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ABSTRACT This study broadens the knowledge of plant-pathogen interactions using phytohormone-deficient and low-sensitivity mutant tomatoes. The experiment was performed under greenhouse conditions and the mutants used were provided by the HCPD laboratory, at ESALQ/USP. The hormone mutant Never ripe showed a larger final lesion size when inoculated with Botrytis cinerea, whereas curl3 was relatively resistant to this pathogen and more susceptible when inoculated with Phytophthora infestans. The transgenic 35S :: nahG was more susceptible to Oidium lycopersici, and the double mutant dgt, Nr more resistant. The low sensitivity to brassinosteroids confers greater susceptibility to P. infestans and higher resistance to B. cinerea. The deficiency of the hormone auxin promotes high resistance to diseases caused by O. lycopersici, P. infestans and B. cinerea.

O presente artigo apresenta os resultados iniciais da cooperação entre o estado de Minas Gerais e os municípios mineiros na implementação do Ensino Fundamental de nove anos, o que resultou em números bastante expressivos para as escolas públicas do estado nas avaliações externas.

The classification of the type of trapped particles is a crucial task for an efficient integration of optical-tweezers in intelligent microfluidic devices. In the recent years, the use of the temporal scattering signal of the trapped particle paved for the use of versatile optical fiber solutions for performing such tasks, a feature previously unavailable as most methods required conventional optical tweezer setups. This work presents a comprehensive comparison of performances achieved with two distinct implementations – i)optical fiber and ii)conventional optical tweezers – for the classification of the material of particles through the analysis of the scattering signal with machine learning algorithms. The results suggest that while micron-sized particles can be accurately classified using the forward scattering information in conventional optical tweezers, when equipped with a quadrant photodetector, the optical fiber tweezers solutions can easily surpass its performance using the back-scattered information if the laser is modulated. Together with the advantages of being simpler, less expensive and more versatile, the results presented suggest that optical fiber solutions can become a valuable tool for miniaturization and integration of intelligent microfluidic devices working towards nanoscopic scales.

The invasion of natural communities by non-indigenous species represents one of the most serious threats to biodiversity. Understanding the ecophysiology of invasive species can provide insights into potential physiological handicaps relative to native species. By doing so, we can leverage the development of ecoengineering solutions for the removal of non-indigenous species, preferably using non-chemical methods. Spartina patens is a known invasive species of cordgrass aggressively proliferating in Mediterranean salt marshes, producing impenetrable monospecific stands. As its occurrence is delimited by the upper high tide water level, we hypothesized that S. patens is intolerant to waterlogging. Therefore, we developed a field experiment where strands of S. patens were kept waterlogged over the entire tidal cycle for 30 days. At the end of the experimental period, plants in the trial plots exhibited severe stress symptoms at different physiological levels compared with control plots (no intervention). At the photobiological level, intervened plants exhibited lower efficiency in producing chemical energy from light, whilst at the biochemical level waterlogging impaired the antioxidant system and increased lipid peroxidation products. Furthermore, the application of chlorophyll a pulse amplitude modulated (PAM) fluorometry, a non-invasive technique, allowed us to evaluate the effectiveness of the implemented measures, being the tool that provided the best separation between the control and intervened population. Considering the physiological traits observed here, ecoengineering solutions based on increased waterlogging of S. patens stands, can be a low-cost and efficient measure to reduce the spreading and growth of this invasive species in the Mediterranean and other salt marshes worldwide with little disturbance.

Outer membrane vesicles (OMVs) are small vesicles constitutively shed by all Gram-negative bacterium, which have been proposed to play a role in Helicobacter pylori persistence and pathogenesis. The methods currently available for the isolation of H. pylori OMVs are diverse and time-consuming, raising the need for a protocol standardization, which was the main aim of this study. Here, we showed that the chemically defined F12 medium, supplemented with cholesterol, nutritionally supports bacterial growth and maintains H. pylori viability for at least 72 h. Additionally, we developed an abridged protocol for isolation of OMVs from these bacterial cultures, which comprises a low-speed centrifugation, supernatant filtration through a 0.45 μm pore, and two ultracentrifugations for OMVs’ recovery and washing. Using this approach, a good yield of highly pure bona fide OMVs was recovered from cultures of different H. pylori strains and in different periods of bacterial growth, as assessed by nanoparticle tracking analysis, transmission electron microscopy (TEM), and proteomic analyses, confirming the reliability of the protocol. Analysis of the proteome of OMVs isolated from H. pylori F12-cholesterol cultures at different time points of bacterial growth revealed differentially expressed proteins, including the vacuolating cytotoxin VacA. In conclusion, this work proposes a time- and cost-efficient protocol for the isolation of H. pylori OMVs from a chemically defined culture medium that is suitable for implementation in research and in the biopharmaceutical field.

