The Addition of Pinto Bean Flour and Margarine in the Development of Red Rice-Based Novel Gluten-Free Cookies to Improve the Technological, Sensory and Physicochemical Properties.

This study aimed to understand how the addition of pinto bean flour (PBF) and margarine affected the technological, sensory, and physicochemical properties of red rice-based gluten-free cookies. PBF addition (25-75 g/100 g) and margarine content (13.3-19.3 g/100 g dough) were varied according to a central composite rotatable design. Cookies selection was performed by overlaid contour and principal component analysis (PCA) for technological properties and sensory evaluation, respectively. Chemical composition analyses, bioactive compounds, and microstructure were carried out in the selected formulations. In addition, other physicochemical analyses were carried out during storage time. PBF addition affected the technological properties and increased the nutritional content of proteins (up to 13%), fibers (8.28%), iron (2.13%), zinc (1.54%), and phenolic compounds (139.46 mg gallic acid equivalent/100 g), but it negatively affected sensory acceptance. However, margarine's addition improved all the sensory attributes in all the trials, showing an acceptance greater than 70%. Cookies with 50 g PBF, 50 g red rice flour, and 16.3 g margarine/100 g dough showed better technological, nutritional characteristics, and physicochemical quality up to 60 days of storage. This work contributed to the incorporation of mixtures of red rice and pinto bean for developing more nutritious cookies for celiac patients or even those who wish to consume gluten-free products.

Development and Characterization of Powdered Antioxidant Compounds Made from Shiraz (Vitis vinifera L.) Grape Peels and Arrowroot (Maranta arundinacea L.).

Grapevine (Vitis vinifera L.) is a plant containing many phenolic compounds, mostly distributed in the peel, pulp, and seeds. This study evaluates the centesimal composition and bioactive compounds in Shiraz grape (Vitis vinifera) peels using spectrophotometric and UHPLC techniques and develops different formulations of compound powders from the peels and arrowroot using conventional drying technology. The results demonstrate that Shiraz grape skin contains significant amounts of insoluble fiber (15.3%), phenolics (157.09 ± 6.96-149.11 ± 9.27 mg GAE g-1), and flavonoids (0.75 ± 0.50-2.00 ± 0.50 mg QE g-1), with excellent antioxidant capacity observed in the alcoholic extracts. The phenolic content in the developed powdered compounds ranged from 128.32 to 139.70 mg GAE g-1. In general, the compounds showed good antioxidant capacity (IC50 = 0.17 to 0.19 µg mL-1). According to the chromatographic evaluation, it was possible to quantify gallic acid, catechin, and epicatechin, the latter of which was found in the largest quantities in the six formulations. The EV5 formulation was the most efficient in terms of phenolic compounds and protein amounts. This formulation's composition and low cost could make it viable for use in the food industry.

Outcomes of one-step incomplete and complete excavation in primary teeth: a 24-month randomized controlled trial.

AIM: To compare 24-month pulp health outcomes of partial caries removal (PCR) and total caries removal (TCR) with composite restoration in primary molars. METHODS: 48 children aged 3-8 years with at least one molar with a deep carious lesion were included. 120 teeth were randomized to control (TCR; n = 54; 69% class II) and test (PCR; n = 66; 63% class II) groups. Total absence of carious tissue was confirmed using a blunt-tipped probe in the TCR group. For PCR, excavation was stopped when hardened, dried dentin with a leathery consistency was achieved. Pulpotomy was performed in cases of pulp exposure. RESULTS: Pulp exposure occurred in 2 and 27.5% of teeth treated with PCR and TCR, respectively (p < 0.01). The operative time was significantly higher for TCR than PCR. Success rates were 92 and 96% in the PCR and TCR groups, respectively (p = 0.34). The success rate tended to be lower in occlusoproximal (92%) than in occlusal (100%) lesions (p = 0.08). CONCLUSION: The clinical and radiographic success rates of PCR and TCR in primary teeth with deep carious lesions were high and did not differ significantly, indicating that PCR is a reliable minimally invasive approach in primary teeth and that the retention of carious dentin does not interfere with pulp vitality. Moreover, PCR provided other clinically relevant advantages over TCR, especially lower incidence of pulp exposure and lower operative time.

