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In the present study, the biological activity of an extract of the secondary metabolites (E-G6-32) produced by the Curvularia sp. G6-32 endophyte (isolated from the medicinal plant Sapindus saponaria L.) was investigated. The antioxidant potential was confirmed by the DPPH (22.5%) and ABTS (62.7%) assays, and the total phenolic compound content was 40 µg gallic acid equivalents/mg. The extract E-G6-32 displayed good inhibitory activity toward butyrylcholinesterase (BuChE; IC50 = 110 ± 0.05 µg mL−1). The extract E-G6-32 was subjected to spectroscopic and mass spectrometry analyses. Comparison with the literature data confirmed that (-)-asperpentyn (1) was a major component. Asperpentyn belongs to the epoxyquinone family, which has attractive structural complexity, diverse functional groups, and a broad range of biological activities, including specific enzyme inhibitory activity. Our results suggest that Curvularia sp. G6-32 is a promising source of bioactive secondary metabolites and contains (-)-asperpentyn, which has potential pharmaceutical interest.

Despite early reperfusion, patients with ST segment elevation myocardial infarction (STEMI) may present large myocardial necrosis and significant impairment of ventricular function. The present study aimed to evaluate the role of subtypes of B lymphocytes and related cytokines in the infarcted mass and left ventricular ejection fraction obtained by cardiac magnetic resonance imaging performed after 30 days of STEMI. This prospective study included 120 subjects with STEMI submitted to pharmacoinvasive strategy. Blood samples were collected in subjects in the first (D1) and 30th (D30) days post STEMI. The amount of CD11b+ B1 lymphocytes (cells/ml) at D1 were related to the infarcted mass (rho = 0.43; P=0.033), measured by cardiac MRI at D30. These B1 cells were associated with CD4+ T lymphocytes at D1 and D30, while B2 classic lymphocytes at day 30 were related to left ventricular ejection fraction (LVEF). Higher titers of circulating IL-4 and IL-10 were observed at D30 versus D1 (P=0.013 and P

Made available in DSpace on 2020-12-12T01:35:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-09-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) We have previously reported that β-(1→3,1→6)-ᴅ-glucans produced by endophytes Diaporthe sp. G27-60 and G65-65 (GenBank accession codes JF766998 and JF767007, respectively) are promising anti-proliferation agents against human breast carcinoma (MCF-7) and hepatocellular carcinoma (HepG2-C3A) cells. However, the literature fails to describe the effects of Diaporthe exopolysaccharides (EPS) on eukaryotic healthy cells. The fungus Metarhizium anisopliae has been employed as model-system to evaluate the toxicity of pharmaceutical and agricultural-interest substances, taking into account, among other parameters, the speed of conidia germination. Current study verified the effect of different concentrations of Diaporthe β-glucans on the germination speed of M. anisopliae. Conidia were incubated with β-glucans treatments (50, 200 and 400 μg/mL) at 28ºC, sampled during 24 h and analyzed by light microscopy. At the end of a 24-h incubation, the amount of germinated conidia reached ≈99% for controls and ranged between 97.7 and 98.6% for treatments. Bayesian analysis indicated that Diaporthe glucans had no toxicity on M. anisopliae and the curve of germination occurred as expected for this fungal strain. Considering the validity of filamentous fungi as model-systems, results are important data on the toxicity of endophytic EPS on healthy cells and may be associated with our previous results obtained for these polymers against tumor cells. Universidade Estadual de Maringá (UEM) Universidade Católica Dom Bosco (UCDB) University of Georgia (UGA) Faculdade de Ciências e Tecnologia Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) Escola Superior de Agricultura Luiz de Queiroz (ESALQ/USP) Núcleo Permanente Docente do Programa de Pós-graduação em Biotecnologia Ambiental da Universidade Estadual de Maringá (UEM) Faculdade de Ciências e Tecnologia Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) CNPq: 311534/2014-7 CNPq: 447265/2014-8

Endophytic microorganisms live inside the plants without causing any damage to their hosts. In the agricultural field, these endophytes might be a strategy of biological control for phytopathogens. We aimed to isolate endophytic fungi from yellow passion fruit (Passiflora edulis) leaves, evaluating its biocontrol capacity by in vitro antagonism against phytopathogen Colletotrichum sp. CNPU378. We also carried out greenhouse experiments in bean seedlings. A high colonization frequency was obtained (89%), and the molecular identification based on DNA sequencing attested Colletotrichum as the most frequent genus and minor occurrence of Curvularia endophytes. The endophytes tested showed different types of competitive interactions in in vitro antagonism inhibition rate ranging from 28.8 to 48.8%. There were 10 promising antagonists tested for their antagonist activity of crude extracts of secondary metabolites, in which strain PE-36 (20.8%) stood out among the other strains evaluated. In the greenhouse assay, plants inoculated only with endophyte Colletotrichum sp. PE-36 was symptomless and suggest that the endophyte strengthened the growth promotion in common bean plants, especially in the root length and number of leaves when compared to control plants and other treatments. Despite many fungi of Colletotrichum genus being described as causative agents of anthracnose, in this study, the plant sampled was colonized predominantly by Colletotrichum endophytes living in asymptomatic relationship. By the way, we come across a Colletotrichum sp. endophyte able to antagonize a Colletotrichum sp. pathogen.

Made available in DSpace on 2019-10-06T17:00:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-03-05 Made available in DSpace on 2019-10-06T17:00:02Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-03-05 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The aims of this research were to evaluate the safety and probiotic potential of Enterococcus spp. strains and select novel strains for future development of new functional fermented products. Bile salt hydrolase (BSH) activity, capacity of auto-aggregation and co-aggregation, hydrophobicity, tolerance to different pH values and NaCl content, mucin degradation, and antibiotic susceptibility were evaluated. Considering the preliminary probiotic features and safety, the strains were selected for complementary tests: tolerance to gastrointestinal tract (GIT) conditions, adhesion to Caco-2 cells and β-galactosidase activity, and presence of genes encoding virulence factors, antibiotic resistance, and biogenic amines were also performed for the selected strains. Enterococcus faecium SJRP20 and SJRP65 resisted well to the GIT conditions, presented low adhesion property, produced β-galactosidase although they did not present genes implicated in adhesion, aggregation, and colonization. Enterococcus faecium SJRP65 showed fewer genes related to antibiotic resistance and virulence factors and presented good functional properties, with interesting features for future application in dairy products. Federal Institute of Maranhão – IFMA Institute of Natural and Exact Sciences Federal University of Mato Grosso – UFMT Department of Food Science and Experimental Nutrition São Paulo University – USP Department of Food Engineering and Technology São Paulo State University – UNESP Department of Food Engineering and Technology São Paulo State University – UNESP FAPESP: 2014/02131-8 CNPq: 307155/2015-3

