Exposure to Pfaffia glomerata causes oxidative stress and triggers hepatic changes.

Medicinal plant species are genetically engineered to obtain higher production of biomass and specific secondary metabolites, which can be used in the pharmaceutical industry. The aim of the present study was to evaluate the effect of Pfaffia glomerata (Spreng.) Pedersen tetraploid hydroalcoholic extract on the liver of adult Swiss mice. The extract was prepared from the plant roots and given to the animals by gavage, for 42 days. The experimental groups were treated with water (control), Pfaffia glomerata tetraploid hydroalcoholic extract (100, 200 and 400 mg/kg) and Pfaffia glomerata tetraploid hydroalcoholic extract discontinuously (200 mg/kg). The last group received the extract every 3 days, for 42 days. The oxidative status, mineral dynamics, and cell viability were analysed. The liver weight and the number of viable hepatocytes were reduced, despite the increased cell's number. Increased levels of malondialdehyde and nitric oxide, and changes in iron, copper, zinc, potassium, manganese and sodium levels were observed. aspartate aminotransferase levels were increased while alanine aminotransferase levels were decreased due to BGEt intake. Our results showed that BGEt induced alterations of oxidative stress biomarkers leading to liver injury, which was associated with a reduction in the number of hepatocytes.

Effects of the juçara fruit (Euterpe edulis Martius) pulp and lyophilized extract on NRF2, KEAP1, SOD1, and GPX2 expression in human colorectal cancer cell lines.

We investigated the effects of the juçara fruit (Euterpe edulis Martius) pulp and lyophilized extract on the expression of cytoprotective genes nuclear factor erythroid 2 (NF-E2)-related factor 2 (NRF2), kelch-like ECH-associated protein 1 (KEAP1), superoxide dismutase (SOD1), and glutathione peroxidase (GPX2) in human colorectal cancer cell lines (HT-29 and Caco-2). Cells were cultured for 24 h in Dulbecco's Modified Eagle's Medium containing juçara fruit pulp (5, 10, or 50 mg/mL) or lyophilized extract (0.05, 0.1, or 0.5 mg/mL), and gene expression was quantified using real-time quantitative reverse transcription polymerase chain reaction. All studied genes showed significant variation in gene expression among different concentrations of pulp or lyophilized extract. Overall, the expression of the selected genes decreased in both cell lines following exposure to the pulp or lyophilized extract in a dose-dependent manner for most of the concentrations studied. In summary, our study showed that the compounds in juçara fruit inhibited the expression of cytoprotective genes associated with the antioxidant response and that, although not cytotoxic at the concentrations studied, they could potentially block the activation of the NRF2/KEAP1 pathway.

A Physiological behavior and tolerance of Lactuca sativa to lead nitrate and silver nitrate heavy metals / Comportamento fisiológico e tolerância de Lactuca sativa aos metais pesados nitrato de chumbo e nitrato de prata

This study goal to evaluate the effects of different concentrations of lead (Pb) and silver (Ag) on germination, initial growth and anatomical alterations of Lactuca sativa L. Plants use various mechanisms to reduce the impacts caused by anthropic action, such as xenobiotic elements of soils and water contaminated by heavy metals. These metals were supplied as lead nitrate and silver nitrate and the following treatments were established: control for both metals, maximum dose of heavy metals, for arable soils, allowed by the National Council of the Environment (Ag = 25 mg. Kg-¹, Pb = 180 mg. Kg-¹), double (Ag = 50 mg. Kg-¹, Pb = 360 mg. Kg-¹) and triple (Ag = 75 mg. Kg-¹, Pb = 540 mg. Kg-¹) of this dosage. Vigor and germination tests of the seeds and possible anatomical changes in the leaves and roots of lettuce plants were performed. The species showed a high capacity to germinate under Pb and Ag stress, and the germination was never completely inhibited; however, the germination decreased with increasing Pb concentrations, but not under Ag stress. The use of increasing doses of metals reduced seed vigor and increased chlorophyll content. An increase in biomass was also observed in plants from treatments submitted to Pb. The phytotoxic effects of metals were more pronounced at 15 days after sowing. Anatomically, L. sativa was influenced by metal concentrations, and had a reduction of up to 79.9% in root epidermis thickness at the highest Pb concentration, although some structures did not suffer significant changes. The results suggest that L. sativa presents tolerance to high concentrations of heavy metals, showing possible mechanisms to overcome the stress caused by these metals. In this research lettuce possibly used the mechanism of exclusion of metals retaining Pb and Ag in the roots preserving the photosynthetic apparatus in the aerial part of the plants. In general, the chemical element Pb was more toxic than Ag, in these experimental conditions.

