Targeting the mitochondrial VDAC in hepatocellular carcinoma using a polyclonal antibody-conjugated to a nitrosyl ruthenium complex

The rational design of anti-cancer agents includes a new approach based on ruthenium complexes that can act as nitric oxide (NO) donor agents against specific cellular targets. One of the most studied classes of those compounds is based on bis(bipyridine) ruthenium fragment and its derivative species. In this work, we present the chemical and cytotoxicity properties against the liver hepatocellular carcinoma cell line HepG2 of cis-[RuII(NO+)Cl(dcbpy)2]2− conjugated to a polyclonal antibody IgG (anti-VDAC) recognizing a cell surface marker. UV–visible bands of the ruthenium complex were assigned with the aid of density functional theory, which also allowed estimation of the structures that explain the biological effects of the ruthenium complex–IgG conjugate. The interaction of cis-[RuII(NO+)Cl(dcbpy)2]3− with mitochondria was evaluated due to the potential of these organelles as anti-cancer targets, and considering they interact with the anti-VDAC antibody. The cytotoxicity of cis-[RuII(NO+)Cl(dcbpy)2]3−-anti-VDAC antibody was up to 80% greater in comparison to the free cis-[RuII(NO+)Cl(dcbpy)2]3− complex. We suggest that this effect is due to site-specific interaction of the complex followed by NO release.Fil: Ramos, Loyanne C. B.. Universidade de Sao Paulo; BrasilFil: Rodrigues, Fernando P.. Universidade de Sao Paulo; BrasilFil: Biazzotto, Juliana C.. Universidade de Sao Paulo; BrasilFil: de Paula Machado, Sergio. Universidade Federal do Rio de Janeiro; BrasilFil: Slep, Leonardo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; ArgentinaFil: Hamblin, Michael R.. Harvard Medical School; Estados UnidosFil: da Silva, Roberto S.. Harvard Medical School; Estados Unidos. Universidade de Sao Paulo; Brasi

Acute effect of aerobic exercise in different intensities in mucociliary clearance of patients with COPD

Design of the Study: Clinical Trial. Objective (s): To analyze the acute effect of aerobic exercise at different intensities in mucociliary clearance in patients with COPD, and to investigate possible associations of the autonomic nervous system in this response. Methods: 22 COPD patients underwent an initial evaluation for collecting personal data and spirometry to assess lung function. It was performed a progressive treadmill test for aerobic exercise prescription. Finally two randomized sessions of aerobic exercise with intensity of 60 % and 90 % of peak speed reached during the incremental test ( vVO2peack ) were performed with at least 24 hours of rest between them. The mucociliary clerance was assessed before and after the exercise sessions by testing the saccharin transit time (STT). Assessment of autonomic modulation was performed by heart rate variability (HRV) which continued throughout the protocol. Results: The values obtained in the STT test after aerobic exercise at 60 % of vVO2peack (9,08 minutes ± 4,96 ) was lower when compared to the STT before exercise ( 11,96 ± 6,31, p = 0,005 ) . That response also occurred after aerobic exercise at 90% of vVO2peack ( 8,90 ± 4,21 min ) compared to baseline ( 12,94 ± 7,22 , p = 0,023 ). Correlation analysis between the final values of STT test and HRV indexes did not show significant differences. Conclusions: Patients with COPD showed acceleration of mucociliary clerance right after a session of aerobic exercise. It was not possible to observe the association of autonomic modulation in this responseModelo do Estudo: Experimental. Objetivo(s) do estudo: Analisar o efeito agudo do exercício aeróbio em diferentes intensidades no transporte mucociliar de pacientes com DPOC, bem como investigar possíveis associações do sistema nervoso autônomo nesta resposta. Metodologia: Foram analisados 22 pacientes com DPOC que realizaram avaliação inicial para coleta de dados pessoais e espirometria a fim de avaliar a função pulmonar. Realizou-se um teste progressivo em esteira ergométrica para prescrição do exercício aeróbio. Por fim foram realizadas duas sessões de exercício aeróbio randomizadas em esteira ergométrica com intensidade de 60% e 90% do pico da velocidade atingida no teste incremental (vVO2pico) com pelo menos 24 horas de descanso entre elas. O transporte mucociliar foi avaliado antes e após realização do exercício por meio do teste do tempo de trânsito da sacarina (TTS). A avaliação da modulação autonômica foi realizada por meio da variabilidade da frequência cardíaca (VFC) a qual prosseguiu durante todo o protocolo. Resultados: Os valores obtidos no teste de TTS dos pacientes com DPOC após exercício aeróbio a 60% da vVO2pico (9,08 ± 4,96 minutos) foi menor comparado ao TTS antes do exercício (11,96 ± 6,31; p = 0,005). O que também ocorreu após exercício aeróbio a 90% da vVO2pico (8,90 ± 4,21 minutos) quando comparado ao momento basal (12,94 ± 7,22; p = 0,023). As análises de correlação entre os valores finais de TTS e índices da VFC não apontaram diferenças significativas. Conclusões: Pacientes com DPOC apresentaram aceleração da transportabilidade mucociliar frente a uma sessão de exercício aeróbio. Não foi possível observar associação da modulação autonômica nesta resposta após o exercíci

