•Brewers’ spent grain protein hydrolysates present in vitro chemical antioxidant activity.•Protein hydrolysates protected against oxidative stress in Caco-2 and HepG2 cells.•Hydrolysates from ...Brewers’ spent grain and spent yeast had strongest protective ability.•A potential functional ingredient was obtained from two major brewing by-products.
The protein fraction of Brewers’ spent grain (BSG) was used as substrate to obtain hydrolysates with antioxidant activity. Three enzymatic approaches were applied: brewer’s spent yeast (BSY) proteases, Neutrase® and Alcalase®, at the same proteolytic activity (1U/mL), using an enzyme/substrate ratio of 10:100 (v/v), at 50°C, 4h. Total Phenolic Content (TPC) and Ferric Ion Reducing Antioxidant Power (FRAP) of hydrolysates and fractions <10kDa and <3kDa were assayed. Additionally, the protective ability of <10kDa fractions against oxidative stress on Caco-2 and HepG2 cells was investigated. Alcalase® hydrolysate presented significantly (p<0.05) higher TPC and FRAP (0.083mgGAE/mgdw; 0.101mgTE/mgdw, respectively) than Neutrase® and BSY hydrolysates. The three BSG protein hydrolysates (fraction <10kDa) exerted protective effect against free-radical induced cytotoxicity in Caco-2 and HepG2 cell lines, but the strongest effect was observed for BSY hydrolysates, therefore, it presents greater potential as functional ingredient.
Coffee, an agronomical crop of great economic importance, is also among the most commonly traded commodities in worldwide markets. Antimicrobial peptides, which play a role in plant defense, have ...been identified and isolated particularly from seeds. We isolated and immunolocalized Cc-LTP2, a new lipid transfer protein (LTP) from Coffea canephora seeds. We report its antimicrobial activity against various phytopathogenic fungi of economic importance, and against the bacterium Xanthomonas euvesicatoria. Peptides from C. canephora seeds were initially extracted using acid buffer and subjected to ion-exchange and reverse-phase chromatographies. A purified peptide of approximately 9 kDa, which we named Cc-LTP2, was then subjected to amino acid sequencing. The analyses showed that it was similar to LTPs isolated from various plants. The tissue and subcellular localization of C. canephora LTPs indicated that they were located in cell walls and intracellular palisade parenchyma, mainly in large vacuoles. The results of immunohistochemistry and histochemistry superposed from C. canephora seed tissues showed that LTPs and lipid bodies are present in organelles, supporting the hypothesis that LTPs from seeds are involved in lipid mobilization during germination. Cc-LTP2 did inhibit the development of the phytopathogenic fungi Colletotrichum lindemuthianum, Colletotrichum gloeosporioides, Fusarium solani, Fusarium lateritium, and Colletotrichum sp, but did inhibit X. euvesicatoria. Cc-LTP2 also increased membrane permeability and induced endogenous production of reactive oxygen species in all the fungi tested.
Leptospira is a genus of bacteria that includes free-living saprophytic species found in water or soil, and pathogenic species, which are the etiologic agents of leptospirosis. Besides all the ...efforts, there are only a few proteins described as virulence factors in the pathogenic strain L. interrogans. This work aims to perform L. biflexa serovar Patoc1 strain Paris global proteome and to compare with the proteome database of pathogenic L. interrogans serovar Copenhageni strain Fiocruz L1–130. We identified a total of 2327 expressed proteins of L. biflexa by mass spectrometry. Using the Get Homologues software with the global proteome of L. biflexa and L. interrogans, we found orthologous proteins classified into conserved, low conserved, and specific proteins. Comparative bioinformatic analyses were performed to understand the biological functions of the proteins, subcellular localization, the presence of signal peptide, structural domains, and motifs using public softwares. These results lead to the selection of 182 low conserved within the saprophyte, and 176 specific proteins of L. interrogans. It is anticipated that these findings will indicate further studies to uncover virulence factors in the pathogenic strain. This work presents for the first time the global proteome of saprophytic strain L. biflexa serovar Patoc, strain Patoc1.
The comparative analysis established an array of specific proteins in pathogenic strain that will narrow down the identification of immune protective proteins that will help fight leptospirosis.
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•The first global proteome of the saprophytic species L. biflexa published.•Comparative proteomic analysis between saprophytic L. biflexa and pathogenic L. interrogans species.•Identification of orthologous proteins and specific proteins in the two leptospiral species.•Selection of possible virulence factors of L. interrogans by bioinformatics analysis.
