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The Lassa virus is endemic in parts of West Africa, and it causes hemorrhagic fever with high mortality. The development of a recombinant protein vaccine has been hampered by the instability of soluble Lassa virus glycoprotein complex (GPC) trimers, which disassemble into monomeric subunits after expression. Here, we use two-component protein nanoparticles consisting of trimeric and pentameric subunits to stabilize GPC in a trimeric conformation. These GPC nanoparticles present twenty prefusion GPC trimers on the surface of an icosahedral particle. Cryo-EM studies of GPC nanoparticles demonstrated a well-ordered structure and yielded a high-resolution structure of an unliganded GPC. These nanoparticles induced potent humoral immune responses in rabbits and protective immunity against the lethal Lassa virus challenge in guinea pigs. Additionally, we isolated a neutralizing antibody that mapped to the putative receptor-binding site, revealing a previously undefined site of vulnerability. Collectively, these findings offer potential approaches to vaccine and therapeutic design for the Lassa virus.

RESUMO Objetivo: Avaliar as modificações nas características das paradas cardíacas no hospital após a implantação de um Time de Resposta Rápida. Métodos: Este foi um estudo observacional prospectivo de paradas cardíacas ocorridas no hospital entre janeiro de 2013 e dezembro de 2017. O critério de exclusão foi parada cardíaca na unidade de terapia intensiva, na emergência ou na sala cirúrgica. O Time de Resposta Rápida foi introduzido no hospital do estudo em julho de 2014. Os pacientes foram classificados em dois grupos: Pré-Time de Resposta Rápida (parada cardíaca no hospital antes da implantação do Time de Resposta Rápida) e Pós- Time de Resposta Rápida (parada cardíaca no hospital após a implantação do Time de Resposta Rápida). Os pacientes foram seguidos até a alta hospitalar ou óbito. Resultados: Ocorreram 308 paradas cardíacas (64,6 ± 15,2 anos; 60,3% homens; 13,9% com ritmo inicial chocável). Houve diminuição de 4,2 para 2,5 no índice de parada cardíaca no hospital por 1.000 admissões após o início da atuação do Time de Resposta Rápida, além de cerca de 124 chamados por 1.000 admissões. A parada antes da implantação do Time de Resposta Rápida se associou com hipóxia (29,4 versus 14,3%; p = 0,006) e alteração da frequência respiratória (14,7 versus 4,2%; p = 0,004) em comparação aos dados referentes à parada cardíaca após a implantação do Time de Resposta Rápida. Parada cardíaca por hipóxia foi mais comum antes da implantação do Time de Resposta Rápida (61,2 versus 38,1%; p < 0,001). Na análise multivariada, o retorno à circulação espontânea se associou com ritmo chocável (RC 2,97; IC95% 1,04 - 8,43) e parada cardíaca testemunhada (RC 2,52; IC95% 1,39 - 4,59) mas não com a implantação do Time de Resposta Rápida (RC 1,40; IC95% 0,70 - 2,81) ou sinais premonitórios (RC 0,71; IC95% 0,39 - 1,28). Na análise multivariada, a mortalidade hospitalar se associou com ritmo não chocável (RC 5,34; IC95% 2,28 - 12,53) e idade (RC 1,03; IC95% 1,01 - 1,05), porém não com a implantação do Time de Resposta Rápida (RC 0,89; IC95% 0,40 - 2,02). Conclusão: Apesar de a implantação de um Time de Resposta Rápida se associar com redução na incidência de parada cardíaca no hospital, ela não se associou com a redução da mortalidade das vítimas de parada cardíaca no hospital. Observou-se significante diminuição nas paradas cardíacas devidas a causas respiratórias após a implantação do Time de Resposta Rápida. ABSTRACT Objective: To evaluate changes in the characteristics of in-hospital cardiac arrest after the implementation of a Rapid Response Team. Methods: This was a prospective observational study of in-hospital cardiac arrest that occurred from January 2013 to December 2017. The exclusion criterion was in-hospital cardiac arrest in the intensive care unit, emergency room or operating room. The Rapid Response Team was implemented in July 2014 in the study hospital. Patients were classified into two groups: a Pre-Rapid Response Team (in-hospital cardiac arrest before Rapid Response Team implementation) and a Post-Rapid Response Team (in-hospital cardiac arrest after Rapid Response Team implementation). Patients were followed until hospital discharge or death. Results: We had a total of 308 cardiac arrests (64.6 ± 15.2 years, 60.3% men, 13.9% with initial shockable rhythm). There was a decrease from 4.2 to 2.5 in-hospital cardiac arrest/1000 admissions after implementation of the Rapid Response Team, and we had approximately 124 calls/1000 admissions. Pre-Rapid Response Team cardiac arrest was associated with more hypoxia (29.4 versus 14.3%; p = 0.006) and an altered respiratory rate (14.7 versus 4.2%; p = 0.004) compared with post-Rapid Response Team cardiac arrest. Cardiac arrest due to hypoxia was more common before Rapid Response Team implementation (61.2 versus 38.1%, p < 0.001). In multivariate analysis, return of spontaneous circulation was associated with shockable rhythm (OR 2.97; IC95% 1.04 - 8.43) and witnessed cardiac arrest (OR 2.52; IC95% 1.39 - 4.59) but not with Rapid Response Team implementation (OR 1.40; IC95% 0.70 - 2.81) or premonitory signs (OR 0.71; IC95% 0.39 - 1.28). In multivariate analysis, in-hospital mortality was associated with non-shockable rhythm (OR 5.34; IC95% 2.28 - 12.53) and age (OR 1.03; IC95% 1.01 - 1.05) but not with Rapid Response Team implementation (OR 0.89; IC95% 0.40 - 2.02). Conclusion: Even though Rapid Response Team implementation is associated with a reduction in in-hospital cardiac arrest, it was not associated with the mortality of in-hospital cardiac arrest victims. A significant decrease in cardiac arrests due to respiratory causes was noted after Rapid Response Team implementation.

