Intravital microscopy: imaging host‐parasite interactions in lymphoid organs

Intravital microscopy allows imaging of biological phenomena within living animals, including host‐parasite interactions. This has advanced our understanding of both, the function of lymphoid organs during parasitic infections, and the effect of parasites on such organs to allow their survival. In parasitic research, recent developments in this technique have been crucial for the direct study of host –parasite interactions within organs at depths, speeds, and resolution previously difficult to achieve. Lymphoid organs have gained more attention as we start to understand their function during parasitic infections and the effect of parasites on them. In this review, we summarize technical and biological findings achieved by intravital microscopy with respect to the interaction of various parasites with host lymphoid organs, namely the bone marrow, thymus, lymph nodes, spleen, and the mucosa‐associated lymphoid tissue, and present a view into possible future applications

Identification of S alleles of different pear cultivars of the "Oeste" region and evaluation of their gametophytic compatibility with 'Rocha' pear

In order to select pear cultivar which are genetically compatible with ‘Rocha’ (Sa, Sj), the identification of S alleles sequences of commercial cultivars with bloom periods overlapping (partially or totally) the bloom period of ‘Rocha’ was performed. Partial sequences of the S alleles from the different cultivars were amplified with specific primers, cloned and sequenced. ‘Pêra De Água’ (S22, Sb) ‘Général Leclerc’ (Sl, Sq) and ‘Alexandrine Douillard’ (Sb, Sk) should be totally compatible. It was identified a Sj allele in the cultivars ‘Amêndoa’, ‘Beurré Precoce Morettini’, ‘Clapp’s Rouge’ e ‘Clapp’s Favourite’ and an allele Sa in ‘Beurré Clairgeau’, indicating that these cultivars are semi-compatible with ‘Rocha’. ‘Carapinheira’ (Sb) and ‘Pérola’ (Sk) carry a different allele from the ones of ‘Rocha’ and an amplification pattern completely dissimilar suggesting full compatibility. In ‘Passe Crassane’ (Sr) and ‘Beurré D’Avril’ (S4), a different allele from the S alleles of ‘Rocha’ was identified indicating compatibility, but total or partial compatibility remained unclear, since the amplification pattern does not exclude that the unknown allele might be Sa from ‘Rocha’

Analysis of the expression of S alleles in "Rocha" pear

Analysis of the expression of S alleles in ‘Rocha’ pear. In this work, the expression of incompatibility genes (S-alleles) in ‘Rocha’ pear was investigated. The comparison of these alleles with the alleles of other cultivars may constitute a tool to help the technicians and farmers to choose pollinators when planning a new ‘Rocha’ orchard. We have used molecular techniques that include pistil RNA extraction from three clones of cultivar ‘Rocha’ (R1, R2 and R4C), fragment amplification by RT-PCR, cloning and sequencing. The flowers utilized to extract RNA were isolated before anthesis to guarantee the absence of pollen from other cultivars in their pistils, which could interfere with the rejection process of the auto-pollen. Analysing the genetic sequences obtained from the three clones, no differences between clones were detectable and the sequences corresponded to Sj and Sa alleles. ‘Rocha’ pear is semicompatible with cultivars which do not present Sj or Sa, totally incompatible with cultivars that carry Sj and Sa, and totally compatible with cultivars that do not carry Sj and Sa

Methodology evaluation of pin microrelief meter

The effects of natural weathering and different managements performed in agriculture may best be understood by studying the soil roughness. The aim of this study was to evaluate the optimization of the use of pin microrelief meter, an instrument used to determine the soil surface roughness, as the number of readings collected over traditional methodology proposed in the bibliography. The study was conducted in Rio Paranaiba (MG), in a Haplustox soil. The experimental design was completely randomized in a 2×3 factorial design with four replications. There were combined two types of primary tillage: conventional tillage with disc plow (PCAD) and harrow (PCGA), and three amounts of readings (100, 200, and 300 reading points) sampled in each experimental unit. Independently of the soil tillage, disc plow and harrow, the collection of 100 readings using a pin microrelief meter of a square meter, was sufficient to determine the surface roughness before and after soil preparation, without accuracy loss compared with the traditional method

S6Ks isoforms contribute to viability, migration, docetaxel resistance and tumor formation of prostate cancer cells

