Maternal exposure to glyphosate-based herbicide causes vascular dysfunction in offspring female rats.

This study analyzed how glyphosate exposure in the gestational period affects vascular function in their female offspring and whether oxidative stress is involved in this effect. To this, pregnant Wistar rats were exposed through drinking water to 0.2% of a glyphosate commercial formulation, and we analyzed the response to acetylcholine and phenylephrine in the aorta from offspring of Glyphosate-based herbicide (O-GBH) and controls (O-CON) rats at six months of age. Relaxation to acetylcholine was reduced in O-GBH than in O-CON. Acute Indomethacin and Apocynin increased relaxation to acetylcholine in O-GBH. The aorta from O-GBH was hyperactive to phenylephrine; the preincubation with N-nitro-L-arginine methyl ester (L-NAME) increased contraction to phenylephrine more in O-CON than O-GBH. TEMPOL similarly reduced phenylephrine response, and L-NAME prevented this effect. The TBARS and GSH levels were increased in O-GBH than in O-CON. Results reinforce the concept that oxidative stress during the perinatal period contributes to the development of vascular changes in adulthood. Results also reveal that oxidative stress parameters altered, and the current levels considered safe for exposure to Glyphosate deserve further investigation, especially in the female gender.

Maternal exposure to glyphosate-based herbicide causes changes in the vascular function of offspring adult rats.

This study analyzed how glyphosate exposure in the gestational period affects vascular function in their offspring, focusing on the influence of age and whether oxidative stress is involved in this effect. To this, pregnant Wistar rats were exposed through drinking water to 0.2% of a glyphosate commercial formulation, and we analyzed the response to acetylcholine and phenylephrine in the aorta from offspring of glyphosate herbicide-based (O-GHB) and controls (O-CON) rats at 3, 6, and 12 months of age. O-GHB groups showed no changes in arterial blood pressure or aorta histological analysis. Relaxation to acetylcholine was reduced in O-GHB than O-CON. Acute TEMPOL increased relaxation to acetylcholine in O-GHB at 6 and 12 months of age. The aorta from O-GHB was hyperactive to phenylephrine only at 6 months of age. Preincubation with N-nitro-L-arginine methyl ester (L-NAME) increased contraction to phenylephrine more in O-CON than O-GHB. TEMPOL similarly reduced phenylephrine response. This effect was prevented by L-NAME. Results reinforce the concept that oxidative stress during the perinatal period contributes to the development of vascular changes in adulthood. Results also reveal that although no changes in cardiac or histological parameters have been demonstrated, the current levels considered safe for exposure to glyphosate deserve further investigation, especially during pregnancy.

Targeting Ca2+ and Mitochondrial Homeostasis by Antipsychotic Thioridazine in Leukemia Cells.

Mitochondria have pivotal roles in cellular physiology including energy metabolism, reactive oxygen species production, Ca2+ homeostasis, and apoptosis. Altered mitochondrial morphology and function is a common feature of cancer cells and the regulation of mitochondrial homeostasis has been identified as a key to the response to chemotherapeutic agents in human leukemias. Here, we explore the mechanistic aspects of cytotoxicity produced by thioridazine (TR), an antipsychotic drug that has been investigated for its anticancer potential in human leukemia cellular models. TR exerts selective cytotoxicity against human leukemia cells in vitro. A PCR array provided a general view of the expression of genes involved in cell death pathways. TR immediately produced a pulse of cytosolic Ca2+, followed by mitochondrial uptake, resulting in mitochondrial permeabilization, caspase 9/3 activation, endoplasmic reticulum stress, and apoptosis. Ca2+ chelators, thiol reducer dithiothreitol, or CHOP knockdown prevented TR-induced cell death. TR also exhibited potent cytotoxicity against BCL-2/BCL-xL-overexpressing leukemia cells. Additionally, previous studies have shown that TR exhibits potent antitumor activity in vivo in different solid tumor models. These findings show that TR induces a Ca2+-mediated apoptosis with involvement of mitochondrial permeabilization and ER stress in leukemia and it emphasizes the pharmacological potential of TR as an adjuvant in antitumor chemotherapy.

Disruption of mitochondrial quality control genes promotes caspase-resistant cell survival following apoptotic stimuli.