ABSTRACT: The differentiation of seedling growth and development is the result of the vigor level of the seed lot. Because of this, knowing the seedling growth curve is essential for the evaluation of the seedling length test, to be carried out at the moment when any differences in vigor really manifest, thus being able to reduce the time of evaluation of the test. In this research, the objective was to determine the most efficient time interval to perform the seedling length test in soybean seeds, depending on the vigor levels. The evaluated characteristics were percentage of seed germination, first germination count, emergency, days for emergence, aerial, root and total length (evaluated every 12 hours), aerial, root and total dry mass in completely randomized design. High vigor seeds originated seedlings with higher growth rates (cm. h-1), with vigor being one of the factors that interfere in the seedling length test. After 96 hours of the seedling length test, there was a statistical differentiation between the three vigor levels tested. The soybean seedling length test between lots, with different vigor, can be performed 96 hours after sowing.

Forecasting COVID-19 disease severity is key to supporting clinical decision making and assisting resource allocation, particularly in intensive care units (ICUs). Here, we investigated the utility of time- and frequency-related features of the backscattered signal of serum patient samples to predict COVID-19 disease severity immediately after diagnosis. ICU admission was the primary outcome used to define disease severity. We developed a stacking ensemble machine learning model including the backscattered signal features (optical fingerprint), patient comorbidities, and age (AUROC = 0.80), which significantly outperformed the predictive value of clinical and laboratory variables available at hospital admission (AUROC = 0.71). The information derived from patient optical fingerprints was not strongly correlated with any clinical/laboratory variable, suggesting that optical fingerprinting brings unique information for COVID-19 severity risk assessment. Optical fingerprinting is a label-free, real-time, and low-cost technology that can be easily integrated as a front-line tool to facilitate the triage and clinical management of COVID-19 patients.

Objetivou-se avaliar o efeito do fosfito de potássio como indutor de resistência no manejo de Phytophthora infestans, agente causal da requeima, em mutantes de tomateiro para tricomas e hormônios, para identificar seu modo de ação e as possíveis vias de sinalização envolvidas. Foram utilizados onze genótipos mutantes (hair absent, Wooly, hairless, Galapagos, notabilis, Never ripe, epinastic, procera, curl3, 35S::nahG e dgt, Nr), que estão no mesmo background genético que o parental selvagem, cultivar Micro-Tom. O experimento foi realizado em delineamento inteiramente casualizado, em esquema bifatorial, com quatro repetições. O fator A correspondeu aos genótipos utilizados e o fator B a aplicação ou não do indutor. As variáveis analisadas foram: período de incubação (PI), tamanho final da lesão (TFL) e taxa de expansão da lesão (r). Para todas as variáveis houve interação significativa entre os genótipos e a aplicação do fosfito de potássio. Nas plantas tratadas com fosfito de potássio, o PI no genótipo Galapagos foi significativamente menor de 52 até 70% em relação aos genótipos mutantes, exceto Never ripe e 35S::nahG. A aplicação do fosfito de potássio aumentou significativamente em 63, 53, e 70% o TFL para os genótipos Wooly, hairless e procera, respectivamente. A r no genótipo Procera foi significativamente menor em 76% à observada no Micro-Tom, e de 50 até 76% em relação aos genótipos mutantes, quando as plantas não foram tratadas com fosfito de potássio. A rota de sinalização mais provável para indução de resistência por fosfito de potássio é a do ácido salicílico.

