Digestive enzyme activity in juvenile Nile tilapia (Oreochromis niloticus, L) submitted to different dietary levels of shrimp protein hydrolysate

The effect of different dietary concentrations of shrimp protein hydrolysate (SPH) on digestive enzyme activity of Nile tilapia juveniles was evaluated. SPH concentrations in diets were 0, 15, 30 and 60 g kg^?1 (treatments SPH0, SPH15, SPH30 and SPH60, respectively). Hemoglobin, azocasein, BApNA (Nα-benzoyl-dl-arginine-p-nitroanilide), SApNA (Suc-Ala-Ala-Pro-Phe p-nitroanilide), aminoacyl of β-naphthylamide and starch were used as substrates for enzyme activity determinations. The activity of total alkaline protease was significantly higher (P < 0.05) in fish under SPH15 and SPH60 treatments than in the control (SPH0). However, the effect was not dose-dependent. Substrate-SDS-PAGE was also performed to evaluate changes in the profile of Nile tilapia digestive proteases caused by SPH. Substrate-SDS-PAGE revealed 12 active proteolytic bands, eight of which responded to SPH dietary incorporation. Inhibition substrate-SDS-PAGE indicated a decrease in the activity of three enzymes, with trypsin activity decreasing with the increase of SPH concentration, whereas the opposite occurred for two aminopeptidases. Distinct protease profiles were also found for each treatment, suggesting adaptability of digestive proteases from Nile tilapia to the different diets.     

Genetic progress during 22 years of black bean improvement

The common bean breeding program coordinated by Embrapa Arroz e Feijão released 50 new cultivars from 1984 to 2010. The aim of the present study was to estimate genetic progress in terms of seed yield and other agronomic traits achieved by the black-type common bean breeding program in Brazil over the 22-year period from 1985 to 2006. Two types of field experiments were performed to assess genetic progress. In regard to experiments on lines, the three best breeding lines from each of 11 evaluation cycles constituted 33 treatments. These evaluations were made in 20 environments and served to estimate progress in seed yield and seed size because these were the criteria applied in selecting the elite lines. Cultivar evaluations included nine cultivars recommended by Embrapa during the period under investigation, and the evaluations were established in 21 environments. These evaluations served to estimate improvements in reaction to angular leaf spot, plant architecture and tolerance to lodging because cultivars usually exhibit superior phenotypes for such characteristics. Evaluations were performed from 2008 to 2010. Estimated improvement in seed yield was 25.2 kg ha^?1 or 1.1 % per year. Improvements were also found in tolerance to lodging (1.7 % per year) and 100-seed weight (0.65 % per year). There was no significant progress for the plant architecture and resistance to angular leaf spot traits.     

Chemical Diversity and Biological Properties of Secondary Metabolites from Sea Hares of Aplysia Genus

The marine environment is an important source of structurally-diverse and biologically-active secondary metabolites. During the last two decades, thousands of compounds were discovered in marine organisms, several of them having inspired the development of new classes of therapeutic agents. Marine mollusks constitute a successful phyla in the discovery of new marine natural products (MNPs). Over a 50-year period from 1963, 116 genera of mollusks contributed innumerous compounds, Aplysia being the most studied genus by MNP chemists. This genus includes 36 valid species and should be distinguished from all mollusks as it yielded numerous new natural products. Aplysia sea hares are herbivorous mollusks, which have been proven to be a rich source of secondary metabolites, mostly of dietary origin. The majority of secondary metabolites isolated from sea hares of the genus Aplysia are halogenated terpenes; however, these animals are also a source of compounds from other chemical classes, such as macrolides, sterols and alkaloids, often exhibiting cytotoxic, antibacterial, antifungal, antiviral and/or antifeedant activities. This review focuses on the diverse structural classes of secondary metabolites found in Aplysia spp., including several compounds with pronounced biological properties.     

The effects of <Emphasis Type="Italic">Gibberella zeae</Emphasis>, <Emphasis Type="Italic">Barley Yellow Dwarf Virus</Emphasis>, and co-infection on <Emphasis Type="Italic">Rhopalosiphum padi</Emphasis> olfactory preference and performance

Insect-borne viruses promote several changes in plant phenotype, which can modify plant-vector interactions in favor of virus survival and dissemination. Although co-infections commonly occur in the field, little is known about their effects on interactions with the vector. The ecological interactions between Barley Yellow Dwarf Virus (BYDV) and its aphid vector, Rhopalosiphum padi, have been investigated extensively, but the vector’s behavior in more complex scenarios has yet to be examined. We assessed olfactory response and performance of R. padi to wheat singly and doubly infected by the pathogenic fungus Giberella zeae and BYDV. Non-viruliferous aphids preferred odors of BYDV-infected wheat over healthy wheat, as previously reported in the literature, and they were still preferentially attracted to BYDV-infected plant during co-infection. However, around 35% more non-viruliferous aphids chose healthy wheat over G. zeae-infected wheat. Viruliferous aphids did not show any preference to the treatments. BYDV-infected wheat was a superior host than healthy wheat for the aphids whose population increased in 25%. We observed a synergistic effect of the co-infected wheat, which was the best host for aphids, and promoted an elevation of 42% on population growth. Our results indicate that co-infection might be beneficial for virus spread as does not interfere with aphid olfactory preference and provides greater colony growth than in singly infected plants.     