Este estudo teve como objetivo avaliar os efeitos de diferentes concentrações de chumbo (Pb) e prata (Ag) na germinação, crescimento inicial e alterações anatômicas de Lactuca sativa L. As plantas utilizam vários mecanismos para reduzir os impactos causados pela ação antrópica, como elementos xenobióticos de solos e água contaminada por metais pesados. Esses metais foram fornecidos como nitrato de chumbo e nitrato de prata e foram estabelecidos os seguintes tratamentos:controle para ambos os metais, dose máxima de metais pesados, para solos cultiváveis, permitida pelo Conselho Nacional do Meio Ambiente (Ag = 25mg.Kg-¹, Pb = 180mg.Kg-¹), dobro (Ag = 50mg.Kg-¹, Pb = 360mg.Kg-¹) e triplo (Ag = 75mg.Kg-¹, Pb = 540 mg.Kg-¹) desta dosagem. Foram realizados testes de vigor e germinação das sementes e possíveis alterações anatômicas nas folhas e raízes das plantas de alface. A espécie apresentou alta capacidade de germinar sob estresse de Ag e Pb, e a germinação nunca foi completamente inibida; entretanto, a germinação diminuiu com o aumento das concentrações de Pb, mas não sob estresse de Ag. O uso de doses crescentes dos metais, reduziu o vigor das sementes e aumentou o teor de clorofila. Também foi observado aumento da biomassa nas plantas a partir dos tratamentos submetidos ao Pb. Os efeitos fitotóxicos dos metais foram mais acentuados aos 15 dias após a semeadura. Anatomicamente, L. sativa foi influenciada pelas concentrações de metais, e teve uma redução de até 79,9% na espessura da epiderme radicular na maior concentração de Pb, embora algumas estruturas não tenham sofrido alterações significativas. Os resultados sugerem que L. sativa apresenta tolerância a altas concentrações de metais pesados, mostrando possíveis mecanismos para superar o estresse causado por esses metais. Nesta pesquisa a alface possivelmente utilizou o mecanismo de exclusão de metais retendo Pb e Ag nas raízes preservando o aparato [...].

A Physiological behavior and tolerance of Lactuca sativa to lead nitrate and silver nitrate heavy metals / Comportamento fisiológico e tolerância de Lactuca sativa aos metais pesados nitrato de chumbo e nitrato de prata

This study goal to evaluate the effects of different concentrations of lead (Pb) and silver (Ag) on germination, initial growth and anatomical alterations of Lactuca sativa L. Plants use various mechanisms to reduce the impacts caused by anthropic action, such as xenobiotic elements of soils and water contaminated by heavy metals. These metals were supplied as lead nitrate and silver nitrate and the following treatments were established: control for both metals, maximum dose of heavy metals, for arable soils, allowed by the National Council of the Environment (Ag = 25 mg. Kg-1, Pb = 180 mg. Kg-1), double (Ag = 50 mg. Kg-1, Pb = 360 mg. Kg-1) and triple (Ag = 75 mg. Kg-1, Pb = 540 mg. Kg -1) of this dosage. Vigor and germination tests of the seeds and possible anatomical changes in the leaves and roots of lettuce plants were performed. The species showed a high capacity to germinate under Pb and Ag stress, and the germination was never completely inhibited; however, the germination decreased with increasing Pb concentrations, but not under Ag stress. The use of increasing doses of metals reduced seed vigor and increased chlorophyll content. An increase in biomass was also observed in plants from treatments submitted to Pb. The phytotoxic effects of metals were more pronounced at 15 days after sowing. Anatomically, L. sativa was influenced by metal concentrations, and had a reduction of up to 79.9% in root epidermis thickness at the highest Pb concentration, although some structures did not suffer significant changes. The results suggest that L. sativa presents tolerance to high concentrations of heavy metals, showing possible mechanisms to overcome the stress caused by these metals. In this research lettuce possibly used the mechanism of exclusion of metals retaining Pb and Ag in the roots preserving the photosynthetic apparatus in the aerial part of the plants. In general, the chemical element Pb was more toxic than Ag, in these experimental conditions.

