Excessive hypercholesterolemia in pregnancy impairs rat uterine artery function via activation of Toll-like receptor 4.

Hypercholesterolemia in pregnancy is a physiological process required for normal fetal development. In contrast, excessive pregnancy-specific hypercholesterolemia increases the risk of complications, such as preeclampsia. However, the underlying mechanisms are unclear. Toll-like receptor 4 (TLR4) is a membrane receptor modulated by high cholesterol levels, leading to endothelial dysfunction; but whether excessive hypercholesterolemia in pregnancy activates TLR4 is not known. We hypothesized that a high cholesterol diet (HCD) during pregnancy increases TLR4 activity in uterine arteries, leading to uterine artery dysfunction. Sprague Dawley rats were fed a control diet (n=12) or HCD (n=12) during pregnancy (gestational day 6-20). Vascular function was assessed in main uterine arteries using wire myography (vasodilation to methacholine and vasoconstriction to phenylephrine; with and without inhibitors for mechanistic pathways) and pressure myography (biomechanical properties). Exposure to a HCD during pregnancy increased maternal blood pressure, induced proteinuria, and reduced the fetal-to-placental weight ratio for both sexes. Excessive hypercholesterolemia in pregnancy also impaired vasodilation to methacholine in uterine arteries, whereby at higher doses, methacholine caused vasoconstriction instead of vasodilation in only the HCD group, which was prevented by inhibition of TLR4 or prostaglandin H synthase 1. Endothelial nitric oxide synthase expression and nitric oxide levels were reduced in HCD compared with control dams. Vasoconstriction to phenylephrine and biomechanical properties were similar between groups. In summary, excessive hypercholesterolemia in pregnancy impairs uterine artery function, with TLR4 activation as a key mechanism. Thus, TLR4 may be a target for therapy development to prevent adverse perinatal outcomes in complicated pregnancies.

Stimulating cardiac glucose oxidation lessens the severity of heart failure in aged female mice.

Heart failure is a prevalent disease worldwide. While it is well accepted that heart failure involves changes in myocardial energetics, what alterations that occur in fatty acid oxidation and glucose oxidation in the failing heart remains controversial. The goal of the study are to define the energy metabolic profile in heart failure induced by obesity and hypertension in aged female mice, and to attempt to lessen the severity of heart failure by stimulating myocardial glucose oxidation. 13-Month-old C57BL/6 female mice were subjected to 10 weeks of a 60% high-fat diet (HFD) with 0.5 g/L of Nω-nitro-L-arginine methyl ester (L-NAME) administered via drinking water to induce obesity and hypertension. Isolated working hearts were perfused with radiolabeled energy substrates to directly measure rates of myocardial glucose oxidation and fatty acid oxidation. Additionally, a series of mice subjected to the obesity and hypertension protocol were treated with a pyruvate dehydrogenase kinase inhibitor (PDKi) to stimulate cardiac glucose oxidation. Aged female mice subjected to the obesity and hypertension protocol had increased body weight, glucose intolerance, elevated blood pressure, cardiac hypertrophy, systolic dysfunction, and decreased survival. While fatty acid oxidation rates were not altered in the failing hearts, insulin-stimulated glucose oxidation rates were markedly impaired. PDKi treatment increased cardiac glucose oxidation in heart failure mice, which was accompanied with improved systolic function and decreased cardiac hypertrophy. The primary energy metabolic change in heart failure induced by obesity and hypertension in aged female mice is a dramatic decrease in glucose oxidation. Stimulating glucose oxidation can lessen the severity of heart failure and exert overall functional benefits.

Proteomic identification of metabolic changes in Paracoccidioides brasiliensis induced by a nitroheteroarylchalcone.

