Impact of different activation wavefronts on ischemic myocardial scar electrophysiological properties during high-density ventricular tachycardia mapping and ablation.

INTRODUCTION: Scar-related ventricular tachycardia (VT) usually results from an underlying reentrant circuit facilitated by anatomical and functional barriers. The later are sensitive to the direction of ventricular activation wavefronts. We aim to evaluate the impact of different ventricular activation wavefronts on the functional electrophysiological properties of myocardial tissue. METHODS: Patients with ischemic heart disease referred for VT ablation underwent high-density mapping using Carto®3 (Biosense Webster). Maps were generated during sinus rhythm, right and left ventricular pacing, and analyzed using a new late potential map software, which allows to assess local conduction velocities and facilitates the delineation of intra-scar conduction corridors (ISCC); and for all stable VTs. RESULTS: In 16 patients, 31 high-resolution substrate maps from different ventricular activation wavefronts and 7 VT activation maps were obtained. Local abnormal ventricular activities (LAVAs) were found in VT isthmus, but also in noncritical areas. The VT isthmus was localized in areas of LAVAs overlapping surface between the different activation wavefronts. The deceleration zone location differed depending on activation wavefronts. Sixty-six percent of ISCCs were similarly identified in all activating wavefronts, but the one acting as VT isthmus was simultaneously identified in all activation wavefronts in all cases. CONCLUSION: Functional based substrate mapping may improve the specificity to localize the most arrhythmogenic regions within the scar, making the use of different activation wavefronts unnecessary in most cases.

Myrciaria Genus: Bioactive Compounds and Biological Activities.

The Myrtaceae family is of angiosperms, imposing its size and economic, cultural, and scientific importance. The genus Myrciaria, belonging to this family, has 33 species currently accepted, many of which are research targets aimed at elucidating their bioactive compounds and biological activities. Most species of the Myrciaria genus have terpenes in their composition, mainly mono and sesquiterpenes, and phenolic compounds such as tannins, phenolic acids, and flavonoids. Other secondary metabolites are also observed, such as alkaloids, steroids, coumarins, saponins, and naphthoquinones. These bioactive compounds are closely related to these species' most diverse biological activities: antioxidant, anti-inflammatory, analgesic, antiproliferative, antimicrobial, antiparasitic, insecticide, metabolic, protective, and nutraceutical. This work aims to provide a review of secondary metabolites and medicinal properties related to the genus Myrciaria, thus stimulating further studies on the species of this genus.

Evaluation of the Antiproliferative Potential of Eugenia pyriformis Leaves in Cervical Cancer Cells.

Eugenia pyriformis, typically known as uvaia, ubaia, uvaieira, uvalha or uvalha-do-campo, is a plant representative of the Myrtaceae family. E. pyriformis decreased HeLa cells proliferation, can induce cell death and reduce cell migration that may be related to metastasis and induction of cell death by apoptosis in vitro assays. Its leaves are used in folk medicine for hypertension control, decreased cholesterol and uric acid, slimming, astringent, and digestive. In this work, the evaluation of the in vitro anticancer potential Cervical Cancer (HeLa cells) and phytochemical analysis in E. pyriformes was performed. It was possible to quantify phenolic compounds and total flavonoids and identify Chlorogenic acid, Quercetrin, and Myricitrin in this species. The crude extract and ethyl acetate fraction inhibited cell viability by 50 % in the dose of 44.42 µg/mL and 40.39 µg/mL, respectively. The induced effector caspase 3/7 activation, which results in apoptosis and the ethyl acetate fraction, decreases cell migration of cancer cell lines; it is responsible for the cleavage of several cellular proteins that will result in the classic phenotype of the apoptotic cell.

Genome Sequence and Analysis of the Flavinogenic Yeast Candida membranifaciens IST 626.

