Leishmania amazonensis promastigotes in 3D Collagen I culture: an in vitro physiological environment for the study of extracellular matrix and host cell interactions

Leishmania amazonensis is the causative agent of American cutaneous leishmaniasis, an important neglected tropical disease. Once Leishmania amazonensis is inoculated into the human host, promastigotes are exposed to the extracellular matrix (ECM) of the dermis. However, little is known about the interaction between the ECM and Leishmania promastigotes. In this study we established L. amazonensis promastigote culture in a three-dimensional (3D) environment mainly composed of Collagen I (COL I). This 3D culture recreates in vitro some aspects of the human host infection site, enabling the study of the interaction mechanisms of L. amazonensis with the host ECM. Promastigotes exhibited “freeze and run” migration in the 3D COL I matrix, which is completely different from the conventional in vitro swimming mode of migration. Moreover, L. amazonensis promastigotes were able to invade, migrate inside, and remodel the 3D COL I matrix. Promastigote trans-matrix invasion and the freeze and run migration mode were also observed when macrophages were present in the matrix. At least two classes of proteases, metallo- and cysteine proteases, are involved in the 3D COL I matrix degradation caused by Leishmania. Treatment with a mixture of protease inhibitors significantly reduced promastigote invasion and migration through this matrix. Together our results demonstrate that L. amazonensis promastigotes release proteases and actively remodel their 3D environment, facilitating their migration. This raises the possibility that promastigotes actively interact with their 3D environment during the search for their cellular “home”—macrophages. Supporting this hypothesis, promastigotes migrated faster than macrophages in a novel 3D co-culture model

Application of cupuassu shell as biosorbent for the removal of textile dyes from aqueous solution

AbstractThe cupuassu shell (Theobroma grandiflorum) which is a food residue was used in its natural form as biosorbent for the removal of C.I. Reactive Red 194 and C.I. Direct Blue 53 dyes from aqueous solutions. This biosorbent was characterized by infrared spectroscopy, scanning electron microscopy, and nitrogen adsorption/desorption curves. The effects of pH, biosorbent dosage and shaking time on biosorption capacities were studied. In acidic pH region (pH 2.0) the biosorption of the dyes were favorable. The contact time required to obtain the equilibrium was 8 and 18 h at 298 K, for Reactive Red 194 and Direct Blue 53, respectively. The Avrami fractionary-order kinetic model provided the best fit to experimental data compared with pseudo-first-order, pseudo-second-order and chemisorption kinetic adsorption models. The equilibrium data were fitted to Langmuir, Freundlich, Sips and Radke–Prausnitz isotherm models. For both dyes the equilibrium data were best fitted to the Sips isotherm model

By-products of camu-camu [Myrciaria dubia (Kunth) McVaugh] as promising sources of bioactive high added-value food ingredients: functionalization of yogurts

Camu-camu (Myrciaria dubia (Kunth) McVaugh) is a fruit economically relevant to the Amazon region, mostly consumed in the form of processed pulp. Our aim was to perform an unprecedented comparative study on the chemical composition and bioactivities of the camu-camu pulp and industrial bio-residues (peel and seed), and then the most promising fruit part was further explored as a functionalized ingredient in yogurt. A total of twenty-three phenolic compounds were identified, with myricetin-O-pentoside and cyanindin-3-O-glucoside being the main compounds in peels, followed by p-coumaroyl hexoside in the pulp, and ellagic acid in the seeds. The peel displayed the richest phenolic profile among samples, as well as the most significant antibacterial (MICs = 0.625–10 mg/mL) and anti-proliferative (GI50 = 180 g/mL against HeLa cells) activities. For this reason, it was selected to be introduced in a food system (yogurt). Taken together, our results suggest the possibility of using the camu-camu peel as a source of food additives.Part of this work was supported by the Instituto Federal do Rondônia (IFRO) (Grant nº 81/PROPESP/IFRO). The authors are grateful to FCT—Portugal and FEDER, under Programme PT2020, for financial support to CIMO (UID/AGR/00690/2019) and B. Albuquerque research grant (SFRH/BD/136370/2018); national funding by FCT, P.I., through the institutional scientific employment program-contract for L. Barros and R.C. Calhelha contracts; C. Pereira contract though the celebration of program-contract foreseen in No. 4, 5, and 6 of article 23º of Decree-Law No. 57/2016, of 29 August, amended by Law No. 57/2017, of 19 July; to FEDER-Interreg España-Portugal for financial support through the projects 0377_Iberphenol_6_E and 0612_TRANS_CO_LAB_2_P; FEDER through the Regional Operational Program North 2020: project Mobilizador Norte-01-0247-FEDER-024479 (ValorNatural®).info:eu-repo/semantics/publishedVersio