In cells undergoing cell-intrinsic apoptosis, mitochondrial outer membrane permeabilization (MOMP) typically marks an irreversible step in the cell death process. However, in some cases, a subpopulation of treated cells can exhibit a sublethal response, termed "minority MOMP." In this phenomenon, the affected cells survive, despite a low level of caspase activation and subsequent limited activation of the endonuclease caspase-activated DNase (DNA fragmentation factor subunit beta). Consequently, these cells can experience DNA damage, increasing the probability of oncogenesis. However, little is known about the minority MOMP response. To discover genes that affect the MOMP response in individual cells, we conducted an imaging-based phenotypic siRNA screen. We identified multiple candidate genes whose downregulation increased the heterogeneity of MOMP within single cells, among which were genes related to mitochondrial dynamics and mitophagy that participate in the mitochondrial quality control (MQC) system. Furthermore, to test the hypothesis that functional MQC is important for reducing the frequency of minority MOMP, we developed an assay to measure the clonogenic survival of caspase-engaged cells. We found that cells deficient in various MQC genes were indeed prone to aberrant post-MOMP survival. Our data highlight the important role of proteins involved in mitochondrial dynamics and mitophagy in preventing apoptotic dysregulation and oncogenesis.

AMPK activation induced by promethazine increases NOXA expression and Beclin-1 phosphorylation and drives autophagy-associated apoptosis in chronic myeloid leukemia.

Relapse and drug resistance is still major challenges in the treatment of leukemia. Promethazine, an antihistaminic phenothiazine derivative, has been used to prevent chemotherapy-induced emesis, although there is no report about its antitumor potential. Thus, we evaluated the promethazine cytotoxicity against several leukemia cells and the underlying mechanisms were investigated. Promethazine exhibited potent and selective cytotoxicity against all leukemia cell types in vitro at clinically relevant concentrations. Philadelphia positive chronic myeloid leukemia (CML) K562 cells were the most sensitive cell line. The cytotoxicity of promethazine in these cells was triggered by the activation of AMPK and inhibition of PI3K/AKT/mTOR pathway. The subsequent downstream effects were NOXA increase, MCL-1 decrease, and Beclin-1 activation, resulting in autophagy-associated apoptosis. These data highlight targeting autophagy may represent an interesting strategy in CML therapy, and also the antitumor potential of promethazine by acting in AMPK and PI3K/AKT/mTOR signaling pathways. Since this drug is currently used with relative low side effects, its repurposing may represent a new therapeutic opportunity for leukemia treatment.

Mycological quality of pecan nuts from Brazil: absence of aflatoxigenic fungi and aflatoxins / Qualidade micológica de nozes pecan do Brasil: ausência de fungos aflatoxigênicos e aflatoxinas

ABSTRACT: In recent years, Brazil has encouraged the cultivation of pecans to meet both demands of domestic and international market of nuts. New genetic varieties of pecans have been selected in recent years, but available scientific information on the occurrence of fungi and aflatoxins in the international literature is out of date. Therefore, the present study aimed to quantify and identify fungal microbiota and the presence of aflatoxins in pecan nuts cultivated in southern Brazil. Fifty-two pecan nut lots (Barton variety) were obtained from producers from 19 cities of Rio Grande do Sul State and analyzed by direct plating in Agar Dichloran Glycerol 18% (DG18) and Aspergillus Flavus and Parasiticus Agar (AFPA), following incubation at 25 °C for 7 days. Aflatoxins analyses were carried out using HPLC coupled with a mass spectrometer. Results revealed at least 10 different genera of fungi. Aspergillus, Penicillium, Fusarium, and Cladosporium were predominant. Xerophilic species of Aspergillus (A. wentii, A. ruber, A. pseudoglaucus, and A. chevalieri) were commonly reported in the samples. No potential aflatoxin-producing species was isolated and no aflatoxins were detected (LOQ=1 μg/kg and LOD=0.1 μg/kg for AFB1 and AFB2, and 0.3 μg/kg for AFG1 and AFG2) in the evaluated samples. The absence of this carcinogenic group of mycotoxins is highly positive and could boost the investments in the sector, as well as stimulate the commercialization and consumption of this variety of nut.