A third mRNA-based “booster” vaccination is the favored strategy to maintain protection against SARS-CoV-2 infection. Yet, significant waning of specific immunity within six months after 2nd vaccination, along with higher incidence of breakthrough infections associated with the time elapsed since 2nd vaccination raises concerns regarding the durability of immunity also after 3rd vaccination. We assessed virus-specific serum antibody and T cell response in the blood after vaccination with the mRNA vaccine BNT162b2 in more than 50 individuals older than 80 years. All old adults demonstrated a strong humoral response to 3rd vaccination which was at average higher and waned slower than the response to 2nd vaccination, indicative of enhanced humoral immunity. In contrast, their respective T cell response quantitatively limited to the level obtained after 2nd vaccination, with similar waning over time and no evidence for enhanced IFNg production. Because BNT162b2-mediated protection from the Omicron variant relies more on T cells than antibodies, our findings raise concern on the durability of protection from the Omicron variant by BNT162b2 in the senior population.

Space is a key element of human life that holds significance across the life course. Spaces, territories and symbolic arrangements are elements of social reality. This chapter examines the role of space in the context of transitions. Our conceptualization of space does not reduce it to qualities of an external environment but views it as a relational arrangement of people and features that are crucial for understanding transitions across the life course instead. Drawing from two empirical studies anchored in dispositive analysis and environmental psychology, we examine (a) transitions to children’s independent mobility, and (b) transitions into multigenerational cohousing. We ask: How can spatial sensitivity shape, challenge and enrich research on transitions in childhood and later life?

Here we compared SARS-CoV-2-specific antibody and T-cell responses between older adults (80 years old, n = 51) and a younger control group (20-53 years old, n = 46) after receiving two doses of BNT162b2. We found that responses in older adults were generally lower, and we identified 10% low-/non-responders. After receiving a third vaccination with BNT162b2, 4 out of 5 low-/non-responders showed antibody and T-cell responses similar to those of responders after two vaccinations.

The current Severe acute respiratory syndrome-related coronavirus 2 (SARS-CoV-2) pandemic is a public health emergency of international concern. Sensitive and precise diagnostic tools are urgently needed. In this study, we developed a SARS-CoV-2 spike (S1) protein enzyme-linked immunosorbent assay (ELISA) to detect SARS-CoV-2-specific antibodies. The SARS-CoV-2 S1 ELISA was found to be specific [97.8% (95% CI, 96.7% - 98.5%)], reproducible and precise (intra-assay coefficient of variability (CV) 5.3%, inter-assay CV 7.9%). A standard curve and the interpolation of arbitrary ELISA units per milliliter served to reduce the variability between different tests and operators. Cross-reactivity to other human coronaviruses was addressed by using sera positive for MERS-CoV- and hCoV HKU1-specific antibodies. Monitoring antibody development in various samples of twenty-three and single samples of twenty-nine coronavirus disease 2019 (COVID−19) patients revealed seroconversion and neutralizing antibodies against authentic SARS-CoV-2 in all cases. The comparison of the SARS-CoV-2 (S1) ELISA with a commercially available assay showed a better sensitivity for the in-house ELISA. The results demonstrate a high reproducibility, specificity and sensitivity of the newly developed ELISA, which is suitable for the detection of SARS-CoV-2 S1 protein-specific antibody responses., Highlights • A highly sensitive and specific SARS-CoV-2 S1 ELISA was developed. • A standard curve is included to reduce variability between assays and operators. • Excellent intra-assay coefficient of variability (CV) 5.3%, inter-assay CV 7.9%. • Good correlation to the virus neutralization test VNT100.

The emergence of SARS-CoV-2 highlights the global need for platform technologies to enable the rapid development of diagnostics, vaccines, treatments, and personal protective equipment (PPE). However, many current technologies require the detailed mechanistic knowledge of specific material-virion interactions before they can be employed, for example, to aid in the purification of vaccine components or in the design of a more effective PPE. Here, we show that an adaption of a polymer microarray method for screening bacterial-surface interactions allows for the screening of polymers for desirable material-virion interactions. Nonpathogenic virus-like particles including fluorophores are exposed to the arrays in an aqueous buffer as a simple model of virions carried to the surface in saliva/sputum. Competitive binding of Lassa and Rubella virus-like particles is measured to probe the relative binding properties of a selection of copolymers. This provides the first step in the development of a method for the discovery of novel materials with promise for viral binding, with the next being development of this method to assess absolute viral adsorption and assessment of the attenuation of the activity of live virus, which we propose would be part of a material scale up step carried out in high containment facilities, alongside the use of more complex media to represent biological fluids.

The emergence of SARS-CoV-2 highlights the global need for platform technologies to enable rapid development of diagnostics, vaccines, treatments, and personal protective equipment (PPE). However, many current technologies require the detailed mechanistic knowledge of specific material-virion interactions before they can be employed, for example to aid in the purification of vaccine components, or in design of more effective PPE. Here we show that an adaption of polymer micro array method for screening bacterial-surface interactions allows for screening of polymers for desirable material-viron interactions. Non-pathogenic virus like particlesincluding fluorophores are exposed to the arrays in aqueous buffer as a simple model of virons carried to the surface in saliva/sputum. Competitive binding of Lassa and Rubella particles is measured to probe the relative binding properties of a selection of copolymers. This provides the first step in the development of a method for discovery of novel materials with promise for viral binding, with the next being development of this method to assess absolute viral adsorption and assessment of the attenuation of the activity of live virus which we propose would be part of a material scale up step carried out in biological laboratory safety level 4 facilities and the use of more complex media to represent biological fluids.

DNA analysis can yield information usable to identify individuals or to associate suspects, victims, and crime scene to one another but does not support a contextualization of trace material. Knowing the composition of biological material from a crime scene, e.g. whether a stain contains saliva or semen, is complementary to DNA-based identification and can be essential in reconstructing the course of events in criminal investigations. Research in the forensic applications of RNA analysis has surged considerably during the last two decades and provided recent advancements for forensic casework. Therefore, as a proof of concept, different commercially available DNA extraction kits were compared to assess their potential to simultaneously co-isolate messengerRNA and microRNA in DNA eluates of forensically relevant materials. The results showed that only one extraction kit elutes enough RNA molecules to perform messengerRNA body fluid identification whereas for microRNA-specific expression two of the tested kits yielded sufficient RNA concentrations.

