Biomechanical gait analysis in sheep: kinematic parameters.

Animals have been used as models to help to better understand biological and anatomical systems, and pathologies in both humans and non-human species, and sheep are often used as an in vivo experimental model for orthopedic research. Gait analysis has been shown to be an important tool in biomechanics research with clinical applications. The purpose of this study was to perform a kinematic analysis using a tridimensional (3D) reconstruction of the sheep hindlimb. Seven healthy sheep were evaluated for natural overground walking, and motion capture of the right hindlimb was collected with an optoelectronic system while the animals walked in a track. The analysis addressed gait spatiotemporal variables, hip, knee and ankle angle and intralimb joint angle coordination measures during the entire walking cycle. This study is the first that describes the spatiotemporal parameters from the hip, knee and ankle joints in a tridimensional way: flexion/extension; abduction/adduction and inter/external rotation. The results of this assessment can be used as an outcome indicator to guide treatment and the efficacy of different therapies for orthopedic and neurological conditions involving the locomotor system of the sheep animal model.

CAR-T cell manufacturing landscape-Lessons from the past decade and considerations for early clinical development.

CAR-T cell therapies have consolidated their position over the last decade as an effective alternative to conventional chemotherapies for the treatment of a number of hematological malignancies. With an exponential increase in the number of commercial therapies and hundreds of phase 1 trials exploring CAR-T cell efficacy in different settings (including autoimmunity and solid tumors), demand for manufacturing capabilities in recent years has considerably increased. In this review, we explore the current landscape of CAR-T cell manufacturing and discuss some of the challenges limiting production capacity worldwide. We describe the latest technical developments in GMP production platform design to facilitate the delivery of a range of increasingly complex CAR-T cell products, and the challenges associated with translation of new scientific developments into clinical products for patients. We explore all aspects of the manufacturing process, namely early development, manufacturing technology, quality control, and the requirements for industrial scaling. Finally, we discuss the challenges faced as a small academic team, responsible for the delivery of a high number of innovative products to patients. We describe our experience in the setup of an effective bench-to-clinic pipeline, with a streamlined workflow, for implementation of a diverse portfolio of phase 1 trials.

Exploring clinical variability in gelsolin amyloidosis: Brazilian family case study with confocal microscopy.

INTRODUCTION: Genetic mutations or inflammatory, degenerative, or neoplastic conditions can trigger amyloidosis. Hereditary gelsolin amyloidosis is a genetic disorder primarily marked by amyloid fibrils composed of misfolded gelsolin fragments. CASE REPORT: We present three sisters with AGel amyloidosis, illustrating its clinical diversity. Patient 1, a 51-year-old, had bilateral ptosis, ocular discomfort, and dry eye syndrome due to cranial nerve involvement. Patient 2, a 53-year-old, experienced progressive bilateral visual impairment. Patient 3, a 50-year-old, exhibited right eye ectropion. Genetic analysis, with the identical mutation, heterozygous c.640G > A (p.Asp214Asn) mutation, confirmed AGel amyloidosis diagnoses, with common findings including lattice corneal amyloidosis, reduced corneal sensitivity, and recurrent corneal erosions. Neurological manifestations included ataxia and peripheral neuropathy, with skin abnormalities observed in patient 1. Ocular involvement severity and distribution varied among patients. DISCUSSION: Common ocular and neurological manifestations validated AGel amyloidosis diagnoses, reinforcing its hereditary basis. Neurological symptoms highlighted the disorder's impact on various organ systems, while skin abnormalities contributed to ocular discomfort. Variable ocular involvement emphasized the disorder's heterogeneity. These patients emphasize hereditary gelsolin amyloidosis's clinical diversity and suggest potential environmental influences on disease expression. Genetic confirmation and confocal microscopy findings reaffirm the genetic basis while raising questions about assessing systemic disease severity, necessitating further investigation in larger cohorts. Ophthalmologists' specialized care is crucial for managing ocular symptoms, given the absence of a universal cure.

Experimental warming promotes phytoplankton species sorting towards cyanobacterial blooms and leads to potential changes in ecosystem functioning.