RESUMO: Nos últimos anos, o Brasil tem incentivado o cultivo de pecãs para atender a demanda do mercado nacional e internacional de nozes. Novas variedades genéticas de pecãs foram selecionadas nos últimos anos, mas informações científicas disponíveis sobre a ocorrência de fungos e aflatoxinas na literatura internacional estão desatualizadas. Portanto, o presente estudo objetivou quantificar e identificar a microbiota fúngica e a presença de aflatoxinas em nozes cultivadas no sul do Brasil. Cinquenta e dois lotes de nozes (variedade Barton) foram obtidos de produtores de 19 municípios do Estado do Rio Grande do Sul e analisados ​​por meio de Ágar Dicloran Glicerol 18% (DG18) e Aspergillus Flavus e Parasiticus Agar (AFPA), após incubação em 25 °C durante 7 dias. Análises de aflatoxinas foram realizadas usando HPLC acoplado a um espectrômetro de massa. Os resultados revelaram pelo menos 10 gêneros diferentes de fungos. Aspergillus, Penicillium, Fusarium e Cladosporium foram predominantes. Espécies xerofílicas de Aspergillus (A. wentii, A. ruber, A. pseudoglaucus e A. chevalieri) foram comumente encontradas nas amostras. Nenhuma espécie potencial produtora de aflatoxinas foi isolada e nenhuma aflatoxina foi detectada (LOQ=1 μg/kg e LOD=0,1 μg/kg para AFB1 e AFB2; e 0,3 μg/kg para AFG1 e AFG2) nas amostras avaliadas. A ausência desse grupo carcinogênico de micotoxinas é altamente positiva e pode impulsionar os investimentos no setor, além de estimular a comercialização e o consumo dessa variedade de nozes.

The PERK Arm of the Unfolded Protein Response Regulates Mitochondrial Morphology during Acute Endoplasmic Reticulum Stress.

Endoplasmic reticulum (ER) stress is transmitted to mitochondria and is associated with pathologic mitochondrial dysfunction in diverse diseases. The PERK arm of the unfolded protein response (UPR) protects mitochondria during ER stress through the transcriptional and translational remodeling of mitochondrial molecular quality control pathways. Here, we show that ER stress also induces dynamic remodeling of mitochondrial morphology by promoting protective stress-induced mitochondrial hyperfusion (SIMH). ER-stress-associated SIMH is regulated by the PERK arm of the UPR and activated by eIF2α phosphorylation-dependent translation attenuation. We show that PERK-regulated SIMH is a protective mechanism to prevent pathologic mitochondrial fragmentation and promote mitochondrial metabolism in response to ER stress. These results identify PERK-dependent SIMH as a protective stress-responsive mechanism that regulates mitochondrial morphology during ER stress. Furthermore, our results show that PERK integrates transcriptional and translational signaling to coordinate mitochondrial molecular and organellar quality control in response to pathologic ER insults.

Acute Hypotension After Moderate-Intensity Handgrip Exercise in Hypertensive Elderly People.

Souza, LR, Vicente, JB, Melo, GR, Moraes, VC, Olher, RR, Sousa, IC, Peruchi, LH, Neves, RV, Rosa, TS, Ferreira, AP, and Moraes, MR. Acute hypotension after moderate-intensity handgrip exercise in hypertensive elderly people. J Strength Cond Res 32(10): 2971-2977, 2018-Isometric handgrip (IHG) training is effective in reducing blood pressure (BP), but little is known about the occurrence of acute hypotension by postisometric exercise hypotension (PIEH) and the underlying mechanisms. Ten sedentary hypertensive elderly people (7 women and 3 men) individuals, with a mean age of 73.2 ± 2.2 years and systolic BP (SBP) of 135.1 ± 6.5, were included; they were hypertensive for 13.2 ± 3.1 years and were receiving medications. These patients underwent 2 experimental sessions of isometric exercise using a portable hydraulic handgrip dynamometer: (a) sham session with 3% of maximal voluntary isometric contraction (MVIC) and (b) experimental isometric session with 30% MVIC, completing a total of 8 sets of 1-minute contraction and 1-minute rest-pause (overload of work = 51.7 kgf·min). Blood pressure and heart rate were evaluated at rest and 1, 5, 10, 15, 30, 45, and 60 minutes postexercise. Blood lactate (Lac) and salivary nitric oxide (NO) were collected at rest, 0, 30, and 60 minutes postexercise. The SBP presented a reduction as of the tenth minute postexercise to session 30% MVIC (Δ = -14.4 at -18.7 mm Hg, p < 0.05). At 60-minute postexercise, the SBP was 30% lower vs. 3% MVIC (-20.2 mm Hg, p = 0.006). There were no differences for the other cardiovascular parameters and vasoactive substances for both sessions (p > 0.05). These results demonstrated that the IHG exercise with a higher overload of work induced PIEH in hypertensive elderly people, but there was no association with Lac and NO. Therefore, this IHG model with easy execution, quick adherence, short duration, and a portable equipment can be an excellent adjuvant strategy for the control and prevention of hypertension.