Free flaps in combination with arterial reconstruction by means of arteriovenous loops or bypass have, meanwhile, been established as a therapeutic option in defect reconstruction for areas without recipient vessels. Our aim was to analyse the long‐term performance, flap autonomy, and the flap perfusion. Patients receiving this combined reconstruction at a single‐centre institution were included. During follow‐up examination, the patency of arterial reconstruction was investigated by duplex ultrasound. Flap micro‐circulation was assessed by laser Doppler flowmetry and white light tissue spectrometry (O2C) as well as by indocyanine green fluorescence angiography. Twenty‐three patients could be clinically followed up. Duplex ultrasound showed, in four cases, arterial pedicle occlusion in spite of vital flap. Comparison of the O2C perfusion parameters between flaps with occluded pedicles and those with intact inflow showed no significant difference (parameters sO2: P = .82; Flow: P = .31). Similar results were obtained by fluorescence angiography; no significant difference could be detected between both groups (parameters Ingress P = .13; Ingressrate P = .54). Combined vascular reconstruction with free tissue transfer is associated with a good long‐term outcome and wound closure. Even after flap transplantation to areas with critical tissue perfusion, the flap can develop autonomy and thus survive after pedicle occlusion.

Lassa mammarenavirus (LASV) is a rodent-borne arenavirus endemic to several West African countries. It is the causative agent of human Lassa fever, an acute viral hemorrhagic fever disease. To date, no therapeutics or vaccines against LASV have obtained regulatory approval. Polyclonal neutralizing antibodies derived from hyperimmunized animals may offer a useful strategy for prophylactic and therapeutic intervention to combat human LASV infections. The LASV envelope surface glycoprotein complex (GP) is the major target for neutralizing antibodies, and it is the main viral antigen used for the design of an LASV vaccine. Here, we assessed the immunogenic potential of mammalian cell-derived virus-like particles (VLPs) expressing GP from the prototypic LASV strain Josiah in a native-like conformation as the sole viral antigen. We demonstrate that an adjuvanted prime-boost immunization regimen with GP-derived VLPs elicited neutralizing antibody responses in rabbits, suggesting that effective antigenic epitopes of GP were displayed. Notably, these antibodies exhibited broad reactivity across five genetic lineages of LASV. VLP-based immunization strategies may represent a powerful approach for generating polyclonal sera containing cross-reactive neutralizing antibodies against LASV.

Introdução: A ressuscitação fluídica agressiva, com volume superior ao preconizado por Baxter, com objetivo de evitar a hipoperfusão de órgãos e restabelecer a volemia rapidamente, se tornou uma tendência nos últimos anos. Esta prática vem sendo discutida mais recentemente após a caracterização do fenômeno fluid creep e pode ser causa de aumento da morbidade e mortalidade em crianças queimadas. A hiperhidratação impacta negativamente a evolução desses pacientes e já se estabeleceram associações do fenômeno fluid creep com diversas complicações, como, por exemplo, a síndrome compartimental abdominal. Pacientes queimados estão sob alto risco para infecção e sepse devido à perda da integridade da pele e à exposição prolongada ao ambiente nosocomial. A principal causa de óbito, se o paciente sobreviver à fase inicial do choque do queimado é a disfunção de múltiplos órgãos e sistemas secundária a sepse. Objetivos: Avaliar a associação entre o fluid creep e episódios infecciosos em crianças queimadas submetidas a duas estratégias diferentes de reanimação hídrica envolvendo albumina. Realizar análises de curvas de probabilidade livre de infecção, além de comparar o tempo de internação e número de procedimentos cirúrgicos realizados nestes grupos de pacientes. Metodologia: Estudo de coorte histórica envolvendo 46 crianças queimadas admitidas no Centro de Tratamento de Queimados do Hospital Universitário de Londrina entre junho de 2012 e janeiro de 2014 e que incluiu casos oriundos de um ensaio clínico randomizado controlado realizado no mesmo período. Foram incluídas crianças entre 1 e 12 anos, com queimaduras entre 15% e 45% de Superfície Corporal Queimada (SCQ) e profundidade de 2º e 3º graus, admitidas até a 12ª hora após o acidente. Todos os pacientes receberam reanimação fluídica com Ringer Lactato (RL), nas primeiras 24 horas, segundo a Fórmula de Parkland modificada (3ml/kg/%SCQ) ajustada de acordo com o débito urinário (DU). Os pacientes do grupo A = Exposto (n=23) receberam albumina 5% entre 8 e 12 horas do momento do acidente, e os do Grupo B= Não Exposto (n=23) receberam albumina 5% partir da 24a hora do acidente. Os grupos foram comparados quanto ao surgimento do fenômeno fluid creep, tempo de internação, número de procedimentos cirúrgicos realizados e infecção até a alta hospitalar. Resultados: Os pacientes do grupo A tiveram menos dias de internação comparadamente ao grupo B (p=0,007). O surgimento do fenômeno fluid creep foi maior em crianças no grupo B (p < 0,001). O tempo de internação maior de 14 dias assim como a presença de fluid creep foi associado com a presença de infecção, respectivamente p=0,002 e p=0,020. O tempo de internação bem como a presença de fluid creep apresentaram diferenças nas curvas de probabilidade livre de infecção (p < 0,05). Todas as crianças do grupo B receberam debridamento comparado com 73,9% daquelas incluídas no grupo A (p=0,022), assim como os enxertos foram mais frequentes no grupo B (p=0,032). Conclusões: Crianças queimadas que receberam albumina entre 8 e 12 horas após o acidente apresentaram significativamente menos fluid creep, menor tempo de internação e menor número de procedimentos cirúrgicos (debridamentos e enxertos) comparativamente ao grupo que recebeu albumina após 24 horas. A presença do fluid creep e o tempo de internação superior a 14 dias apresentaram diferenças nas curvas de probabilidade livre de infecção, demonstrando pior prognóstico infeccioso Introduction: Aggressive fluid resuscitation, defined as the administration of a superior volume to the parameters recommended by Baxter, intend to rapidly set the euvolemia and avoid organ hypoperfusion in burn individuals. The practice became a trend during the last years and has been extensively discussed after the establishment of the fluid creep phenomenon and has been considered as a possible mortality and morbidity factor in burn children. A strong association of fluid creep development has already been established with many conditions, as abdominal compartment syndrome. Burn patients are at high risk of infection and subsequent progression to sepsis due to the loss of skin integrity and long-term exposure to hospital environment. If those patients survive to the burn shock, their main cause of death is the multiple organ failure secondary to sepsis, what highlights the importance of comprehensive knowledge of infectious complications in burn children. Objectives: Assess the association between fluid creep and infection episodes in burn children that were exposed to two different fluid resuscitation strategies with the use of albumin. Analyze the patients infection-free probability curves, length of stay and number of surgery procedures needed. Methods: We developed a historic cohort of 46 burn children admitted at the Burn Treatment Center, State University of Londrina. The same patients were simultaneously enrolled in a parallel clinical trial. In this study, were included patients from 1 to 12 years old, with 15% to 45% of the body surface area (BSA) compromised, second or third-degree burns, admitted up to 12 hours after the accident. Both groups were compared according to the exposure. All the patients received fluid resuscitation with Ringer Lactate (RL) in the first 24 hours, respecting the modified Parkland formula (3 ml/kg/% TBSA) adjusted according to the urine output (UO). Patients from group A were exposed to 5% albumin between 8 and 12 hours from the accident moment and the patients from group B received 5% albumin after 24 hours from the accident. The outcomes observed were development of fluid creep phenomenon, length of stay, number of surgery procedures needed and infection until the hospital discharge. Results: Compared to the group B, the group A had a shorter median length of stay (p=0,007). Fluid creep development had higher rates on group B compared to group A (p < 0,001). The length of stay for more than 14 days, as the fluid creep arising, was associated with infection (respectively, p=0,002 and p=0,02). Both the length of stay and the fluid creep presence showed difference at the infection-free probability curves (p < 0,05). All the patients from group B were submitted to debridement compared to 73,9 from group A (p=0,022). The need of skin graft procedure was also more frequent in patients from group A (p=0,032). Conclusion: Burn children that received albumin between 8 and 12 hours from the accident moment presented significant less fluid creep, shorter length of stay, and smaller number of surgery procedures (as debridement and skin graft) compared to the group that received albumin after 24 hours from the accident. Fluid creep development and length of stay superior to 14 days presented differences at the infection-free probability curves, providing a negative infectious prognosis