Substance Use and Risky Sexual Behavior in the PrEP Outpatient Clinic at the University Hospital of Brasília.

(1) Background: To evaluate the epidemiological profile of people who use drugs at the PrEP outpatient clinic of the University Hospital of Brasília; (2) Methods: Cross-sectional study with a review of data from medical records referring to the first medical consultation. The prevalence ratio was calculated using a Poisson regression model with robust variance; (3) Results: A total of 53% of subjects reported drug use in the last 3 months. The unadjusted prevalence ratio of drug use in trans women was PR: 9.0 (95%CI: 1.4-57.5). people who use drugs have a 1.9 times higher prevalence of STI diagnosis, and a 2.4 times higher prevalence of partners compared to non-users; (4) Conclusions: Substance use was associated with a higher STI prevalence ratio and number of sexual partners.

Maternal Toxoplasma gondii infection affects proliferation, differentiation and cell cycle regulation of retinal neural progenitor cells in mouse embryo.

Background: Toxoplasmosis affects one third of the world population and has the protozoan Toxoplasma gondii as etiological agent. Congenital toxoplasmosis (CT) can cause severe damage to the fetus, including miscarriages, intracranial calcification, hydrocephalus and retinochoroiditis. Severity of CT depends on the gestational period in which infection occurs, and alterations at the cellular level during retinal development have been reported. In this study, we proposed a mouse CT model to investigate the impact of infection on retinal development. Methods: Pregnant females of pigmented C57BL/6 strain mice were infected intragastrically with two T. gondii cysts (ME49 strain) at embryonic day 10 (E10), and the offspring were analyzed at E18. Results: Infected embryos had significantly smaller body sizes and weights than the PBS-treated controls, indicating that embryonic development was affected. In the retina, a significant increase in the number of Ki-67-positive cells (marker of proliferating cells) was found in the apical region of the NBL of infected mice compared to the control. Supporting this, cell cycle proteins Cyclin D3, Cdk6 and pChK2 were significantly altered in infected retinas. Interestingly, the immunohistochemical analysis showed a significant increase in the population of ß-III-tubulin-positive cells, one of the earliest markers of neuronal differentiation. Conclusions: Our data suggests that CT affects cell cycle progression in retinal progenitor cells, possibly inducing the arrest of these cells at G2/M phase. Such alterations could influence the differentiation, anticipating/increasing neuronal maturation, and therefore leading to abnormal retinal formation. Our model mimics important events observed in ocular CT.

PPARα polymorphisms as risk factors for dyslipidemia in a Brazilian population.

BACKGROUND AND AIMS: Peroxisome proliferator-activated receptor α is a nuclear receptor involved in the regulation of several biochemical pathways. Polymorphisms within its gene have been associated with several metabolic traits. We aimed to investigate the association of L162V and Intron 7G>C polymorphisms with serum level markers and common morbidities affecting an older adult/elderly cohort from Cuiaba City, Mato Grosso State, Brazil, as well as to compare the results with a previously studied population from São Paulo City, Brazil. METHODS AND RESULTS: The studied population consisted of 570 subjects from Cuiaba City, Brazil, who were subjected to clinical interviews and blood collection for laboratory examinations and DNA extraction. Dyslipidemia was defined when participants were taking oral hypolipemiants or those with total cholesterol above 200mg/dL, HDL-c below 40 mg/dL, LDL-c above 130 mg/dL and TG above 150 mg/dL. Restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR) was used for polymorphism genotyping. Individual polymorphism and haplotype data were available for analyses. In the studied sample, allele frequencies were 0.052 and 0.292 for 162V and Intron 7C, respectively. In brief, 162V allele was associated with dyslipidemia (p=0.025), and after correction for alcohol consumption and waist-to-rip ratio, a tendency of association could still be observed (p=0.050). In addition, Intron 7C allele was associated with dyslipidemia even after correction for the same variables (p=0.029). When compared to our previous study from São Paulo, we found some divergences regarding these results, which may be explained by differences between the two populations. Haplotype association analyses revealed an association between L/C haplotype and dyslipidemia (p=0.021) and between V/C haplotype and lower LDL-c levels when compared to L/G haplotype (p=0.044). CONCLUSION: These results may help to clarify the role of PPARα gene in lipid and lipoprotein metabolism and the evaluation of its polymorphisms and haplotypes as being characterized as genetic risk factors for metabolic disturbances.