O programa de controle de infecção relacionada à assistência à saúde em meio ambiente hospitalar e o dever de fiscalização da agência nacional de vigilância sanitária* / The infection control program regarding health care in hospital environments and the duty of surveillance of the Brazilian Health Regulatory Agency

O presente artigo estuda a função de fiscalização da Agência Nacional de Vigilância Sanitária nos casos de infecção hospitalar e, em especial, no que tange ao programa de Controle de Infecção relacionada à assistência à saúde, utilizado pelos estabelecimentos de saúde para minimizar os riscos de infecção por patógenos. Trata-se de artigo teórico-documental, com a análise da legislação pertinente e os riscos existentes para pacientes e profissionais da saúde no ambiente hospitalar. O estudo traz como proposta uma reflexão sobre a necessidade de fiscalização efetiva por parte da agência nos estabelecimentos de saúde e esboça formas de buscar a efetivação do poder de polícia. (AU)

This article studies the surveillance role of the Brazilian Health Regulatory Agency in cases of hospital infection, and in particular to the Program of Hospital Infection Control in health care, implemented by health institutions to minimize the risks of pathogen infection. It is a theoretic-documental article, with an analysis of the pertinent legislation and the existing risks to patients and health professionals in hospital environments. The study proposes a reflection on the need for the Brazilian agency's effective inspection actions in healthcare establishments and describes ways of pursuing the implementation of policing powers.(AU)

Enhancement of chlorpromazine antitumor activity by Pluronics F127/L81 nanostructured system against human multidrug resistant leukemia.

The development of specific tyrosine kinase inhibitors (TKIs) revolutionized the treatment of chronic myeloid leukemia (CML). However, chemoresistance of tumor cells to TKIs has already been described, and several mechanisms account for the multidrug resistance (MDR) phenotypes, including the overexpression of P-glycoprotein (P-gp). This decreases the rate of healing and complete tumor remission. Nanotechnological tools have been studied to allow advances in this field. Poloxamers (Pluronics(®)) have been proposed as drug carriers to improve therapeutic efficacy and decrease side effects, even in cancer therapy, due to their ability to inhibit P-gp. Antipsychotic phenothiazines have been described as potent cytotoxic drugs against several types of tumor cells in vitro. Here, we show that nanostructured micellar systems containing the phenothiazine derivative chlorpromazine (CPZ) potentiated the cytotoxicity of free CPZ and increased the selectivity against CML tumor cells, demonstrating the pharmacological potential of these poloxamer-based nanostructured systems containing CPZ in cancer therapy.

Protective Effect of Plantago major Extract against t-BOOH-Induced Mitochondrial Oxidative Damage and Cytotoxicity.

Plantago major L. produces several chemical substances with anti-inflammatory and analgesic activities and its use in the treatment of oral and throat inflammation in popular medicine is well described. In this study, the antioxidant potential of the Plantago major hydroethanolic extract was screened and its protective action was evaluated against t-BOOH-induced oxidative stress. The extract was obtained by fractionated percolation using 50% ethanolic solution and, after drying, suspended in dimethyl sulfoxide. The chromatographic profile of crude extract was obtained with the identification of some phytochemical markers and the total phenols and flavonoids were quantified. The scavenger activity against DPPH (1,1-diphenyl-2-picrylhydrazyl) radicals was determined and the antioxidant activity in biological systems was evaluated in isolated rat liver mitochondria and HepG2 cells. The extract exhibited a significant free radical scavenger activity at 0.1 mg/mL, and decreased the ROS (reactive oxygen species) generation in succinate-energized mitochondria. Such an effect was associated with the preservation of the intrinsic antioxidant defenses (reduced glutathione and NAD(P)H) against the oxidation by t-BOOH, and also to the protection of membranes from lipid oxidation. The cytoprotective effect of PmHE against t-BOOH induced cell death was also shown. These findings contribute to the understanding of the health benefits attributed to P. major.

Comparative cytogenetics between two species of the family Pseudopimelodidae (Siluriformes): occurrence of natural triploidy and supernumerary chromosomes.