The ascomycetous yeast Candida membranifaciens has been isolated from diverse habitats, including humans, insects, and environmental sources, exhibiting a remarkable ability to use different carbon sources that include pentoses, melibiose, and inulin. In this study, we isolated four C. membranifaciens strains from soil and investigated their potential to overproduce riboflavin. C. membranifaciens IST 626 was found to produce the highest concentrations of riboflavin. The volumetric production of this vitamin was higher when C. membranifaciens IST 626 cells were cultured in a commercial medium without iron and when xylose was the available carbon source compared to the same basal medium with glucose. Supplementation of the growth medium with 2 g/L glycine favored the metabolization of xylose, leading to biomass increase and consequent enhancement of riboflavin volumetric production that reached 120 mg/L after 216 h of cultivation. To gain new insights into the molecular basis of riboflavin production and carbon source utilization in this species, the first annotated genome sequence of C. membranifaciens is reported in this article, as well as the result of a comparative genomic analysis with other relevant yeast species. A total of 5619 genes were predicted to be present in C. membranifaciens IST 626 genome sequence (11.5 Mbp). Among them are genes involved in riboflavin biosynthesis, iron homeostasis, and sugar uptake and metabolism. This work put forward C. membranifaciens IST 626 as a riboflavin overproducer and provides valuable molecular data for future development of superior producing strains capable of using the wide range of carbon sources, which is a characteristic trait of the species.

Prediction of Drug Synergism between Peptides and Antineoplastic Drugs Paclitaxel, 5-Fluorouracil, and Doxorubicin Using In Silico Approaches.

Chemotherapy is the main treatment for most early-stage cancers; nevertheless, its efficacy is usually limited by drug resistance, toxicity, and tumor heterogeneity. Cell-penetrating peptides (CPPs) are small peptide sequences that can be used to increase the delivery rate of chemotherapeutic drugs to the tumor site, therefore contributing to overcoming these problems and enhancing the efficacy of chemotherapy. The drug combination is another promising strategy to overcome the aforementioned problems since the combined drugs can synergize through interconnected biological processes and target different pathways simultaneously. Here, we hypothesized that different peptides (P1-P4) could be used to enhance the delivery of chemotherapeutic agents into three different cancer cells (HT-29, MCF-7, and PC-3). In silico studies were performed to simulate the pharmacokinetic (PK) parameters of each peptide and antineoplastic agent to help predict synergistic interactions in vitro. These simulations predicted peptides P2-P4 to have higher bioavailability and lower Tmax, as well as the chemotherapeutic agent 5-fluorouracil (5-FU) to have enhanced permeability properties over other antineoplastic agents, with P3 having prominent accumulation in the colon. In vitro studies were then performed to evaluate the combination of each peptide with the chemotherapeutic agents as well as to assess the nature of drug interactions through the quantification of the Combination Index (CI). Our findings in MCF-7 and PC-3 cancer cells demonstrated that the combination of these peptides with paclitaxel (PTX) and doxorubicin (DOXO), respectively, is not advantageous over a single treatment with the chemotherapeutic agent. In the case of HT-29 colorectal cancer cells, the combination of P2-P4 with 5-FU resulted in synergistic cytotoxic effects, as predicted by the in silico simulations. Taken together, these findings demonstrate that these CPP6-conjugates can be used as adjuvant agents to increase the delivery of 5-FU into HT-29 colorectal cancer cells. Moreover, these results support the use of in silico approaches for the prediction of the interaction between drugs in combination therapy for cancer.

Citral Effects on the Expression Profile of Brain-Derived Neurotrophic Factor and Inflammatory Cytokines in Status Epilepticus-Induced Rats Using the Lithium-Pilocarpine Model.

