Screening Brazilian Macrophomina phaseolina isolates for alkaline lipases and other extracellular hydrolases

Macrophomina phaseolina, phylum Ascomycota, is a phytopathogenic fungus distributed worldwide in hot dry areas. There are few studies on its secreted lipases and none on its colony radial growth rate, an indicator of fungal ability to use nutrients for growth, on media other than potato-dextrose agar. In this study, 13 M. phaseolina isolates collected in different Brazilian regions were screened for fast-growth and the production of hydrolases of industrial interest, especially alkaline lipases. Hydrolase detection and growth rate determination were done on citric pectin, gelatin, casein, soluble starch, and olive oil as substrates. Ten isolates were found to be active on all substrates tested. The most commonly detected enzymes were pectinases, amylases, and lipases. The growth rate on pectin was significantly higher (P < 0.05), while the growth rates on the different media identified CMM 2105, CMM 1091, and PEL as the fastest-growing isolates. The lipase activity of four isolates grown on olive oil was followed for 4 days by measuring the activity in the cultivation broth. The specific lipolytic activity of isolate PEL was significantly higher at 96 h (130 mU mg protein–1). The broth was active at 37 °C, pH 8, indicating the potential utility of the lipases of this isolate in mild alkaline detergents. There was a strong and positive correlation (0.86) between radial growth rate and specific lipolytic activity. [Int Microbiol 2012; 15(1):1-7

Nova proposta para uso de bacteriófagos no controle de mancha foliar em repolho, causada por Xanthomonas campestris pv. Campestris.

Um novo método de produção e identificação de bacteriófagos em suspensão, e seu potencial como controlador biológico. A suspensão obtida por esta forma foi eficaz para controlar tanto in vitro quanto in vivo a bactéria Xanthomonas campestris pv. campestris. Evidenciou-se uma diferença entre a suspensão criada do patógeno (PLP) e outra que não foi (PLN), pois havia halos no crescimento bacteriano em 48 h/ 28º C produzido pelas suspensões PLS e PLP, a suspensão PLN não apresentou halo. Apesar dos tratamentos PLP e PLS não diferirem estatisticamente em relação às UFCs, atingindo valores de 5x 105 e 1x105 respectivamente, diferiram da suspensão bruta (PLN) e da testemunha. Procedeu-se então avaliação in vivo com as suspensões que criaram halo, quando foram incluídos os tratamentos (PLPS) e (PLPP), que consistiram de inóculo adicionado nas suspensões matrizes (PLS e PLP), para o aumento de bacteriófagos. Verificou-se, então, a redução do índice de doença para a testemunha, mostrando que o tratamento com suspensão de bacteriófagos protege a planta. Os índices mais baixos de doenças foram (PLP) e (PLS), com valores de 1,46 e 1,40, respectivamente

Estudo da utilização da fase de evaporação constante da secagem na concentração de liquidos

Orientador : Roberto H. MorettiDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de AlimentosResumo: Dos métodos disponíveis para a concentração de alimentos líquidos, destacam-se a evaporação, osmose reversa e crio-concentração, por serem técnicas de concentração ã baixa temperatura mui to bem estudadas, apresentando cada uma, desvantagens devido ao alto custo de investimento e/ou operação que representam. Foi estudada uma técnica alternativa de concentração onde são utilizadas as propriedades desidratantes de um gás seco em movimento, quando em contato com um líquido, para remoção de água. Verificou-se que tanto a temperatura do líquido quanto do gás influenciam a evaporação de água, sendo essa também dependente da velocidade do gás. Verificou-se que das diversas-formas de contato entre gás e líquido estudadas, a mais elevada evaporação de água foi obtida quando o líquido foi distribuído na forma de "spray" na corrente do gás. Na concentração de solução de sacarose e vinhoto, foi verifica da a diminuição da quantidade de água evaporada em função do aumento da concentração do produto, tal como ocorre em qualquer outro processo convencional de concentração. Na concentração de vinhoto, a técnica "mostrou-se viável economicamente, podendo ser utilizada na remoção de 35-40% da água do mesmo, utilizando unicamente a energia residual de uma destilaria de álcoolAbstract: Of the methods available for concentration of liquid foods, special emphasis should be given to evaporation, reverse osmosis and crio-concentration, because they are very well known techniques for concentration at low temperature, although each of them presents disadvantages due to their high investment and/or operation costs. An alternative technique of concentration has been studied, whereas the desidration properties of a dry gas "in motion were used when in contact with a liquid, for the removal of water. It was observed that the temperature of the liquid and gas had great influence on the evaporation of water, the last one being also dependent of the gas velocity. It was also noted that from the different forms of contact between liquid and gas studied, the highest evaporation was achieved when the liquid was sprayed in the gas current. By concentrating sugar solution and vinasse, it was observed that a substancial reduction in water evaporation occurred with an increase of the product concentration, like as in any other conventional concentration technique. For the concentration of vinasse the technique showed to be economically feasable, being able to evaporate 35-40% of its water by the use of the residual energy of an alcohol distillery onlyMestradoMestre em Tecnologia de Alimento

Antibacterial compounds from marine bacteria, 2010–2015

This review summarizes the reports on antibacterial compounds that have been obtained from marine-derived bacteria during the period 2010–2015. Over 50 active compounds were isolated during this period, most of which (69%) were obtained from Actinobacteria. Several compounds were already known, such as etamycin A (11) and nosiheptide (65), and new experiments with them showed some previously undetected antibacterial activities, highlighting the fact that known natural products may be an important source of new antibacterial leads. New broad-spectrum antibacterial compounds were reported with activity against antibiotic resistant Gram-positive and Gram-negative bacteria. Anthracimycin (33), kocurin (66), gageotetrins A–C (72–74), and gageomacrolactins 1–3 (86–88) are examples of compounds that display promising properties and could be leads to new antibiotics. A number of microbes produced mixtures of metabolites sharing similar chemical scaffolds, and structure–activity relationships are discussed80412151228CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPES3053390/2013-91267382; 130648

The Wnt serpentine receptor Frizzled-9 regulates new bone formation in fracture healing.