Aim: To access the metabolic changes caused by a chalcone derivative (LabMol-75) through a proteomic approach. Methods: Proteomic analysis was performed after 9 h of Paracoccidioides brasiliensis yeast (Pb18) cell incubation with the LabMol-75 at MIC. The proteomic findings were validated through in vitro and in silico assays. Results: Exposure to the compound led to the downregulation of proteins associated with glycolysis and gluconeogenesis, ß-oxidation, the citrate cycle and the electron transport chain. Conclusion: LabMol-75 caused an energetic imbalance in the fungus metabolism and deep oxidative stress. Additionally, the in silico molecular docking approach pointed to this molecule as a putative competitive inhibitor of DHPS.

3-phenacylideneoxindoles as a new class of antifungal compounds against Paracoccidioides spp.

Aims: Considering the need to identify new compounds with antifungal action, the activity of five 3-phenacylideneoxindoles compounds was evaluated. Materials & methods: The compounds were synthesized, and their antifungal activity was elucidated through minimum inhibitory concentration tests and interaction assay with other antifungals. Potential targets of compounds were predicted in silico. Results: 3-phenacylideneoxindoles compounds inhibited fungal growth with minimum inhibitory concentration and minimum fungicidal concentration ranging from 3.05 to 12.26 µM. The compounds demonstrated high selectivity index and presented a synergistic effect with itraconazole. In silico prediction revealed the pentafunctional AROM polypeptide, enolase, superoxide dismutase, catalase and kinases as proteins targets of the compound 4a. Conclusion: The results demonstrate that 3-phenacylideneoxindoles is a potential new class of antifungal compounds for paracoccidioidomycosis treatment.

Patients affected by paracoccidioidomycosis (PCM) require long-term treatment, which commonly influences their adherence. In addition, only three drugs are in clinical use, which indicates the relevance of research in identifying new drugs for treating PCM. Thus, five drugs were tested in the laboratory to verify whether they could prevent the growth of the fungus without being toxic to humans. In addition, whether these compounds in combination with drugs used to treat PCM could be even more potent was evaluated. All compounds tested efficiently inhibited the growth of Paracoccidioides, the fungus that causes PCM. One drug was identified that, combined with itraconazole, decreased the required dose of both the discovered compound and itraconazole needed to inhibit fungal growth. Using computational tools, this work suggests how the new drug could act against the fungus. The results demonstrate a potential new treatment option, but more studies are needed to confirm the safety of these drugs.

Connecting Aortic Stiffness to Vascular Contraction: Does Sex Matter?

This study was designed to connect aortic stiffness to vascular contraction in young male and female Wistar rats. We hypothesized that female animals display reduced intrinsic media-layer stiffness, which associates with improved vascular function. Atomic force microscopy (AFM)-based nanoindentation analysis was used to derive stiffness (Young's modulus) in biaxially (i.e., longitudinal and circumferential) unloaded aortic rings. Reactivity studies compatible with uniaxial loading (i.e., circumferential) were used to assess vascular responses to a selective α1 adrenergic receptor agonist in the presence or absence of extracellular calcium. Elastin and collagen levels were indirectly evaluated with fluorescence microscopy and a picrosirius red staining kit, respectively. We report that male and female Wistar rats display similar AFM-derived aortic media-layer stiffness, even though female animals withstand higher aortic intima-media thickness-to-diameter ratio than males. Female animals also present reduced phenylephrine-induced aortic force development in concentration-response and time-force curves. Specifically, we observed impaired force displacement in both parts of the contraction curve (Aphasic and Atonic) in experiments conducted with and without extracellular calcium. Additionally, collagen levels were lower in female animals without significant elastin content and fragmentation changes. In summary, sex-related functional differences in isolated aortas appear to be related to dissimilarities in the dynamics of vascular reactivity and extracellular matrix composition rather than a direct response to a shift in intrinsic media-layer stiffness.

Age-Related Decline in Vascular Responses to Phenylephrine Is Associated with Reduced Levels of HSP70.