Epilepsy is one of the most common neurological disorders. About one-third of people with epilepsy are refractory to available treatments. Studies suggest that mechanisms linked to the immune response and inflammatory process are related to seizure disorders. Citral is a monoterpene found in the essential oil of several plants, as in Cymbopogon citratus, used to make teas and has been the subject of numerous researches, from which it has been possible to demonstrate antiseizure and anti-inflammatory activities. In this study, the effects of citral on status epilepticus (SE) induced by the lithium-pilocarpine model in rats were investigated. Quantitative reverse transcription PCR (RT-qPCR) evaluated latency for seizure development, neuronal death in the hippocampus, and expression of the brain-derived neurotrophic factor (BDNF), tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), interleukin-1ß ( IL-1ß) and factor nuclear kappa B (NF-κB) genes. The results revealed that citral was able to increase latency until the first seizure, decrease neuronal death 2 h after SE and inhibit overexpression of proinflammatory genes.

Jaboticaba (Myrciaria cauliflora) Peel Supplementation Prevents Hepatic Steatosis Through Hypolipidemic Effects and Cholesterol Metabolism Modulation in Diet-Induced Nonalcoholic Fatty Liver Disease Rat Model.

Jaboticaba (Myrciaria cauliflora), a Brazilian fruit, is a good source of dietary fiber and phenolic compounds, which are concentrated mainly in the peel. These compounds have been considered promising in prevention and treatment of hypercholesterolemia and hepatic steatosis. In this study, we investigated the effects of 4% jaboticaba peel powder (JPP) supplementation on cholesterol metabolism and hepatic steatosis in livers of rats fed a high-fat (HF) diet. The rats were fed a standard AIN-93M (control) diet or an HF diet containing 32% lard and 1% cholesterol, both with and without 4% JPP. The M. cauliflora peel composition revealed a low-lipid high-fiber content and phenolic compounds. The phenolic compounds in JPP, tentatively identified by high-performance liquid chromatography and mass spectrometry (HPLC-MS/MS) analysis, were confirmed to contain phenolic acids, flavonoids, and anthocyanins. Moreover, JPP presented significant antioxidant activity in vitro and was not cytotoxic to HepG2 cells, as determined by the lactate dehydrogenase (LDH) assay. After 6 weeks of treatment, our results showed that JPP supplementation increased lipid excretion in feces, reduced serum levels of total cholesterol and nonhigh-density lipoprotein cholesterol, decreased serum aspartate aminotransferase (AST) activity, and attenuated hepatic steatosis severity in rats fed the HF diet. Furthermore, JPP treatment downregulated expression of ACAT-1, LXR-α, CYP7A1, and ABCG5 genes. Therefore, jaboticaba peel may represent a viable dietary strategy to prevent nonalcoholic fatty liver disease as the JPP treatment alleviated hepatic steatosis through improvement of serum lipid profiles and modulation of mRNA expression of genes involved in cholesterol metabolism.

Heterogeneity of pollen food allergy syndrome in seven Southern European countries: The @IT.2020 multicenter study.

BACKGROUND: Pollen food allergy syndrome (PFAS) is a frequently underdiagnosed disease due to diverse triggers, clinical presentations, and test results. This is especially relevant in geographic areas with a broad spectrum of pollen sensitization, such as Southern Europe. OBJECTIVES: To elucidate similarities and differences of PFAS in nine Southern European centers and identify associated characteristics and unique markers of PFAS. METHODS: As part of the @IT.2020 Multicenter Study, 815 patients with seasonal allergic rhinitis (SAR), aged 10-60 years, were recruited in seven countries. They completed questionnaires regarding SAR, comorbidities, family history, and PFAS, and underwent skin prick testing (SPT) and serum IgE testing. RESULTS: Of the 815 patients, 167 (20.5%) reported PFAS reactions. Most commonly, eliciting foods were kiwi (58, 34.7%), peach (43, 25.7%), and melon (26, 15.6%). Reported reactions were mostly local (216/319, 67.7%), occurring within 5 min of contact with elicitors (209/319, 65.5%). Associated characteristics included positive IgE to at least one panallergen (profilin, PR-10, or nsLTP) (p = 0.007), maternal PFAS (OR: 3.716, p = 0.026), and asthma (OR: 1.752, p = 0.073). Between centers, heterogeneity in prevalence (Marseille: 7.5% vs. Rome: 41.4%, p < 0.001) and of clinical characteristics was apparent. Cypress played a limited role, with only 1/22 SPT mono-sensitized patients reporting a food reaction (p < 0.073). CONCLUSIONS: PFAS is a frequent comorbidity in Southern European SAR patients. Significant heterogeneity of clinical characteristics in PFAS patients among the centers was observed and may be related to the different pollen sensitization patterns in each geographic area. IgE to panallergen(s), maternal PFAS, and asthma could be PFAS-associated characteristics.