The development and validation of a simple, reliable and fast method for the determination of cadmium and lead in fish and shrimp by GF AAS, following sample freeze-drying and tetramethylammonium hydroxide (TMAH) solubilization, is presented. The method development was achieved by sequentially applying factorial designs until optimization within the bilinear approximation of the method was accomplished. As such, seven experimental parameters were initially studied, which were considered important for the determination of cadmium and lead in seafood samples using TMAH solubilization. The validation of the method was completed in order to comply with international food regulations and method accreditation under ISO 17025. In this sense, the following parameters were evaluated: linearity, limits of detection and quantification, precision, recovery (trueness), specificity and robustness. Linearity of response was satisfactory for the concentration ranges of both analytes. The residuals for both elements were homoscedastic and independent, with normal distributions. Limits of quantification (LOQ), based on signal standard deviation for low-in-cadmium and lead samples, were 6.25 μg kg−1 and 31.25 μg kg−1, respectively. These figures are in accordance with performance criteria required by the Commission Regulation (EU) no 836/2011. The repeatability of the method, calculated from the analysis of seven sample replicates at two concentration levels of cadmium and lead, by the same analyst, was usually better than 10%. Recovery was estimated from the repeatability evaluation, lying in the range of 84% to 99% for cadmium, and 94% to 107% for lead. Furthermore, a wide evaluation of the method robustness was performed during the method optimization. Thus, the results showed the suitability of the developed method for the determination of Cd and Pb in fish and shrimp by GF AAS, following TMAH solubilization for food control purposes. Additionally, the use of sequential factorial designs in order to achieve bilinear optimization of the method was proven to be a very valuable tool, by sharply reducing optimization time and the number of experiments.

RESUMO: A presente pesquisa objetivou avaliar a qualidade fisiológica de sementes de pimenta (Capsicum chinense Jacq.) var. Bode Vermelha obtidas de frutos em diferentes estádios de maturação. Os frutos de C. chinense foram separados em cinco estádios de maturação, baseando-se na coloração do pericarpo (frutos verdes - estádio 1, frutos alaranjados - estádio 2, frutos vermelhos-claros - estádio 3, frutos vermelhos - estádio 4 e frutos vermelho-carmim - estádio 5). Para a avaliação da maturação e da qualidade fisiológica das sementes foram determinadas as características biométricas dos frutos e sementes, massa de mil sementes, teor de água das sementes, condutividade elétrica, teste padrão de germinação, tempo médio de germinação, porcentagem de plântulas normais, velocidade de formação de plântulas, além dos testes de envelhecimento acelerado, e comprimento de plântulas. Os caracteres comprimento, diâmetro, massa e número de sementes dos frutos e comprimento, largura, espessura e massa das sementes foram analisados por meio de parâmetros estimados utilizando-se estatística descritiva. Para as demais variáveis foi utilizado delineamento experimental inteiramente casualizado. Os resultados foram submetidos à análise de variância e a comparação de médias pelo teste Scott-Knott, ao nível de 5% de probabilidade. O estádio de maturação influencia na qualidade fisiológica das sementes de Capsicum chinense Jacq. var. Bode Vermelho, sendo as sementes oriundas de frutos de coloração vermelho (estádio 4) a vermelho-carmim (estádio 5) as que apresentaram os melhores desempenhos e, portanto, estão fisiologicamente maduras, sendo os mais indicados para obtenção das sementes. ABSTRACT: This research aimed to evaluate the physiological quality of pepper (Capsicum chinense Jacq.) var. Bode Vermelha seeds, obtained from fruits at different ripening stages. The fruits of C. Chinese were separated according to the ripening stages, based on pericarp color (green fruits - stage 1, orange fruits - stage 2, light red fruits - stage 3, red fruits - stage 4, and carmine-red fruits - stage 5). To evaluate the ripeness and the physiological quality of the seeds were determined the biometric characteristics of the fruits and seeds, mass of thousand seeds, seeds water content, electrical conductivity, standard germinations test, speed of germination index, percentage of normal seedlings, speed of seedlings formation, accelerated aging, and seedling length. The characters length, diameter, weight, and number of seeds of the fruits and length, width, thickness, and mass of the seeds were analyzed by the estimated parameters using descriptive statistics. For the other variables was used the completely randomized design. The results were submitted to variance analysis and comparison of means by the Scott-Knott test at 5% probability. The ripeness stage affects the physiological quality of C. chinense Jacq. var. Bode Red seeds, being the seeds from red (stage 4) to carmine-red (stage 5) fruits those with the best performance thus, they are physiologically ripe, being the most indicated to seeds obtaining.