Endoglucanases are enzymes widely employed in different industrial fields, albeit with high production costs. Studies on new microbial sources and low-cost substrates are highly relevant, including those on agro-industrial. Current analysis evaluates peanut hull (PH) and sawdust (SD) as substrates for submerged cultures of 14 endophytic fungi isolated from grapevine (Vitis labrusca L.) cultivars Bordô and Concord. Endophytes were grown on a carboxymethylcellulose (CMC) medium and the cup plate assay showed that eight strains (belonging to genera Cochliobolus, Diaporthe, Fusarium and Phoma) had positive results: enzymatic halos ranged from 10.8±0.02 to 15.5±0.07 mm in diameter. Diaporthe sp. strains (GenBank accession codes KM362392, KM362368 and KM362378) and Fusarium culmorum KM362384 were highlighted as the most promising sources. Further, PH and SD as substrates for the fermentation of these fungi were evaluated by the cup plate assay and endoglucanase activity assay. Highest halo diameters were obtained for Diaporthe sp. KM362392: 16.1±0.01 mm (CMC), 14.5±0.01 mm (PH) and 14.7±0.03 mm (SD). The fungus also presented the highest levels of endoglucanase activity: analysis of variance revealed that CMC (3.52±0.98 μmol/min), PH (2.93±0.23 μmol/min) and SD (3.26±0.38 μmol/min) were similarly efficient as substrates. Results deepen knowledge on V. labrusca endophytes that may be endoglucanase sources, even though further optimizations in submerged cultures with PH and SD should be undertaken to increase the enzymatic production from these wastes. Endoglucanases são enzimas amplamente empregadas em diferentes setores industriais; embora sua produção apresente custos elevados. Estudos sobre novas fontes microbianas e substratos mais baratos são de grande importância, incluindo os resíduos agroindustriais. Nesse estudo, casca de amendoim (CA) e serragem (SE) foram testadas como substratos para o cultivo submerso de 14 fungos endofíticos isolados das cultivares Bordô e Concord de videira (Vitis labrusca L.) Os endófitos foram crescidos em meio contendo carboximetilcelulose (CMC) e o ensaio cup plate mostrou resultados positivos para oito fungos (pertencentes aos gêneros Cochliobolus, Diaporthe, Fusarium and Phoma); os halos enzimáticos variaram entre 10,8±0,02 e 15,5±0,07 mm de diâmetro. Linhagens de Diaporthe sp. (códigos de acesso no GenBank KM362392, KM362368 e KM362378) e Fusarium culmorum KM362384 se destacaram como produtores mais promissores. Então, o uso de CA e SE como substratos para a fermentação desses fungos foi avaliado pelo ensaio cup plate e pela quantificação da atividade de endoglucanase. Os maiores halos enzimáticos foram obtidos para Diaporthe sp. KM362392: 16,1±0,01 mm (CMC), 14,5±0,01 mm (CA) e 14,7±0,03 mm (SE). Esse fungo também apresentou os maiores níveis de endoglucanase: a análise de variância revelou que CMC (3,52±0,98 μmol/min), CA (2,93±0,23 μmol/min) e SE (3,26±0,38 μmol/min) foram substratos similarmente eficientes. Esses resultados expandem o conhecimento sobre endófitos de V. labrusca que são fontes de endoglucanases; futuras otimizações quanto ao cultivo submerso com CA e SE podem ser utilizadas para aumentar a produção enzimática a partir do uso desses resíduos.

Obesity is currently considered a global epidemic and it leads to several alterations on the human body and its metabolism. There are evidences showing that the intestinal microbiota can influence on the pathogenesis of obesity. Microbiota plays a vital role not only in the digestion and absorption of nutrients, but also in the homeostatic maintenance of host immunity, metabolism, and gut barrier. Its dietary alteration is an important target in the treatment of obesity. Emerging evidence suggests that modifying the composition of the gut microbiota through probiotic, prebiotic, and synbiotic supplementation may be a viable adjuvant treatment option for obese individuals. In this review, the impact of probiotics, prebiotics, and synbiotics on the anthropometric profile, biochemical regulation, clinical, and immunological markers, as well as on the gut microbiota of obese hosts is described. It also emphasizes how changes in the composition and/or metabolic activity of the gut microbiota through the administration of nutrients with probiotic, prebiotic, or synbiotic properties can modulate the host's gene expression and metabolism, and thereby positively influence on the host's adipose tissue development and related metabolic disorders. The beneficial effects on the host's metabolism promoted by prebiotics, probiotics, and synbiotics have been successfully demonstrated by several studies. However, further investigation is needed to fully explain the cellular mechanisms of action of probiotics and prebiotics on human health, and also to elucidate the relationship between microbiota and obesity etiology, using well-designed, long-term, and large-scale clinical interventions.

Made available in DSpace on 2019-10-04T12:38:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-06-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The objective of this study was to evaluate the probiotic properties of Lactobacillus casei and Lactobacillus fermentum strains, as well as to select novel and safe strains for future development of functional fermented products. The in vitro auto-aggregation, co-aggregation, hydrophobicity, -galactosidase production, survival to gastrointestinal tract (GIT), and antibiotic susceptibility were evaluated. The selected strains were additionally tested by the presence of genes encoding adhesion, aggregation and colonization, virulence factors, antibiotic resistance, and biogenic amine production, followed by the evaluation of acidifying kinetic parameters in milk, and survival of the strains under simulated GIT conditions during refrigerated storage of fermented milk. Most strains of both species showed high auto-aggregation; some strains showed co-aggregation ability with other lactic acid bacteria (LAB) and/or pathogens, and both species showed low hydrophobicity values. Seven L. casei and six L. fermentum strains produced -galactosidase enzymes, and ten strains survived well the simulation of the GIT stressful conditions evaluated in vitro. All strains were resistant to vancomycin, and almost all the strains were resistant to kanamycin. L. casei SJRP38 and L. fermentum SJRP43 were distinguished among the other LAB strains by their higher probiotic potential. L. fermentum SJRP43 presented fewer genes related to virulence factors and antibiotic resistance and needed more time to reach the maximum acidification rate (V-max). The other kinetic parameters were similar. Both strains survived well (>8 log(10) CFU/mL) to the GIT-simulated conditions when incorporated in fermented milk. Therefore, these strains presented promising properties for further applications in fermented functional products. Sao Paulo State Univ, Dept Food Engn & Technol, Rua Cristovao Colombo 2265, BR-15054000 Sao Jose Preto, SP, Brazil UFMT Mato Grosso Fed Univ, Dept Food & Nutr, Fac Nutr, Cuiaba, MT, Brazil Univ Sao Paulo, Dept Food Sci & Expt Nutr, Fac Pharmaceut Sci, Sao Paulo, SP, Brazil Sao Paulo State Univ, Dept Food Engn & Technol, Rua Cristovao Colombo 2265, BR-15054000 Sao Jose Preto, SP, Brazil FAPESP: 2014/02131-8 FAPESP: 2014/02132-4 CNPq: 307155/2015-3