Evaluation of the anti-inflammatory potential of Solidago microglossa (Arnica-brasileira) in vivo and its effects on PPARγ activity.

Solidago microglossa is used as an anti-inflammatory agent in traditional Brazilian medicine, and this work evaluated the anti-inflammatory potential of the crude ethanolic extract of the flowers of S. microglossa in vivo, as assayed by paw edema models induced by carrageenan, prostaglandin E2, bradykinin and compound 48/80. In the chemical profile, we identified compounds by electrospray ionization mass spectrometry and quantified them by HPLC-DAD. Additionally, this study analyzed the potential to activate the in vitro transcriptional activity of PPARγ, which is a nuclear receptor linked to the anti-inflammatory response. It was possible to identify five compounds: quinic acid, quercetin, chlorogenic acid, hyperoside, and rutin. In the paw edema evaluation, it was possible to show the potential of reducing edema during the inflammatory process. The crude ethanolic extract of the flowers of S. microglossa activated PPARγ compared to the full agonist rosiglitazone and in a dose-response manner. It is possible to conclude that the extract of the flowers of S. microglossa showed anti-inflammatory activity, and the phenolic compounds present in this species might be responsible for this activity.

"Whole" vs. "fragmented" approach to EAACI pollen season definitions: A multicenter study in six Southern European cities.

BACKGROUND: The adequate definition of pollen seasons is essential to facilitate a correct diagnosis, treatment choice, and outcome assessment in patients with seasonal allergic rhinitis. A position paper by the European Academy of Allergy and Clinical Immunology (EAACI) proposed season definitions for Northern and Middle Europe. OBJECTIVE: To test the pollen season definitions proposed by EAACI in six Mediterranean cities for seven pollen taxa. METHODS: As part of the @IT.2020 multi-center study, pollen counts for Poaceae, Oleaceae, Fagales, Cupressaceae, Urticaceae (Parietaria spp.), and Compositae (Ambrosia spp., Artemisia spp.) were collected from January 1 to December 31, 2018. Based on these data, pollen seasons were identified according to EAACI criteria. A unified monitoring period for patients in AIT trials was created and assessed for feasibility. RESULTS: The analysis revealed a great heterogeneity between the different locations in terms of pattern and length of the examined pollen seasons. Further, we found a fragmentation of pollen seasons in several segments (max. 8) separated by periods of low pollen counts (intercurrent periods). Potential monitoring periods included often many recording days with low pollen exposure (max. 341 days). CONCLUSION: The Mediterranean climate leads to challenging pollen exposure times. Monitoring periods for AIT trials based on existing definitions may include many intermittent days with low pollen concentrations. Therefore, it is necessary to find an adapted pollen season definition as individual solution for each pollen and geographical area.

Modulation of sensory nerve fiber excitability by transcutaneous cathodal direct current stimulation.