Este estudo teve como objetivo avaliar os efeitos de diferentes concentrações de chumbo (Pb) e prata (Ag) na germinação, crescimento inicial e alterações anatômicas de Lactuca sativa L. As plantas utilizam vários mecanismos para reduzir os impactos causados pela ação antrópica, como elementos xenobióticos de solos e água contaminada por metais pesados. Esses metais foram fornecidos como nitrato de chumbo e nitrato de prata e foram estabelecidos os seguintes tratamentos: controle para ambos os metais, dose máxima de metais pesados, para solos cultiváveis, permitida pelo Conselho Nacional do Meio Ambiente (Ag = 25mg.Kg-1, Pb = 180mg.Kg-1), dobro (Ag = 50mg.Kg-1, Pb = 360mg.Kg-1) e triplo (Ag = 75mg.Kg-1, Pb = 540 mg.Kg -1) desta dosagem. Foram realizados testes de vigor e germinação das sementes e possíveis alterações anatômicas nas folhas e raízes das plantas de alface. A espécie apresentou alta capacidade de germinar sob estresse de Ag e Pb, e a germinação nunca foi completamente inibida; entretanto, a germinação diminuiu com o aumento das concentrações de Pb, mas não sob estresse de Ag. O uso de doses crescentes dos metais, reduziu o vigor das sementes e aumentou o teor de clorofila. Também foi observado aumento da biomassa nas plantas a partir dos tratamentos submetidos ao Pb. Os efeitos fitotóxicos dos metais foram mais acentuados aos 15 dias após a semeadura. Anatomicamente, L. sativa foi influenciada pelas concentrações de metais, e teve uma redução de até 79,9% na espessura da epiderme radicular na maior concentração de Pb, embora algumas estruturas não tenham sofrido alterações significativas. Os resultados sugerem que L. sativa apresenta tolerância a altas concentrações de metais pesados, mostrando possíveis mecanismos para superar o estresse causado por esses metais. Nesta pesquisa a alface possivelmente utilizou o mecanismo de exclusão de metais retendo Pb e Ag nas raízes preservando o aparato fotossintético na parte aérea das plantas. De forma geral o elemento químico Pb se mostrou mais tóxico que Ag, nestas condições experimentais.

Efficacy of the laser at low intensity on primary burning oral syndrome: a systematic review.

BACKGROUND: Primary burning mouth syndrome (BMS) is a chronic clinical condition of idiopathic mainly characterized by pain and a burning sensation in the oral cavity. The application of laser at low intensity therapy is a treatment option. This systematic review evaluated the efficacy of laser therapy in treating symptoms of burning mouth syndrome. MATERIAL AND METHODS: The study was formulated according to the PRISMA and Cochrane guidelines. Seven databases were used as primary sources of research. Only randomized controlled clinical trials were included. The efficacy of the therapy was estimated comparing the values of the visual and numerical scales of pain before and after laser treatment, through qualitative analysis. RESULTS: The search resulted in 348 records and only eight filled the eligibility criteria and were included. All studies evaluated pain and / or a burning sensation considering a time interval of two to ten weeks. The total sample consisted of 314 patients submitted to treatment: 123 from the control group, who participated with laser off or with the tip blocked, and 191 from the intervention group, treated with low-level laser therapy. The female gender stood out and the average age of the participants was 60.89 years. The main symptoms reported were pain and a burning sensation in the oral mucosa and tongue. The parameters adopted by the authors for laser treatment were diverse and the variables were not fully described in the published studies. Visual analog and numerical scales were used to assess symptoms and only three studies showed statistical significance. CONCLUSIONS: It is suggested that laser therapy may be an effective alternative in the treatment of BMS. New randomized clinical trials should consider well-established protocols to better understand the efficacy of laser therapy without confounding the effects.

Anti-Candida activity of the water-soluble lectin from Moringa oleifera seeds (WSMoL).

This work reports the effects of the water-soluble lectin from Moringa oleifera seeds (WSMoL) on growth and survival of Candida species. In addition, cellular alterations linked to the antifungal effect were investigated. The minimal inhibitory (MIC) and fungicidal (MFC) concentrations were determined and 24-h growth curves in absence and presence of lectin were established. Flow cytometry was used to evaluate the induction of apoptosis/necrosis, alterations in mitochondrial membrane potential (ΔΨm), and occurrence of lysosomal damage. WSMoL inhibited the growth of C. albicans, C. glabrata, C. krusei and C. parapsilosis with MIC of 20µg/mL. The lowest MFC (20µg/mL) was detected for C. glabrata and the highest (80µg/mL) for C. albicans and C. parapsilosis. The inhibitory effect started from the ninth to nineteenth hour of incubation depending on the fungal species. Incubation with the lectin at the MIC for 24h increased the number of cells undergoing apoptosis and necrosis. Hyperpolarization of the mitochondrial membrane was detected after 12-h treatment, followed by reduction of ΔΨm or depolarization after 24h. No lysosomal damage was detected in treated cells. In conclusion, WSMoL is a fungistatic and fungicide agent against Candida with differential effects depending on the species.