Mineralization of sheep manure and its influence on lettuce production

Diversos resíduos orgânicos são utilizados na agricultura sem o adequado conhecimento da sua dinâmica de mineralização. Avaliou-se a mineralização de esterco de ovinos e sua influência na produção de alface. Utilizou-se o delineamento experimental de blocos ao acaso com três repetições. Foram utilizadas 25 t ha-1 como dose de esterco para cada um dos seguintes tratamentos: 1) esterco de ovinos que se alimentaram de feno de mandioca (PAM); 2) esterco de ovinos que se alimentaram de subproduto de ervilha (ERV); 3) esterco de ovinos que se alimentaram de feno de capim coast-cross (FCC); 4) esterco de ovinos que se alimentaram de subproduto de saccharina (SAC) e 5) solo sem aplicação de esterco (testemunha). Foi determinada semanalmente a respiração basal do solo, utilizada como indicador de mineralização da matéria orgânica. A massa fresca de alface foi avaliada como medida de produção. Os tratamentos ERV, FCC e SAC apresentaram ganhos de massa fresca na ordem de 68, 65 e 62% em relação à testemunha e de 43, 39 e 33% em relação ao PAM, respectivamente. A produção menor promovida pelo PAM, em relação às demais, pode ser explicada pela forma de mineralização da matéria orgânica que apresentou elevada respiração microbiana cinco dias após o transplantio, com acentuado declínio, nas medições subseqüentes, ao longo do ciclo da cultura. Os demais tratamentos apresentaram mineralização sincronizada com conseqüente aumento na produção de massa fresca. ____________________________________________________________________________________________________________ ABSTRACTSeveral organic wastes are used in agriculture with no precise knowledge about the mineralization dynamics of these materials. In this study the sheep manure mineralization and its influence on the lettuce production was evaluated. A randomized block design with three replications was used. Five treatments were studied using 25 t ha-1 as dose of manure: 1) sheep manure obtained from animals fed with cassava straw (PAM); 2) sheep manure obtained from animals fed with residue of pea crop (ERV); 3) sheep manure obtained from animals fed with Coast-Cross hay (FCC), 4) sheep manure obtained from animals fed with saccharin residue (SAC) and 5) soil without application of manure (control). Weekly the basal respiration was determined and used as an indicator of organic matter mineralization. Lettuce fresh mass was evaluated as a measure of production. Treatments ERV, FCC and SAC showed superior weight gains of 68, 65 and 62% compared to the control and 43, 39 and 33% compared to MAP, respectively. Lower production promoted by the MAP in relation to the other treatments can be explained by organic matter mineralization that showed a high microbial respiration five days after transplanting, with marked decline in subsequent measurements during the crop cycle. The other systems showed mineralization synchronized with the production increase of lettuce fresh mass

In vivo and in vitro anti-inflammatory activity of red clover Trifolium pratense dry extract

Red clover Trifolium pratense L., Fabaceae, contains four isoflavones, mainly formononetin and biochanin A, and in smaller concentrations, daidzein and genistein. These compounds have gained a lot interest due to its human health benefits, such as estrogenic and progestogenic activities, antioxidant, anti-cancer and others. The objective of this study was to determine in vivo and in vitro anti-inflammatory activity of red clover dry extract. The in vitro anti-inflammatory activity was assayed by the technique using the Boyden chamber method, evaluating the leukocyte migration inhibition (chemotaxis). The in vivo anti-inflammatory activity was tested by a carrageenan-induced rat paw edema test. The results of anti-inflammatory in vitro test showed that there was a significant inhibition of leukocyte migration at the concentrations of 100, 50, 25, 10 and 5 µg/mL of red clover dry extract, these doses resulted in 94.73, 95.39, 94.73, 84.68 and 78.75% of inhibition for each dose, respectively. The anti-inflammatory in vivo test resulted in a significant activity in both tested doses (100 and 50 mg/kg of red clover dry extract) and at each tested time. The average percentage of edema inhibition was 63.37%. The findings of this study suggested that red clover extract might be suitable for the treatment of inflammatory diseases