The adaptation of microorganisms to different environmental conditions, such as temperature, salinity, extreme pH values, and oscillation in the availability of nutrients (carbon, nitrogen, and phosphate), has a vital role for their survival. The transduction of the signal generated by the sensing of the extracellular pH by fungi occurs through a conserved molecular pathway that involves the activation of PacC transcription factor. This adaptive response is observed in various species and impacts the metabolic processes, pathogenicity, and biotechnological applications in the industry. Variations in pH can influence nearly all physiological processes that lead to the secretion of macromolecules and metabolites. Under the influence of the regulatory circuits of carbon, nitrogen, sulfur, and pH, the regulation of inorganic phosphate (Pi) uptake is performed by several regulatory genes, such as Nuc-1 transcription factor. Nuc-1 is involved with the post-transcriptional control and regulation of the transcription of structural genes that act in the homeostasis system and Pi uptake. Pi is essential for maintaining cell structure and many metabolic processes, being a growth limiter in nature. Thus, the genetic and molecular mechanisms that control the adaptation of microorganisms to the variations of Pi levels are being shown in many model organisms. In this way, the study of the adaptive microbial response to Pi oscillations has high importance, since these investigations may contribute to portray the regulation of molecular mechanisms involved with the Pi system and pH regulator circuit. The pH regulation, once established, will permit the genetic manipulation of economic and medical processes.

Made available in DSpace on 2020-12-12T01:29:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-10-01 This study aimed to evaluate the impact of a two-week treatment period with probiotic low-fat fermented goat milk by Lactobacillus casei Lc-1, supplemented with passion fruit by-product (1%), on the modulation of gut microbiota from obese individuals using the Simulator of Human Intestinal Microbial Ecosystem (SHIME) system. The effects were carried out through the study of gut microbiota composition, using 16S rRNA next generation sequencing, quantification of short-chain fatty acids (SCFA) and ammonium ions. The microbiota composition changed across three vessels representing the colon regions, because of fermented milk treatment. Fermented goat milk administration caused a reduction of bacteria belonging to genera Prevotella, Megamonas and Succinivibrio, which can produce SCFA, and an increase of Lactobacillus and Bifidobacterium genera in all simulated colon regions. There was no effect on SCFA and on ammonium ions concentration during treatment period. Fermented milk shifted the obese donors’ microbiota without changing metabolites production. It happens, possibly, due to a balance in abundances among bacterial genera that can produce or not SCFA, and among bacterial genera with high or low proteolytic activity. Our outcomes help to clarify the effects of the ingestion of a probiotic low-fat fermented goat milk product on colon microbiota composition. Institute of Natural and Exact Sciences UFR - Federal University of Rondonópolis Department of Food Engineering and Technology UNESP - São Paulo State University Department of Food Science UNESP - São Paulo State University Department of Food Engineering and Technology UNESP - São Paulo State University Department of Food Science UNESP - São Paulo State University

Made available in DSpace on 2018-12-11T17:10:08Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-04-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The aim of this study was to evaluate the safety and probiotic potential characteristics of ten Lactobacillus spp. strains (Lactobacillus fermentum SJRP30, Lactobacillus casei SJRP37, SJRP66, SJRP141, SJRP145, SJRP146, and SJRP169, and Lactobacillus delbrueckii subsp. bulgaricus SJRP50, SJRP76, and SJRP149) that had previously been isolated from water buffalo mozzarella cheese. The safety of the strains was analyzed based on mucin degradation, hemolytic activity, resistance to antibiotics and the presence of genes encoding virulence factors. The in vitro tests concerning probiotic potential included survival under simulated gastrointestinal (GI) tract conditions, intestinal epithelial cell adhesion, the presence of genes encoding adhesion, aggregation and colonization factors, antimicrobial activity, and the production of the β-galactosidase enzyme. Although all strains presented resistance to several antibiotics, the resistance was limited to antibiotics to which the strains had intrinsic resistance. Furthermore, the strains presented a limited spread of genes encoding virulence factors and resistance to antibiotics, and none of the strains presented hemolytic or mucin degradation activity. The L. delbrueckii subsp. bulgaricus strains showed the lowest survival rate after exposure to simulated GI tract conditions, whereas all of the L. casei and L. fermentum strains showed good survivability. None of the tested lactobacilli strains presented bile salt hydrolase (BSH) activity, and only L. casei SJRP145 did not produce the β-galactosidase enzyme. The strains showed varied levels of adhesion to Caco-2 cells. None of the cell-free supernatants inhibited the growth of pathogenic target microorganisms. Overall, L. fermentum SJRP30 and L. casei SJRP145 and SJRP146 were revealed to be safe and to possess similar or superior probiotic characteristics compared to the reference strain L. rhamnosus GG (ATCC 53103). Departamento de Engenharia e Tecnologia de Alimentos Instituto de Biociências Letras e Ciências Exatas Universidade Estadual Paulista (UNESP), R. Cristovão Colombo, 2265 Departamento de Alimentos e Nutrição Faculdade de Nutrição Universidade Federal de Mato Grosso (UFMT) Instituto de Ciências Exatas e Naturais Universidade Federal de Mato Grosso (UFMT) Departamento de Veterinária Universidade Federal de Viçosa (UFV) Departamento de Biologia Instituto de Biociências Letras e Ciências Exatas Universidade Estadual Paulista (UNESP), Rua Cristóvão Colombo, 2265 Departamento de Engenharia e Tecnologia de Alimentos Instituto de Biociências Letras e Ciências Exatas Universidade Estadual Paulista (UNESP), R. Cristovão Colombo, 2265 Departamento de Biologia Instituto de Biociências Letras e Ciências Exatas Universidade Estadual Paulista (UNESP), Rua Cristóvão Colombo, 2265 FAPESP: 2014/02131-8 FAPESP: 2014/02132-4

