Recovery of bioactive compounds from Pinus pinaster wood by consecutive extraction stages

Pinus pinaster wood samples, obtained at different positions of three healthy trees, were subjected to two sequential extractions using an Accelerated Solvent Extraction instrument. The first extraction was carried out with hexane (to remove lipophilic extractives) and the second one with acetone/water (95:5 v/v) to recover bioactive phenolic compounds, the target compounds of this study. The extracted fractions were assayed for total yield and composition. The extracts contained a spectrum of phenolic compounds (simple phenolics, phenolic stilbenes, flavonoids and lignans) and non-phenolic components (juvabiones, resin and fatty acids, steryl esters and triglycerides). The fractionation effects achieved by consecutive extractions and the recovery of bioactive phenolics are discussed.     

Analysis of IgG response to experimental infection with RH Toxoplasma gondii in goats.

The IgG response of goats experimentally infected with RH Toxoplasma gondii has been analysed using an indirect ELISA and Western-blot analysis. Specific IgG antibodies were first detected at 14 days post-inoculation (p.i.), reaching a peak by day 35 p.i. and showing slight fluctuations until the end of the experiment (91 p.i.). Specific IgG showed a reactivity over a whole range of peptides (125-24 kDa approximately), but the highest reactivity was observed against a group of antigens with a molecular weight between 34 and 28 kDa, in particular against a 30 kDa fraction which is considered to represent the major surface protein of T. gondii named p30 or SAG-1.     

Immunocytochemical Study of the Ultimobranchial Tubule in Wistar Rats

A systematic immunohistochemical study of the ultimobranchial tubule (UBT) has been carried out in 45 Wistar rats of different ages (0, 5, 10, 15, 20, 25, 30, 60 and 120 days). The existence of calcitonin immunoreactive cells in the UBT wall has been demonstrated in a 5-days old rat. In addition, immunohistochemical studies for thyroglobulin revealed positive staining in follicular cells connected to the UBT and, occasionally, in isolated cells lying within solid clusters from the UBT. These last results together with the continued and repeated existence of numerous mitosis and PAS (+) microfollicles, apparently rising from the UBT, support the hypothesis that the ultimobranchial body (UBB) may contribute partially to the formation of a part of the follicular component.     

Species diversity: Concept,measurement, and response to clearcutting and site-preparation

A review of aspects of diversity recently introduced in the statistical literature by Patil and Tallie is given: species diversity as an average property of a community is defined; the property averaged is identified as species rarity; and relations between common indices of diversity and associated measures of rarity are illuminated. This paper also computes common indices of diversity from substantial empirical data collected before clearcutting and site-preparation under two management regimes and for 3 years following planting of slash pine (Pinus elliottii) to illustrate that the initial response to these forest operations is increased species diversity, i.e., increased species rarity. This response may be expected whenever clearcutting and site preparation reduces but does not eliminate previously abundant, late-successional species and permits the reappearance of early-successional species, including forbs and grasses.     

Vaccination of one-day-old broiler chicks against infectious coryza

In order to evaluate the protection conferred by an experimental inactivated vaccine against infectious coryza, three challenge trials were undertaken using 112 1-day-old broilers. The vaccine "Hepa Inmuno NC" included bacterial antigens of Avibacterium paragallinarum (serogroups A, B, variant B, and C) as well as antigens of Newcastle virus and hepatitis virus. Fifty-six broiler chicks were vaccinated at the first day of life at the hatchery while another 56 chicks were left unvaccinated. Three infection trials were conducted simultaneously using each of the three serogroups A, B, or C of Av. paragallinarum. In each trial, 17 vaccinated and 17 unvaccinated broilers were used. Challenge was performed at day 31 of life by injection, into the left infraorbital sinus, of approximately 1 x 10(5) colony forming units of the corresponding Av. paragallinarum strain. Clinical signs were recorded on day 2 postchallenge. All broilers were euthanatized and both infraorbital sinuses were bacteriologically examined for the presence of Av. paragallinarum on day 5 postchallenge. In comparison with the unvaccinated broilers, the vaccine significantly reduced the number of broilers with clinical signs after challenge with serogroup B, and significantly fewer vaccinated broilers were positive for the presence of Av. paragallinarum after challenge with serogroup C. On the other hand, no significant protection was observed when broilers were challenged with Av. paragallinarum from serogroup A. Despite the high infection rates in vaccinated chicks after an experimental infection with Av. paragallinarum, it was possible to reduce colonization of Av. paragallinarum (serogroup B) and clinical signs (serogroup C) in broiler chicks by vaccination at the first day of life. Further cross-protection trials should be done, including other Av. paragallinarum strains in the vaccine, especially those from serogroup A.     