Efeitos não-alvo de inseticidas utilizados para controle de Bemisia tabaci (Gennadius, 1889) biótipo B (Hemiptera: Aleyrodidae) sobre a mortalidade e biologia de Spodoptera frugiperda (Smith, 1797) (Lepidoptera: Noctuidae) em soja [Glycine max (l.) Merrill]

Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) is a polyphagous pest that has gained importance in soybean cultivation in recent years, due to the damage it causes to leaves and pods. The objective of this work was to evaluate the non-target effect of insecticides recommended and used to control Bemisia tabaci (Genn.) biotype B on S. frugiperda. The experimental design was entirely randomized with five treatments and fifty replications. The treatments were: 1. negative control (distilled water), 2. positive control (lufenuron), 3. buprofezin, 4. pyriproxyfen, and 5. diafenthiuron. In each replicate, one caterpillar was placed in a plastic cup, where sprayed soybean leaves with the maximum dosage of insecticides recommended for B. tabaci were placed.The treatments were evaluated daily during all stages: larval, pupal, and adult emergence and fecundity. The results obtained for larval mortality showed that diafenthiuron aimed 100% control, followed by pyriproxyfen with 28% and buprofezin with 12%. The duration of the larval period of the insects exposed to pyriproxyfen and buprofezin were longer compared to the control. While the biological parameters of pupae were found that pyriproxyfen caused lower pupal formation rate and higher pupal mortality, followed by buprofezin. In adult emergence, pyriproxyfen showed the lowest rate with 13.8% and the highest percentage of deformed adults with 60%. Thus, it was possible to conclude that the insecticides under study cause S. frugiperda mortality or effects on its biological patterns

Profiling microglia in a mouse model of Machado-Joseph disease

Microglia have been increasingly implicated in neurodegenerative diseases (NDs), and specific disease associated microglia (DAM) profiles have been defined for several of these NDs. Yet, the microglial profile in Machado–Joseph disease (MJD) remains unexplored. Here, we characterized the profile of microglia in the CMVMJD135 mouse model of MJD. This characterization was performed using primary microglial cultures and microglial cells obtained from disease-relevant brain regions of neonatal and adult CMVMJD135 mice, respectively. Machine learning models were implemented to identify potential clusters of microglia based on their morphological features, and an RNA-sequencing analysis was performed to identify molecular perturbations and potential therapeutic targets. Our findings reveal morphological alterations that point to an increased activation state of microglia in CMVMJD135 mice and a disease-specific transcriptional profile of MJD microglia, encompassing a total of 101 differentially expressed genes, with enrichment in molecular pathways related to oxidative stress, immune response, cell proliferation, cell death, and lipid metabolism. Overall, these results allowed us to define the cellular and molecular profile of MJD-associated microglia and to identify genes and pathways that might represent potential therapeutic targets for this disorder.This work was supported by Fundação para a Ciência e a Tecnologia (FCT) (PTDC/NEUNMC/3648/2014) and COMPETE-FEDER (POCI-01-0145-FEDER-016818). It was also supported by Portuguese funds through FCT in the framework of the Project POCI-01-0145-FEDER-031987 (PTDC/MED-OUT/31987/2017). A.B.C. was supported by a doctoral fellowship from FCT (PD/BD/ 127828/2016). S.P.N. was also supported by FCT (PD/BD/114120/2015). Work in the JBR laboratory was financed by FEDER—Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020—Operational Programme for Competitiveness and Internationalization (POCI), Portugal 2020, and by Portuguese funds through FCT in the framework of the Project POCI-01-0145- FEDER030647 (PTDC/MED-NEU/31318/2017). This work was funded by ICVS Scientific Microscopy Platform, member of the national infrastructure PPBI (Portuguese Platform of Bioimaging) (PPBIPOCI-01-0145-FEDER-022122), and by National funds, through FCT—project UIDB/50026/2020 and UIDP/50026/2020