In this study, we evaluated the survival of Enterococcus faecium and Enterococcus durans, isolated from cheese, in the presence of medications and under simulated in vitro gastrointestinal conditions. The presence of genes encoding virulence factors, the susceptibility to antimicrobial agents, and adhesion properties were also assessed. Enterococcus faecium and E. durans both exhibited resistance to most of the tested medications but showed a large sensitivity to analgesics and antihypertensives; they also showed wide susceptibility to antimicrobial agents. Enterococcus durans SJRP29 had greater resistance to the presence of medications in comparison with the probiotic Lactobacillus acidophilus La-5. The strains, except for E. durans SJRP05, did not harbor virulence genes. Enterococcus durans SJRP14, SJRP17, and SJRP26 were sensitive to all tested antimicrobial agents. Enterococcus faecium was more stable during the simulation of gastrointestinal tract and showed greater viability. At the end of the assay, except for E. durans SJRP17, all strains showed high viability (>7 log cfu/mL). Enterococcus durans SJRP29 stood out from the other strains and was selected for further evaluation; it tolerated up to 3.0% NaCl at 30 and 37°C, besides having good adhesion properties (high values of auto-aggregation, co-aggregation, and hydrophobicity). Additionally, the microorganism did not show bile salt hydrolase activity or mucin degradation. These results encourage carrying out additional tests to evaluate the probiotic features by using in vitro dynamic models and in vivo tests before applying these strains to a food system.

Endophytic fungi have been described as producers of important bioactive compounds; however, they remain under-exploited as exopolysaccharides (EPS) sources. Therefore, this work reports on EPS production by submerged cultures of eight endophytes isolated from Piper hispidum Sw., belonging to genera Diaporthe, Marasmius, Phlebia, Phoma, Phyllosticta and Schizophyllum. After fermentation for 96 h, four endophytes secreted EPS: Diaporthe sp. JF767000, Diaporthe sp. JF766998, Diaporthe sp. JF767007 and Phoma herbarumJF766995. The EPS from Diaporthe sp. JF766998 differed statistically from the others, with a higher percentage of carbohydrate (91%) and lower amount of protein (8%). Subsequently, this fungus was grown under submerged culture for 72, 96 and 168 h (these EPS were designated EPSD1-72, EPSD1-96 and EPSD1-168) and the differences in production, monosaccharide composition and apparent molecular were compared. The EPS yields in mg/100 mL of culture medium were: 3.0 ± 0.4 (EPSD1-72), 15.4 ± 2.2 (EPSD1-96) and 14.8 ± 1.8 (EPSD1-168). The EPSD1-72 had high protein content (28.5%) and only 71% of carbohydrate; while EPSD1-96 and EPSD1-168 were composed mainly of carbohydrate (≈95 and 100%, respectively), with low protein content (≈5%) detected at 96 h. Galactose was the main monosaccharide component (30%) of EPSD1-168. Differently, EPSD1-96 was rich in glucose (51%), with molecular weight of 46.6 kDa. It is an important feature for future investigations, because glucan-rich EPS are reported as effective antitumor agents., Graphical abstract, Highlights • First report of the EPS production by Diaporthe endophytes. • Diaporthe sp. JF766998 was the best EPS producer. • The time of 96 h was considered excellent to the EPS production. • The EPS from Diaporthe sp. JF766998 showed high contents of glucose.

In Brazil, tomato is one of the most consumed vegetables and the fungus Fusarium oxysporum is one of the most important phytopathogen of tomato plants (Lycopersicon esculentum Mill.). Thus, the search of beneficial microorganisms with growth-promoting and/or biological control properties represent an important tool for agricultural biotechnology. Herein, two Trichoderma endophytes (strains 36b and 164b) associated with Coffea arabica were investigated on their growth-promoting activities on plants and their antagonist effects and interactions against F. oxysporum. Molecular multigene (ITS- TEF-TUB-CAL) identification and phylogenetic analysis allowed the identification of these endophytes as belonging to Trichoderma atroviride species. When inoculated with the endophytic strain 36b, tomato plants reached the highest speed of seedling emergence (83.3%), but both endophytes increased the number of leaves, root length and dry biomass of treated plants. Regarding the in vitro antagonism assay, reduced phytopathogen growth by approximately 70 (strain 36b) and 52% (strain 164b) which indicates a partial replacement of endophytes after initial deadlock with mycelial contact. Scanning electron microscopy allowed to observe the presence of Fusarium macroconidia between endophytic hyphae and conidia, with the helicoidization of endophytic hyphae, which wrapped around the pathogen hyphae, suggesting a mechanical inhibition by strangulation.

Fungus-caused diseases are among the greatest losses in grapevine culture. Biological control of pathogens by endophytes may be used to decrease fungicide application rates and environmental impacts. Previously, Diaporthe sp. B46-64 and C27-07 were highlighted as antagonists of grapevine phytopathogens. Herein, molecular multigene (ITS-TUB-TEF1) identification and phylogenetic analysis allowed the identification of these endophytes as belonging to Diaporthe schini species. Agrobacterium tumefaciens-mediated transformation was employed for obtaining 14 stable and traceable gfp- or DsRed-expressing transformants, with high transformation efficiency: 96% for the pFAT-GFP plasmid and 98% for pCAM-DsRed plasmid. Transformants were resistant to hygromycin B with gene hph confirmed by polymerase chain reaction and proved to be mitotically stable, expressing the fluorescent phenotype, with morphological differences in the colonies when compared with wild strains. In vitro antagonism tests revealed an increased antagonistic activity of some transformant strains. The current genetic transformation of D. schini mediated by A. tumefaciens proved to be an efficient technique within the randomized insertion of reporter genes for the monitoring of the strain in the environment. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s12088-019-00787-0) contains supplementary material, which is available to authorized users.

Made available in DSpace on 2019-10-06T16:20:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-06-01 Made available in DSpace on 2019-10-06T16:20:45Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-06-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The effect of Ln. mesenteroides subsp. mesenteroids SJRP55 on the fatty acid profile, on the content of bioactive unsaturated fatty acids, on the inhibition Listeria monocytogenes and on bacterial viability in fermented cream were evaluated. Fermented cream treatments were produced using Lactococcus spp. alone (T1), in co-culture with L. monocytogenes (T2) or Ln. mesenteroides subsp. mesenteroids SJRP55 (T3), and with both microorganisms simultaneously (T4), followed by 28 day-storage. Ln. mesenteroides subsp. mesenteroides SJRP55 caused a reduction on L. monocytogenes population and had distinguished effect on fatty acid profile, causing a reduction on the monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA), as well as an increase in saturated fatty acids (SFA), only on the first day of storage. Fermentation resulted in increased conjugated linoleic acid (CLA) and decreased α-linolenic (ALA) and oleic (OA) acids contents. The cream fermented by Lactococcus spp. and Ln. mesenteroides subsp. mesenteroides SJRP55 showed potential functional properties in CLA content. The treatments were characterized according to their bioactive unsaturated fatty acids using the principal component analysis (PCA). The first principal component (70.25%) was explained by OA and CLA, and the second principal component (25.79%) was explained by ALA, totaling 96.44%. Ln. mesenteroides subsp. mesenteroides SJRP55 improved the functional properties and safety of fermented cream. São Paulo State University – UNESP Department of Food Engineering and Technology, São José do Rio Preto São Paulo University – USP Department of Biochemical and Pharmaceutical Technology Federal University of Mato Grosso - UFMT Institute of Natural and Exact Sciences São Paulo State University – UNESP Department of Food Engineering and Technology, São José do Rio Preto FAPESP: 2010/09302-1 CNPq: 307155/2015-3

