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Gluten proteins contribute to the rheological properties of dough. Mass spectrometric techniques help to understand the contribution of these proteins to the quality of the end product. This work aimed to apply modern proteomic techniques to characterize and provide a better understanding of gluten proteins in wheat flours of different technological qualities. Nine Brazilian wheat flours (Triticum aestivum) classified by rheological gluten force were used to extract the proteins. Extracts were pooled together by technological qualities in low (LW), medium (MD), and superior (SP). Peptides were analyzed by nanoUPLC and mass spectrometry multiplex method (MSE). Collectively, 3545 peptides and 1297 proteins were identified, and 116 proteins were found differentially abundant. Low molecular weight glutenin subunits (LMW-GS) were found up-regulated only in SP samples. Proteins related to wheat grain hardness, such as puroindoline-A, were found in significant concentration in LW samples. After domain prediction, LW presented a different pattern with a lower abundance of functional domains, and SP presented chaperones, known to be involved in adequate folding of the storage proteins. NanoUPLC-MSE was efficient in analyzing and distinguishing the proteomic pattern of wheat flours from different qualities, pointing out the differentially abundant gluten proteins and providing a better understanding of wheat flour quality. SIGNIFICANCE: Common wheat is one of the most important staple food sources in the world. The improvement and comprehension of wheat quality has been a major objective of plant breeders and cereal chemists. Our findings highlighted the application of a modern proteomic approach to obtain a better understanding of the impact of gluten proteins on the technological quality of different wheat flours. The obtained data revealed different abundances of wheat quality-related proteins in superior quality flours when compared with samples of low rheological properties. In addition, multivariate statistical analysis clearly distinguished the flours of different qualities. This work contributes to the consolidation of research in the field of wheat technological quality.

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Acinetobacter baumannii has often been associated with colonization and/or infection in neonatal intensive care units (NICU). This study describes a clonal spread of carbapenem-resistant A. baumannii (CRAB) isolates in an NICU. In total, 21 CRAB isolates were collected from premature newborns. Only polymyxin B was active against such isolates. Nineteen CRAB isolates were clonally related (Cluster C, which belonged to worldwide-disseminated ST1). All newborns had peripheral access and had received ß-lactam therapy previously. The implementation of strict infection control measures was of fundamental importance to eradicate the clonal type in the study hospital.

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Blood vessel invasion is a key feature of invasive aspergillosis. This angioinvasion process contributes to tissue thrombosis, which can impair the access of leukocytes and antifungal drugs to the site of infection. It has been demonstrated that human umbilical vein endothelial cells (HUVECs) are activated and assume a prothrombotic phenotype following contact with Aspergillus fumigatus hyphae or germlings, a process that is independent of fungus viability. However, the molecular mechanisms by which this pathogen can activate endothelial cells, together with the endothelial pathways that are involved in this process, remain unknown. Using a label-free approach by High Definition Mass Spectrometry (HDMS(E)), differentially expressed proteins were identified during HUVEC-A. fumigatus interaction. Among these, 89 proteins were determined to be up- or down-regulated, and another 409 proteins were exclusive to one experimental condition: the HUVEC control or HUVEC:AF interaction. The in silico predictions provided a general view of which biological processes and/or pathways were regulated during HUVEC:AF interaction, and they mainly included cell signaling, immune response and hemostasis pathways. This work describes the first global proteomic analysis of HUVECs following interaction with A. fumigatus germlings, the fungus morphotype that represents the first step of invasion and dissemination within the host. BIOLOGICAL SIGNIFICANCE: A. fumigatus causes the main opportunistic invasive fungal infection related to neutropenic hematologic patients. One of the key steps during the establishment of invasive aspergillosis is angioinvasion but the mechanism associated with the interaction of A. fumigatus with the vascular endothelium remains unknown. The identification of up- and down-regulated proteins expressed by human endothelial cells in response to the fungus infection can contribute to reveal the mechanism of endothelial response and, to understand the physiopathology of this high mortality disease. This article is part of a Special Issue entitled: Trends in Microbial Proteomics.