OBJECTIVE: To assess the lasting effects on sensory nerve membrane excitability of transcutaneous peripheral nerve stimulation with cathodal direct currents (pDCS). METHODS: We performed pDCS in 10 healthy subjects with the active electrode placed over the distal right forearm and the reference electrode on the back of the right hand. We used 5×5cm rubber electrodes and the current applied was 2.5mA during 15min. Three pDCS sessions were performed on the same day: first, a baseline stimulation was performed, followed by a sham stimulation and lastly a cathodal stimulation. Median sensory nerve excitability measurements were performed at baseline and immediately after each pDCS session using the TRONDNF nerve excitability protocol of the QTRAC program (measurement on the second finger). RESULTS: The protocol was completed and well tolerated in all subjects. RRP (relative refractory period) and refractoriness at 2.5ms were significantly different across the three study conditions, with a significant increase of RRP immediately following cathodal stimulation compared with baseline assessment (mean 4.2 versus 5.3, P=0.002). Other measurements were not modulated by the intervention. Sham-stimulation did not change axonal excitability. CONCLUSIONS: Cathodal pDCS stimulation increased RRP of sensory fibers, but no other consistent long-lasting effect was observed. This finding might suggest a reduction of sensory fiber excitability induced by cathodal pDCS.

Cervical trans-spinal direct current stimulation: a modelling-experimental approach.

BACKGROUND: Trans-spinal direct current stimulation (tsDCS) is a non-invasive technique with promising neuromodulatory effects on spinal cord (SC) circuitry. Computational studies are essential to guide effective tsDCS protocols for specific clinical applications. This study aims to combine modelling and experimental studies to determine the electrode montage that maximizes electric field (E-field) delivery during cervical tsDCS. METHODS: Current and E-field distributions in the cervical SC were predicted for four electrode montages in a human realistic model using computational methods. A double-blind crossover and randomized exploratory study was conducted using the montage that maximized E-field delivery. tsDCS was applied for 15 min in 10 healthy subjects (anodal, cathodal, sham, with polarity assigned to the cervical electrode), with a current intensity of 2.5 mA, resulting in a total current charge density delivery of 90 mC/cm2. Upper limb motor (transcranial magnetic stimulation) and sensory evoked potentials (MEP, SEP), M-waves, H-reflex and F-wave responses were analysed. Central and peripheral conduction times were determined using MEP. Repeated measures ANOVA and Friedman test were used for statistical analysis (significance level α = 0.05). RESULTS: All montages presented higher current density and E-field magnitudes in the cervical SC region between the electrodes. However, electrodes at C3 and T3 spinous processes (C3-T3) originated the highest E-field magnitude (0.50 V/m). Using C3-T3 montage we observed significant changes in N9 SEP latency (p = 0.006), but significance did not persist in pairwise comparisons (sham-anodal: p = 0.022; sham-cathodal: p = 0.619; anodal-cathodal: p = 0.018; α = 0.017, Bonferroni corrected). MEP latency and central motor conduction time (CMCT) modified significantly on stimulation (p = 0.007 and p = 0.015, respectively). In addition, pairwise comparisons confirmed significant differences between sham and cathodal conditions after Bonferroni correction for MEP latency (sham-anodal: p = 0.868; sham-cathodal: p = 0.011; anodal-cathodal: p = 0.023) and CMCT (sham-anodal: p = 0.929; sham-cathodal: p = 0.010; anodal-cathodal: p = 0.034). CONCLUSIONS: Computational models predicted higher E-field delivery in the cervical SC for the C3-T3 montage. Polarity-dependent effects in motor responses were reported using this montage consistent with spinal motor modulation. tsDCS experimental protocol designs should be guided by modelling studies to improve effectiveness.

A multivariate statistical approach to the integration of different land-uses, seasons, and water quality as water resources management tool.