Wnt signaling is a key regulator of bone metabolism and fracture healing. The canonical Wnt/β-catenin pathway is regarded as the dominant mechanism, and targeting this pathway has emerged as a promising strategy for the treatment of osteoporosis and poorly healing fractures. In contrast, little is known about the role of non-canonical Wnt signaling in bone. Recently, it was demonstrated that the serpentine receptor Fzd9, a Wnt receptor of the Frizzled family, is essential for osteoblast function and positively regulates bone remodeling via the non-canonical Wnt pathway without involving β-catenin-dependent signaling. Here we investigated whether the Fzd9 receptor is essential for fracture healing using a femur osteotomy model in Fzd9(-/-) mice. After 10, 24 and 32 days the fracture calli were analyzed using biomechanical testing, histomorphometry, immunohistochemistry, and micro-computed tomography. Our results demonstrated significantly reduced amounts of newly formed bone at all investigated healing time points in the absence of Fzd9 and, accordingly, a decreased mechanical competence of the callus tissue in the late phase of fracture healing. In contrast, cartilage formation and numbers of osteoclasts degrading mineralized matrix were unaltered. β-Catenin immunolocalization showed that canonical Wnt-signaling was not affected in the absence of Fzd9 in osteoblasts as well as in proliferating and mature chondrocytes within the fracture callus. The expression of established differentiation markers was not altered in the absence of Fzd9, whereas chemokines Ccl2 and Cxcl5 seemed to be reduced. Collectively, our results suggest that non-canonical signaling via the Fzd9 receptor positively regulates intramembranous and endochondral bone formation during fracture healing, whereas it does not participate in the formation of cartilage or in the osteoclastic degradation of mineralized matrix. The finding that Fzd9, in addition to its role in physiological bone remodeling, regulates bone repair may have implications for the development of treatments for poorly or non-healing fractures

Immunostaining of β-catenin at day 10 and histochemical TRAP staining indicating osteoclasts at day 24 in the fracture calli.

<p>WT: upper panel, <i>Fdz9</i>^−/−: lower panel. β-catenin was expressed in osteoblasts (OB) and proliferating chondroblasts (CB) but to a lesser extend in hypertrophic chondrocytes (HC). C: cortex. There were no differences between both genotypes. Only TRAP positive cells with ≥2 nuclei were identified as osteoclasts (OC). TRAP-staining either revealed no significant differences between both genotypes.</p

Immunostaining of chemokines Cxcl5 and Ccl2 at day 10.

<p>WT: upper panel, <i>Fdz9</i>^−/−: lower panel. C: cortex. Cxcl5 (<i>left</i>) and Ccl2 (<i>right</i>) were expressed by precursor cells, osteoblasts and chondrocytes in both genotypes, but staining was less intense in the absence of <i>Fzd9</i>. C: cortex, HC: hypertrophic chondrocytes, OB: osteoblasts, OT: osteocytes.</p

Immunostaining of osteoblast differentiation markers Runx2 and Osteocalcin at day 10.

<p>WT: upper panel, <i>Fdz9</i>^−/−: lower panel. Runx2 (<i>left</i>) was expressed mainly in the nucleus of preosteoblastic cells and osteoblasts (OB) located in the fracture callus. Osteocalcin (<i>right</i>) was mainly localized in the cytoplasma of osteoblasts near mineralized matrix and in the bone matrix. There were no differences in staining of Runx2 and Osteocalcin between both genotypes. C: cortex, OT: osteocytes.</p

Histological evaluation of relative amounts of tissues in the fracture calli at different time points post fracture.

<p>WT: white columns, <i>Fzd9</i>^−/−: grey columns. All values are presented as median, interquartile ranges, minimum and maximum. n = 5–10; Mann-Whitney-U-test, *p<0.05. TOT: total osseous tissue, Cg: cartilage, FT: fibrous tissue. A: day 10, WT: n = 6, <i>Fzd9</i>^−/−: n = 5, B: day 24, WT: n = 7, <i>Fzd9</i>^−/−: n = 8, C: day 32, WT: n = 10, <i>Fzd</i>9^−/−: n = 8.</p

µCT evaluation of the fracture calli of WT and <i>Fzd9</i>^{−<b>/</b>−} mice at days 24 and 32 post fracture.

<p>A: Bone volume/total volume (BV/TV). B: Bone mineral density (BMD) C: 3D-models of femurs at day 24 in anterior-posterior (left) and lateral view (right). The upper panel shows WT femurs and the lower panel <i>Fzd9</i>^−/− femurs. <i>Fzd9</i>^−/− mice exhibited a less mineralized callus at both investigated time points. At day 24, the geometry of the calli of <i>Fzd9</i>^−/− mice was different from WT mice, resulting in an increased moment of inertia (<i>I_x</i>) (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084232#pone-0084232-g007" target="_blank">Figure 7C</a>).</p