A positive feedback loop where climate warming enhances eutrophication and its manifestations (e.g., cyanobacterial blooms) has been recently highlighted, but its consequences for biodiversity and ecosystem functioning are not fully understood. We conducted a highly replicated indoor experiment with a species-rich subtropical freshwater phytoplankton community. The experiment tested the effects of three constant temperature scenarios (17, 20, and 23 °C) under high-nutrient supply conditions on community composition and proxies of ecosystem functioning, namely resource use efficiency (RUE) and CO2 fluxes. After 32 days, warming reduced species richness and promoted different community trajectories leading to a dominance by green algae in the intermediate temperature and by cyanobacteria in the highest temperature treatments. Warming promoted primary production, with a 10-fold increase in the mean biomass of green algae and cyanobacteria. The maximum RUE occurred under the warmest treatment. All treatments showed net CO2 influx, but the magnitude of influx decreased with warming. We experimentally demonstrated direct effects of warming on phytoplankton species sorting, with negative effects on diversity and direct positive effects on cyanobacteria, which could lead to potential changes in ecosystem functioning. Our results suggest potential positive feedback between the phytoplankton blooms and warming, via lower net CO2 sequestration in cyanobacteria-dominated, warmer systems, and add empirical evidence to the need for decreasing the likelihood of cyanobacterial dominance.

Rotavirus A and D circulating in commercial chicken flocks in southeastern Brazil.

Rotavirus (RV) outbreaks can cause significant economic losses in the livestock and poultry industries. Stool samples were collected from asymptomatic laying and broiler chickens from commercial poultry farms in the states of Rio de Janeiro and Espírito Santo in southeastern Brazil for detection of RV species A and D (RVA and RVD, respectively) by reverse transcription polymerase chain reaction. RV was detected in 10.5% (34/325) of samples: 22 (64.7%) were positive for RVA and nine (26.5%) for RVD, while three (8.8%) exhibited coinfections with both viruses. Sequence analysis of a VP6 fragment from seven RVA-positive samples identified the I11 genotype in all samples. Information regarding avian RV epidemiology is still scanty, despite the high prevalence of RV infections in several bird species and subsequent economic impact. Consequently, monitoring infections caused by avian RVs, especially in commercial birds, is essential not only to provide new and relevant information regarding the biology, epidemiology, and evolution of these viruses, but also to facilitate the implementation of preventive measures.

Expanding the Potential of Self-Assembled Silk Fibroin as Aerogel Particles for Tissue Regeneration.

A newly produced silk fibroin (SF) aerogel particulate system using a supercritical carbon dioxide (scCO2)-assisted drying technology is herein proposed for biomedical applications. Different concentrations of silk fibroin (3%, 5%, and 7% (w/v)) were explored to investigate the potential of this technology to produce size- and porosity-controlled particles. Laser diffraction, helium pycnometry, nitrogen adsorption-desorption analysis and Fourier Transform Infrared with Attenuated Total Reflectance (FTIR-ATR) spectroscopy were performed to characterize the physicochemical properties of the material. The enzymatic degradation profile of the SF aerogel particles was evaluated by immersion in protease XIV solution, and the biological properties by cell viability and cell proliferation assays. The obtained aerogel particles were mesoporous with high and concentration dependent specific surface area (203-326 m2/g). They displayed significant antioxidant activity and sustained degradation in the presence of protease XIV enzyme. The in vitro assessment using human dermal fibroblasts (HDF) confirm the particles' biocompatibility, as well as the enhancement in cell viability and proliferation.

Levels of angiopoietin-2 are predictive for mortality in yellow fever virus infected patients.

In 2018 there was a large yellow fever outbreak in São Paulo, Brazil, with a high fatality rate. Yellow fever virus can cause, among other symptoms, haemorrhage and disseminated intravascular coagulation, indicating a role for endothelial cells in the disease pathogenesis. Here, we conducted a case-control study and measured markers related to endothelial damage in plasma and its association with mortality. We found that angiopoietin-2 is strongly associated with a fatal outcome and could serve as a predictive marker for mortality. This could be used to monitor severe patients and provide care to improve disease outcome.

Mechanism of RanGTP priming H2A-H2B release from Kap114 in an atypical RanGTP•Kap114•H2A-H2B complex.