Made available in DSpace on 2018-12-11T17:22:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-12-01 Made available in DSpace on 2018-12-11T17:22:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-12-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) This study aimed at characterizing guava, orange and passion fruit by-products and investigating the effect of adding these fruit by-products to probiotic fermented goat milk and cereal-based fermented products. Fruit by-products showed total fiber content, phenolic compounds and antioxidant activity varying from, respectively, 58.20–89.80%, 253.14–420.89 mg GAE/100 g, and 11.38–17.37 μmol TE/g. Most carotenoids were represented by β-carotene, which ranged from 7.91 to 56.07 μg/g. The presence of fruit by-products did not affect the fermentation time of fermented oat beverage and fermented goat milk; however, a significant increase (ranging from 0.28 to 0.91 h) in fermentation time of fermented rice beverages was observed after the addition of by-products. Fruit by-products also resulted in an increase in acidification throughout storage; however, they did not affect the counts of probiotic bacteria. A decrease in probiotic survival during in vitro gastrointestinal simulation was observed in all treatments. Nonetheless, the presence of orange and passion fruit by-products enhanced the resistance of the probiotics to simulated gastrointestinal conditions and resulted in population 2 log CFU/mL higher than the control treatment. Fruit by-products can be considered relevant sources of bioactive compounds useful in raising the functional attributes of probiotic fermented products. São Paulo State University – UNESP Department of Food Engineering and Technology, São José do Rio Preto Federal University of Mato Grosso - UFMT Institute of Natural and Exact Sciences Federal University of Mato Grosso - UFMT Institute of Natural and Exact Sciences, Avenida dos Estudantes, 5055 São Paulo State University – UNESP Department of Food Engineering and Technology, São José do Rio Preto FAPESP: 2014/02131-8 FAPESP: 2014/02132-4 CNPq: 2015-3 CNPq: 307155

Endophytic fungi colonize the interior of plant tissues and organs, establishing an intimate mutualistic association with no visible symptoms. The fungi may help protect the plant against herbivores and pathogens, making them potentially useful endophytes in the biological control of diseases and agricultural pests. The biotechnological interest in these organisms has stimulated research related to the bioprospecting of endophytic fungi. Grapevine is among the oldest of plants cultivated by man, with the grape being one of the most highly consumed fruits in the world. Diseases cause significant damage to grape cultures, making their integrated control important to reduce the use of pesticides and, consequently, environmental and human contamination. The rustic species Vitis labrusca L. (Vitaceae), used in the preparation of juices and wines, is highly resistant to fungal diseases. We isolated leaf endophytic fungi of the Bordo and Concord cultivars (V. labrusca L.), which were ordered into 68 and 62 morpho-groups of the Bordo and Concord cultivars, respectively. We used scanning electron microscopy to confirm the presence of endophytes in the leaves. Endophytic diversity was analyzed based on sequencing the ITS1-5.8S-ITS2 region of rDNA, allowing the identification of fungi belonging to genera including Cochliobolus, Bipolaris, Fusarium, Alternaria, Diaporthe, Phoma and Phomopsis. Phylogenetic analysis confirmed the identity of the endophytes. The biotechnological potential of endophytes was tested in vitro for the control of pathogenic fungi of grapevines including Alternaria sp., Sphaceloma sp. and Glomerella sp. Inhibition percentages above 50 % as demonstrated by some isolates demonstrate their potential for biological control.

In this study, we evaluated the effect of different matrices (MRS, milk and milk with inulin) on the tolerance of probiotic strains (Lactobacillus acidophilus La-5 and Bifidobacterium animalis subsp. lactis BB-12) to simulated conditions similar to those found in the gastrointestinal tract. Both probiotic strains demonstrated a significantly lower viability after exposure to in vitro gastric and intestinal conditions, and B. animalis subsp. lactis BB-12 showed higher survival during the test compared to L. acidophilus La-5 in all tested matrices. Milk and inulin protected probiotics from in vitro gastrointestinal stress. These results suggest that it is critical to formulate the food matrix to be used as probiotic carrier.

Emotion processing impairments are common in patients undergoing brain surgery for fronto-temporal tumour resection, with potential consequences on social interactions. However, evidence is controversial concerning side and site of lesions causing such deficits. This study investigates visual and auditory emotion recognition in brain tumour patients with the aim of clarifying which lesion sites are related to impairments in emotion processing from different modalities. Thirty-four patients were evaluated, before and after surgery, on facial expression and emotional prosody recognition; voxel-based lesion–symptom mapping (VLSM) analyses were performed on patients’ post-surgery MRI images. Results showed that patients’ performance decreased after surgery in both visual and auditory modalities, but, in general, recovered 3 months after surgery. In facial expression recognition, left brain-damaged patients showed greater post-surgery deterioration than right brain-damaged ones, whose performance specifically decreased for sadness and fear. VLSM analysis revealed two segregated areas in the left hemisphere accounting for post-surgery scores for happy (fronto-temporo-insular region) and surprised (middle frontal gyrus and inferior fronto-occipital fasciculus) facial expressions. Our findings demonstrate that surgical removal of tumours in the fronto-temporal region produces impairment in facial emotion recognition with an overall recovery at 3 months, suggesting a partially different representation of positive and negative emotions in the left and right hemispheres for visually – but not auditory – presented emotions; moreover, we show that deficits in specific expression recognition are associated with discrete lesion locations.

The effect of addition of apple, banana and grape flours (1%) on the characteristics of fermented milk by culture ABT-4 was evaluated. Supplementation with fruit flour affected only the acidification rate, and there was no effect on fermentation time. There was a decrease in the pH level and an increase in titratable acidity throughout storage. The level of probiotics was not affected by milk supplementation with fruit flours during 28 days of storage. Decreased probiotic survival during in vitro gastrointestinal simulation was observed in all the treatments. Nonetheless, the supplementation with fruit flours improved Lactobacillus acidophilus tolerance to simulated gastrointestinal conditions, specifically at days 14 and 28 of storage. On the other hand, only banana flour had a protective effect on Bifidobacterium animalis subsp. lactis after 28 days of storage. This is the first report that demonstrated a protective effect of fruit flours on the simulated gastrointestinal resistance of probiotics.