There are currently no reports on the isolation and molecular examination of Toxoplasma gondii from bats. Here, we report the isolation and genotypic characterisation of two T. gondii isolates from ...bats. A total of 369 bats from different municipalities in São Paulo state, southeastern Brazil, were captured and euthanised, and collected tissues (heart and pectoral muscle) were processed for each bat or in pools of two or three bats and bioassayed in mice (a total of 283 bioassays). Eleven PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) markers were used to genotype positive samples: SAG1, SAG2 (5′–3′SAG2 and alt. SAG2), SAG3, BTUB, GRA6, L358, c22-8, c29-2, PK1, CS3 and Apico. The parasite was isolated from two bats from São Paulo city: an insectivorous bat, the velvety free-tailed bat Molossus molossus, and a hematophagous bat, the common vampire bat Desmodus rotundus. Isolates were designated TgBatBr1 and TgBatBr2, respectively. The genotype of the isolate from M. molossus (TgBatBr1) has been previously described in an isolate from a capybara from São Paulo state, and the genotype from the D. rotundus isolate (TgBatBr2) has already been identified in isolates from cats, chickens, capybaras, sheep, a rodent and a common rabbit from different Brazilian states, suggesting that this may be a common T. gondii lineage circulating in some Brazilian regions. Isolation of T. gondii from a hematophagous species is striking. This study reveals that bats can share the same isolates that are found in domesticated and wild terrestrial animals. This is the first report of the isolation and genotyping of T. gondii in chiropterans.
Parasitoids are of great importance to forest ecosystems due to their ecological role in the regulation of the population of other insects. The species richness and abundance of parasitoids in the ...forest canopy and understory, both on the borders and in the interior of a tropical forest reserve in Central Amazonia were investigated. For a 12-month period, specimen collections were made every 15 days from suspended traps placed in the forest canopy and in the understory strata, both on the border and in the interior of forest areas. A total of 12,835 Hymenoptera parasitoids from 23 families were acquired. Braconidae, Diapriidae, Mymaridae, Eulophidae, and Scelionidae were the most represented in the area and strata samples. The results indicate that there were no significant differences in the species richness or abundance of Hymenoptera between the forest borders and the inner forest. The data does show that the presence of Hymenoptera is significantly greater in the understory in both the border and interior areas than in the canopy (vertical stratification). Aphelinidae and Ceraphronidae were significantly associated with the inner forest, while the other seven families with the border of the reserve. The abundance of Hymenoptera parasitoids presented seasonal variations during the year related to the rainy and dry seasons.
Recent studies in patients with previously untreated T1 and T2 squamous cell carcinoma (SCC) of the tongue and floor of the mouth have shown a relationship between tumor thickness, neck metastasis, ...and survival. Our study was conducted to determine the indication of elective neck dissection in patients with early oral cavity SCC.
Sixty-seven patients were stratified by stage (T1 and T2 NO), and those in each stage were randomized to receive one of two types of treatment; resection alone (RA) or resection plus elective supraomohyoid neck dissection (RSOND). Fifty-two patients (78%) were men and 15 (22%) were women. The median age was 57 years old (range 34 to 95).
Twenty-six (39%) patients had tumor in the floor of the mouth and 41 (61%), in the tongue. Using the criteria of the Union Internationale Contre le Cancer (UICC), 1987, we classified 31 tumors (46%) as T1 lesions and 36 (54%) as T2 lesions. Thirty patients had a tumor thickness < or = 4 mm and 37 had a tumor thickness > 4 mm. Thirty-three (49%) patients were treated with RA, and 34 patients (51%) were treated with RSOND. Seven (21%) patients of the RSOND group had occult cervical metastasis. There were recurrences in 14 (42%) patients of the RA group and 8 (24%) patients of the RSOND group. The disease-free survival rates at 3.5 years for RA and RSOND patients were 49%, and 72%, respectively. The impact of sex, age, site, cancer stage, and tumor thickness was assessed by the Mantel-Haenszel chi-square procedure. Later stage (P = 0.05) and increased tumor thickness (P = 0.005) were significantly associated with treatment failures.
Neck dissection remains mandatory in the early stage of oral SCC, because of better survival rates compared to RA and the poor salvage rate. In particular, patients with tumor thickness > 4 mm treated with RSOND had significant benefit on disease-free survival.
Goats are economically important in many countries, and little is known of caprine toxoplasmosis in Brazil. Antibodies to Toxoplasma gondii were assayed in the sera of 143 goats from 3 Brazilian ...states, using modified agglutination test (MAT titer ≥1:25); 46 (32.2%) tested positive. Samples of brain, heart, diaphragm, and masseter of seropositive animals were pooled, digested in pepsin, and bioassayed in mice. Viable T. gondii specimens were isolated from tissue homogenates of 12 goats; the isolates were designated TgGtBr1-12. Ten of the 12 isolates killed 100% of infected mice, indicating that goats can harbor mouse-virulent T. gondii and, hence, can serve as a source of infection for humans.