The externalities generated by disorderly urbanization and lack of proper planning becomes one of the main factors that must be considered in water resource management. To address the multiple uses of water and avoid conflicts among users, decision-making must integrate these factors into quality and quantity aspects. The water quality index (WQI), using the correlation matrix and the multivariate principal component analysis (PCA) and cluster analysis (CA) techniques were used to analyze the surface water quality, considering urban, rural, and industrial regions in an integrated way, even with data gaps. The results showed that the main parameters that impacted the water quality index were dissolved oxygen, elevation, and total phosphorus. The results of PCA analysis showed 86.25% of the variance in the data set, using physicochemical and topographic parameters. In the cluster analysis, the dissolved oxygen, elevation, total coliforms, E. coli, total phosphorus, total nitrogen, and temperature parameters showed a significant correlation between the data's dimensions. In the industrial region, the characteristic parameter was the organic load, in the rural region were nutrients (phosphorus and nitrogen), and in the urban region was E. coli (an indicator of the pathogenic organisms' presence). In the classification of the samples, there was a predominance of "Good" quality, however, samples classified as "Acceptable" and "Bad" occurred during the winter and spring months (dry season) in the rural and industrial regions. Water pollution is linked to inadequate land use and occupation and population density in certain regions without access to sanitation services.

A 15-minute session of direct current stimulation does not produce lasting changes in axonal excitability.

OBJECTIVE: To assess the applicability and lasting effects on axonal membrane excitability of transcutaneous peripheral nerve direct current stimulation (pDCS). METHODS: We included 15 healthy subjects. pDCS was performed with the active electrode placed over the left forearm and the reference electrode on the back of the hand. We used 5×5cm rubber electrodes and the current applied was 2.5mA during 15minutes. Three pDCS sessions were performed on the same day with a 20-minute interval between them: first a sham stimulation, followed by cathodal and anodal stimulations in random order. Motor nerve excitability measurements were performed immediately after each pDCS session using the TRONDNF nerve excitability protocol of the QTRAC program. RESULTS: The protocol was completed and well tolerated in all subjects. There were no consistent significant differences in excitability measurements between the three sessions. CONCLUSIONS: No consistent long-lasting effects were noted on peripheral nerve excitability beyond the period of application of pDCS. We showed that a 15-minute session of DCS is not able to produce lasting changes in axonal excitability, supporting the hypothesis that the functional and clinical impact of DCS protocols applied to the central nervous system is related to long-term synaptic changes rather sustained local changes in axonal rest membrane potential.

Advances and potential application of gold nanoparticles in nanomedicine.

Nanomedicine is an emerging research area which has brought new possibilities and promising applications in image, diagnosis, and treatment. Nanoparticles (NPs) for medicinal purposes can be made of several material types such as silica, carbon, different polymers, and metals as silver, copper, titanium, and gold. Gold NPs (AuNPs) are the most studied and used, mostly due to their characteristics including simple preparation, controllable size and distribution, biocompatibility, good acceptance of surface modifications, and specific surface plasmon resonance (SPR). This study reviews the scientific literature regarding the potential applications of AuNPs in the development of new diagnostic and therapeutic strategies for nanomedicine, including their biomedical use as a drug carrier, as an agent in radio and phototherapy, and bioimaging for image diagnosis. While it becomes clear that much research remains to be done to improve the use of these nanoparticles, with particular concern for safety issues, the evidence from the literature already points to the great potential of AuNPs in nanomedicine.

Systematic review of pathways to mental health care in Brazil: narrative synthesis of quantitative and qualitative studies.

BACKGROUND: Pathways to care are actions and strategies employed by individuals in order to get help for health-related distress and the related processes of care providers. On several systematic reviews regarding pathways to mental health care (PMHC), studies regarding South American countries were not present. This review synthesizes qualitative and quantitative research about PMHC in Brazil. METHODS: LILACS, MEDLINE and SCIELO databases were searched for papers regarding PMHC in Brazil. The results were organized in pathway stages, based on Goldberg and Huxley's 'model of Levels and Filters' and on Kleinman's framework of 'Popular, Folk and Professional health sectors'. Analysis also considered the changes in national mental health policy over time. RESULTS: 25 papers were found, with data ranging from 1989 to 2013. Complex social networks were involved in the initial recognition of MH issues. The preferred points of first contact also varied with the nature and severity of problems. A high proportion of patients is treated in specialized services, including mild cases. There is limited capacity of primary care professionals to identify and treat MH problems, with some improvement from collaborative care in the more recent years. The model for crisis management and acute care remains unclear: scarce evidence was found over the different arrangements used, mostly stressing lack of integration between emergency, hospital and community services and fragile continuity of care. CONCLUSIONS: The performance of primary care and the regulation of acute demands, especially crisis management, are the most critical aspects on PMHC. Although primary care performance seems to be improving, the balanced provision and integration between services for adequate acute and long-term care is yet to be achieved.