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Chromobacterium violaceum is a Gram-negative proteobacteria found in water and soil; it is widely distributed in tropical and subtropical regions, such as the Amazon rainforest. We examined protein expression changes that occur in C. violaceum at different growth temperatures using electrophoresis and mass spectrometry. The total number of spots detected was 1985; the number ranged from 99 to 380 in each assay. The proteins that were identified spectrometrically were categorized as chaperones, proteins expressed exclusively under heat stress, enzymes involved in the respiratory and fermentation cycles, ribosomal proteins, and proteins related to transport and secretion. Controlling inverted repeat of chaperone expression and inverted repeat DNA binding sequences, as well as regions recognized by sigma factor 32, elements involved in the genetic regulation of the bacterial stress response, were identified in the promoter regions of several of the genes coding proteins, involved in the C. violaceum stress response. We found that 30 °C is the optimal growth temperature for C. violaceum, whereas 25, 35, and 40 °C are stressful temperatures that trigger the expression of chaperones, superoxide dismutase, a probable small heat shock protein, a probable phasing, ferrichrome-iron receptor protein, elongation factor P, and an ornithine carbamoyltransferase catabolite. This information improves our comprehension of the mechanisms involved in stress adaptation by C. violaceum.

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Avaliou-se a interação idade da matriz versus peso do ovo sobre o desempenho produtivo de codornas de corte (Coturnix coturnix coturnix) no 21º e 42º dias de idade. Em cada classe de idade da matriz, iniciada a partir de 70, 205 e 280 dias, foram incubados 600 ovos, separados em três categorias de peso: categoria 1 - ovos com 11,0-12,9g; categoria 2 - ovos com 13,0-14,9g e categoria 3 - ovos com 15,0-16,9g. Após a eclosão, em cada idade da matriz, foram utilizadas 225 codornas de um dia, de ambos os sexos, provenientes das três categorias de peso, sendo 75 codornas por categoria. O delineamento experimental foi inteiramente ao acaso, e os tratamentos consistiram das três classes de idades da matriz x três categorias de peso do ovo, com cinco repetições de 15 codornas. A dieta experimental foi única para todos os tratamentos com 28 por cento proteína bruta e 2900kcal de energia metabolizável. A categoria peso do ovo e a classe de idade da matriz influenciaram o peso corporal, o ganho de peso e o consumo da dieta no 21º e 42º dias de idade. Codornas oriundas de ovos mais pesados e de matrizes das classes de 205 e 280 dias de idade apresentaram melhor desempenho (P<0,05). A viabilidade foi influenciada pela categoria peso do ovo, ou seja, codornas provenientes de ovos mais leves apresentaram menor viabilidade aos 21º e 42º dias de idade (P<0,05). Ovos mais pesados originaram codornas mais pesadas ao nascimento (P<0,05).

The effect of female breeder age x egg weight interaction was evaluated on the performance of meat type quails (Coturnix coturnix coturnix) at 21st and 42nd days of age. For every female breeder age class (70, 205, and 280 days of age), 600 eggs were classified according to egg weight class (class 1: 11.0-12.9; class 2: 13.0-14.9, and class 3: 15.0-16.9g). A total of 225 quails of both genders, totaling 75 quails for every egg weight class were used in this study. A completely randomized experimental design with five replicates of 15 quails was used. The experimental diets were formulated to contain 28 percent crude protein and 2,900 kcal of metabolizable energy (ME). Weight gain, body weight, feed consumption, and weight gain: feed consumption ratio from hatch to 21st day and from hatch to 42nd day of age were recorded. Both egg weight and female breeder age classes affected body weight, weight gain, and feed consumption at 21st and 42nd days of age. Quails from heavier egg weight class and female breeder age classes 205 and 280 days of age showed higher performance (P<0.05). Quail viability was affected by egg weight class, i.e. quails from lighter eggs showed lower viability at 21st and 42nd days of life. Heavier eggs originated heavier quails at hatch (P<0.05).

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A new D49 PLA(2) was purified from the venom of Calloselasma rhodostoma after two chromatographic steps. Molecular exclusion chromatography was done through a Protein-Pack 300 SW column (0.78 cm x 30 cm), eluting with 0.25 M ammonium bicarbonate, pH 7.9, at a flow rate of 0.3 ml/min. Reverse-phase HPLC was then performed on mu-Bondapack C-18. The sample was determined to have a molecular mass of 13,870.94 Da MALDI-TOF by mass spectrometry, and the amino acid composition showed that Cr-IV 1 presented a high content of Lys, Tyr, Gly, Pro, and 14 half-Cys residues, typical of a basic PLA(2). Cr-IV 1 presented a sequence of 122 amino acid residues: DLWEFGQMILKETGSLPFPY YTTYGCYCGV GGRGGKPKDA TDRCCFVHDC CYGKLTGCPK TNDRYSYSRL DYTIVCGEGG PCKQICECDK AAAVCFRENL RTYNKKYRYHLKPFCKEPAE TC and a calculated pI value of 8.0. Cr-IV 1 had PLA(2) activity in the presence of a synthetic chromogenic substrate (4-nitro-3-(octanoyloxy)benzoic acid) and showed a rapid cytolytic effect on mouse skeletal muscle myoblasts and myotubes in culture. In mice, Cr-IV 1 induced myonecrosis and edema upon intramuscular and intravenous injections, respectively. The LD(50) of Cr-IV 1 was determined to be 0.07 mg/k body weight by intracerebroventricular (i.c.v.) injection. The combination of structural and functional information obtained herein classifies Cr-IV 1 as a new member of the D49 PLA(2) family, as it presents the typical behavior of a phospholipase A(2) from this family.