One of the roadblocks towards the practical use of antimicrobial peptides for medical use is their relatively high cost when synthesized chemically. Effective recombinant production has only been ...successful in some cases, such as the previously reported production in Pichia pastoris of the antimicrobial plectasin derivative peptide NZ2114. The same production host has also been used extensively to produce so-called protein-polymers: sequences that consist of repetitions of simple amino acid motifs found in structural proteins such as collagen and elastin, and that can be designed to self-assemble in micelles, fibers and hydrogels. With the eventual goal of producing recombinant biomaterials such as antimicrobial protein polymer, we here explore the secreted production in Pichia pastoris of a fusion of NZ2114 with a hydrophilic random coil protein polymer C
. The intact NZ2114-C
fusion copolymer was produced with a yield of purified protein on the order of 1 g.L
supernatant. We find that purified NZ2114-C
has an activity against clinical strain MRSA, but very much lower than activity of chemically synthesized NZ2114. We conclude that possibly, the activity of NZ2114 is impaired by the C-terminal attachment to the protein polymer chain, but other reasons for the low activity cannot yet be excluded either. This article is protected by copyright. All rights reserved.
A novel antifungal protein with a molecular mass around 50 kDa was purified from seeds of Sesbania virgata (Cav.) Pers. using ammonium sulfate fractionation followed by gel filtration on a Sephadex ...G-75 Superfine (Sigma) column and reverse-phase high performance liquid chromatography on a C8 column. The protein, designated FP1-A, with a novel N-terminal sequence AMVHSPGG(S)FS(P), showed growth inhibitory activity of filamentous fungi Aspergillus niger, Cladosporium cladosporioides, Colletotrichum gloeosporioides and Fusarium solani.
ABSTRACT Chondrosarcoma, an unusual malignant neoplasm, develops in cartilaginous tissue and presents low rate of metastasis, mainly affecting the axial skeleton from the adult to senile dogs. In the ...face of unusual occurrence of chondrosarcoma in the long bones of young dogs, the present report aimed to describe it in the right humerus of a two-and-a-half-year-old Siberian Husky, attended at the Veterinary Hospital of the University of Franca, with limping of the right thoracic limb, for 20 days. The radiographic examination of the humerus showed bone lysis and periosteal proliferation. In the incisional biopsy, proliferation of atypical chondrocytes with diffuse distribution, interspersed with compact bone matrix, was observed. The amputation of the limb was performed, and the fragment histopathological analysis showed grade I chondrosarcoma. Periodic returns were made for neoplastic staging, and at 240 days after surgery lung metastases were detected, however, the tutor did not authorize chemotherapy and radiotherapy for financial reasons and due to the absence of respiratory symptoms so far (410 days after surgery). Although uncommon, chondrosarcoma can affect the long bones of young dogs, with clinical signs similar to other bone neoplasms, and, even with the radical limb amputation, can demonstrate systemic metastasis.
RESUMO O condrossarcoma, neoplasia maligna incomum, desenvolve-se em tecido cartilaginoso e apresenta baixo índice de metástases, acometendo principalmente o esqueleto axial de cães adultos a senis. Diante da ocorrência incomum de condrossarcoma em ossos longos de cães jovens, o presente relato teve como objetivo descrevê-lo no úmero direito de um Husky Siberiano de dois anos e meio de idade, atendido no Hospital Veterinário da Universidade de Franca, com claudicação do membro torácico direito, há 20 dias. O exame radiográfico do úmero mostrou lise óssea e proliferação periosteal. Na biópsia incisional, observou-se proliferação de condrócitos atípicos com distribuição difusa, intercalados com matriz óssea compacta. Foi realizada a amputação do membro, e a análise histopatológica do fragmento evidenciou condrossarcoma grau I. Foram feitos retornos periódicos para estadiamento neoplásico e, aos 240 dias após a cirurgia, foram detectadas metástases pulmonares. O tutor não autorizou quimioterapia e radioterapia por motivos financeiros e por ausência de sintomas respiratórios até o momento (410 dias após a cirurgia). Apesar de incomum, o condrossarcoma pode acometer os ossos longos de cães jovens, com sinais clínicos semelhantes a outras neoplasias ósseas, e, mesmo com a amputação radical do membro, pode demonstrar metástase sistêmica.
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