P2Y12 but not P2Y13 Purinergic Receptor Controls Postnatal Rat Retinogenesis In Vivo.

Adenine nucleotides through P2Y1 receptor stimulation are known to control retinal progenitor cell (RPC) proliferation by modulating expression of the p57KIP2, a cell cycle regulator. However, the role of Gi protein-coupled P2Y12 and P2Y13 receptors also activated by adenine nucleotides in RPC proliferation is still unknown. Gene expression of the purinergic P2Y12 subtype was detected in rat retina during early postnatal days (P0 to P5), while expression levels of P2Y13 were low. Immunohistochemistry assays performed with rat retina on P3 revealed P2Y12 receptor expression in both Ki-67-positive cells in the neuroblastic layer and Ki-67-negative cells in the ganglion cell layer and inner nuclear layer. Nonetheless, P2Y13 receptor expression could not be detected in any stratum of rat retina. Intravitreal injection of PSB 0739 or clopidogrel, both selective P2Y12 receptor antagonists, increased by 20 and 15%, respectively, the number of Ki-67-positive cells following 24 h of exposure. Moreover, the P2Y12 receptor inhibition increased cyclin D1 and decreased p57KIP2 expression. However, there were no changes in the S phase of the cell cycle (BrdU-positive cells) or in mitosis (phospho-histone-H3-positive cells). Interestingly, an increase in the number of cyclin D1/TUNEL-positive cells after treatment with PSB 0739 was observed. These data suggest that activation of P2Y12 receptors is required for the successful exit of RPCs from cell cycle in the postnatal rat retina.

Adenine Nucleotides Control Proliferation In Vivo of Rat Retinal Progenitors by P2Y1 Receptor.

Previous studies demonstrated that exogenous ATP is able to regulate proliferation of retinal progenitor cells (RPCs) in vitro possibly via P2Y1 receptor, a G protein-coupled receptor. Here, we evaluated the function of adenine nucleotides in vivo during retinal development of newborn rats. Intravitreal injection of apyrase, an enzyme that hydrolyzes nucleotides, reduced cell proliferation in retinas at postnatal day 2 (P2). This decrease was reversed when retinas were treated together with ATPγ-S or ADPß-S, two hydrolysis-resistant analogs of ATP and ADP, respectively. During early postnatal days (P0 to P5), an increase in ectonucleotidase (E-NTPDase) activity was observed in the retina, suggesting a decrease in the availability of adenine nucleotides, coinciding with the end of proliferation. Interestingly, intravitreal injection of the E-NTPDase inhibitor ARL67156 increased proliferation by around 60 % at P5 rats. Furthermore, immunolabeling against P2Y1 receptor was observed overall in retina layers from P2 rats, including proliferating Ki-67-positive cells in the neuroblastic layer (NBL), suggesting that this receptor could be responsible for the action of adenine nucleotides upon proliferation of RPCs. Accordingly, intravitreal injection of MRS2179, a selective antagonist of P2Y1 receptors, reduced cell proliferation by approximately 20 % in P2 rats. Moreover, treatment with MRS 2179 caused an increase in p57KIP2 and cyclin D1 expression, a reduction in cyclin E and Rb phosphorylated expression and in BrdU-positive cell number. These data suggest that the adenine nucleotides modulate the proliferation of rat RPCs via activation of P2Y1 receptors regulating transition from G1 to S phase of the cell cycle.