Background: The S6 Kinase (S6K) proteins are some of the main downstream effectors of the mammalian Target Of Rapamycin (mTOR) and act as key regulators of protein synthesis and cell growth. S6K is overexpressed in a variety of human tumors and is correlated to poor prognosis in prostate cancer. Due to the current urgency to identify factors involved in prostate cancer progression, we aimed to reveal the cellular functions of three S6K isoforms-p70-S6K1, p85-S6K1 and p54-S6K2-in prostate cancer, as well as their potential as therapeutic targets. Methods: In this study we performed S6K knockdown and overexpression and investigated its role in prostate cancer cell proliferation, colony formation, viability, migration and resistance to docetaxel treatment. In addition, we measured tumor growth in Nude mice injected with PC3 cells overexpressing S6K isoforms and tested the efficacy of a new available S6K1 inhibitor in vitro. Results: S6Ks overexpression enhanced PC3-luc cell line viability, migration, resistance to docetaxel and tumor formation in Nude mice. Only S6K2 knockdown rendered prostate cancer cells more sensitive to docetaxel. S6K1 inhibitor PF-4708671 was particularly effective for reducing migration and proliferation of PC3 cell line. Conclusions: These findings demonstrate that S6Ks play an important role in prostate cancer progression, enhancing cell viability, migration and chemotherapy resistance, and place both S6K1 and S6K2 as a potential targets in advanced prostate cancer. We also provide evidence that S6K1 inhibitor PF-4708671 may be considered as a potential drug for prostate cancer treatment16CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informação2012/13558-

XRD and FTIR analysis of Ti–Si–C–ON coatings for biomedical applications

Ti–Si–C–ON films were deposited by DC reactive magnetron sputtering using different partial pressure ratio of oxygen (pO2) and nitrogen (pN2). Compositional analysis revealed the existence of three different growth zones for the films; (I) N/Ti = 2.1 (high atomic ratio) and low oxygen content; (II) 0.76 < N/Ti < 2.1 (intermediate atomic ratio) and (III) N/Ti ≤ 0.12 (low ratio) and high oxygen content. For high N/Ti atomic ratio (N/Ti = 2.1) the XRD pattern exhibits reflections that correspond to a mixture of two different phases: a metallic-like Ti and a fcc NaCl type structure. Its electrical resistivity presents a metallic character and, consequently, has high infrared reflectivity. For the intermediate N/Ti ratio (0.76 < N/Ti < 2.1), the films crystallize in a B1-NaCl crystal structure typical for TiC0.2N0.8. Their FTIR spectra present C–N modes, besides the TiN ones, that indicate a progressive substitution of nitrogen by carbon atoms with increasing oxygen content (and lowering N/Ti ratio). For the highest oxygen content (and lower N/Ti ratio) the presence of the Ti–O–Ti stretching mode shows the formation of highly resistive Ti–O compounds consistent with the semiconductor character of this film. Biofilm formation as well as material cytotoxicity seemed to be related with the presence of the Ti

The step of incorporation of Bacillus coagulans GBI-30 6086 into “requeijão cremoso” processed cheese does not affect metabolic homeostasis of rats

Dairy product consumption is a common habit in Brazil. These products present a good matrix for probiotic incorporation. Thus, in this study the feasibility of producing a probiotic "requeijao cremoso" incorporated with Bacillus coagulans GBI-30 6086 in three different steps and its metabolic effect in an animal model for 2 weeks has been evaluated. Wistar adult health rats were randomized into one to five groups (n = 8 for each group): Control (C); "requeijao cremoso" without probiotic (RC); probiotic inoculated in the milk before pasteurization at 65 degrees C/30 min (RPP); "requeijao cremoso" inoculated before the fusion step and consequently exposed to 90 degrees C/5 min (RPF); and "requeijao cremoso" inoculated after fusion step, i.e., once the product temperature reached 50 degrees C (RPAF). At the end of treatment, analysis of molecular markers of proteins of stress and antioxidant system, HSP 25, 60, 70 and 90, SOD and catalase were performed in the animals' muscles by Western Blot technique. The HSP25, HSP90 and catalase levels of C, RPP, RPF, and RPAF were similar, indicating that the homeostasis remained unchanged. The incorporation of B. coagulans GBI-30 6086 in the "requeijao cremoso" was shown to be stable and the microorganism remained viable in all steps tested. The incorporation of the probiotic strain in the fusion stage facilitated the technological process, since it allowed a better homogenization of the product and did not affect the maintenance of the metabolic homeostasis of rats10CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPsem informação302763/2014-7; 305804/2017-013/21544-9; 18/24540-8; 2019/21188-