Aging impairs the expression of HSP70, an emergent player in vascular biology. However, it is unknown if age-related alterations in HSP70 are linked to a decline in arterial function. In this study, we test the hypothesis that the contributions of HSP70 to vascular contraction are diminished in middle-aged animals. We determined the basal levels of HSP70 in the aorta of young and middle-aged Sprague Dawley male rats using Western blotting. Functional studies were performed in a wire myograph system. Force development in response to phenylephrine was assessed in the presence or absence of extracellular calcium (Ca2+), and in aortic rings treated or non-treated with an HSP70 inhibitor. Fluorescent probes were used to evaluate vascular oxidative stress and nitric oxide levels. We report that middle-aged rats have significantly lower levels of HSP70. Blockade of HSP70 attenuated vascular phasic and tonic contraction in isolated aortas. It appears that a functional HSP70 is required for proper Ca2+ handling as inhibition of this protein led to reduced force-displacement in response to Ca2+ dynamics. Furthermore, middle-aged aortic rings exposed to the HSP70 inhibitor display higher reactive oxygen species levels without changes in nitric oxide. In summary, we show that middle-aged animals have lower levels of HSP70 in aortas, which associates with an age-related decline in vascular responses to α-1 adrenergic stimulation.

Apelin pathway in cardiovascular, kidney, and metabolic diseases: Therapeutic role of apelin analogs and apelin receptor agonists.

The apelin/apelin receptor (ApelinR) signal transduction pathway exerts essential biological roles, particularly in the cardiovascular system. Disturbances in the apelin/ApelinR axis are linked to vascular, heart, kidney, and metabolic disorders. Therefore, the apelinergic system has surfaced as a critical therapeutic strategy for cardiovascular diseases (including pulmonary arterial hypertension), kidney disease, insulin resistance, hyponatremia, preeclampsia, and erectile dysfunction. However, apelin peptides are susceptible to rapid degradation through endogenous peptidases, limiting their use as therapeutic tools and translational potential. These proteases include angiotensin converting enzyme 2, neutral endopeptidase, and kallikrein thereby linking the apelin pathway with other peptide systems. In this context, apelin analogs with enhanced proteolytic stability and synthetic ApelinR agonists emerged as promising pharmacological alternatives. In this review, we focus on discussing the putative roles of the apelin pathway in various physiological systems from function to dysfunction, and emphasizing the therapeutic potential of newly generated metabolically stable apelin analogs and non-peptide ApelinR agonists.

Dissecting the interaction between HSP70 and vascular contraction: role of [Formula: see text] handling mechanisms.

Heat-shock protein 70 (HSP70) is a ubiquitously expressed molecular chaperone with various biological functions. Recently, we demonstrated that HSP70 is key for adequate vascular reactivity. However, the specific mechanisms targeted by HSP70 to assist in this process remain elusive. Since there is a wealth of evidence connecting HSP70 to calcium ([Formula: see text]), a master regulator of contraction, we designed this study to investigate whether blockade of HSP70 disrupts vascular contraction via impairment of [Formula: see text] handling mechanisms. We performed functional studies in aortas isolated from male Sprague Dawley rats in the presence or absence of exogenous [Formula: see text], and we determined the effects of VER155008, an inhibitor of HSP70, on [Formula: see text] handling as well as key mechanisms that regulate vascular contraction. Changes in the intracellular concentration of [Formula: see text] were measured with a biochemical assay kit. We report that blockade of HSP70 leads to [Formula: see text] mishandling in aorta stimulated with phenylephrine, decreasing both phasic and tonic contractions. Importantly, in [Formula: see text] free Krebs' solution, inhibition of HSP70 only reduced the [Formula: see text] of the phasic contraction if the protein was blocked before IP3r-mediated [Formula: see text] release, suggesting that HSP70 has a positive effect towards this receptor. Corroborating this statement, VER155008 did not potentiate an IP3r inhibitor's outcomes, even with partial blockade. In another set of experiments, the inhibition of HSP70 attenuated the amplitude of the tonic contraction independently of the moment VER155008 was added to the chamber (i.e., whether it was before or after IP3r-mediated phasic contraction). More compelling, following re-addition of [Formula: see text], VER155008 amplified the inhibitory effects of a voltage-dependent [Formula: see text] channel blocker, but not of a voltage-independent [Formula: see text] channel inhibitor, indicating that HSP70 has a positive impact on the latter. Lastly, the mechanism by which HSP70 modulates vascular contraction does not involve the [Formula: see text] sensitizer protein, Rho-kinase, nor the SERCA pump, as blockade of these proteins in the presence of VER155008 almost abolished contraction. In summary, our findings shed light on the processes targeted by HSP70 during vascular contraction and open research avenues for potential new mechanisms in vascular diseases.