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The anti-inflammatory and antinociceptive effects of the benzylated cubebin derivative, obtained by reaction of (-)-cubebin with benzyl bromide, were investigated using different animal models. The (-)-o-benzyl cubebin showed a low anti-inflammatory effect (16.2%) in relation to cubebin (57%) and indomethacin (77%) in the carrageenin-induced paw edema in rats, but on the other hand it was more effective (80%) than (-)-cubebin (41%) in inhibiting acetic acid-induced writhing in mice, producing dose-response correlation with doses of 10, 20 and 40 mg/kg, respectively. Moreover, this derivative compound did not show activity in both the hot plate and the cell migration test in rats. Overall, the results showed that the benzylation of cubebin were efficient in enhancing only its analgesic activity.

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The anti-inflammatory and antinociceptive effects of the acetylated (2), methylated (3) and aminated (4) derivatives of cubebin (1), obtained by its reaction with acetic anhydride, methyl iodide and dimethylethylamine chloride, respectively, were investigated, using different animal models. The compound (2) was the most effective anti-inflammatory one in the carrageenin-induced paw edema in rats and was the only one which showed dose-response correlation for this assay with r = 0.993 and Y = 64.58x + 0.22. Besides, compounds (2) and (4) were more effective than cubebin in inhibiting acetic acid-induced writhing in mice, producing dose-response correlation with doses of 10, 20 and 30 mg/kg, respectively. Regarding the hot plate and the cell migration tests in rats, none of the four tested compounds showed activity. Overall, the results showed that the acetylation and amination of cubebin were efficient in enhancing its analgesic activity, as well as its anti-inflammatory activity.

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Three different concentrations (1, 10 and 50 microg/ml) of lyophilized hydroalcoholic crude extract of Pfaffia glomerata roots were assayed in vitro against strains of Trypanosoma cruzi (Y) and Leishmania braziliensis. It was observed that P. glomerata hydroalcoholic extract was relatively active within the tested concentrations for L. (V) braziliensis, but inactive against T. cruzi. Despite the fact that both protozoans belong to the Trypanosomatidae family, we suggest that the difference observed for activity should be related to the biological differences between the two parasite species.

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Foram utilizadas técnicas de análise multivariada para comparar dois híbridos obtidos de linhas de frango de corte desenvolvidas pela Universidade Federal de Viçosa com dois híbridos comerciais. As características avaliadas foram: peso ao abate (PAB), peso da carcaça (PC), peso do peito (PP), peso da contra-coxa (PCC), peso da coxa (PCX) e rendimento de carcaça (RCA). A análise de variância multivariada indicou diferença significativa entre os vetores de médias das características. Pelo teste de Roy, observou-se que para PC e PP os produtos comerciais foram superiores e para PCX e RCA näo houve diferenças significativas entre os produtos comercias e os cruzamentos. Quanto à funçäo discriminante linear de Fisher, observou-se superioridade significativa dos produtos comerciais pelo teste de Roy

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Cubebin, a dibenzylbutyrolactone lignan isolated from the crude hexane extract of the leaves of Zanthoxyllum naranjillo, showed a significant anti-inflammatory activity by using the paw edema induced by carrageenin in rats, but did not provide a significant reduction in the cell migration for the acute carrageenin-induced inflammatory reaction in the peritoneal cavity of rats. Neither was it effective in reducing the edema induced by dextran nor the edema induced by histamine. It partially reduced the edema induced by serotonin. Moreover, it significantly reduced the edema induced by prostaglandin PGE2 and the number of writhings induced by both acetic acid and PGI2 in mice. Therefore, it may be suggested that the mechanism of action of cubebin is similar to that observed for most of the non-steroidal drugs.

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