Effect of the subdose of human chorionic gonadotropin applied in the Hou Hai acupoint on ovulation induction in mares / [Efeito da subdose de gonadotrofina coriônica humana aplicada no acuponto Hou Hai sobre a indução da ovulação em éguas]

The objective of this study was to evaluate the effects of an hCG sub dose applied at the Hou Hai acupoint on corpus luteum (CL) quality and ovulation induction in mares. Fifteen crossbred mares were distributed in randomized blocks and used in three periods with each period employed as the blocking factor in three treatments: T1 = 1500 IU of hCG via intravenous (IV); T2 = 450 IU of hCG applied at the false acupoint (IV); and T3 = 450 IU of hCG applied at the Hou Hai acupoint. Mean diameter of the CL, serum concentration of progesterone (P4), vascularization of the pre-ovulatory follicle and CL were evaluated. Females administered 450 IU of hCG at the Hou Hai acupoint exhibited greater ovulation rates (33.33%) 48h after induction; The minimum number of colored pixel (NCP) of the pre-ovulatory follicle of control females was superior (40.33) to that of mares administered 450 IU of hCG IV at the false acupoint (36.84) and similar to that of those administered hCG at the Hou Hai acupoint (39.31). Further, moderately positive correlations were found between the CL diameter and the P4 concentration on D8 (P<0.05). IV administration of 450 IU of hCG or at the Hou Hai acupoint was efficient at inducing ovulation and ensuring the quality of CL in mares.(AU)

O objetivo foi avaliar os efeitos de uma subdose de hCG aplicada no acuponto Hou Hai na qualidade do corpo lúteo (CL) e na indução da ovulação em éguas. Quinze éguas mestiças foram distribuídas em blocos ao acaso, sendo o período utilizado como fator de blocagem, em: T1 = 1500 UI de hCG por via intravenosa (IV); T2 = 450 UI de hCG aplicado no falso acuponto (IV) e T3 = 450 UI de hCG aplicada no acuponto Hou Hai. Avaliou-se diâmetro médio do CL, concentração sérica de progesterona (P4), vascularização do folículo pré-ovulatório e do CL. As fêmeas que receberam 450 UI de hCG no acuponto Hou Hai apresentaram maiores taxas de ovulação (33,33%) 48h após a indução. O número de pixels coloridos (NPC) mínimo do folículo pré-ovulatório das fêmeas do grupo controle foi superior (40,33) ao das éguas que receberam 450 UI de hCG IV no falso acuponto (36,84) e semelhante ao das éguas que receberam hCG no acuponto Hou Hai (39,31); correlações moderadamente positivas foram encontradas entre o diâmetro do CL e a concentração de P4, ambos no D8 (P <0,05). A administração IV de 450 UI de hCG ou no acuponto Hou Hai foi eficiente na indução da ovulação e na garantia da qualidade do CL nas éguas.(AU)

Latrodectus geometricus (Aranea: Theridiidae) envenoming: Rapid resolution of symptoms following F(ab')2 antivenom therapy.

The Brown Widow spider (Latrodectus geometricus) is an invasive species whose geographic range has been expanding worldwide. It is a relative species of the Black Widow and Red-backed spiders of the genus Latrodectus. Despite its broad geographic distribution cases of Brown Widow envenomation have rarely been documented. The venom of L. geometricus is similar to the venom of L. mactans with the primary venom component being alpha-latrotoxin, and consequent envenoming by L. geometricus to humans has resulted in symptoms similar to those reported for other Latrodectus spp. Specific FDA approved Latrodectus antivenom (IgG) available in North America has been effectively used in treating venom-induced symptoms following L. mactans envenoming. The patient reported here involved a confirmed L. geometricus envenoming who was efficaciously treated with an alternately available F(ab')2 antivenom from Mexico.

Furoxan derivatives demonstrated in vivo efficacy by reducing Mycobacterium tuberculosis to undetectable levels in a mouse model of infection.

OBJECTIVES: The most recent survey conducted by the World Health Organization described Tuberculosis (TB) as one of the top 10 causes of death and the leading cause of death from a single infectious agent. The increasing number of TB-resistant cases has contributed to this scenario. In light of this, new strategies to control and treat the disease are necessary. Our research group has previously described furoxan derivatives as promising scaffolds to be explored as new antitubercular drugs. RESULTS: Two of these furoxan derivatives, (14b) and (14c), demonstrated a high selectivity against Mycobacterium tuberculosis. The compounds (14b) and (14c) were also active against a latent M. tuberculosis strain, with MIC90 values of 6.67 µM and 9.84 µM, respectively; they were also active against monoresistant strains (MIC90 values ranging from 0.61 to 20.42 µM) and clinical MDR strains (MIC90 values ranging from 3.09 to 42.95 µM). Time-kill experiments with compound (14c) showed early bactericidal effects that were superior to those of the first- and second-line anti-tuberculosis drugs currently used in therapy. The safety of compounds (14b) and (14c) was demonstrated by the Ames test because these molecules were not mutagenic under the tested conditions. Finally, we confirmed the safety, and high efficacy of compounds (14b) and (14c), which reduced M. tuberculosis to undetectable levels in a mouse aerosol model of infection. CONCLUSION: Altogether, we have identified two advanced lead compounds, (14b) and (14c), as novel promising candidates for the treatment of TB infection.