Neuromodulation of lower limb motor responses with transcutaneous lumbar spinal cord direct current stimulation.

OBJECTIVE: Trans-spinal direct current stimulation (tsDCS) is a promising technique to modulate spinal circuits. Combining clinical with modelling studies can improve effectiveness of tsDCS protocols. The aim of this study is to measure the effects of lumbar tsDCS on motor spinal responses and observe if these are consistent with the electric field (E-field) predicted from a computational model. METHODS: The exploratory study design was double-blind crossover and randomized. tsDCS was delivered for 15 min (anodal, cathodal, sham) at L2 vertebra level (2.5 mA, 90 C/cm2) in 14 healthy subjects. F-wave, H-reflex, cortical silent period, motor evoked potential and sympathetic skin response were analyzed. Statistical methods were applied with Bonferroni correction for multiple comparisons, a p < 0.05 was set as significant. A human volume conductor model was obtained from available databases. E-field distributions in the spinal grey matter (GM) and white matter (WM) were calculated. RESULTS: No tsDCS effects were observed. E-field magnitude predicted in the lumbosacral spinal GM and WM was <0.15 V/m, insufficient to ensure neuromodulation, which is consistent with the absence of effects. CONCLUSION: The tsDCS protocol applied did not change motor response to delivered stimulus, thus we observed no effect on motor spinal circuits. SIGNIFICANCE: Future tsDCS protocols should be supported by computational models.

Hepatoprotective Properties of a Polyphenol-Enriched Fraction from Annona crassiflora Mart. Fruit Peel against Diabetes-Induced Oxidative and Nitrosative Stress.

A polyphenol-enriched fraction from Annona crassiflora fruit peel (Ac-Pef) containing chlorogenic acid, (epi)catechin, procyanidin B2, and caffeoyl-glucoside was investigated against hepatic oxidative and nitrosative stress in streptozotocin-induced diabetic rats. Serum biochemical parameters, hepatic oxidative and nitrosative status, glutathione defense system analysis, and in silico assessment of absorption, distribution, metabolism, excretion, and toxicity (ADMET) of the main compounds of Ac-Pef were carried out. Ac-Pef treatment during 30 days decreased serum alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase activities, as well as hepatic lipid peroxidation, protein carbonylation and nitration, inducible nitric oxide synthase level, and activities and expressions of glutathione peroxidase, superoxide dismutase, and catalase. There were increases in antioxidant capacity, glutathione reductase activity, and reduced glutathione level. ADMET predictions of Ac-Pef compounds showed favorable absorption and distribution, with no hepatotoxicity. A. crassiflora fruit peel showed hepatoprotective properties, indicating a promising natural source of bioactive molecules for prevention and therapy of diabetes complications.

Investigation, expression, and molecular modeling of ORF2, a metagenomic lipolytic enzyme.