Cross-reactivity of commercially available anti-human monoclonal antibodies with canine cytokines: establishment of a reliable panel to detect the functional profile of peripheral blood lymphocytes by intracytoplasmic staining.

BACKGROUND: The process for obtaining monoclonal antibodies against a specific antigen is very laborious, involves sophisticated technologies and it is not available in most research laboratories. Considering that most cytokines remain partially conserved among species during evolution, the search for antibody cross-reactivity is an important strategy for immunological studies in veterinary medicine. In this context, the amino acid sequence from human and canine cytokines have demonstrated 49-96 % homology, suggesting high probability of cross-reactivity amongst monoclonal antibodies. For this, 17 commercially available anti-human monoclonal antibodies [IL-1α, IL-1ß, IL-2, IL-4, IL-5, IL-6, IL-8 (#1, #2), IL-10, IL-12, IL-13, IL-17A, IFN-γ (#1, #2), TNF-α (#1, #2) and TGF-ß], were evaluated in vitro for intracellular cytokine detection in a stimulated canine blood culture by flow cytometry and confocal microscopy. Lymphocytes from peripheral blood of healthy and two unhealthy dogs were analyzed. RESULTS: Eleven anti-human mAbs [IL-1α, IL-4, IL-5, IL-6, IL-8 (#1, #2), IL-12, IL-17A, TNF-α (#1, #2) and TGF-ß] cross-reacted against canine intracellular cytokines. The specificity of the assays was not affected after Fc-blocking. Three anti-human cytokine mAbs [IL-4, IL-8 (#2) and TGF-ß] when evaluated by confocal microscopy also cross-reacted with intracellular canine cytokines. The identification of human mAbs that cross-reacted with canine cytokines may support their use as immunological biomarkers in veterinary medicine studies. CONCLUSION: The identification of these 11 anti-human cytokine mAbs that cross-reacted with canine cytokines will be useful immunological biomarkers for pathological conditions by flow cytometry and fluorescence microscopy in dogs.

Receptor-ligand molecular docking.

Docking methodology aims to predict the experimental binding modes and affinities of small molecules within the binding site of particular receptor targets and is currently used as a standard computational tool in drug design for lead compound optimisation and in virtual screening studies to find novel biologically active molecules. The basic tools of a docking methodology include a search algorithm and an energy scoring function for generating and evaluating ligand poses. In this review, we present the search algorithms and scoring functions most commonly used in current molecular docking methods that focus on protein-ligand applications. We summarise the main topics and recent computational and methodological advances in protein-ligand docking. Protein flexibility, multiple ligand binding modes and the free-energy landscape profile for binding affinity prediction are important and interconnected challenges to be overcome by further methodological developments in the docking field.

Immunological changes in canine peripheral blood leukocytes triggered by immunization with first or second generation vaccines against canine visceral leishmaniasis.

In this study, we summarized the major phenotypic/functional aspects of circulating leukocytes following canine immunization with Leishvaccine and Leishmune®. Our findings showed that Leishvaccine triggered early changes in the innate immunity (neutrophils and eosinophils) with late alterations on monocytes. Conversely, Leishmune(®) induced early phenotypic changes in both, neutrophils and monocytes. Moreover, Leishvaccine triggered mixed activation-related phenotypic changes on T-cells (CD4+ and CD8+ and B-lymphocytes, whereas Leishmune(®) promoted a selective response, mainly associated with CD8+ T-cell activation. Mixed cytokine profile (IFN-γ/IL-4) was observed in Leishvaccine immunized dogs whereas a selective pro-inflammatory pattern (IFN-γ/NO) was induced by Leishmune® vaccination. The distinct immunological profile triggered by Leishvaccine and Leishmune® may be a direct consequence of the distinct biochemical composition of these immunobiological, i.e. complex versus purified Leishmania antigen along with Bacillus Calmette-Guérin (BCG) versus saponin adjuvant. Both immunobiologicals are able to activate phagocytes and CD8+ T-cells and therefore could be considered as a putative vaccines against canine visceral leishmaniasis (CVL).