Endophytic fungi live in the interior of healthy plants without causing them any damage. These fungi are of biotechnological interest; they may be used in the biological control of pests and plant diseases, and in the pharmaceutical industry. The aquatic macrophytes Eichhornia azurea (Kunth) and Eichhornia crassipes (Mart.) belong to the Pontederiaceae family. The first is a fixed-floating species and the second is a free-floating species that is known for its phytoremediation potential. The fungal endophytes associated with the leaves of E. azurea and E. crassipes, native to the Upper Paraná River floodplain, Brazil, were isolated. The sequencing of the ITS1-5.8S-ITS2 region of ribosomal DNA was performed and the nucleotide sequences obtained were compared with those available in the GenBank database for the molecular identification of the isolates. The construction of phylogenetic trees was performed using the MEGA5 software. The results showed that high colonization frequencies were obtained from the 610 foliar fragments sampled from each plant: 87.86% for E. azurea and 88.85% for E. crassipes. At the genus level, it was possible to identify 19 fungal endophytes belonging to the genera Alternaria, Bipolaris, Cercospora, Diaporthe, Gibberella, Pestalotiopsis, Plectosphaerella, Phoma, and Saccharicola. Two other endophytes were identified at the species level (Microsphaeropsis arundinis). Genera Bipolaris, Cercospora, Microsphaeropsis, and Phoma were found as endophytes in the two macrophytes and the other genera were host-specific, being isolated from only one macrophyte, proving that there is a small difference in the endophytic diversity of the two Eichhornia species analyzed.

The objective of this study was to evaluate the influence of different combinations of cultures, Lactobacillus acidophilus — La and Bifidobacterium animalis subsp. lactis — Bb as pure cultures or in co-culture with Streptococcus thermophilus — St, on fermented milk during fermentation, including changes to the acidification profile, organic acid production and lactose consumption and during 28-day storage at 4 °C, in terms of bacteria viability, syneresis, sensory properties, organic acid content and viability under simulated in vitro gastrointestinal conditions. La culture was showed the lowest acidification rate (V max ) values, whereas the pure St culture showed the highest V max values. During fermentation, Bb produced the largest amount of acetic acid, and only La was able to metabolize citric acid. Syneresis decreased during storage for all treatments. Counts of S. thermophilus and B. animalis subsp. lactis remained stable during the storage period in all treatments, while the counts of L. acidophilus decreased over time only in the case of the La treatment. The simulation of probiotic resistance to gastrointestinal conditions indicated that bifidobacteria possess a greater tolerance to acid and bile than the lactobacilli strain. The La treatment resulted in lower scores for all attributes in both periods of sensory analysis. When lactic acid was present in smaller quantities and citric acid was present in larger amounts, the scores regarding flavor and overall acceptability attributes were higher. Depending on the combination of microorganisms used in fermented milk manufacturing, it had positive or negative impacts on the product's characteristics.

Pelargonium sidoides, known as umckaloabo, is a plant originally from South Africa and its extract is used in popular medicine. The fungus Metarhizium anisopliae, an important entomopathogen used in biological control of pests, is also employed as model for the evaluation of toxicity and compatibility of different products, considering, among other parameters, conidia germination speed. Therefore, this study aimed to evaluate the sensibility of M. anisopliae var. anisopliae in the presence of the extract of P. sidoides, at a concentration of 20.625 and 2.0625 mg/ml. During incubation at 28°C, samples were collected at 0, 6, 8, 10, 12 and 24 h and analyzed by light microscopy, with observation of 300 conidia, in triplicate, for both treatments and negative controls (in a completely randomized design). This study shows that the two concentrations of P. sidoides extract tested are capable of delaying the M. anisopliae conidia germination speed, in comparison to the controls, although the germination frequency has been restored after about 12 h of conidia incubations, with did not lack conidia viability, indicating no toxicity.

Background Improving the knowledge of angiomyolipoma physiopathology might help in refining its pharmacological treatment. We investigated if angiomyolipoma cells have migratory properties, how their growth and motility can be influenced by the hormonal milieu, and if this can be related to a specific gender. Methods Primary cells were isolated from angiomyolipomas surgically resected for therapeutical reasons in a female and in a male patient. The genetic control demonstrated no TSC2 deletion. Bi- (wound healing) and three-dimensional (transwell assay) migration were analyzed in vitro in basal conditions and under the influence of 17- β-estradiol and SDF-1α. Results Treatment up to 72 hours with 17-β-estradiol (0.1-100 nM), tamoxifen (0.2-20 μM) or with both, does not modify angiomyolipoma cells proliferation. On the other hand, SDF-1α and 17-β-estradiol treatment induce a significant motility increase (both bi- and three-dimensional) which becomes evident already after 2 hours of incubation. Angiomyolipoma cells express mRNA coding for SDF-1α and 17-β-estradiol receptors and secrete both the metalloproteases principally involved in malignant phenotype acquisition, i.e. MMP-2 and MMP-9. Conclusion Angiomyolipoma cells behave similarly, despite their different source. Primary angiomyolipoma cells migrate in response to hormonal milieu and soluble factors, and produce active metalloproteases, both aspects being consistent with the theory claiming they can migrate to the lungs (and/or other organs) and colonizing them. No main feature, among the aspects we analyzed, seems to be referable to the gender of origin.

20 p.-2 fig.-3 tab., This study reports the characterization and antiproliferative activity of exopolysaccharides (EPS) produced by submerged cultures of the endophytes Diaporthe sp. JF766998 and Diaporthe sp. JF767007 isolated from the medicinal plant Piper hispidum Sw. Both strains secreted a crude EPS that, upon size exclusion chromatography, showed to contain a heteropolysaccharide (galactose, glucose and mannose) and a high-molecular weight glucan. Data from methylation analysis, FTIR and NMR spectroscopy (1H, COSY, TOCSY and HSQC-DEPT) indicated that the purified glucan consisted of a main chain of glucopyranosyl β-(1 → 3) linkages substituted at O-6 by glucosyl residues. According to MTT assay, some treatments of both β-glucans have antiproliferative activity against human breast carcinoma (MCF-7) and hepatocellular carcinoma (HepG2-C3A) cells after 24 and 48 h of treatment, exhibiting a degree of inhibition ratio that reached the highest values at 400 μg/mL: 58.0% (24 h) and 74.6% (48 h) for MCF-7 cells, and 61.0% (24 h) and 83.3% (48 h) for HepG2-C3A cells. These results represent the first reports on the characterization and antiproliferative effect of β-glucans from Diaporthe species and also expand the knowledge about bioactive polysaccharides from endophytic sources., The authors are grateful to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; 311534/2014-7 and 447265/2014-8), Fundação Araucária (276/2014) and Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP; 05/53879-3) for financial support.