The Climate-Driven Genetic Diversity Has a Higher Impact on the Population Structure of Plasmopara viticola Than the Production System or QoI Fungicide Sensitivity in Subtropical Brazil.

Downy mildew, caused by Plasmopara viticola, is the main disease affecting vineyards in subtropical Brazil. Here, we collected 94 P. viticola isolates from four organic and conventional vineyards in the two main grape-growing states of Brazil to evaluate the sensitivity to the quinone outside inhibitor (QoI) azoxystrobin by pheno- and genotyping assays. The impact of location, production system and sensitivity to QoI fungicides on the population genetics and structure of P. viticola was determined using 10 microsatellite markers. Cytochrome b sequencing revealed that 28 and 100% of the isolates from vineyards under organic and conventional management carried the G143A mutation, respectively. The G143A mutation was associated with high levels of azoxystrobin resistance. Three out of the 94 isolates analyzed carried the M125I alteration, not previously described in P. viticola, which was associated with a five-fold reduction in azoxystrobin sensitivity compared to wild-type isolates. Haplotype network analysis based on cytochrome b gene sequences suggested that the Brazilian populations are more closely related to the European than the North American population. A total of six haplotypes were identified, with two of them carrying the G143A mutation. Microsatellite analysis revealed high allelic and genotypic variation among the four populations. Population differentiation analyses indicated that state of origin directly influences the population biology of P. viticola, while production system and QoI sensitivity have little effect. Great genetic diversity, sexual reproduction and high levels of admixture were observed in Rio Grande do Sul State. In contrast, populations in São Paulo State were dominated by a few clonal genotypes, and no admixed genotype was detected between the two genetic pools identified in the state. This study raises the hypothesis that winter weather conditions influence the overwinter survival strategy with profound effects in the population biology of P. viticola.

Technology Incorporation in Primary Teeth Endodontics: Case Reports.

BACKGROUND: Radical pulp therapy is an important and a relatively frequent procedure in pediatric dental offices. AIM: This study presents two clinical cases of pulpectomies in deciduous teeth using electronic apex locator and rotary instrumentation. CASE DESCRIPTIONS: In Case 1, pulpectomy was performed in a deciduous incisor with diagnosis of pulp necrosis in a 3-year-old patient using an electronic apical locator and manual instrumentation. In Case 2, it was performed in a primary molar with diagnosis of irreversible acute pulpitis in a 9-year-old patient with the use of electronic apical locator and rotational instrumentation. The used technologies contributed to favorable final result of the endodontic treatments in deciduous teeth in both cases. CONCLUSION: The incorporation of technology to the endodontic treatment of primary teeth favored the gain of time and conicity of the root canal preparation. However, studies still need to be performed to consolidate the technique. CLINICAL SIGNIFICANCE: Endodontic treatment of primary teeth still generates much discussion about the materials and techniques employed. The use of electronic apical locator and rotational instrumentation is successful treatment alternatives for teeth with pulp necrosis, with the advantage of limiting clinical time and presenting a favorable outcome. HOW TO CITE THIS ARTICLE: Gomes LAS, Oliveira AA, de Campos Neves ATS, et al. Technology Incorporation in Primary Teeth Endodontics: Case Reports. Int J Clin Pediatr Dent 2020;13(2):180-185.