Antibacterial activity of Barbatimão (Stryphnodendron adstringens) against Staphylococcus aureus: in vitro and in silico studies.

We evaluated the activity of the aqueous fraction and the ethyl acetate fraction of Stryphnodendron adstringens against Staphylococcus aureus and proposed their mechanism of action. The antibacterial activity of S. adstringens fractions was evaluated against S. aureus and the cell targets were rated by docking. The fractions showed moderate antibacterial activity against S. aureus without toxicity on two mammalian cell lines. They also showed synergistic antibacterial activity with tannic acid (TA). In silico assays indicated FabG, FabZ and FabI as probable targets. The metabolic pathway for fatty acid biosynthesis in S. aureus was affected by components of S. adstringens. The synergistic effect when combining TA with S. adstringens fractions suggests a natural alternative to S. aureus control. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study describing the possible targets of action of Stryphnodendron adstringens on Staphylococcus aureus. Molecular dynamics simulations showed that the components of S. adstringens affected the metabolic pathway for fatty acid biosynthesis (FAS II) in S. aureus, inhibiting the FabI, FabG and FabZ enzymes. As tannic acid (TA) is a known inhibitor of some targets identified, we showed synergistic antibacterial activity of S. adstringens in combination with TA. This combination did not show toxicity against HaCaT and Vero cells and based on all these results we suggest that S. adstringens can be a natural and sustainable alternative to S. aureus control.

Terapias biológicas en embarazo y lactancia / Biological Therapies in Pregnancy and lactation

Las terapias biológicas son una herramienta terapéutica fundamental en cierto grupo de pacientes, sin embargo, su seguridad en el embarazo y lactancia no está aún dilucidada. El objetivo de esta revisión es actualizar los conocimientos y recomendaciones respecto al uso de fármacos biológicos durante el embarazo y lactancia. En relación al embarazo, las recomendaciones actuales sugieren que los anti TNF se podrían continuar o mantener hasta el segundo trimestre del embarazo. La terapia anti CD20 (Rituximab) y otros biológicos se recomiendan suspender en el embarazo. Durante la lactancia, se puede iniciar o continuar cualquier terapia biológica. En la primera mitad del embarazo se pueden utilizar algunos biológicos con relativo buen perfil de seguridad y en la lactancia la evidencia sugiere que no producirían mayor riesgo en el niño. Sería ideal contar con estudios randomizados controlados, pero esto podría generar un dilema ético.

Biological therapies are a fundamental therapeutic tool in a certain group of patients. However, the safety of these drugs during pregnancy and lactation remains unclear. Current recommendations regarding anti TNF use during pregnancy suggest these could be maintained until the second trimester of pregnancy. Anti CD20 therapy (Rituximab) and other biologics should not be used in pregnancy. Patients can safely initiate or continue to use biological therapy during lactation. Some biologicals are relatively safe to use during pregnancy and recent studies show that they can be used during lactation without being a major threat to the child. It would be ideal to have randomized controlled studies, but this could create an ethical dilemma.

Mitos y realidades sobre el aporte de lácteos y calcio / Myths and facts about dairy foods and calcium supplementation

Desde una perspectiva histórica, el consumo de leche de otros mamíferos y sus derivados ha jugado un rol importante en el desarrollo de la sociedad humana. Sin embargo, en los últimos años se han planteado dudas respecto al real beneficio del consumo de productos lácteos y suplementos de calcio en la salud ósea. Más aún, se les han atribuido potenciales efectos adversos como el incremento del riesgo cardiovascular, aumento en la incidencia de cáncer, trastornos digestivos, nefrolitiasis y una mayor mortalidad. Si bien la evidencia disponible puede ser controversial, en la mayoría de los casos desmiente categóricamente estas afirmaciones.En esta revisión de la literatura, intentamos despejar las inquietudes respecto al beneficio y riesgo del consumo de lácteos, suplementos de calcio y Vitamina D.

From the historical perspective, milk and dairy product consumption has played an important role in the development of human society. However, in the recent years, some doubts have been raised regarding the benefits of dairy food consumption and calcium supplements on bone health. Additionally, potential adverse effects have been attributed to their use, such as increased cardiovascular risk, increased incidence of cancer, digestive troubles, nephrolithiasis and increased mortality rate. Although the available evidence may be controversial, in most cases it categorically refutes these statements.In this review of the literature, we try to address concerns regarding benefit and risks of consumption of dairy products, calcium and vitamin D supplements.

Copper(II) complex-loaded castor oil-based nanostructured lipid carriers used against Mycobacterium tuberculosis : Development, characterisation, in vitro and in vivo biological assays.