Previously, we showed that the nuclear import receptor Importin-9 wraps around the H2A-H2B core to chaperone and transport it from the cytoplasm to the nucleus. However, unlike most nuclear import systems where RanGTP dissociates cargoes from their importins, RanGTP binds stably to the Importin-9•H2A-H2B complex, and formation of the ternary RanGTP•Importin-9•H2A-H2B complex facilitates H2A-H2B release to the assembling nucleosome. It was unclear how RanGTP and the cargo H2A-H2B can bind simultaneously to an importin, and how interactions of the three components position H2A-H2B for release. Here, we show cryo-EM structures of Importin-9•RanGTP and of its yeast homolog Kap114, including Kap114•RanGTP, Kap114•H2A-H2B, and RanGTP•Kap114•H2A-H2B, to explain how the conserved Kap114 binds H2A-H2B and RanGTP simultaneously and how the GTPase primes histone transfer to the nucleosome. In the ternary complex, RanGTP binds to the N-terminal repeats of Kap114 in the same manner as in the Kap114/Importin-9•RanGTP complex, and H2A-H2B binds via its acidic patch to the Kap114 C-terminal repeats much like in the Kap114/Importin-9•H2A-H2B complex. Ran binds to a different conformation of Kap114 in the ternary RanGTP•Kap114•H2A-H2B complex. Here, Kap114 no longer contacts the H2A-H2B surface proximal to the H2A docking domain that drives nucleosome assembly, positioning it for transfer to the assembling nucleosome or to dedicated H2A-H2B chaperones in the nucleus.

Release Assays and Potency Assays for CAR T-Cell Interventions.

Chimeric antigen receptor (CAR) T-cells are considered "living drugs" and offer a compelling alternative to conventional anticancer therapies. Briefly, T-cells are redirected, using gene engineering technology, toward a specific cancer cell surface target antigen via a synthetic chimeric antigen receptor (CAR) protein. CARs have a modular design comprising four main structures: an antigen-binding domain, a hinge region, a transmembrane domain, and one or more intracellular signaling domains for T-cell activation. A major challenge in the CAR T-cell manufacturing field is balancing product quality with scalability and cost-effectiveness, especially when transitioning from an academic clinical trial into a marketed product, to be implemented across many collection, manufacturing, and treatment sites. Achieving product consistency while circumnavigating the intrinsic variability associated with autologous products is an additional barrier. To overcome these limitations, a robust understanding of the product and its biological actions is crucial to establish a target product profile with a defined list of critical quality attributes to be assessed for each batch prior to product certification. Additional challenges arise as the field progresses, such as new safety considerations associated with the use of allogenic T-cells and genome editing tools. In this chapter, we will discuss the release and potency assays required for CAR T-cell manufacturing, covering their relevance, current challenges, and future perspectives.

Molecular Characterization of Avian Rotaviruses F and G Detected in Brazilian Poultry Flocks.

Avian rotaviruses (RVs) are important etiologic agents of gastroenteritis in birds. In general, avian RVs are understudied; consequently, there is a paucity of information regarding these viruses. Therefore, the characterization of these viral species is highly relevant because more robust information on genetic, epidemiologic, and evolutionary characteristics can clarify the importance of these infections, and inform efficient prevention and control measures. In this study, we describe partial genome characterizations of two avian RV species, RVF and RVG, detected in asymptomatic poultry flocks in Brazil. Complete or partial sequences of at least one of the genomic segments encoding VP1, VP2, VP4, VP6, VP7, NSP1, NSP4, NSP4, or NSP5 of 23 RVF and 3 RVG strains were obtained, and demonstrated that multiple variants of both RVF and RVG circulate among Brazilian poultry. In this study, new and important information regarding the genomic characteristics of RVF and RVG is described. In addition, the circulation of these viruses in the study region and the genetic variability of the strains detected are demonstrated. Thus, the data generated in this work should help in understanding the genetics and ecology of these viruses. Nonetheless, the availability of a greater number of sequences is necessary to advance the understanding of the evolution and zoonotic potential of these viruses.

Development of a chlorhexidine delivery system based on dental reline acrylic resins.