Squamous cell carcinoma arising from chronic lymphedema: case report and review of the literature.

CONTEXT: Squamous cell carcinoma arising from chronic lymphedema has only been reported in the literature 11 times (12 cases). Some aspects of its pathogenesis remain unclear and, for the first time, attention has been drawn to epidemiological data. CASE REPORT: A 90-year-old white female with chronic unilateral lower-limb lymphedema, secondary to trauma 20 years earlier, presented with a three-month history of a vegetating cutaneous lesion. There had not been any previous local ulceration. The tumor was completely excised and the histopathological analysis showed that it was an infiltrating squamous cell carcinoma. A literature review in the Medline (Medical Literature Analysis and Retrieval System Online) and Lilacs (Literatura Latino-Americana e do Caribe em Ciências da Saúde) databases using the MeSH (Medical Subject Heading) terms "Carcinoma, Squamous Cell" AND "Lymphedema" identified 112 references and found 12 similar case reports.

Synthesis, antimalarial evaluation and molecular modeling studies of hydroxyethylpiperazines, potential aspartyl protease inhibitors, part 2.

The antimalarial acitivity of hydroxyethylamines, synthesized from the reaction of intermediated hydroxyethypiperazines with benzenesulfonyl chlorides or benzoyl chlorides, has been evaluated in vitro against a W2 Plasmodium falciparum clone. Some of the nineteen tested derivatives showed a significant activity in vitro, thus turning into a promising new class of antimalarials. In addition, a molecular modeling study of the most active derivative (5l) was performed and its most probable binding modes within plasmepsin II enzyme were identified.

Treinamento muscular respiratório em pacientes com lesão raqui-medular dependentes de ventilação mecânica / Respiratory muscle training in patients with injury spinal-medullary dependent on mechanical ventilation

As lesões medulares freqüentemente acarretam complicações respiratórias. O comportamento da função pulmonar em pacientes com trauma raquimedular (TRM) é causado pela alteração da biomecânica da caixa toráxica devido à paralisia da musculatura abdominal, intercostal e pela perda parcial ou total da função do músculo diafragma. Essas disfunções trazem complicações como a fadiga dos músculos respiratórios. Devido a essa restrição ventilatória, os pacientes tornam-se dependentes da ventilação mecânica. Baseando-se em resultados de outros estudos na literatura, conclui-se que o tratamento pode ser realizado com o treinamento da musculatura respiratória. Entretanto essa revisão da literatura demonstra apenas trabalhos realizados em adultos, sendo assim necessário à realização de estudos mais amplos quanto ao tipo de treinamento muscular inspiratório em crianças com TRM.

Squamous cell carcinoma arising from chronic lymphedema: case report and review of the literature / Surgimento de carcinoma escamocelular a partir de linfedema crônico: relato de caso e revisão da literatura

CONTEXT: Squamous cell carcinoma arising from chronic lymphedema has only been reported in the literature 11 times (12 cases). Some aspects of its pathogenesis remain unclear and, for the first time, attention has been drawn to epidemiological data. CASE REPORT: A 90-year-old white female with chronic unilateral lower-limb lymphedema, secondary to trauma 20 years earlier, presented with a three-month history of a vegetating cutaneous lesion. There had not been any previous local ulceration. The tumor was completely excised and the histopathological analysis showed that it was an infiltrating squamous cell carcinoma. A literature review in the Medline (Medical Literature Analysis and Retrieval System Online) and Lilacs (Literatura Latino-Americana e do Caribe em Ciências da Saúde) databases using the MeSH (Medical Subject Heading) terms "Carcinoma, Squamous Cell" AND "Lymphedema" identified 112 references and found 12 similar case reports.