Gerenciamento de resíduos químicos como uma proposta didática na disciplina de físico-química aplicada à biotecnologia

O gerenciamento e o reaproveitamento de resíduos químicos é uma atividade de grande importância para a formação acadêmica do graduando na área de ciências. Tendo em vista essa preocupação, no laboratório de Físico-Química/Química Analítica do Instituto de Saúde e Biotecnologia foram desenvolvidas algumas ações pelos acadêmicos do curso de Bacharelado em Biotecnologia durante as aulas experimentais da disciplina de Físico-Química. O resultado desse trabalho gerou o presente relato de experiência que tem como objetivo promover entre os acadêmicos o desenvolvimento da conscientização ambiental através do gerenciamento de resíduos químicos. No primeiro momento foi feito a triagem dos reagentes e dos resíduos químicos e o levantamento da carga horária semestral de aulas no referido laboratório, visando o desenvolvimento de metodologias para o tratamento, descarte e reaproveitamento adequado em aulas práticas. Foram analisadas dezoito soluções ociosas. Dessas soluções, foram aferidos pH, volume e verificado a toxicidade. Os resultados das substâncias avaliadas apontaram que 20% da substâncias são tóxicas, 27% são oxidantes, 5% são cancerígenas, 24% corrosivas e 24% irritantes, sendo o volume total 5,541 L. Além disso, o levantamento da carga horária semestral apontou 90 horas para o semestre par e 120 horas para o semestre ímpar distribuídas nas disciplinas de Química Analítica e Físico-Química. A partir desses resultados foi proposto requisitar bombonas para armazenar os descartes de maneira correta e um formulário de prática experimental conforme os princípios da química verde, visando mitigar o descarte incorreto e com a finalidade de reutilização ou doação de reagentes ociosos. Ainda que o projeto esteja em fase de aperfeiçoamento, a experiência dos acadêmicos com os procedimentos deste trabalho piloto no Instituto de Saúde e Biotecnologia promoveram o desenvolvimento da conscientização do controle de resíduos químicos, a consolidação da utilização de técnicas analíticas e maior conhecimento sobre as regras de segurança do laboratório por parte dos discentes da disciplina de Físico-Química aplicada à Biotecnologia

GQ-16, a TZD-derived partial PPARγ agonist, induces the expression of thermogenesis- related genes in brown fat and visceral white fat and decreases visceral adiposity in obese and hyperglycemic mice

Background Beige adipocytes comprise a unique thermogenic cell type in the white adipose tissue (WAT) of rodents and humans, and play a critical role in energy homeostasis. In this scenario, recruitment of beige cells has been an important focus of interest for the development of novel therapeutic strategies to treat obesity. PPARγ activation by full agonists (thiazolidinediones, TZDs) drives the appearance of beige cells, a process so-called browning of WAT. However, this does not translate into increased energy expenditure, and TZDs are associated with weight gain. Partial PPARγ agonists, on the other hand, do not induce weight gain, but have not been shown to drive WAT browning. The present study was designed to investigate the effects of GQ-16 on BAT and on browning of WAT in obese mice. Methods Male Swiss mice with obesity and hyperglycemia induced by high fat diet were treated with vehicle, rosiglitazone (4 mg/kg/d) or the TZD-derived partial PPARγ agonist GQ-16 (40 mg/ kg/d) for 14 days. Fasting blood glucose, aspartate aminotransferase, alanine aminotransferase and lipid profile were measured. WAT and brown adipose tissue (BAT) depots were excised for determination of adiposity, relative expression of Ucp-1, Cidea, Prdm16, Cd40 and Tmem26 by RT-qPCR, histological analysis, and UCP-1 protein expression analysis by immunohistochemistry. Liver samples were also removed for histological analysis and determination of hepatic triglyceride content. Results GQ-16 treatment reduced high fat diet-induced weight gain in mice despite increasing energy intake. This was accompanied by reduced epididymal fat mass, reduced liver triglyceride content, morphological signs of increased BAT activity, increased expression of thermogenesis- related genes in interscapular BAT and epididymal WAT, and increased UCP-1 protein expression in interscapular BAT and in epididymal and inguinal WAT. Conclusion This study suggests for the first time that a partial PPARγ agonist may increase BAT activity and induce the expression of thermogenesis-related genes in visceral WAT. General Significance These findings suggest that PPARγ activity might be modulated by partial agonists to induce WAT browning and treat obesity