The high recurrence rate of common denture stomatitis after antifungal treatment is still concerning. This condition is caused by low patient compliance and incomplete local elimination of the main etiological factor - Candida albicans, often associated with other microorganisms, such as Streptococcus species. Impregnating denture materials with antimicrobials for local delivery is a strategy that can overcome the side effects and improve the efficacy of conventional treatments (topical and/or systemic). In this work, we describe the development of three hard autopolymerizing reline acrylic resins (Kooliner, Ufi Gel Hard, and Probase Cold) loaded with different percentages of chlorhexidine (CHX). The novel formulations were characterized based on their antimicrobial activity, mechanical, morphological and surface properties, in-vitro drug release profiles, and cytotoxicity. The addition of CHX in all resins did not change their chemical and mechanical structure. Among all the tested formulations, Probase Cold loaded with 5 wt% CHX showed the most promising results in terms of antimicrobial activity and lack of serious detrimental mechanical, morphological, surface, and biological properties.

Viral Kinetics in Sylvatic Yellow Fever Cases.

BACKGROUND: Yellow fever is a mosquito-borne zoonotic disease caused by yellow fever virus (YFV). Between 2017 and 2019, more than 504 human cases and 176 deaths were confirmed in the outskirts of São Paulo city. Throughout this outbreak, studies suggested a potential association between YFV viremia and mortality. METHODS: Viral ribonucleic acid was measured using reverse-transcription quantitative polymerase chain reaction in plasma samples collected at up to 5 time points, between 3 and 120 days after symptoms onset. RESULTS: Eighty-four patients with confirmed YFV infection were included. Most were males, median age was 42, and 30 (36%) died. Deceased patients were older than survivors (P = .003) and had a higher viremia across all time points (P = .0006). Mean values of viremia had a positive, statistically significant correlation with peak values of neutrophils, indirect bilirubin, aspartate transaminase, international normalized ratio, and creatinine. Finally, a Cox proportional hazards model adjusted for age and laboratory variables showed that viremia is independently associated with death, with a mean 1.84-fold increase (84%) in the hazard of death (P < .001) for each unit increase in mean log10 viremia. CONCLUSIONS: Our results raise the importance of monitoring YFV viremia and suggest a potential benefit of antiviral drugs or neutralizing monoclonal antibodies early in the course of this infection to improve disease outcomes.

Autologous stem cell transplantation improves cardiopulmonary exercise testing outcomes in systemic sclerosis patients.

OBJECTIVES: Autologous haematopoietic stem cell transplantation (AHSCT) is a disease-modifying treatment for patients with severe SSc. Here, we aimed at assessing cardiopulmonary function outcomes of SSc patients after AHSCT. METHODS: Twenty-seven SSc adult patients treated with AHSCT were included in this retrospective study. Most had the diffuse cutaneous subset (93%) and pulmonary involvement (85%). Before and 12 months after AHSCT, patients underwent cardiopulmonary exercise testing, transthoracic echocardiography, pulmonary function test with diffusing capacity for carbon monoxide (DLCO), 6-min walk test (6MWT) and quality of life evaluations. RESULTS: After AHSCT, the peak VO2 increased from 954 to 1029 ml/min (P = 0.02), the percentage of predicted peak VO2 increased from 48.9 to 53.5 m (P = 0.01), and the distance measured by the 6MWT increased from 445 to 502 m (P = 0.01), compared with baseline. Improvements in peak VO2 correlated positively with improvements in 6MWT distance, and negatively with a decrease in resting heart rate. At baseline, patients with DLCO >70% had higher peak VO2 values than those with DLCO <70% (P = 0.04), but after AHSCT all patients showed improved VO2 values, regardless of baseline DLCO levels. Increases in VO2 levels after AHSCT positively correlated with increases in the physical component scores of the Short Form-36 quality of life questionnaire (r = 0.70; P = 0.0003). CONCLUSION: AHSCT improves the aerobic capacity of SSc patients probably reflecting combined increments in lungs, skeletal muscle and cardiac function.