O estudo teve como objetivo descrever e analisar as notificações de suspeitas de Reações Adversas a Medicamentos (RAM) do serviço de transplante de um hospital sentinela de Fortaleza, Ceará. Tratou-se de uma pesquisa descritiva, transversal e observacional que utilizou, como fonte secundária, as fichas de notificação de suspeita de RAM da gerência de riscos do ano de 2014. No período analisado, foram realizadas 35 notificações envolvendo pacientes transplantados, 52 medicamentos e 50 reações na gerência de risco do Hospital Universitário Walter Cantídio. Todas as notificações foram realizadas de forma espontânea, sendo 94,5% feitas pelo farmacêutico. Os grupos de idade dos pacientes mais acometidos foram os de 50-60 anos e de >60, com uma média de 51,8 (± 14,1) anos. As RAM acometeram principalmente homens (55,0%), transplantados renais (69,7%). Os principais medicamentos envolvidos foram ganciclovir (21,2%), tacrolimus (13,5%) e micofenolato de sódio (11,5%). A maioria das RAM estava relacionada às afecções da pele e distúrbios afins (23,0%), sendo 77,0% prurido e 23,0% eritema. Analisando a classificação de causalidade das RAM, 46,0% foram classificadas como possível e 88,0% foram classificadas quanto à gravidade como moderada. Conclui-se que as suspeitas de reações adversas notificadas em pacientes transplantados estavam relacionadas ao tratamento farmacoterapêutico comumente estabelecido no pós-transplante, levando a necessidade da monitorização.

Objetivo: Contribuir para a formação do conhecimento, no Brasil, sobre o impacto do papel do farmacêutico clínico no cuidado ao paciente pós-transplante internado, analisando os resultados das intervenções farmacêuticas realizadas. Método: Foi realizado um estudo descritivo de janeiro a julho/2014. Os dados foram coletados dos registros do Serviço de Farmácia Clínica (conciliação medicamentosa, análise de prescrição e acompanhamento farmacoterapêutico), incluindo todos os pacientes com problemas relacionados a medicamentos e intervenções farmacêuticas documentadas. Variáveis epidemiológicas e relacionadas às atividades clínicas do farmacêutico foram descritas. Resultados: Um total de 131 pacientes apresentaram problemas relacionados a medicamentos, especialmente durante o "Acompanhamento Farmacoterapêutico". As categorias mais frequentes foram: "medicamento não prescrito" (125; 21.7%), "sobredose" (97; 16.8%) e "subdose" (93; 16.1%). Os principais fármacos envolvidos foram "ganciclovir" (81; 13.4%) e "tacrolimus" (33; 5.5%). A intervenção farmacêutica incluiu "dose (adequação)" (193; 33.4%), "inclusão" (122; 21.1%) ou "suspensão" (122; 21.1%) do medicamento. Os principais resultados clínicos foram "prevenção" (481; 83.4%) ou "melhora" (59; 10.2%) de problemas de saúde, como "efeitos adversos" (170; 29.5%), “infecção” (133; 23%) e "rejeição" (41; 7.1%), sendo verificada uma relação significativa (p

This paper is the story of a critical archival pedagogy that emerged through the undergraduate course Radical Lesbian Thought. As teachers and students, we dialogically co-constructed the praxis and content of the course throughout the semester. We employed archives throughout the course as theory, site, and pedagogy. In this paper we identify three archival frameworks: dialogue and difference, collaborative knowledge production, and archival methodology; and detail how they informed three course activities: reading and writing archival letters, visiting the Lesbian Herstory Archives, and completing final archival projects. We argue that archives provide theoretical and practical opportunities, in the tradition of critical pedagogy, to challenge and rearrange powered classroom structures and practices of thought.

A CAPN1 missense mutation in Parson Russell Terrier dogs is associated with spinocerebellar ataxia. We now report that homozygous or heterozygous CAPN1-null mutations in humans result in cerebellar ataxia and limb spasticity in four independent pedigrees. Calpain-1 knockout (KO) mice also exhibit a mild form of ataxia due to abnormal cerebellar development, including enhanced neuronal apoptosis, decreased number of cerebellar granule cells, and altered synaptic transmission. Enhanced apoptosis is due to absence of calpain-1-mediated cleavage of PH domain and leucine-rich repeat protein phosphatase 1 (PHLPP1), which results in inhibition of the Akt pro-survival pathway in developing granule cells. Injection of neonatal mice with the indirect Akt activator, bisperoxovanadium, or crossing calpain-1 KO mice with PHLPP1 KO mice prevented increased postnatal cerebellar granule cell apoptosis and restored granule cell density and motor coordination in adult mice. Thus, mutations in CAPN1 are an additional cause of ataxia in mammals, including humans.

Multiple myeloma (MM) is the second most common blood cancer. Treatments for MM include corticosteroids, alkylating agents, anthracyclines, proteasome inhibitors, immunomodulatory drugs, histone deacetylase inhibitors and monoclonal antibodies. Survival outcomes have improved substantially due to the introduction of many of these drugs allied with their rational use. Nonetheless, MM patients successively relapse after one or more treatment regimens or become refractory, mostly due to drug resistance. This review focuses on the main drugs used in MM treatment and on causes of drug resistance, including cytogenetic, genetic and epigenetic alterations, abnormal drug transport and metabolism, dysregulation of apoptosis, autophagy activation and other intracellular signaling pathways, the presence of cancer stem cells, and the tumor microenvironment. Furthermore, we highlight the areas that need to be further clarified in an attempt to identify novel therapeutic targets to counteract drug resistance in MM patients.