A copper(II) complex-loaded castor oil-based nanostructured lipid carrier was evaluated to enhance the poor water solubility of antimicrobial compounds, improving their biological properties and antimicrobial activity against Mycobacterium tuberculosis. Nanostructured lipid carriers were composed of the castor oil, polyoxyethylene 40 stearate and caprylic/capric triglyceride, poloxamer 407, cetyltrimethylammonium bromide and three different copper(II) complexes. The systems were ultrasonicated at an amplitude of 8% for 20 min and an ice bath was used throughout the procedure. The blank nanostructured lipid carrier (F5) and nanostructured lipid carriers loaded with copper(II) complex 1, 2 and 3 (F5.1, F5.2 and F5.3, respectively) for 45 days presented values of mean diameter, poly dispersity index and zeta potential ranging from 186 to 199 nm, 0.14 to 0.2 and 24 to 30 mV, respectively. Atomic force microscopy indicated that the nanostructured lipid carriers were distributed at the nanoscale, corroborating the mean diameter data. Differential scanning calorimetry determined the melting points of the constituents of the nanostructured lipid carriers. The antimicrobial activity of copper(II) complexloaded F5 against M. tuberculosis H37Rv showed better anti-tuberculosis activity than the free complexes. In vivo biological assays of complex-loaded F5 demonstrated reduced toxicity. Our results suggest that nanostructured lipid carriers could be a potential nanotechnological strategy to optimise tuberculosis treatment.

Direct and transdentinal biostimulatory effects of grape seed extract rich in proanthocyanidin on pulp cells.

AIM: To evaluate the potential biostimulatory effects of grape seed extract (GSE) on a primary culture of human pulp cells. METHODOLOGY: Human molars were used to obtain the primary pulp cell culture and 0.5-mm dentine discs. For GSE direct exposure, dose-response (0.0065-6.5%) and time response (1-60 min of contact) were examined. For transdentinal exposure, 0.65% of GSE was tested for 24 h. Cellular metabolism, nitric oxide and collagen production, and cell morphology alterations were assessed at periods of 24 and 72 h. After cell differentiation and direct exposure to GSE, the total protein production (TP), alkaline phosphatase activity (ALP) and formation of mineralization nodules (MN) were assessed. The results were analysed by parametric tests or non-parametric tests (α = 0.05). RESULTS: The lower concentration of GSE tested (0.0065%) was associated with an increase in cellular metabolism, a reduction in the production of nitric oxide and an increase in extracellular matrix synthesis (collagen). Distinct behaviours were observed for the different concentrations, without a reduction of cellular metabolism >10% compared with the control, either when applied directly or transdentinally. SEM revealed no significant change in cell morphology, except for the positive control (H2 O2 ). There was no difference in TP, ALP or MN between the control group and the group exposed to GSE. CONCLUSIONS: Treatment with grape seed extract, even at the highest concentration and longest period, caused neither direct nor transdentinal cytotoxic effects on human pulp cells. Grape seed extract components may play a biostimulatory role and protect dental pulp cells when in direct contact.

The impact of operational strategies on the performance of a photo-EBPR system.

A novel Phototrophic - Enhanced Biological Phosphorus Removal (Photo-EBPR) system, consisting of a consortium of photosynthetic organisms and polyphosphate accumulating organisms (PAOs), was studied in this work. A sequencing batch reactor was fed with a mixture of acetate and propionate (75%-25%) and subjected to dark/light cycles in order to select a photo-EBPR system containing PAOs and photosynthetic organisms, the latter likely providers of oxygen to the system. The results from the selection period (stage 1) showed that the photo-EBPR culture was capable of performing P release in the dark and P uptake in the presence of light, under limited oxygen concentrations. During the optimization period, the aeration period, which was initially provided at the end of the light phase, was gradually reduced until a non-aerated system was achieved, while the light intensity was increased. After optimization of the operational conditions, the selected consortium of photosynthetic organisms/PAOs showed high capacity of P removal in the light phase in the absence of air or other electron acceptor. A net P removal of 34 ± 3 mg-P/L was achieved, with a volumetric P removal rate of 15 ± 2 mg-P/L.h, and 79 ± 8% of P removal from the system. Also, in limiting oxygen conditions, the P uptake rate was independent of the PHA consumption, which demonstrates that the organisms obtained energy for P removal from light. These results indicated that a photo-EBPR system can be a potential solution for P removal with low COD/P ratios and in the absence of air, prospecting the use of natural sunlight as illumination, which would reduce the costs of EBPR operation regarding aeration.

Drug repurposing strategy against Trypanosoma cruzi infection: In vitro and in vivo assessment of the activity of metronidazole in mono- and combined therapy.