The Pseudopimelodidae family comprises 35 species however, cytogenetic studies have been performed in only six species. This study uncovered karyotypic data on Pseudopimelodus pulcher and Microglanis cottoides. Both species possessed 2n = 54, with 20m + 16sm + 10st + 8a and FN = 100 for P. pulcher and 30m + 14sm + 6st + 4a and FN = 104 for M. cottoides. A female of M. cottoides with 45m + 21sm + 9st + 6a (2n = 81) plus two extra small chromosomes was found, indicating a natural triploidy with supernumerary chromosomes. The formation of the polyploid individual seems to have come from a diploid female gamete, due to the presence of a marker chromosome pair partially heterochromatic presents only in females and common to that exemplar. This triploid female showed three chromosomes with nitrate staining (AgNOR), 18S rDNA probe and chromomycin A3 (CMA3) staining. AgNORs were observed on pairs 12 and 23 in P. pulcher and pair 24 in M. cottoides, results that were confirmed with an 18S rDNA probe and CMA3 fluorochrome. These are the first chromosomal data for P. pulcher and provide the first description of natural triploidy with the presence of supernumerary chromosomes in this family and emphasizing well the chromosomal rearrangements diversification between this species.

Pluronics f-127/l-81 binary hydrogels as drug-delivery systems: influence of physicochemical aspects on release kinetics and cytotoxicity.

We investigated the structure of the binary mixture of Pluronic F-127 (PL F-127) and Pluronic L-81 (PL L-81), as hydrogels for sumatriptan delivery and investigated the mixture possible use via subcutaneous route for future applications as a long-acting antimigraine formulation. We studied the drug-micelle interaction by dynamic light scattering and differential scanning calorimetry, sol-gel process by rheology, and small-angle X-ray scattering (SAXS). We also employed pharmaceutical formulation aspects by dissolution rate, release profile, and cytotoxicity studies for apoptosis and/or necrosis in fibroblasts (3T3) and neural cells (Neuro 2a). Micellar hydrodynamic diameter studies revealed the formation of binary PL-micelles by association of PL F-127/PL L-81. The mixed micelle and binary hydrogels formation was also verified by only one phase transition temperature for all formulations, even in the presence of sumatriptan. The characterization of the hydrogel supramolecular organization by SAXS, rheology studies, and in vitro dissolution/release results showed a probable relationship between the transition of the lamellar to the hexagonal phase and the lower release constant values observed, indicating that PL L-81 participates in micelle-hydrogel formation and aggregation processes. Furthermore, the reduced cytotoxicity (annexin V-fluorescein isothiocyanate positive staining), with minor PL L-81 concentration, points to its potential use for the development of binary PL-systems containing sumatriptan capable of modulating the gelation process. This use may employ the minimum PL concentration and be interesting for pharmaceutical applications, particularly for migraine treatment.

Associação entre recessão gengival e hipersensibilidade dentinária / Association between gingival recession and dentine hypersensitivity - a pilot study

O presente estudo teve por objetivo avaliar a associação entre experiência de recessão gengival e hipersensibilidade dentinária em alunos do curso de Odontologia da Universidade Luterana do Brasil (Ulbra). Vinte estudantes (n = 20), de ambos os sexos, com idades entre 20 e 24 anos, responderam um questionário e, posteriormente, foram examinados clinicamente quanto a presença de recessão gengival, através de exame visual, e hipersensibilidade dentinária, por meio de estímulo com jato de ar. Ambas as variáveis foram tratadas de forma dicotômica (presença/ausência) e analisadas através do teste qui-quadrado (α = 0,05). Os resultados em relação à autopercepção demonstraram prevalência de 35% para recessão gengival e de 40% para hipersensibilidade. A análise clínica revelou prevalência de 100% de recessão gengival e de 55% de hipersensibilidade dentinária, em relação aos indivíduos. A associação entre os eventos foi estatisticamente significativa (p < 0,001). Concluiu-se que recessão gengival apresenta-se de forma frequente e está associada ao aparecimento de hipersensibilidade dentinária em jovens

B chromosomes in four different populations of Rhamdia quelen (Siluriformes, Heptapteridae): a comparative study of frequency and distribution.

In the present study 25 specimens of Rhamdia quelen from four different localities were analyzed cytogenetically. All showed a diploid number of 58 chromosomes, with a karyotypic formula of 36m + 16sm + 6st and FN = 116. Metacentric B chromosomes showing inter- and intraindividual variation were observed in all populations. C-banding revealed differences in the heterochromatin distribution pattern, with evidence of completely heterochromatic B chromosomes in three populations: Agua das Pedras, Agua dos Patos and Taquari rivers, and partially heterochromatic B chromosomes in the population at the fish farm station in Timbó/SC. The occurrence of B chromosomes in such distinct populations suggests that they could have arisen from the same ancestral state, before the geographic dispersal of this species.