Large numbers of well-characterized clinical samples are fundamental to establish relevant associations between the microbiota and disease. Formalin-fixed and paraffin-embedded (FFPE) tissues are routinely used and are widely available clinical materials. Since current approaches to study the microbiota are based on next-generation sequencing (NGS) targeting the bacterial 16S rRNA gene, our aim was to evaluate the feasibility of FFPE gastric tissues for NGS-based microbiota characterization. Analysis of sequencing data revealed the presence of bacteria in the paraffin control. After the subtraction of the operational taxonomic units (OTUs) present in the paraffin control to the FFPE tissue sample dataset, we evaluated the microbiota profiles between paired FFPE and frozen gastric tissues, and between different times of archiving. Compared with frozen gastric tissues, we detected a lower number of OTUs in the microbiota of paired FFPE tissues, regardless of the time of archiving. No major differences in microbial diversity were identified, but taxonomic variation in the relative abundance of phyla and orders was observed between the two preservation methods. This variation was also evident in each case and throughout the times of FFPE archiving. The use of FFPE tissues for NGS-based microbiota characterization should be considered carefully, as biases can be introduced by the embedding process and the time of tissue archiving.

ABSTRACT Objective: To describe and analyze the pharmaceutical orientation given at hospital discharge of transplant patients. Methods: This was a cross-sectional, descriptive and retrospective study that used records of orientation given by the clinical pharmacist in the inpatients unit of the Kidney and Liver Transplant Department, at Hospital Universitário Walter Cantídio, in the city of Fortaleza (CE), Brazil, from January to July, 2014. The following variables recorded at the Clinical Pharmacy Database were analyzed according to their significance and clinical outcomes: pharmaceutical orientation at hospital discharge, drug-related problems and negative outcomes associated with medication, and pharmaceutical interventions performed. Results: The first post-transplant hospital discharge involved the entire multidisciplinary team and the pharmacist was responsible for orienting about drug therapy. The mean hospital discharges/month with pharmaceutical orientation during the study period was 10.6±1.3, totaling 74 orientations. The prescribed drug therapy had a mean of 9.1±2.7 medications per patient. Fifty-nine drug-related problems were identified, in which 67.8% were related to non-prescription of medication needed, resulting in 89.8% of risk of negative outcomes associated with medications due to untreated health problems. The request for inclusion of drugs (66.1%) was the main intervention, and 49.2% of the medications had some action in the digestive tract or metabolism. All interventions were classified as appropriate, and 86.4% of them we able to prevent negative outcomes. Conclusion: Upon discharge of a transplanted patient, the orientation given by the clinical pharmacist together with the multidisciplinary team is important to avoid negative outcomes associated with drug therapy, assuring medication reconciliation and patient safety.

AIRmax and AIRmin mice strains were selected according to the intensity of their acute inflammatory responsiveness. Previous studies have shown that AIR mice differ in their resistance to chemically induced skin tumors and in the development of melanoma metastases, in addition to differences in neutrophil and NK cells activity. In the present work, we aimed to evaluate whether the difference of susceptibility to murine melanoma is associated with NK cytotoxic activity against Yac.1 cells and lymphocyte subsets. Mice were subcutaneously inoculated with B16F10 or S91 melanoma cells. After 7, 14, or 30 days, the animals were euthanized to analyze the number of lymphocyte subsets, cytotoxic activity, and number of cytokine-producing spleen cells. AIRmax mice presented a higher number of CD4+/CD25+ cells than that of AIRmin mice following inoculation of B16F10 cells, whereas inoculation of S91 cells reduced CD4+/CD25+ and increased TCD8+ cell subsets in the AIRmax mice. AIRmax mice had a higher number of interleukin (IL)-10- and IL-12-producing cells and a lower number of interferon-γ–producing cells than those of AIRmin mice at 30 days. The cytotoxic activity of nonadherent spleen cells was similar in both the AIR strains. These results suggest that melanoma cells can induce different responses in AIR mice, possibly owing to alterations in regulatory mechanisms, such as the action of CD4+/CD25+ regulatory T cells and IL-10, in AIRmax mice.