Localization of coated iron oxide (Fe3O4) nanoparticles on tomato seeds and their effects on growth

IFood demand due to the growing population globally has been stretching the agriculture sector to the limit. This demands the cultivation of plants in shrinking land areas which makes the search for highly effective systems for plant nutrition and pest control important. In this context, the application of nanoparticles (NPs) in agriculture can have a transformative effect on food production techniques as it can enable the delivery of bioactive agents (including growth factors, pesticides and fungicides) directly to plants. Herein we report the application of unfunctionalized as well as amine-functionalized and polycaprolactone-coated Fe3O4 nanoparticles (NP) to seed treatment and plant growth. The study reveals that the treatment has no side effects on plant germination and development. Furthermore, the penetration of NPs in tomato seeds (Solanum Lycopersicum) and their translocation in tomato seedlings post-treatment depends on the level of the NPs functionalization. The research also demonstrates that a fraction of NP is preferentially translocated to tomato seedling roots and shoots, however, uptake is lower for functionalized NPs (both amine and PCL). Thus, the results suggest that the functionalization of NPs can act as a versatile platform for delivering of active compounds, such as fungicides and growth factor agents

Occurrence of postural deviations in children of a school of Jaguariúna, São Paulo, Brazil

OBJECTIVE: Evaluate the posture of students from a public school in Brazil and to identify the differences between normal deviation during growth and compensatory alterations. METHODS: Students from first to fourth grade of a public school in the city of Jaguariúna, São Paulo, Brazil, were evaluated. The students were positioned at sagital anterior and posterior coronal planes for postural assessment. Kendall points were used as the normal reference. RESULTS: 247 students were evaluated, 131 boys and 116 girls, and the main postural deviations found were: shoulder unbalance (50.2%), protracted shoulder (39.7%), abducted scapula (40.5%), knock-knee (29.6%), pelvic unbalance (21.5%), pelvic anteversion (19%), knee hyperextension (19%), medial rotation of hip (12.9%), protracted cervical (11.7%), head tilt (15.4%), thoracic hyperkyphosis (9.7%) and lumbar hyperlordosis (26.3%). CONCLUSIONS: A high incidence of postural alterations was detected in school children. Some of the postural alterations, such as abducted scapula, unbalance and protraction of the shoulders, knock-knee and lumbar hiperlordosis, normally occur and they are naturally corrected during growth. However, some postural problems, such as protraction and inclination of the cervical spine, were also prevalent and they require early intervention.OBJETIVO: Avaliar a postura de escolares do ensino público fundamental e diferenciar as alterações que fazem parte do crescimento normal das compensatórias. MÉTODOS: Trata-se de um estudo analítico, descritivo, de corte transversal, no qual se realizou avaliação postural em escolares de primeira a quarta série de uma escola pública da cidade de Jaguariúna, situada no interior do Estado de São Paulo. Cada aluno foi avaliado nos planos coronal-anterior, coronal-posterior e sagital, utilizando-se o método de Kendall como referência de alinhamento postural normal. RESULTADOS: A avaliação postural de 247 escolares (131 masculinos e 116 femininos) identificou a seguinte incidência de alterações: desnível (50,2%) e protrusão de ombro (39,7%), escápula alada (40,5%), aumento do ângulo valgo de joelho (29,6%), inclinação (21,5%) e anteroversão pélvica (19%), hiperextensão de joelho (19%), rotação de fêmur (12,9%), protrusão (11,7%) e inclinação cervical (15,4%), cifose torácica (9,7%) e hiperlordose lombar (26,3%). CONCLUSÕES: Houve elevada incidência de escápula alada, desnível e protrusão de ombro, aumento do ângulo valgo de joelho e hiperlordose lombar, consideradas normais durante o desenvolvimento da criança. As inclinações e a protrusão cervical, alterações que necessitam de intervenção precoce, também foram identificadas.748