The objective of this study was to evaluate the spatial variability in soybean seeds quality and in soil chemical attributes of a production field. Data were collected at 138 georeferenced points of a soybean production property located in Santo Antônio de Leverger - MT. Soil related variables, such as phosphorus, potassium, calcium, magnesium, and organic matter (OM) contents, pH, cation exchange capacity (CEC) and base saturation (V%) were evaluated. On the other hand, yield, one thousand seed mass, size, germination, emergence in seedbed, electrical conductivity, accelerated aging and tetrazolium reaction were evaluated as seed variables. The data were submitted to descriptive and geostatistical analysis, and the fit semivariogram parameters were used to elaborate spatial distribution maps of the analyzed variables. After the analysis, it was possible to conclude that there was spatial variability in the evaluated attributes for both seeds and soil related variables, indicating that the soybean seed production area can be divided into management zones, which allows the definition of areas to be harvested or discarded within a field of seed production. Resumo: Objetivou-se avaliar a variabilidade espacial da qualidade de sementes de soja e dos atributos químicos do solo em campo de produção. Os dados do experimento foram coletados em 138 pontos georreferenciados em propriedade produtora de sementes de soja localizada no município de Santo Antônio de Leverger - MT. Para as variáveis relacionadas ao solo, avaliou-se os teores de fósforo, potássio, cálcio, magnésio, matéria orgânica (MO), o pH, a capacidade de troca catiônica (CTC) e a saturação por bases (V%). Nas sementes, avaliou-se produtividade, massa de 1000 sementes, tamanho, germinação, emergência em canteiro, condutividade elétrica, envelhecimento acelerado e tetrazólio. Os dados foram submetidos à análise descritiva e geoestatística, e os parâmetros dos semivariogramas ajustados foram utilizados para elaboração de mapas de distribuição espacial das variáveis analisadas. Após análise, conclui-se que há variabilidade espacial nos atributos avaliados nas sementes e no solo, indicando que a área para produção de sementes de soja pode ser dividida em zonas de manejo, que permite a definição de áreas a serem colhidas e descartadas dentro de um campo de produção de sementes.

We have developed a microfluidic chip-based qualitative assay for sensitive (10 RNA copies) detection of multiple transcripts in single cells. We determined the expression patterns of 17 developmentally important genes and isoforms in individual mouse preimplantation embryos from superovulated matings and blastomeres. The ubiquitously expressed histone variant H3f3a and the transcription factor Pou5f1 generated mRNA-derived products in all analyzed (1-cell, 2-cell, 4-cell, and morula stage) embryos and in all analyzed blastomeres from 16-cell embryos, indicating a uniform reactivation of pluripotency gene expression during mouse preimplantation development. In contrast, mRNA expression of different methyltransferases for DNA methylation, methylcytosine-binding proteins for chromatin modification, and base excision repair enzymes, which may provide a mechanism for active demethylation, varied considerably between individual cells from the same embryo and even more dramatically between cells from different embryos. We conclude that at a given point in time the transcriptome encoding the reprogramming machinery and, by extrapolation, genome reprogramming differs between blastomeres. By studying the cell-to-cell variability in gene expression, we can distinguish the following two classes: mouse 16-cell embryos in which most cells express the reprogramming machinery and embryos in which most cells do not contain detectable mRNA levels of DNA and chromatin modification genes. Immunolocalization of DNMT3A, MBD3, APEX1, and LIG3 in most or all nuclei of 40-60-cell embryos is a good indicator of functional activity of genes that are activated by the 16-cell stage.