The interest on novel biological preservation methods has been increasing during recent years, supported by research indicating that antagonistic microorganisms and their antimicrobial metabolites may have some potential use as natural preservatives as a way not only to control the growth but also to inactivate undesired microorganisms in food. Biopreservation using lactic acid bacteria (LAB) and/or their antimicrobial metabolites represents an alternative for improving food safety. These antimicrobial properties of LAB were derived from competition for nutrients and the production of one or more antimicrobial active metabolites such as organic acids (mainly lactic and acetic acid), hydrogen peroxide and also other compounds, such as bacteriocins and antifungal peptides. The important contribution of probiotic LAB in food preservation has been attracting much attention because of the nutritional qualities of the raw material through an extended shelf life of food and their ability to inhibit spoilage and foodborne pathogens, which is interesting for the food industry. In this review, we compiled the available data on the commonly used preservative and present the current knowledge regarding the antimicrobial compounds, especially bacteriocins, the mechanisms of the action and recent applications of antimicrobial compounds in food. The use of competitive microbiota as a biotechnological tool for food preservation may lead to improve the optimization and quality assurance of food products while at the same time retaining the sensory qualities of the product such as color, flavor, texture and nutritional value.

Tropical and subtropical plants are rich in endophytic community diversity. Endophytes, mainly fungi and bacteria, inhabit the healthy plant tissues without causing any damage to the hosts. These fungi can be useful for biological control of pathogens and plant growth promotion. Some plants of the genus Piper are hosts of endophytic microorganisms; however, there is little information about endophytes on Piper hispidum, a medicinal shrub used as an insecticide, astringent, diuretic, stimulant, liver treatment, and for stopping hemorrhages. We isolated the fungal endophyte community associated with P. hispidum leaves from plants in a Brazilian forest remnant. The endophytic diversity was examined based on sequencing of the ITS1-5.8S-ITS2 region of rDNA. A high colonization frequency was obtained, as expected for tropical angiosperms. Isolated endophytes were divided into 66 morphogroups, demonstrating considerable diversity. We identified 21 isolates, belonging to 11 genera (Alternaria, Bipolaris, Colletotrichum, Glomerella, Guignardia, Lasiodiplodia, Marasmius, Phlebia, Phoma, Phomopsis, and Schizophyllum); one isolate was identified only to the order level (Diaporthales). Bipolaris was the most frequent genus among the identified endophytes. Phylogenetic analysis confirmed the molecular identification of some isolates to genus level while for others it was confirmed at the species level.

Metarhizium anisopliae has been considered a promising alternative with low environmental impacts for the biological control of a variety of insect-pests. Another alternative is the use of biological pesticides such as insect growth regulators, including lufenuron. An assessment of the potential impact of fungicides on M. anisopliae is of critical importance to integrated pest management, permitting the compatible use of this entomopathogen with chemical defensives. Based on this, this study aimed to evaluate the effect of different concentrations of lufenuron on the conidia germination speed of M. anisopliae . Conidia were incubated at 28°C and sampled throughout 12 h. Bayesian analysis showed an inhibition of conidia germination in the presence of 2.0 mg/ml of lufenuron, whereas their compatibility was observed in the concentrations of 1.0 mg/ml and 700 µg/ml. It indicates that in these last two concentrations, the fungicide has no toxicity on M. anisopliae , what suggests that it can be employed in biological-chemical combinations, maintaining viable the fungal inoculums after its application in the field, with a low environmental impact.

The aim of this study was to evaluate the measurement of C-peptide, insulin, sex hormone binding globulin (SHBG), and dehydroepiandrosterone sulfate (DHEAS), tests of specific clinical value that are not frequently measured in routine clinical laboratories, carried out using an automated system, AIA 2000 (Tosoh). The obtained data demonstrated that the evaluated system, characterized by fluorescence system detector provides satisfactory analytical performance comparable to those of different instrument that use the well known and widely diffused chemiluminescent principle (Immulite 2000). These characteristics allow an excellent comparability between methods for the measurement of same specific hormones not frequently used in clinical practice.

Millions of tons of corn, pineapple, sugarcane and wheat are annually produced and their industrial processing generates large amounts of wastes. Current study evaluated the use of corncob (CC), pineapple peel (PP), sugarcane bagasse (SB) and wheat bran (WB) as substrates for α-amylase production by submerged cultures of nine endophytes from Piper hispidum Sw. Initially, fungi were grown on a starch medium and the cup plate assay showed that five strains were amylase-positive: Bipolaris sp. JF767001, Marasmius cladophyllus JF767003, Phlebia sp. JF766997, Phyllosticta capitalensis JF766988 and Schizophyllum commune JF766994, with enzymatic halos ranging between 6.30 ± 0.08 and 8.87 ± 0.68 mm in diameter. Further, the use of agro-industrial wastes was evaluated by the cup plate assay, where the highest halo diameters were obtained from fungi grown on PP or SB: up to 15.00 ± 0.16 mm ( Phlebia sp.) and 14.80 ± 0.18 mm ( S. commune ), respectively. Submerged cultures with PP or SB showed the highest levels of α-amylase activity according to the starch-iodine assay, or rather, up to 4.14 ± 0.02 U mL -1 ( Bipolaris sp.) and 4.09 ± 0.02 U mL -1 ( Phlebia sp.), respectively. Results give an in-depth knowledge on tropical endophytes that might be α-amylase sources, and indicate the suitability of these agro-industrial wastes as substrates for fungal enzymatic production.

In this work, we investigated the effect of supplementing fermented milk with quinoa flour as an option to increase probiotic activity during fermented milk production and storage. Fermented milk products were produced with increasing concentrations of quinoa flour (0, 1, 2, or 3g/100g) and submitted to the following analyses at 1, 14, and 28 d of refrigerated storage: postacidification, bacterial viability, resistance of probiotics to simulated gastrointestinal (GI) conditions, and adhesion of probiotics to Caco-2 cells in vitro. The kinetics of acidification were measured during the fermentation process. The time to reach maximum acidification rate, time to reach pH 5.0, and time to reach pH 4.6 (end of fermentation) were similar for all treatments. Adding quinoa flour had no effect on fermentation time; however, it did contribute to postacidification of the fermented milk during storage. Quinoa flour did not affect counts of Bifidobacterium animalis ssp. lactis BB-12 or Lactobacillus acidophilus La-5 during storage, it did not protect the probiotic strains during simulated GI transit, and it did not have a positive effect on the adhesion of probiotic bacteria to Caco-2 cells in vitro. Additionally, the adhesion of strains to Caco-2 cells decreased during refrigerated storage of fermented milk. Although the addition of up to 3% quinoa flour had a neutral effect on probiotic activity, its incorporation to fermented milk can be recommended because it is an ingredient with high nutritive value, which may increase the appeal of the product to consumers.