Percepções de acadêmicos de farmácia sobre a homeopatia / Perceptions of undergraduate pharmacy students on homeopathy

A homeopatia define princípios que buscam interpretar os seres vivos como um todo, envolve a arte da análise de sinais e sintomas patológicos e da manipulação de medicamentos específicos para o restabelecimento da saúde integral dos seres vivos. Existe uma resolução específica do Conselho Federal de Farmácia (2005) que estabelece os requisitos mínimos para que o profissional farmacêutico possa atuar como responsável técnico em homeopatia. Com o intuito de compreender o entendimento dos graduandos em farmácia que já cursaram ou cursarão a disciplina de homeopatia oferecida pelo curso de farmácia no Centro Universitário Luterano de Ji- Paraná (CEULJI/ULBRA), buscou-se avaliar por meio de um questionário o entendimento sobre algumas definições e conceitos relacionados à homeopatia e ao universo das práticas integrativas. Do total de participantes (n=120) um número considerável (96%) declarou conhecer a homeopatia, sendo que aproximados 30% obtiveram informações prévias de pessoas próximas, menos da metade dosparticipantes conseguiram caracterizá-la ou defini-la de maneira correta como instrumento terapêutico e alguns a confundiram com terapia floral (28,8%). Dos que declararam já terem se tratado com homeopatia, 81% o fizeram sem prescrição médica e grande parte não sabia da existência da Política Nacional de Práticas Integrativas e Complementares (PNPIC) do Sistema Único de Saúde (SUS). Concluiu-se que há necessidade de intervenção didática sobre o tema para maior esclarecimento sobre as definições e conceitos relacionados à prática da homeopatia e das terapias integrativas, para os futuros profissionais farmacêuticos. (AU)

Homeopathy establishes principles to understand living beings as wholes; this art involves the analysis of pathological signs and symptoms, and the manipulation of specific drugs to induce integral recovery of health in living beings. A resolution by the Brazilian Federal Council of Pharmacy (2005) establishes the minimum requirements for pharmacists to assume the task of preparing homeopathic medicines. To understand the ideas of undergraduate pharmacy students at Lutheran University Center of Ji-Paraná, Brazil (Centro Universitário Luterano de Ji,-Paraná (CEULJI/ULBRA), a questionnaire was applied to assess their understanding relative to some definitions and concepts used in homeopathy and integrative medicine as a whole. Out of the full sample (n = 120), a considerable proportion of participants (96%) declared to know what homeopathy is, and 30% had obtained this information from people close to them. less than half of the participants were able to characterize or define homeopathy correctly as a therapeutic tool, while some mistook it for Bach flower therapy (28.8%). Among the participants who reported having used homeopathy, 81% did it without a medical prescription, and a considerable part of them was unaware of the existence of the National Policy for Integrative and Complementary Practices (PNPIC) of the Unified Health System (SUS). We conclude that didactic interventions on this subject are needed looking to a better explanation about the definitions and concepts related to the practice of the homeopathy and the integrative therapies, for those future pharmacists. (AU)

Percepções de acadêmicos de farmácia sobre a homeopatia / Perceptions of undergraduate pharmacy students on homeopathy

A homeopatia define princípios que buscam interpretar os seres vivos como um todo, envolve a arte da análise de sinais e sintomas patológicos e da manipulação de medicamentos específicos para o restabelecimento da saúde integral dos seres vivos. Existe uma resolução específica do Conselho Federal de Farmácia (2005) que estabelece os requisitos mínimos para que o profissional farmacêutico possa atuar como responsável técnico em homeopatia. Com o intuito de compreender o entendimento dos graduandos em farmácia que já cursaram ou cursarão a disciplina de homeopatia oferecida pelo curso de farmácia no Centro Universitário Luterano de Ji- Paraná (CEULJI/ULBRA), buscou-se avaliar por meio de um questionário o entendimento sobre algumas definições e conceitos relacionados à homeopatia e ao universo das práticas integrativas. Do total de participantes (n=120) um número considerável (96%) declarou conhecer a homeopatia, sendo que aproximados 30% obtiveram informações prévias de pessoas próximas, menos da metade dosparticipantes conseguiram caracterizá-la ou defini-la de maneira correta como instrumento terapêutico e alguns a confundiram com terapia floral (28,8%). Dos que declararam já terem se tratado com homeopatia, 81% o fizeram sem prescrição médica e grande parte não sabia da existência da Política Nacional de Práticas Integrativas e Complementares (PNPIC) do Sistema Único de Saúde (SUS). Concluiu-se que há necessidade de intervenção didática sobre o tema para maior esclarecimento sobre as definições e conceitos relacionados à prática da homeopatia e das terapias integrativas, para os futuros profissionais farmacêuticos.