Introdução: A reanimação fluídica da criança queimada é um desafio devido à intolerância à insuficiente ou excessiva oferta de líquidos. Há dúvidas em relação à utilização de solução coloide na ressuscitação volêmica e também quanto ao melhor momento a ser administrada. O momento ideal para a administração de albumina permanece em foco de debate, se deveria ser utilizada como estratégia de resgate, quando o volume de cristaloide infundido se torna excessivo, ou rotineiramente, como intervenção primária em pacientes com queimaduras extensas. Objetivos: Avaliar e comparar quanto a evolução clínica de crianças com lesões térmicas que receberam abordagem de infusão precoce (entre 8 e 12 horas do acidente) de solução coloide natural versus crianças que receberam abordagem de infusão tardia (após 24 horas do acidente) da mesma solução para reanimação na fase aguda. Metodologia: Ensaio Clínico Randomizado Controlado, realizado no Centro de Tratamento de Queimados do Hospital Universitário de Londrina. Foram estudadas 46 crianças (1 a 12 anos), apresentando entre 15% e 45% de Superfície Corporal Queimada, admitidas até a 12a hora após o acidente. Intervenção: Para a ressuscitação hídrica dos pacientes, foi utilizada solução cristaloide baseada na Fórmula de Parkland modificada, ajustada de acordo com o débito urinário. O Grupo Intervenção (23 pacientes) foi randomizado para receber solução de albumina entre 8 e 12 horas do acidente, e o Grupo Controle (23 pacientes) recebeu a mesma solução após 24 horas do acidente. Resultados: Houve possibilidade de redução de infusão de solução cristaloide durante o período de ressuscitação dos pacientes. O grupo Intervenção recebeu um volume de solução cristaloide com uma mediana de -31,99% (P=0,025) no 1º dia, -19,37% (P=0,002) no 2º dia e -45,3% (P=0,002) no 3º dia de ressuscitação. Não foram observadas diferenças significantes entre os grupos em relação à diurese. A incidência acumulada de fluid creep na população estudada foi de 30,43% (n=14). O RR para o desenvolvimento do fluid creep no Grupo Intervenção foi de 0,0769 (IC 95% 0,0109 a 0,5407). A mediana do tempo de internação dos pacientes do Grupo Controle foi de 18 (15-21) dias e a do Grupo Intervenção foi de 14 (10-17) dias, P=0,004. Conclusões: A infusão precoce de solução de albumina em crianças com queimaduras entre 15% e 45% de superfície corporal reduziu a necessidade de infusão de solução cristaloide no período de ressuscitação. Foram identificados significativamente menos casos de fluid creep e observado menor tempo de internação entre pacientes que receberam albumina precocemente Introduction: The fluidic resuscitation of burned children is a challenge due to intolerance to insufficient or excessive supply of liquids. There are questions regarding the use of colloids in fluid resuscitation solution as well as the timing of the administration. The ideal time for the administration of albumin, in other words whether it should be used as a rescue strategy when the volume of crystalloid infused becomes excessive, or routinely, as primary intervention for patients with extensive burns, remains in the focus of discussion. Objectives: Evaluate the clinical outcomes of children with burn injuries who have received early infusion treatment (between 8 and 12 hours after the accident) with natural colloids solution compared to the children who have received late infusion treatment (24 hours after the accident) with the same solution, in order to resuscitate them in the acute phase. Methods: Randomized Controlled Trial carried out at the Burn Treatment Center, State University of Londrina. Forty-six children (1 to 12 year olds) who had between 15% and 45% total body surface area and were admitted up to 12 hours after the accident were studied. Intervention: For the fluid resuscitation of patients, a crystalloid solution based on modified Parkland Formula was adjusted according to urine output. The Intervention Group (n= 23) were randomized to receive albumin solution between 8 and 12 hours after the accident and the Control Group (n= 23) received the same solution later than 24 hours after the burn injury. Results: During the resuscitation of patients, it was possible to reduce the infusion of crystalloid solution. The Intervention Group required a volume of crystalloid solution with a median of -31.99% (P = 0.025) on day 1, - 19.37% (P = 0.002) on day 2 and -45.3% (P = 0.002) on day 3 of resuscitation. No significant differences were observed in the groups in relation to diuresis. The cumulative incidence of fluid creep in the population studied was 30.43% (n=14). The RR for the development of fluid creep in the Intervention Group was 0.0769 (IC 95% 0.0109 to 0.5407). The patients in the Control Group spent an average time of 18 (15-21) days in hospital, while the patients in the Intervention Group spent 14 (10-17) days, P=0,004. Conclusions: The early infusion of albumin solution in children with 15% to 45% total body surface area reduced the need of crystalloid infused during the resuscitation period. Significantly lower cases of fluid creep were identified and lower length of stay was observed among patients who were treated earlier with albumin

Objetivou-se comparar a similaridade qualitativa das caracteristicas fenotipicas de plantas de alface cv. Marisa, desenvolvidas a partir de lotes de sementes produzidas nos Estados Unidos da America (EUA) e no Chile. O delineamento experimental foi em blocos ao acaso, com dois tratamentos (lotes de sementes provenientes dos EUA – Lote 1 e Chile – Lote 2) e seis repeticoes, sendo cada unidade experimental constituida por uma parcela de 3,0 x 1,2 m (3,6 m²) e a area util para avaliacao composta de 10 plantas de alface, excluindo-se as primeiras e as ultimas plantas de cada linha de cultivo.Avaliou-se nas mudas, 26 dias apos a semeadura, o numero de folhas; comprimento da maior raiz (cm); massa da materia seca da parte aerea (MS aerea) e das raizes (MS raizes) (g). No ponto de colheita, antes do corte das plantas, avaliou-se o diâmetro e a altura (cm) e; apos a colheita, avaliou-se a massa da materia fresca total da parte aerea (g planta -1 ); a massa da materia fresca comercial (g planta -1 ); o numero de folhas comerciais, contando-se aquelas maiores que 3 cm de comprimento, iniciando-se pelas folhas basais ate a ultima folha aberta; a massa da materia fresca do caule (g) e; o comprimento do caule (cm), apos a retirada das folhas. Verificou-se diferenca nas caracteristicas agronomicas avaliadas na fase de producao de mudas e no ponto de colheita, destacando-se aquelas provenientes de sementes dos EUA. DOI: 10.12957/ric.2015.19093

Phasins play an important role in the formation of poly(3-hydroxybutyrate) [poly(3HB)] granules and affect their size. Recently, three homologues of the phasin protein PhaP1 were identified inRalstonia eutrophastrain H16. The functions of PhaP2, PhaP3 and PhaP4 were examined by analysis ofR. eutrophaH16 deletion strains (ΔphaP1, ΔphaP2, ΔphaP3, ΔphaP4, ΔphaP12, ΔphaP123and ΔphaP1234). When cells were grown under conditions permissive for poly(3HB) accumulation, the wild-type strain and all single-phasin negative mutants (ΔphaP2, ΔphaP3and ΔphaP4), with the exception of ΔphaP1, showed similar growth and poly(3HB) accumulation behaviour, and also the size and number of the granules were identical. The single ΔphaP1mutant and the ΔphaP12, ΔphaP123and ΔphaP1234mutants showed an almost identical growth behaviour; however, they accumulated poly(3HB) at a significantly lower level than wild-type and the single ΔphaP2, ΔphaP3or ΔphaP4mutants. Gel-mobility-shift assays and DNaseI footprinting experiments demonstrated the capability of the transcriptional repressor PhaR to bind to a DNA region +36 to +46 bp downstream of thephaP3start codon. The protected sequence exhibited high similarity to the binding sites of PhaR upstream ofphaP1, which were identified recently. In contrast, PhaR did not bind to the upstream or intergenic regions ofphaP2andphaP4, thus indicating that the expression of these two phasins is regulated in a different way. Our current model for the regulation of phasins inR. eutrophastrain H16 was extended and confirmed.