Endophytes are being considered for use in biological control, and the enzymes they secrete might facilitate their initial colonization of internal plant tissues and direct interactions with microbial pathogens. Microbial proteases are also biotechnologically important products employed in bioremediation processes, cosmetics, and the pharmaceutical, photographic and food industries. In the present study, we evaluated antagonism and competitive interactions between 98 fungal endophytes and Alternaria alternata, Colletotrichum sp., Phyllosticta citricarpa and Moniliophthora perniciosa. We also examined the proteolytic activities of endophytes grown in liquid medium and conducted cup plate assays. The results showed that certain strains in the assemblage of P. hispidum endophytes are important sources of antifungal properties, primarily Lasiodiplodia theobromae JF766989, which reduced phytopathogen growth by approximately 54 to 65%. We detected 28 endophytes producing enzymatic halos of up to 16.40 mm in diameter. The results obtained in the present study highlight the proteolytic activity of the endophytes Phoma herbarum JF766995 and Schizophyllum commune JF766994, which presented the highest enzymatic halo diameters under at least one culture condition tested. The increased activities of certain isolates in the presence of rice or soy flour as a substrate (with halos up to 17.67 mm in diameter) suggests that these endophytes have the potential to produce enzymes using agricultural wastes.

Biopsies of human endometrium were studied for the presence of basic fibroblast growth factor (bFGF). An immunoreactive Mr 18 000 bFGF-like molecule was detected at high levels both in ovulatory cycle and postmenopausal endometrium. This molecule was identified as bFGF on the basis of its molecular weight, its affinity for heparin, its capacity to induce plasminogen activator production and cell proliferation in endothelial GM 7373 cells, and its cross-reactivity with various anti-bFGF antibodies. The levels of endometrial bFGF do not change during the menstrual cycle but they increase significantly after menopause, as evaluated both by biological and immunological assays. Lower levels of an acidic FGF-like activity were also evident in ovulatory cycle endometrium but, at variance with bFGF, no significant increase of this activity was observed in postmenopausal endometrium. These data represent the first characterization of a polypeptide growth factor present in human endometrium.

Hepatoblastomas (HBs) recapitulate liver development. It is possible that HBs result from malignant transformation of hepatic precursor cells, and they may reflect a blockage in normal development. Here we study the expression of cytokeratins (CKs) in order to delineate the immunoprofile and relationship with liver development, as well as vimentin and alphafetoprotein (AFP), of HBs. Immunohistochemistry was performed in a tissue microarray (TMA) containing representative areas of 18 HBs (fetal and/or embryonal and/or mesenchymal); we also reviewed 11 cases not included in the TMA. No cases stained for CKs 1, 5/6, 7, 10, 13, 15, 16, 20, and 34βE12. CK8 stained 73.07% of fetal, 50% of embryonal, and 18% of mesenchymal areas. CK18 stained 100% of epithelial areas. CK19 staining was intense and diffuse in 100% of embryonal samples, but it was weaker in fetal areas (66.66%). AE1 stained epithelial areas in all cases, and it stained 29.41% of mesenchymal areas. AE3 stained 84.61% of embryonal and 60% of fetal components. AE1/AE3 showed stronger staining in embryonal (100%) than in fetal areas (76.92%). Vimentin staining was strong in embryonal (66.66%) and mesenchymal (84.61%) components but weak in fetal areas (8%). Alphafetoprotein was positive in only 20% of fetal and 70% of embryonal areas. Our results support the hypothesis that immunoexpression of HBs follows the stages of normal liver development. Embryonal areas look less differentiated, expressing vimentin and biliary epithelium CKs, whereas fetal areas display a more developed phenotype, similar to that of mature hepatocytes. These data aid in understanding the ontogenesis of HBs and may be used in histopathological diagnosis.

Gladiolus hortulanus, também conhecida como palma-de-santa-rita, é uma planta originária da África tropical e do sul, sendo utilizada para ornamentação. O objetivo do presente trabalho foi avaliar o desenvolvimento de Gladiolus hortulanus L. H. Bailey var. White Friendship, com a utilização de diferentes fertilizantes. O trabalho foi desenvolvido na Casa de Vegetação climatizada, (Pad & Fan e temperatura ambiente de 25° C +/- 3, lux = 7,16X10) da Faculdade de Engenharia – Unesp, Campus de Ilha Solteira, no período de 25 de agosto a 04 de dezembro de 2007. Foram utilizados 3 gramas dos fertilizantes Osmocote (19-06-10), NPK 04-14-08, Basacote 3M e Floranid N32, em vasos de 1,3 litros, preenchidos com substrato comercial Plantmax. Foram também avaliados o número de botões florais, número de flores, altura da haste floral, perímetro da haste floral, teor de clorofila das folhas e análise de macro e micronutrientes nas folhas. Concluiu-se que, para o padrão comercial recomendado pela Cooperativa Holambra (categoria III) e Ceagesp (categoria longa), os melhores tratamentos foram Basacote 3M e NPK 04-14-08.

Com o objetivo de determinar a soroprevalência da infecção por Ehrlichia canis, bem como identificar fatores de risco associados à soropositividade, foi realizado um inquérito sorológico em 109 cães atendidos no Hospital Veterinário/Centro de Saúde e Tecnologia Rural (CSTR)/Universidade Federal de Campina Grande(UFCG), Campus de Patos, Estado da Paraíba. O diagnóstico sorológico da erliquiose foi realizado por meio da reação de imunofluorescência indireta (RIFI) e as amostras que apresentaram títulos de anticorpos >; 40 foram consideradas positivas. Das 109 amostras, 72,5% foram positivas (IC 95% = 63,1% - 80,6%). Animais que tiveram contato com cães (odds ratio = 3,59; IC 95% = 1,41 - 9,12), contato com açudes (odds ratio = 8,39; IC 95% = 1,01 - 69,87) ou com algum acesso à rua (odds ratio = 6,24; IC 95% = 1,28 - 30,39) estiveram mais expostos ao risco de infecção.