Immunocytochemical study of Ki-67 as a prognostic marker in canine mammary neoplasia

Background: Canine mammary tumors are challenging for clinicians and pathologists because of complex histologic classification, low specificity of cytologic diagnosis, and unpredictable biological behavior. In histologic specimens, expression of tumor proliferation marker Ki‐67, a nuclear nonhistone protein, has been shown to have prognostic value for canine mammary tumors and to correlate with malignancy and low survival rates. Objective: The objective of this study was to measure the proliferation index of canine mammary tumors by immunochemical detection of Ki‐67 in cytologic specimens and to determine its relationship to clinical and pathologic variables and patient outcome. Methods: Spontaneous mammary tumors from 31 female dogs were surgically excised. Imprint specimens for cytologic evaluation were wet‐fixed in ethanol; histologic specimens were prepared routinely. Immunostaining was performed with the PH 177 monoclonal antibody against Ki‐67; proliferation index was graded from negative to +++. Dogs were followed for 18 months. Multivariate logistic regression analysis was used to determine correlations between immunocytochemical results, tumor and clinical variables, and patient outcome. Results: Ki‐67 proliferation indices in cytologic specimens were significantly lower for nonmalignant tumors than for malignant tumors. High index values of Ki‐67 were positively correlated with metastasis, death from neoplasia, low disease‐free survival rates, and low overall survival rate. With the exception of 4 specimens for which cellularity was insufficient, positive expression of Ki‐67 in cytologic specimens correlated with that of histologic specimens. Conclusions: The prognostic value of the Ki‐67 index in canine mammary tumors by using wet‐fixed cytology imprint specimens was similar to that observed previously for histologic specimens. Immunocytochemical detection of Ki‐67 could improve the accuracy and value of cytology by providing safe and rapid information about malignancy and patient outcome.     

Feline plasma cell pododermatitis: a study of 8 cases

Eight cases of feline plasma cell pododermatitis, collected over a 3-year period, were studied using histological (haematoxylin and eosin), histochemical (methyl green-pyronin) and immunohistochemical (antibody against lambda light chains of immunoglobulins) techniques. No sex, breed or age predisposition, or paw predilection was observed. The disease began as a swelling of the footpads, followed by ulceration. Histologically, lesions were characterized by the presence of a large number of plasma cells in a predominantly perivascular pattern. Binucleated plasma cells and mitotic figures were observed. Clinical follow-up for over 1 year revealed total remission of the lesions both after glucocorticoid therapy in four cases, and total surgical excision in two animals. Follow-up studies were not possible in one case.     

Tumor slices as a model to evaluate doxorubicin in vitro treatment and expression of trios of genes PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2 in canine mammary gland cancer

Background

In women with breast cancer submitted to neoadjuvant chemotherapy based in doxorubicin, tumor expression of groups of three genes (PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2) have classified them as responsive or resistant. We have investigated whether expression of these trios of genes could predict mammary carcinoma response in dogs and whether tumor slices, which maintain epithelial-mesenchymal interactions, could be used to evaluate drug response in vitro.

Methods

Tumors from 38 dogs were sliced and cultured with or without doxorubicin 1 μM for 24 h. Tumor cells were counted by two observers to establish a percentage variation in cell number, between slices. Based on these results, a reduction in cell number between treated and control samples ≥ 21.7%, arbitrarily classified samples, as drug responsive. Tumor expression of PRSS11, MTSS1, CLPTM1 and SMYD2, was evaluated by real time PCR. Relative expression results were then transformed to their natural logarithm values, which were spatially disposed according to the expression of trios of genes, comprising PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2. Fisher linear discrimination test was used to generate a separation plane between responsive and non-responsive tumors.

Results

Culture of tumor slices for 24 h was feasible. Nine samples were considered responsive and 29 non-responsive to doxorubicin, considering the pre-established cut-off value of cell number reduction ≥ 21.7%, between doxorubicin treated and control samples. Relative gene expression was evaluated and tumor samples were then spatially distributed according to the expression of the trios of genes: PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2. A separation plane was generated. However, no clear separation between responsive and non-responsive samples could be observed.

Conclusion

Three-dimensional distribution of samples according to the expression of the trios of genes PRSS11, MTSS1, CLPTM1 and PRSS11, MTSS1, SMYD2 could not predict doxorubicin in vitro responsiveness. Short term culture of mammary gland cancer slices may be an interesting model to evaluate chemotherapy activity.     

Composition and structure of organosolv lignins from four eucalypt species

Lignins were extracted from different species using organosolv process (ethanol/water). Obtained organosolv lignins were characterised by various methods to determine their composition, structure and functional groups with the purpose of evaluating their potential use for obtaining value-added compounds. The purity of organosolv lignins was determined. The total phenols content and the antioxidant power were analysed in order to know the reactivity of lignins. The ratio S/G was determined by nitrobenzene oxidation. In addition, molecular weight distribution, infrared spectroscopy and thermo gravimetrical analysis were carried out in order to determine the physical and chemical properties of organosolv lignins. Obtained organosolv lignins have very high purity and low sugar and inorganic contamination. All organosolv lignins samples have high polydispersity, and lignin from grey ironbark wood had the highest molecular weight average. Among the organosolv lignins, lemon-scented gum showed the highest average value of S/G ratio and the lowest average value of total phenols.