Caracterização do ambiente alimentar de uma universidade pública do estado do Rio de Janeiro / Characterization of the food environment in a public university of Rio de Janeiro State

Resumo Introdução O ambiente alimentar pode influenciar as escolhas alimentares e de bebidas pela população e seu estado nutricional. Objetivo Caracterizar o ambiente alimentar dos campi de universidade federal do Rio de Janeiro. Método Estudo transversal com estabelecimentos que comercializavam alimentos e bebidas dentro dos campi da universidade. A avaliação do ambiente alimentar foi realizada com a aplicação de um instrumento para coleta das informações: tipo de estabelecimento, alimentos/preparações, modalidade, características, cardápio e informações nutricionais, promoções, propagandas, disponibilidade de alimentos/bebidas, variedade e preço. Os alimentos e preparações observados, a presença de estratégias como informação nutricional, propagandas e promoções foram classificados em facilitadores/ barreiras para alimentação saudável, de acordo com o Guia Alimentar para a População Brasileira. Resultados Participaram do estudo cinco estabelecimentos, quatro eram comerciais e um o restaurante universitário (RU). Todos os estabelecimentos aceitavam dinheiro e cartão, havia cardápio e preço na maioria. Nenhum apresentava informação nutricional. A oferta de frutas/hortaliças foi encontrada em quatro estabelecimentos. A disponibilidade de edulcorante artificial foi encontrada em todos os locais. Observou-se grande variedade de balas (8 ± 7,1), refrigerantes (9 ± 3,7), chocolate (8 ± 7,1), biscoito salgado (8 ± 12,3) e doce (7 ± 9,1). Frutas apresentaram preço médio superior, R$ 3,83, quando comparadas aos salgados, R$ 2,85 e biscoitos, R$ 1,50. Conclusão O ambiente alimentar universitário apresentou maior oferta de alimentos não saudáveis, apresentando barreiras na promoção de práticas alimentares mais adequadas.

Abstract Background The food environment can influence food and beverage choice by the population and their nutritional status. Objective To characterize the food environment of campuses at the Federal University of Rio de Janeiro. Method A cross-sectional study of food and beverage establishments within the university was carried out. The assessment of the food environment was performed with the application of an instrument to collect information: type, food or preparations, mode, characteristics, menu and nutritional information, promotions, advertisements, availability of food or drinks, variety, and price. The observed foods and preparations, the presence of an approach such as nutritional information, advertisements, and promotions were classified as facilitators/barriers for healthy eating, according to the Food Guide for the Brazilian Population. Results Of the 5 establishments that participated, 4 were commercial and 1 was a university restaurant (RU). All establishments accepted cash and cards, and most had a menu and price. None had nutritional information. The supply of fruits and vegetables was found in 4 establishments. The availability of artificial sweetener was found in all locations. A wide variety of candies (8 ± 7.1), soft drinks (9 ± 3.7), chocolate (8 ± 7.1), salted biscuits (8 ± 12.3), and sweets (7 ± 9.1) were observed. Fruits had a higher average price, R$3.83, when compared to savory foods, R$ 2.85, and biscuits, R$ 1.50. Conclusion The university food environment presented a greater offer of unhealthy foods, presenting barriers to the promotion of more appropriate eating practices.

Plastic debris decrease fish feeding pressure on tropical reefs.

Fish feeding on the benthos mediate important ecological processes in reef ecosystems that are threatened by several anthropogenic activities, including plastic pollution. The impacts of plastic pollution on reef ecological processes, such as herbivory, is still unclear. We experimentally tested in the field how the presence of plastic on the benthos affects fish feeding by using a blocked design to compare fish feeding within 1m2 plastic-free areas and areas with either clean plastic or plastic colonized with biofilm in reefs of NE Brazil. Each area was videoed for 10 min, through which we identified fish species, estimated sizes and quantified the number of bites and feeding frequency. The presence of plastic reduced fish feeding on the benthos, regardless of the biofilm presence, and fish barely interacted with plastic debris. Our results demonstrate that the impact of plastic goes beyond entanglement and ingestion, affecting species behavior and ecological role.

Humoral and cellular immune responses to CoronaVac up to one year after vaccination.