Amiloidose é caracterizada por deposição extracelular anormal de amilóide em diferentes tecidos e órgãos, geralmente associada com disfunção do tecido ou órgão envolvido, cuja causa não é ainda conhecida. Pode ser dividida em amiloidose sistêmica e localizada. A forma sistêmica é dividida em: (1) primária; (2) amiloidose associada ao mieloma múltiplo; (3) secundária; (4) amiloidose heredofamiliar. Há uma considerável diferença de sobrevida entre os pacientes com forma localizada e sistêmica de amiloidose, e também com mieloma múltiplo. Não há tratamento satisfatório para amiloidose sistêmica. O presente estudo relata um caso de amiloidose sistêmica primária, com revisão de literatura. O diagnóstico de amiloidose foi conseguido com biópsia de língua e o envolvimento sistêmico confirmado através de aspirado de gordura abdominal. Apesar de a amiloidose não ser a principal hipótese diagnóstica que deva ser aventada quando nos deparamos com um paciente com macroglossia, esta doença não pode ser esquecida, uma vez que o suporte clínico é essencial para o controle das doenças que podem estar associadas, como falência renal, cardíaca e o mieloma múltiplo.Amyloidosis is characterized by the abnormal, extracelular deposition of one of a family of unrelated proteins, amyloid, in different tissues or organs; it is usually associated with tissue or organ disfunction. The cause is not yet known. Amyloidosis can be divided in systemic or localized form. The sistemic form can be divided in: (1) primary; (2) amyloidosis associated with multiple myeloma; (3) secondary; (4) heredofamilial amyloidosis. There is a considerable difference in the survival of patients with localized and systemic amyloidosis, and also with or without multiple myeloma. Satisfactory treatment for systemic amyloidosis does not exist. The present study was undertaken in order to report a case of primary sistemic amyloidosis as well as literature review. The amyloidosis diagnosis was achieved by tongue biopsy and its sistemic involvement by fat abdominal aspirate. Even amyloidosis should not be the first diagnostic hypothesis when macroglossia is present, this disease can not be forgotten since clinical attendance is essential to control the associated disorders as renal, cardiac failure and myeloma multiple.

O objetivo deste trabalho foi avaliar a influencia de adubos orgânicos na populacao microbiana e sua atividade no solo. Para tal, foram coletadas, em dois periodos, amostras de solo na Fazenda Experimental da Universidade Federal de Mato Grosso, no Municipio de Santo Antonio de Leverger. O delineamento experimental adotado foi em blocos casualizados, com quatro repeticoes de quatro plantas cada. Retirou-se amostras de solo na profundidade de 0-20 cm, de areas adubadas com seis fontes de adubos orgânicos, aplicados em cobertura, nas quantidades determinadas pelo teor de N nos materiais orgânicos: esterco de bovinos, esterco de aves, efluente de suinos, composto de esterco de bovinos, composto de esterco de aves e composto de efluente de suinos. Avaliou-se o carbono da biomassa microbiana (CBM), a respiracao microbiana (RM) e o quociente metabolico (qCO 2 ). Os valores de carbono da biomassa microbiana dos estercos de aves e bovinos foram maiores e, portanto estimulam o aporte de microrganismos no solo. Os adubos orgânicos, exceto o esterco de aves curtido, nao alteram a atividade dos microrganismos. DOI: http://dx.doi.org/10.12957/ric.2014.12008

O líquen plano é uma desordem comum do epitélio escamoso estratificado que acomete as mucosas oral e genital, a pele, as unhas e o couro cabeludo. O líquen plano oral (LPO) afeta mulheres de meiaidade e apresenta padrões e distribuição característicos, como estriações brancas, pápulas ou placas brancas, eritema, erosões e bolhas, que podem estar associadas a medicações e/ou materiais dentários no paciente. O diagnóstico clínico somente poderá ser feito se a doença apresentar padrões clássicos, como lesões concomitantes na mucosa oral e na pele. O diagnóstico laboratorial por meio do exame histopatológico se caracteriza pela presença de projeções do epitélio em forma de dentes de serra e corpos de Civatte, e possibilita excluir condições de displasia e malignidade. A imunofluorescência direta é utilizada em suspeita de outras doenças, como pênfigo e penfigoide. O LPO é tratado com agentes anti-inflamatórios, principalmente, corticosteroides tópicos, e novos agentes e técnicas têm-se demonstrado eficazes. A transformação maligna do LPO e sua incidência exata permanecem controversas. Este trabalho tem como objetivo apresentar, com base na revisão da literatura, a etiopatogenia, o diagnóstico clínico, exames complementares e complicações do LPO.

Lichen planus is a common disorder of the stratified squamous epithelium that affects oral and genital mucous membranes, skin, nails, and scalp. Oral Lichen Planus (OLP) affects middle-aged women and shows distribution patterns and characteristics such as white striations, white plaques or papules, erythema, blisters and erosions, and may be associated with medication and/or dental materials used by the patient. The clinical diagnosis can only be made if the disease presents classical patterns such as concomitant lesions in the oral mucosa and skin. The laboratory diagnosis is histopathologically characterized by the presence of projections of the epithelium in the form of sawtooth and Civatte bodies and allows the exclusion of dysplasia and malignancy. Direct immunofluorescence is used when there is suspicion of other diseases, such as pemphigus and pemphigoid. OLP is treated with anti-inflammatory agents, particularly topical corticosteroids; new agents and techniques have proved effective. The malignant transformation of OLP and its exact incidence remain controversial. This work aims at presenting, through literature review, the etiopathogenesis, clinical diagnosis, laboratory tests, and complications of OLP.