Metronidazole (Mtz) is a commercial broad-spectrum nitroimidazolic derivative with relevant antimicrobial activity and relative safety profile. Therefore, it is fair to consider Mtz a candidate for drug repurposing for other neglected conditions such as Chagas disease (CD), a parasitic pathology caused by Trypanosoma cruzi. CD is treated only with benznidazole (Bz) and nifurtimox, both introduced in clinics decades ago despite important limitations, including low efficacy on the later disease stage (chronic form) and severe side effects. New cheap and fast alternative treatments for CD are needed, thus the repurposing of Mtz was assessed in vitro and in vivo in mono- and combined therapy. In vitro assays demonstrated EC50>200µM for Mtz, while for Bz the values ranged from 2.51µM (intracellular forms) to 11.5µM (bloodstream trypomastigotes). When both drugs were combined in fixed-ratio proportions, Mtz promoted Bz potency (lower EC50 values). In vivo toxicity assays for Mtz in mice showed no adverse effects neither histopathological alterations up to 2000mg/kg. Regarding experimental T. cruzi infection, Bz 100mg/kg suppressed parasitemia while Mtz (up to 1000mg/kg) in monotherapy did not, but prolonged animal survival at 250 and 500 regimen doses. The combination of both drugs (Bz 10+Mtz 250) prevented mortality (70%) besides protected against electric cardiac alterations triggered by the parasite infection. Although not able to reduce parasite load, the combination therapy prevented animal mortality; this was possibly due to a protection of the electric cardiac physiology that is normally altered in experimental infection of T. cruzi. It also suggested that the interaction with Mtz could have improved the pharmacokinetics of Bz. Our study emphasizes the importance of drug repurposing and combined therapy for CD to contribute to alternative therapies for this neglected and silent pathology.

Epitelização de enxertos cutâneos em feridas recentes de coelhos tratados com membrana amniótica canina e/ou laserterapia / Skin graft epithelialization in rabbit fresh wounds treated with amniotic membrane and/or laser therapy

A enxertia cutânea é uma técnica cirúrgica simples e bastante útil para o reparo de feridas, principalmente aquelas onde existe dificuldade da aplicação do fechamento primário ou de outras técnicas reconstrutivas. Entretanto, para a sobrevivência do enxerto, é necessário que o leito da ferida esteja saudável e com presença de tecido de granulação exuberante. O objetivo do presente trabalho foi avaliar a aplicação da membrana amniótica e da laserterapia como potenciais estimulantes da cicatrização em enxertos aplicados em feridas sem tecido de granulação. Foram utilizados 42 coelhos, divididos em quatro grupos: grupo controle (GC), grupo membrana (GM), grupo laser (GL) e grupo membrana e laser (GML), submetidos a avaliações macro e microscópicas. Na avaliação macroscópica, foi possível notar que os pacientes dos grupos nos quais a membrana amniótica foi utilizada (GM e GML) apresentaram evidências associadas à acentuada reação inflamatória, à falha de integração do enxerto e à consequente necrose dele. Já os pacientes do GL apresentaram melhor aspecto do enxerto no último dia de avaliação. Na análise microscópica, observou-se intensa integração do enxerto à derme, reepitelização acentuada e escassas células inflamatórias no local do enxerto no GL. O contrário foi observado nos pacientes do GM e GML, nos quais aparentemente houve rejeição da membrana. A formação de colágeno não se correlacionou com outros fatores, como inflamação e necrose, em nenhum dos grupos de tratamento. Dessa forma, é possível afirmar que a laserterapia mostrou ser efetiva, contribuindo para o processo cicatricial e a integração do enxerto. Já a membrana amniótica canina não deve ser utilizada para esse fim, pois provoca intensa reação inflamatória, além de impedir a nutrição do enxerto.(AU)

Skin grafting is a simple surgical technique and useful to repair wounds, especially those where there is a difficulty to apply primary closure skin or other reconstructive techniques. However, for graft survival a healthy wound bed and the presence of an exuberant granulation tissue are necessary. This study aimed to evaluate the application of amniotic membrane and laser therapy as potential healing stimulants in grafts applied in wounds without granulation tissue. For this, we used 42 rabbits divided into four treatment groups, control group (CG), membrane group (MG), laser group (LG) and membrane and laser group (MLG), submitted to macroscopic and microscopic evaluation. The macroscopic examination showed that the patients of the groups where the amniotic membrane was used in order to stimulate re-epithelialization (MG and MLG) presented evidences associated with severe inflammatory reaction, graft integration failure and consequent necrosis. LG patients apparently had the best graph aspect in the last valuation date. Microscopic examination showed intense integration of the graft to the dermis, high re-epithelialization level, and scarce inflammatory cells in the graft site of LG patients. The opposite was observed in patients in the MG and MLG groups, where a rejection of the membrane was observed. Finally, collagen formation was not correlated with other factors such as inflammation and necrosis in any of the treatment groups. We can conclude that laser therapy was effective, contributing to the healing process and integration of the graft. Thus, canine amniotic membrane should not be used for this purpose because it causes intense inflammatory reaction besides avoiding graft nutrition.(AU)

Global volatile profile of virgin olive oils flavoured by aromatic/medicinal plants.