Coronavac is a widely used SARS-CoV-2 inactivated vaccine, but its long-term immune response assessment is still lacking. We evaluated SARS-CoV-2-specific immune responses, including T cell activation markers, antigen-specific cytokine production and antibody response following vaccination in 53 adult and elderly individuals participating in a phase 3 clinical trial. Activated follicular helper T (Tfh), non-Tfh and memory CD4+ T cells were detected in almost all subjects early after the first vaccine dose. Activated memory CD4+ T cells were predominantly of central and effector memory T cell phenotypes and were sustained for at least 6 months. We also detected a balanced Th1-, Th2- and Th17/Th22-type cytokine production that was associated with response over time, together with particular cytokine profile linked to poor responses in older vaccinees. SARS-CoV-2-specific IgG levels peaked 14 days after the second dose and were mostly stable over one year. CoronaVac was able to induce a potent and durable antiviral antigen-specific cellular response and the cytokine profiles related to the response over time and impacted by the senescence were defined.

Arginine-Modified 3D-Printed Chromatographic Supports.

The increasing progression of biopharmaceutical-based therapies highlights the demand for efficient chromatographic methods that can be used to purify the desired biomolecules (e.g., nucleic acids, enzymes, or monoclonal antibodies) which are presently under consideration in clinical trials or approved by the Food and Drug Administration. These molecules present distinct chemical and structural properties, which are critical cues for the development and production of adequate chromatographic supports. Until now, it has not been possible to fully control the characteristics of the chromatographic matrices to assure the total reproducibility of their structure and packing. Meanwhile, three-dimensional printing (3DP) is in the early stage of its use in the production of chromatographic supports as a fast, very precise, and reproducible methodology. Although 3DP can provide excellent performance properties to the chromatographic structures, it cannot, per se, lead to high-quality pharmaceutical products. However, the association of affinity ligands, such as amino acids, which is possible in 3DP, could enable the attainment of high-purity yields of the desired molecules. Beyond the amino acids most widely studied as chromatographic ligands, arginine has been successfully immobilized on different chromatographic supports (namely, agarose bead matrices, macroporous matrices, and monoliths) to achieve extra-pure gene therapy products. In this research, we studied the immobilization of arginine on 3DP chromatographic supports, evaluating the stability of the ligand/chromatographic support linkage under different chromatographic conditions to determine the robustness of these new prototypes. Moreover, we also applied plasmid DNA samples to these supports to observe the practical behaviour of the developed arginine 3DP chromatographic structures.

Evaluation of an EPID in vivo monitoring system using local and external independent audit measurements.

PURPOSE: The aim of this work was to evaluate the SunCHECK PerFRACTION, the software for in vivo monitoring using EPID images. MATERIALS/METHODS: First, the PerFRACTION ability to detect errors was investigated simulating two situations: (1) variation of LINAC output and (2) variation of the phantom thickness. An ionization chamber was used as reference to measure the introduced dose variations. Both tests used EPID in integrated mode (absolute dose). Second, EPID measurements in integrated mode were carried out during an independent Brazilian governmental audit that provided four phantoms and TLDs. PerFRACTION calculated the absolute dose on EPID plane, and it compared with predicted calculated dose for every delivered plan. The dose deviations reported using PerFRACTION were compared with dose deviations reported by the independent audit. Third, an end-to-end test using a heterogeneous phantom was performed. A VMAT plan with EPID in cine mode was delivered. PerFRACTION calculated the mean dose on CBCT using EPID information and log files. The calculated doses at four different points were compared with ionization chambers measurements. RESULTS: About the first test, the largest difference found was 1.2%. Considering the audit results, the variations detected by TLD measurements and by PerFRACTION dose calculation on EPID plane were close: 12 points had variations less than 2%, 2 points with variation between 2% and 3%, and 2 points with deviations greater than 3% (max 3.7%). The end-to-end tests using a heterogeneous phantom achieved dose deviation less than 1.0% in the water-equivalent region. In the mimicking lung region, the deviations were higher (max 7.3%), but in accordance with what is expected for complex situations. CONCLUSION: The tests results indicate that PerFRACTION dose calculations in different situations have good agreement with standard measurements. Action levels were suggested for absolute dose on EPID plane as well as 3D dose calculation on CBCT using PerFRACTION.