Molecular determinants of the SARS-CoV-2 fusion peptide activity

The COVID-19 pandemic, caused by the SARS-CoV-2 virus, emerged in late 2019 and quickly spread worldwide, resulting in over 125 million infections and 2.7 million deaths as of March 2021 accordingly to the World Health Organization. Despite the great advances achieved by the scientific community in providing crucial information about this virus, we are still far from completely understanding it. SARS-CoV-2 is an enveloped virus, meaning that it is encapsulated by a lipid membrane, which needs to be fused to the host membrane to begin the infection process. Fusion between viral and host membrane is catalyzed by the spike (S) glycoprotein. The S-protein is composed of essential elements for the infection mechanism, namely the receptor-binding domain known to bind to angiotensin-converting enzyme 2 during the viral entry pathway. Another important region, known as the fusion peptide (FP), plays an essential part in the fusion mechanism, by inserting into and disturbing the host membrane. There is still not a consensus among scientists in terms of the fusion peptide location on the S-protein sequence, with two major candidate regions having been proposed. We recently used a machine learning-based tool developed by us to identify viral FPs with accuracies over 85%. With this tool a putative FP, previously suggested in the literature, has been identified, as well as other proposals including the requirement of more than one FP. To further address this question, we are performing a systematic analysis of the SARS-CoV-2 putative FPs, using Molecular Dynamics (MD) simulations, which provide a detailed perspective of how these peptides insert and interact with the membrane. In parallel, we are characterizing these systems experimentally. Additionally we are exploring therapeutic strategies targeting these regions. Given the major role of the FP in the virus infection process, this work provides relevant insights and contributes to the fight against COVID-19.info:eu-repo/semantics/publishedVersio

Proposta de modelo multicritério para avaliação de áreas potenciais para investimento industrial: abordagem MCDA-C: Proposal of a multicriteria model for evaluating potential areas for industrial investment: MCDA-C approach

A presente pesquisa propõe-se a elaborar um modelo multicritério de apoio à tomada de decisão para avaliação de áreas potenciais para investimento industrial observando-se possíveis impactos causados ao público-alvo bem como questões referentes ao desenvolvimento sustentável econômico, social e ambiental da região. Assim, a região analisada, situada ao nordeste catarinense, vem recebendo demanda crescente de potenciais investidores que buscam a melhor opção para viabilizar instalações fabris. Nesta mesma região encontra-se o Porto de Itapoá, cuja atividade está relacionada com o desenvolvimento de apoios estruturais industriais, logísticos, ambientais e de sistemas de informações integradas. Tendo em vista que vários fatores são necessários para a avaliação de áreas potenciais para o investimento industrial, optou-se na presente pesquisa pela elaboração de um modelo de apoio à decisão baseado na abordagem multicritério. Para avaliação foram definidas cinco regiões, das quais duas apresentaram índices considerados de ótimo potencial para instalação industrial. Também foi possível identificar os pontos fortes, fracos, ameaças e oportunidades, e relacionar com a preocupação voltada ao planejamento estratégico e a preservação dos recursos naturais da região analisada. Ao final da pesquisa apresenta-se recomendações para embasamento de planejamento estratégico voltado ao desenvolvimento urbano e industrial

An optimized nanoparticle delivery system based on chitosan and chondroitin sulfate molecules reduces the toxicity of amphotericin B and is effective in treating tegumentary leishmaniasis

Amphotericin B (AmpB) is active against leishmaniasis, but its use is hampered due to its high toxicity observed in patients. In this study, a nanoparticles-delivery system for AmpB (NQC-AmpB), containing chitosan and chondroitin sulfate molecules, was evaluated in BALB/c mice against Leishmania amazonensis. An in vivo biodistribution study, including biochemical and toxicological evaluations, was performed to evaluate the toxicity of AmpB. Nanoparticles were radiolabeled with technetium-99m and injected in mice. The products presented a similar biodistribution in the liver, spleen, and kidneys of the animals. Free AmpB induced alterations in the body weight of the mice, which, in the biochemical analysis, indicated hepatic and renal injury, as well as morphological damage to the kidneys of the animals. In general, no significant organic alteration was observed in the animals treated with NQC-AmpB. Mice were infected with L. amazonensis and treated with the nanoparticles or free AmpB; then, parasitological and immunological analyses were performed. The NQC-AmpB group, as compared to the control groups, presented significant reductions in the lesion size and in the parasite burden in all evaluated organs. These animals presented significantly higher levels of IFN-γ and IL-12, and low levels of IL-4 and IL-10, when compared to the control groups. The NQC-AmpB system was effective in reducing the infection in the animals, and proved to be effective in diminishing the toxicity evoked by AmpB, which was observed when it was administered alone. In conclusion, NQC-AmpB could be considered a viable possibility for future studies in the treatment of leishmaniasisThis work was supported by grants from Pró-Reitoria de Pesquisa from UFMG (Edital 01/2014), Instituto Nacional de Ciência e Tecnologia em Nano-biofarmacêutica (INCT-Nanobiofar), FAPEMIG (CBB-APQ-00496-11 and CBB-APQ-00819-12), and CNPq (APQ-472090/2011-9 and APQ-482976/2012-8). MACF is a grant recipient of FAPEMIG/CAPES. EAFC, VNC, and AAGF are grant recipients of CNPq. Eduardo AF Coelho and André AG Faraco are co-senior authors of this stud