The global volatile profile of commercial virgin olive oils and flavoured olive oils with aromatic/medicinal plants, was established using liquid-liquid microextraction (LLME) and headspace solid-phase microextraction (HS-SPME) combined with gas chromatography-quadrupole mass spectrometry (GC-qMS). More than 60 volatile organic compounds (VOCs belonging to different groups were identified using both methods. Olive oils volatile profile was slightly influenced by maceration process, which occurred at room temperature (20±2°C) for 15days. The predominant differences were observed in terpenoids group, since some of them were only identified in the flavoured olive oils, while others showed an increase with the maceration process. VOCs mass transfer from plants to olive oils could explain the observed results. Principal components analysis (PCA) applied to LLME/GC-qMS data allowed to distinguish the olive oils. The flavoured oils would increase the use of olive oil among consumers as consequence of the improvement of its aromatic profile and healthy properties.

Venous flow during manual lymphatic drainage applied to different regions of the lower extremity in people with and without chronic venous insufficiency: a cross-sectional study.

OBJECTIVES: To evaluate the effect of manual lymphatic drainage (MLD) on venous flow when applied to the medial and lateral aspects of the thigh and leg in patients with chronic venous insufficiency (CVI) and healthy subjects. DESIGN: Cross-sectional study. SETTING: Participants were assessed in a school-based health community attendant service. PARTICIPANTS: Fifty-seven subjects participated in this study {mean age: 43 [standard deviation (SD) 14] years, 38 women and 19 men}. Of these, 28 subjects had CVI [mean age 47 (SD 12) years] and 29 subjects did not have CVI [mean age 39 (14) years]. INTERVENTION: MLD was applied by a certificated physical therapist to the medial and lateral aspects of the thigh and leg. MAIN OUTCOME MEASUREMENTS: Cross-sectional area; blood flow velocities in the femoral vein, great saphenous vein, popliteal vein and small saphenous vein at baseline and during MLD, measured by duplex ultrasound. RESULTS: Flow volume in the femoral vein increased from baseline [5.19 (SD 3.25)cm3/second] when MLD was applied to the medial [7.03 (SD 3.65)cm3/second; P≤0.001; mean difference -1.69; 95% confidence interval (CI) -2.42 to -0.97] and lateral [6.16 (SD 3.35)cm3/second; P≤0.001; mean difference -1.04; 95% CI -1.70 to -0.39] aspects of the thigh. Venous flow augmentation in the femoral vein and great saphenous vein was higher when MLD was applied to the medial aspect of the thigh (P<0.001), while MLD had a similar effect on venous blood flow regardless of whether it was applied to the medial or the lateral aspect of the leg (P=0.731). CONCLUSIONS: MLD increases blood flow in deep and superficial veins. MLD should be applied along the route of the venous vessels for improved venous return.

Acrocomia aculeata prevents toxicogenetic damage caused by the antitumor agent cyclophosphamide.

Acrocomia aculeata is a plant rich in antioxidant compounds. Studies suggest that this plant has anti-inflammatory, antidiabetic, and diuretic potential. We assessed the antigenotoxic, antimutagenic, immunomodulation, and apoptotic potentials of A. aculeata alone and in combination with an antitumor agent, cyclophosphamide. Swiss male mice (N = 140) were used. The animals were divided into 14 experimental groups as follows: a negative group, a positive group (100 mg/kg cyclophosphamide), groups that only received the oil extracted from the almond (AO) and from the pulp (PO) of A. aculeata at doses of 3, 15, and 30 mg/kg, and the associated treatment groups (oils combined with cyclophosphamide) involving pretreatment, simultaneous, and post-treatment protocols. Data suggest that both oils were chemopreventive at all doses, based on the tested protocols. The highest damage reduction percentages, observed for AO and PO were 88.19 and 90.03%, respectively, for the comet assay and 69.73 and 70.93%, respectively, for the micronucleus assay. Both AO and PO demonstrated immunomodulatory activity. The oils reduced the capacity of cyclophosphamide to trigger apoptosis in the liver, spleen, and kidney cells. These results suggest that A. aculeate AO and PO can be classified as a functional food and also enrich other functional foods and nutraceuticals with chemopreventive features. However, they are not appropriate sources for chemotherapeutic adjuvants, in particular for those used in combination with cyclophosphamide.