One clone exhibiting lipolytic activity was selected among 30 positives from a metagenomic library of a microbe consortium specialized in petroleum hydrocarbon degradation. From this clone, a sublibrary was constructed and a metagenome contig was assembled and analyzed using the ORF Finder; thus, it was possible to identify a potential ORF that encodes a lipolytic enzyme, denoted ORF2. This ORF is composed of 1035-bp 345 amino acids and displayed 98 % identity with an alpha/beta hydrolase from Pseudomonas nitroreducens (accession number WP024765380.1). When analyzed against a metagenome database, ORF2 also showed 76 % of sequence identity with a hypothetical protein from a marine metagenome (accession number ECT55726.1). The ProtParam analyses indicated that the recombinant protein ORF2 has a molecular mass approximately 39 kDa, as expected from its amino acid sequence, and based on phylogenetic analysis and molecular modeling, it was possible to suggest that ORF2 is a new member from family V. This enzyme exhibits the catalytic triad and conserved motifs typical from this family, wherein the serine residue is located in the central position of the conserved motif GASMGG. The orf2 gene was cloned in the expression vector pET28a, and the recombinant protein was superexpressed in Escherichia coli BL21(DE3) cells. The lipolytic activity of protein bands presented in a SDS-PAGE gel was confirmed by zymogram analyses, indicating ORF2 activity. These discoveries raise the possibility of employing this protein in biotechnological applications, such as bioremediation.

Avaliação do efeito hipolipemiante do extrato seco de Solanum Melongena L. (berinjela) versus placebo e sinvastatina: ensaio clínico randomizado / Evaluation of lipid-lowering effect of dry extract of Solanum melongena L. (eggplant) versus placebo and simvastatin: a randomized clinical trial

JUSTIFICATIVA E OBJETIVO: As dislipidemias são uma das principais causas de morbidade e mortalidade por doenças car- diovasculares. O objetivo deste estudo foi avaliar o efeito hipo- lipemiante do extrato seco de Solanum melongena L. (berinjela) versus placebo e sinvastatina, em pacientes com hipercolesterole- mia. MÉTODOS: Foi realizado um ensaio clínico randomiza- do com 36 indivíduos adultos, hipercolesterolêmicos, alocados em quatro grupos: Grupo 1, ao qual foi administrado placebo (n=10); Grupo 2, que recebeu extrato seco de berinjela (n=10; colesterol total=200 a 239mg/dL); Grupo 3 que também recebeu extrato seco de berinjela (n=9; colesterol total ?240mg/dL); e Grupo 4, que foi tratado com sinvastatina (n=7; colesterol total ?240mg/dL). Foram realizados exames laboratoriais na linha de base, após 30, 60 e 90 dias de tratamento. RESULTADOS: Houve redução significativa de colesterol total e LDL no Grupo 4 (p<0,001 e p=0,001, respectivamente). Os demais valores e grupos não apresentaram diferença no tratamento. CONCLU- SÃO: O extrato seco de Solanum melongena L. não apresentou diferença estatisticamente significativa na redução dos níveis do colesterol total, frações e triglicerídeos quando comparado ao uso de sinvastatina ou placebo.(AU)

BACKGROUND AND OBJETIVE: Dyslipidemia is the leading cause of morbidity and mortality from cardiovascular disease. The aim of this study was to evaluate the lipid-lowering effect of dry extracts of Solanum melongena L. (eggplant) versus placebo and simvastatin in patients with hypercholesterolemia. METHODS: A randomized clinical trial was performed on 36 adult subjects with hypercholesterolemia. They were divided into four groups: Group 1, which was treated with placebo (n=10); Group 2, that received dry extract of eggplant (n=10; total cholesterol=200 to 239mg/dL); Group 3, that also received dry extract of eggplant (n=9; total cholesterol ?240mg/dL) and Group 4, treated with simvastatin (n=7; total cholesterol ?240mg/dL). Laboratory tests were carried out at baseline and 30, 60 and 90 days after the start of treatment. RESULTS: There was a significant reduction in total cholesterol and LDL in Group 4 (p<0.001 and 0.001, respectively). There was no significant difference in cholesterol levels in the other groups because of treatment. CONCLUSION: The dry extract of Solanum melongena L. showed no statistically significant difference in reducing the levels of total cholesterol, fractions, and triglycerides, when compared with the use of simvastatin or placebo.(AU)