CONTEXTO: O crescimento de carcinoma de células escamosas a partir de linfedema crônico foi relatado apenas 11 vezes na literatura (12 casos). Aspectos sobre sua patogênese ainda não foram completamente elucidados e pela primeira vez chama-se atenção para os dados epidemiológicos. RELATO DE CASO: Paciente do sexo feminino, branca, 90 anos de idade, portadora de linfedema crônico unilateral de membro inferior secundário a trauma sofrido há 20 anos, apresentou história de lesão cutânea vegetante de três meses. Não havia relato de lesão local prévia. O tumor foi completamente excisado e sua análise histopatológica mostrou tratar-se de carcinoma escamocelular infiltrante. Uma revisão da literatura nos bancos de dados Medline (Medical Literature Analysis and Retrieval System Online) e Lilacs (Literatura Latino-Americana e do Caribe em Ciências da Saúde), utilizando-se dos termos MeSH (Medical Subject Heading) "Carcinoma, Squamous Cell" AND "Lymphedema", identificou 112 citações e encontrou relatos de 12 casos semelhantes.

Aplicação das técnicas de retalho tarsal lateral e Kuhnt-Szymanowski modificada no tratamento do ectrópio senil / The use of tarsal strip and Kuhnt-Szymanowski techniques for senile ectropion treatment

Ectrópio caracteriza-se pela eversão da margem palpebral. As quatro formas clínicas principais são: involucional, cicatricial, congênita e paralítica, sendo a primeira a mais freqüente. Geralmente, cursa com eversão do ponto lacrimal ou de toda a pálpebra, epífora, hiperemia conjuntival e sensação de ardência ocular, podendo resultar em dano corneano permanente, com cegueira potencial. Muitas opções de tratamento foram descritas para cada uma das formas clínicas. O objetivo deste trabalho é demonstrar a utilização das técnicas de Retalho Tarsal Lateral e Kuhnt-Szymanowski modificada no tratamento do ectrópio senil, comparando com dados da literatura. Foi realizada análise retrospectiva dos prontuários de 35 pacientes, sendo 19 (54,3%) operados pela técnica de retalho tarsal lateral e 16 (45,7%) pela técnica de Kuhnt-Szymanowski modificada. Dezenove pacientes eram do sexo feminino e 16 do masculino, com idade variando entre 56 e 92 anos, com média de 75 anos. Todos os pacientes submetidos à cirurgia obtiveram remissão dos sintomas a partir do primeiro dia de pós-operatório. Na série estudada, ambas as técnicas apresentaram resultados satisfatórios, com melhora dos sintomas e ausência de recidiva.

Ectropion is an outward turning of the eyelid. The four main types are involutional, cicatricial, congenital and paralytic. Most often it presents as an eversion of the lid margin, epiphora, conjunctival hyperemia and ocular irritation, which may lead to permanent corneal damage. Several treatment options have been described for each clinical form. The purpose of this paper is to demonstrate the utilization of two operative techniques for the management of involutional ectropion. A retrospective analysis of 35 patients' charts has been performed. 19 patients underwent surgical correction by tarsal strip technique and 16, by the Kuhnt Szymanowski technique. Nineteen patients were female and 16 male. Their age ranged from 56 to 92 years. All operated patients showed complete disappearance of all symptoms. In this survey, both procedures showed satisfactory results, with symptom regression and lack of recurrence up to the present date.

A genetic algorithm for the ligand-protein docking problem

We analyzed the performance of a real coded "steady-state" genetic algorithm (SSGA) using a grid-based methodology in docking five HIV-1 protease-ligand complexes having known three-dimensional structures. All ligands tested are highly flexible, having more than 10 conformational degrees of freedom. The SSGA was tested for the rigid and flexible ligand docking cases. The implemented genetic algorithm was able to dock successfully rigid and flexible ligand molecules, but with a decreasing performance when the number of ligand conformational degrees of freedom increased. The docked lowest-energy structures have root mean square deviation (RMSD) with respect to the corresponding experimental crystallographic structure ranging from 0.037 Å to 0.090 Å in the rigid docking, and 0.420 Å to 1.943 Å in the flexible docking. We found that not only the number of ligand conformational degrees of freedom is an important aspect to the algorithm performance, but also that the more internal dihedral angles are critical. Furthermore, our results showed that the initial population distribution can be relevant for the algorithm performance.