Homeopathy establishes principles to understand living beings as wholes; this art involves the analysis of pathological signs and symptoms, and the manipulation of specific drugs to induce integral recovery of health in living beings. A resolution by the Brazilian Federal Council of Pharmacy (2005) establishes the minimum requirements for pharmacists to assume the task of preparing homeopathic medicines. To understand the ideas of undergraduate pharmacy students at Lutheran University Center of Ji-Paraná, Brazil (Centro Universitário Luterano de Ji,-Paraná (CEULJI/ULBRA), a questionnaire was applied to assess their understanding relative to some definitions and concepts used in homeopathy and integrative medicine as a whole. Out of the full sample (n = 120), a considerable proportion of participants (96%) declared to know what homeopathy is, and 30% had obtained this information from people close to them. less than half of the participants were able to characterize or define homeopathy correctly as a therapeutic tool, while some mistook it for Bach flower therapy (28.8%). Among the participants who reported having used homeopathy, 81% did it without a medical prescription, and a considerable part of them was unaware of the existence of the National Policy for Integrative and Complementary Practices (PNPIC) of the Unified Health System (SUS). We conclude that didactic interventions on this subject are needed looking to a better explanation about the definitions and concepts related to the practice of the homeopathy and the integrative therapies, for those future pharmacists.

Antimicrobial activity of amazonian medicinal plants.

OBJECTIVES: The aqueous extracts of currently utilized Amazonian medicinal plants were assayed in vitro searching for antimicrobial activity against human and animal pathogenic microorganisms. METHODS: Medium resuspended lyophilized aqueous extracts of different organs of Amazonian medicinal plants were assayed by in vitro screening for antimicrobial activity. ATCC and standardized microorganisms obtained from Oswaldo Cruz Foundation/Brazil were individually and homogeneously grown in agar plate, and holes previously perforated in the gel were filled with diluted plant aqueous extracts. Inhibition halos were evaluated and controlled by the use of the fluoroquinolone ciprofloxacin. RESULTS: The Amazonian medicinal plants, Hymenelobium petraeum showed inhibitory activity over Staphylococcus aureus, Enterococcus faecalis, Salmonella enterica serovar Typhi, Acinetobacter baumannii and Candida albicans, while Vatairea guianensis and Symphonia globulifera presented inhibitory activity exclusively for Staphylococcus aureus. Also, Ptychopetalum olacoides and Pentaclethra macroloba inhibited the growth of Klebsiella ozaenae and Acinetobacter baumannii. CONCLUSION: The aqueous botanic extracts that showed activity against microroganisms of ATTC and Osvaldo Cruz strains had at least 40% of antimicrobial activity when compared to halo inhibition produced by the commercial antibiotic ciprofloxacin utilized as a control. Of all plants extracts assayed, the Hymenelobium petraeum had the best performance, sometimes exhibiting higher activity than ciprofloxacin. It is not well-defined by the physicians the exact indication of the majority of medicinal plants in the Amazon area in Brazil. Natives utilize the plants according to their symptoms, based on the traditional knowledge transmitted orally from generation to generation, among Amerindians, Afrodescendents and ethnic mixed populations. A significant number of Amazonian medicinal plants are totally unknown related to their medicinal properties including mechanism of action and therapeutic effects, as very few information is reported in the scientific literature. A tiny amount of data is presented, as the preliminary antimicrobial properties of the medicinal plants here accessed, under the urgent necessity of new antibiotics in the market and in face of the increased resistance of infectious microorganisms to antimicrobials.