Identification of new targets of S-nitrosylation in neural stem cells by thiol redox proteomics

Nitric oxide (NO) is well established as a regulator of neurogenesis. NO increases the proliferation of neural stem cells (NSC), and is essential for hippocampal injury-induced neurogenesis following an excitotoxic lesion. One of the mechanisms underlying non-classical NO cell signaling is protein S-nitrosylation. This post-translational modification consists in the formation of a nitrosothiol group (R-SNO) in cysteine residues, which can promote formation of other oxidative modifications in those cysteine residues. S-nitrosylation can regulate many physiological processes, including neuronal plasticity and neurogenesis. In this work, we aimed to identify S-nitrosylation targets of NO that could participate in neurogenesis. In NSC, we identified a group of proteins oxidatively modified using complementary techniques of thiol redox proteomics. S-nitrosylation of some of these proteins was confirmed and validated in a seizure mouse model of hippocampal injury and in cultured hippocampal stem cells. The identified S-nitrosylated proteins are involved in the ERK/MAPK pathway and may be important targets of NO to enhance the proliferation of NSC.PTDC/QUI-QFI/29319/2017; UID/BIM/04773/2019info:eu-repo/semantics/publishedVersio

Effect of maternal heat-stress on follicular growth and oocyte competence in Bos indicus cattle.

The objective was to determine whether exposure of Gir (Bos indicus) cows to heat-stress (HS) causes immediate and delayed deleterious effect on follicular dynamics, hormonal profile and oocyte competence. The cows were kept in tie-stalls for an adaptive thermoneutral period of 28 days (Phase I, Days −28 to −1). In Phase II (Days 0–28) cows were randomly allocated into control (CG, n = 5) and HS (HS, n = 5) treatments. The HS cows were placed in an environmental chamber at 38 °C and 80% relative humidity (RH) during the day and 30 °C, 80% RH during the night for 28 days. The CG group was maintained in shaded tie-stalls (ambient temperature) for 28 days. During Phase III (Days 28–147) animals were placed in tie-stalls (Days 28–42) followed by pasture (Days 42–147) under thermoneutrality. In each phase, weekly ovum pick up (OPU) sessions were to evaluate follicular development, morphology of cumulus–oocyte complexes (COCs), and developmental competence after in vitro maturation, fertilization, and culture. Serum concentrations of progesterone (P4) and cortisol were evaluated by radioimmunoassay. Exposure of Gir cows to HS had no immediate effect on reproductive function, but exerted a delayed deleterious effect on ovarian follicular growth, hormone concentrations, and oocyte competence. Heat-stress increased the diameter of the first and second largest follicles from Days 28 to 49. Indeed, HS increased the number of >9 mm follicles (characterized as follicular codominance) during this phase. Cows exposed to HS had longer periods of non-cyclic activity (P4 < 1 ng/mL), as well as shorter estrous cycles. However, HS did not affect cortisol concentration as compared to CG. Although HS had no significant effect on cleavage rate, it reduced blastocyst development during Phase III. In conclusion, long-term exposure of B. indicus cattle to HS had a delayed deleterious effect on ovarian follicular dynamics and oocyte competence

Mycorrhizal association in wheat genotypes submitted to variable irrigation in the Brazilian Cerrado.

Mycorrhizal association contributes to plant growth, influencing tolerance to abiotic stresses such as water deficit. There is considerable variation in infection by arbuscular mycorrhizal fungi (AMF) in cultivars of the same crop, but there is little information regarding these differences in wheat. The objective of this work was to evaluate the influence of water deficit on the arbuscular mycorrhizal association in wheat genotypes in the Cerrado region and the association between soil attributes and mycorrhizal colonization. The experiment was conducted in a no-till system, using different water regimes. The experimental design was a randomized block with subdivided plots scheme, with 12 treatments and 3 repetitions. The plots consisted of 4 wheat genotypes and the subplots included 3 water regimes. Mycorrhizal colonization, soil microbial biomass carbon, total soil organic carbon, easily extractable glomalin-related soil protein, spore number and AMF species diversity were evaluated. Mycorrhizal colonization was not influenced by wheat genotypes, but it was favored by the higher water regime, being 44.8% higher when compared to the lower water regime. The soil moisture was positively correlated with the soil attributes with the exception of the number of AMF spores. The community of AMF associated with wheat genotypes was similar, comprising of 12 species, predominantly Claroideoglomus etunicatum and Glomus macrocarpum. The low variation among wheat genotypes for AMF diversity suggests no selective influence of the plants on the AMF community in the area of the study. Water regime was shown to be a dominant factor in mycorrhizal association