Acute lung injury (ALI) still poses a major challenge in critical care medicine. Neutrophils, platelets, and chemokines are all considered key components in the development of ALI. The Duffy antigen receptor for chemokines (DARC ) is thought to be involved in scavenging, transendothelial transport, and presentation of neutrophil-specific chemokines. DARC is expressed on endothelial cells and erythrocytes but not on leukocytes. Here, we show that DARC is crucial for chemokine-mediated leukocyte recruitment in vivo. However, we also demonstrate that changes in chemokine and chemokine receptor homeostasis, associated with Darc gene deficiency, exert strong anti-inflammatory effects. Neutrophils from Darc gene-deficient ( Darc−/ −) mice display a more prolonged downregulation of CXCR2 during severe inflammation than neutrophils from wild-type mice. In a CXCR2-dependent model of acid-induced ALI, Darc gene deficiency prevents ALI. Darc−/ −mice demonstrate fully preserved oxygenation, only a small increase in vascular permeability, and a complete lack of pulmonary neutrophil recruitment. Further analysis reveals that only neutrophils but neither endothelial cells nor erythrocytes from Darc−/ −mice confer protection from ALI. The protection appears to be due to abolished pulmonary recruitment of neutrophils from Darc−/ −mice. The generation of neutrophil-platelet aggregates, a key mechanism in both pulmonary neutrophil recruitment and thrombus formation, is also affected by altered CXCR2 homeostasis in Darc−/ −mice. CXCR2 blockade enhances the formation of platelet-neutrophil aggregates and thereby corrects a formerly unknown bleeding defect in Darc−/ −mice. In summary, our study suggests that chemokine/chemokine receptor homeostasis plays a previously unrecognized and crucial role in severe ALI.

Review on PSGL-1 effects on signaling in myeloid leukocytes activates following selectin engagement. Cell-cell interactions mediating leukocyte recruitment and inflammation are crucial for host defense. Leukocyte recruitment into injured tissue proceeds in a multistep process. The first contact of leukocytes with endothelial cells (“capturing” or “tethering”) is mediated by selectins and their counter-receptor P-selectin glyco-protein ligand (PSGL)-1. During capture and rolling, leukocytes collect different inflammatory signals, which can activate various pathways. Integration of these signals leads to leukocyte activation, integrin-mediated arrest, cytoskeleton rearrangement, polarization, and transmigration. PSGL-1 on leukocytes also binds to activated platelets, where P-selectin is expressed at locally high site densities following α-granule fusion with the plasma membrane. Here, we review the signaling functions of PSGL-1 and speculate how the different known signaling events might relate to different phases of leukocyte recruitment.

We have developed a microfluidic chip-based qualitative assay for sensitive (10 RNA copies) detection of multiple transcripts in single cells. We determined the expression patterns of 17 developmentally important genes and isoforms in individual mouse preimplantation embryos from superovulated matings and blastomeres. The ubiquitously expressed histone variant H3f3a and the transcription factor Pou5f1 generated mRNA-derived products in all analyzed (1-cell, 2-cell, 4-cell, and morula stage) embryos and in all analyzed blastomeres from 16-cell embryos, indicating a uniform reactivation of pluripotency gene expression during mouse preimplantation development. In contrast, mRNA expression of different methyltransferases for DNA methylation, methylcytosine-binding proteins for chromatin modification, and base excision repair enzymes, which may provide a mechanism for active demethylation, varied considerably between individual cells from the same embryo and even more dramatically between cells from different embryos. We conclude that at a given point in time the transcriptome encoding the reprogramming machinery and, by extrapolation, genome reprogramming differs between blastomeres. By studying the cell-to-cell variability in gene expression, we can distinguish the following two classes: mouse 16-cell embryos in which most cells express the reprogramming machinery and embryos in which most cells do not contain detectable mRNA levels of DNA and chromatin modification genes. Immunolocalization of DNMT3A, MBD3, APEX1, and LIG3 in most or all nuclei of 40-60-cell embryos is a good indicator of functional activity of genes that are activated by the 16-cell stage.

Paquidermoperiostose é genodermatose autossômica dominante rara, que afeta pele e ossos, porém sua prevalência e incidência permanecem desconhecidas. Ocorre predominantemente em homens, que também mostram fenótipo mais grave da doença. Descrevese o caso de paciente do sexo masculino, de 60 anos.

Amyloidosis is characterized by an abnormal extracellular deposition of amyloid in different tissues and organs, where it usually causes some type of dysfunction. Its cause is unknown. The two main forms of amyloidosis are systemic and localized; the latter is rare. No satisfactory treatment for systemic amyloidosis has been discovered, and mean survival is poor, ranging from 5 to 15 months depending on the presence or absence of multiple myeloma. We report a case of primary systemic amyloidosis in a 71-year-old man. The diagnosis of amyloidosis was established by tongue biopsy, and its systemic nature was identified by analysis of aspirated abdominal fat. At the 1-year follow-up, the patient's clinical condition had not changed, and he was thereafter lost to follow-up.

We have developed a microfluidic device operating at a planar surface instead of a closed channel network. The fluid is transported in single droplets using surface acoustic waves (SAW) on a piezoelectric LiNbO(3) substrate. The surface of the piezo is chemically structured to induce high contact angles of the droplets or enclose areas where the liquid can wet the substrate. Combining the SAW technique with thin film resistance heaters, a biological analysis chip with integrated DNA amplification by PCR and hybridization was designed. To prevent evaporation of the PCR reagents at high temperatures the sample is enclosed in droplets of mineral oil. On this chip the SAW resolves dried primers, shifts the oil capped liquid between the two heaters and mixes during hybridization. The chip is able to perform a highly sensitive, fast and specific PCR with a volume as low as 200 nl. During the temperature cycles an online monitoring of the DNA concentration is feasible with an optical unit, providing a sensitivity of 0.1 ng. After PCR the product is moved to the second heater for the hybridization on a spotted DNA array. With our chip we were able to detect a single nucleotide polymorphism (SNP) responsible for the Leiden Factor V syndrome from human blood.