mediated cleavage of Ret51 was also observed in hippocampal neurons subjected to transient oxygen and glucose deprivation (OGD), a model of global brain ischemia, as well as in the ischemic region in the cerebral cortex of mice exposed to transient middle cerebral artery occlusion. Although the reduction of Ret51 protein levels decreased the total GDNF-induced receptor activity (as determined by assessing total phospho-Ret51 protein levels) and their downstream signaling activity, the remaining receptors still showed an increase in phosphorylation after incubation of hippocampal neurons with GDNF. Furthermore, GDNF protected hippocampal neurons when present before, during or after OGD, and the effects under the latter conditions were more significant in neurons transfected with human Ret51. These results indicate that the loss of Ret51 in brain ischemia partially impairs the neuroprotective effects of GDNF.

Gamma oscillations in the somatosensory cortex of newborn rats.

Here we addressed a question of whether gamma oscillations previously described in the whisker-related barrel cortex are a universal pattern of activity in the somatosensory cortex of newborn rats. Intracortical recording of local field potentials and action potentials in neurons using multisite silicon electrodes in 2-7-day-old rats showed that mechanical stimulation of single fingers or specific areas on the plantar or back side of the foot evoked early gamma oscillations followed by spindle-burst oscillations in the corresponding regions of the somatosensory cortex. Early gamma oscillations had maximum amplitude in layer IV of the somatosensory cortex and effectively synchronized action potentials in layer IV neurons. It was concluded that early gamma oscillations evoked by activation of the topographic sensory input are a universal activity pattern of the entire somatosensory cortex of newborn rats.

Brain injury activates microglia that induce neural stem cell proliferation ex vivo and promote differentiation of neurosphere-derived cells into neurons and oligodendrocytes.

Brain damage, such as ischemic stroke, enhances proliferation of neural stem/progenitor cells (NSPCs) in the subventricular zone (SVZ). To date, no reliable in vitro systems, which can be used to unravel the potential mechanisms underlying this lesion-induced effect, have been established. Here, we developed an ex vivo method to investigate how the proliferation of NSPCs changes over time after experimental stroke or excitotoxic striatal lesion in the adult rat brain by studying the effects of microglial cells derived from an injured brain on NSPCs. We isolated NSPCs from the SVZ of brains with lesions and analyzed their growth and differentiation when cultured as neurospheres. We found that NSPCs isolated from the brains 1-2 weeks following injury consistently generated more and larger neurospheres than those harvested from naive brains. We attributed these effects to the presence of microglial cells in NSPC cultures that originated from injured brains. We suggest that the effects are due to released factors because we observed increased proliferation of NSPCs isolated from non-injured brains when they were exposed to conditioned medium from cultures containing microglial cells derived from injured brains. Furthermore, we found that NSPCs derived from injured brains were more likely to differentiate into neurons and oligodendrocytes than astrocytes. Our ex vivo system reliably mimics what is observed in vivo following brain injury. It constitutes a powerful tool that could be used to identify factors that promote NSPC proliferation and differentiation in response to injury-induced activation of microglial cells, by using tools such as proteomics and gene array technology.

An easier method to identify the individual genomic composition of allopolyploid complexes.

A new method for the fast identification of the genomic composition of the cyprinid Squalius alburnoides is presented. The method is based on a length polymorphism detected in the beta-actin gene, which serves as the basis for the development of a semi-quantitative PCR.

Análise comparativa da eficácia do alongamento manual entre a técnica de energia muscular e o alongamento estático / Comparative analysis of the effectiveness of manual stretching between muscle energy technique and static stretching

Os alongamentos são usados no processo de prevenção e reabilitação. Tendo em vista a grande variedade de técnicas empregadas objetivando o ganho de flexibilidade, foi observada a necessidade da realização de um estudo que compare a eficácia entre algumas dessas técnicas, facilitando a eleição no momento de sua utilização. As técnicas de energia muscular (TEM) utilizam a contração voluntária, já o alongamento estático, segundo Kisner, é um método em que os tecidos moles são submetidos a um alongamento mantido, além de sua resistência. A eficácia das manobras empregadas foi avaliada quanto ao ganho imediato de flexibilidade em 40 indivíduos que se enquadraram nos critérios de inclusão, separados em 2 grupos (A e B) sendo aplicado no grupo A, TEM e no grupo B, alongamento estático. Constatou-se que apesar das duas apresentarem ganhos estatisticamente significativos, o alongamento com a TEM foi mais eficaz.

The stretching exercises are often used for injury prevention and rehabilitation process. There are a wide variety of techniques aiming to gain flexibility; however, we observed the need of studies to compare the efficacy of these techniques, to guide the election of their use. The muscle energy techniques (MET) use active muscle contraction, and the static stretching, according to Kisner, is a method based on stretching/maintenance of soft tissues, over their resistance. The effectiveness of the maneuvers employed was evaluated by immediate flexibility gain in 40 individuals, who fulfilled the criterion for inclusion, divided into 2 groups (A and B), group A used MET technique, and group B static stretching. We observed that although both groups showed statistic significant gains, the MET was more effective as a method to promote muscle stretching.

Changes in calcium dynamics following the reversal of the sodium-calcium exchanger have a key role in AMPA receptor-mediated neurodegeneration via calpain activation in hippocampal neurons.

Proteolytic cleavage of the Na(+)/Ca(2+) exchanger (NCX) by calpains impairs calcium homeostasis, leading to a delayed calcium overload and excitotoxic cell death. However, it is not known whether reversal of the exchanger contributes to activate calpains and trigger neuronal death. We investigated the role of the reversal of the NCX in Ca(2+) dynamics, calpain activation and cell viability, in alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptor-stimulated hippocampal neurons. Selective overactivation of AMPA receptors caused the reversal of the NCX, which accounted for approximately 30% of the rise in intracellular free calcium concentration ([Ca(2+)](i)). The NCX reverse-mode inhibitor, 2-[2-[4-(4-nitrobenzyloxy)phenyl]ethyl]isothiourea (KB-R7943), partially inhibited the initial increase in [Ca(2+)](i), and prevented a delayed increase in [Ca(2+)](i). In parallel, overactivation of AMPA receptors strongly activated calpains and led to the proteolysis of NCX3. KB-R7943 prevented calpain activation, cleavage of NCX3 and was neuroprotective. Silencing of NCX3 reduced Ca(2+) uptake, calpain activation and was neuroprotective. Our data show for the first time that NCX reversal is an early event following AMPA receptor stimulation and is linked to the activation of calpains. Since calpain activation subsequently inactivates NCX, causing a secondary Ca(2+) entry, NCX may be viewed as a new suicide substrate operating in a Ca(2+)-dependent loop that triggers cell death and as a target for neuroprotection.