Squamous cell carcinoma of the vagina and cervix in sheep - case report

This report describes the clinical and histopathological characteristics of a squamous cell carcinoma infiltrating the cervix and the vaginal wall, producing reproductive symptoms and subnormal fertility in an adult ewe. Necropsy showed a large (15-cm-long) neoplastic mass infiltrating the vaginal wall and the cervix. Histopathological examination revealed atypical squamous epithelial cords invading the basal membrane and dermis, round anaplastic cells, focal areas of necrosis, keratinisation of isolated cells, and pronounced infiltration by mononuclear cells around the cords. No squamous cell carcinoma of such localisation has been reported from sheep before. In humans, this tumour is the most common gynaecological malignancy in the world.     

Waxes composition of reproduction cork from Quercus suber and its variability throughout the industrial processing

Summary The chemical composition of waxes was studied in cork planks from three different trees of Spanish Quercus suber at four different stages of the in‐dustrial processing of the first transformation: stripping, first rest, boiling fol‐lowed by open air rest and boiling followed by store-room rest. Waxes were extracted with chloroform (CHCl₃) and the extract was submitted to saponification in order to obtain the neutral and acid fractions. The trimethylsilyl derivatives of both fractions were analysed by GC-MS. An important decrease of the total content of waxes -CHCl₃ extract-(average values: 10.13 to 4.91%) and of the acid fraction (average values: 2.51 to 0.96%) was observed throughout the industrial processing. Some differences were also found among trees, two of them being richer in total waxes (average, 8.14 and 7.71%) than the other one (average, 4.67%). Concerning the three groups of identified components (fatty acids, fatty alcohols and triterpenes), the contents of the fatty alcohols and acids suffer an important reduction during the first rest (average values: 30.39 to 17.53% and 58.73 to 31.48%, respectively), while after boiling, there is a decrease of the triterpenes concentrations (79.16 to 58.00% -neutral fraction- and 40.23 to 23.20% -acid fraction-), being greater when the second rest is carried out in a store room. Significant differences among trees were only found in the fatty alcohols contents. The neutral fraction was mainly composed by fatty alcohols (all the even members from C₁₈ to C₂₆, with traces of intermediate odd members and some unsaturated groups, C₂₀ and C₂₁) and triterpenes (14 components, among them friedelin, betulin and cerin were identified), and a very small amount of monocarboxylic fatty acids (C₁₆, C₁₈, C₂₂ and C₂₄ members) was also present. The main group of components of the acid fraction was that of fatty acids (saturated even C₁₂–C₂₄ and odd C₁₅, C₁₇, C₂₁ members, accompanied by a great amount of unsaturated terms and some ω-hydroxyacids, 18-hydroxy-9,12-octadecadienoic and 18-hydroxy-9-octadecenoic acids). Four triterpenes, the C₂₀, C₂₄ and C₂₈ alcohols and ferulic acid were also detected in the acid fraction. Some of these individual components of each fraction were also affected by the industrial processing and presented significant differences among the studied trees. Received 12 December 1997     

Staling of Bread: Role of Amylose and Amylopectin and Influence of Starch‐Degrading Enzymes

The present investigation aims at understanding the mechanism of bread firming during staling. Changes in the starch fraction due to the addition of amylases and their influence on the texture of bread crumb were studied during aging and after rebaking of stale bread. Pan bread was prepared by a conventional baking procedure. The influence of three different starch‐degrading enzymes, a conventional α‐amylase, a maltogenic α‐amylase, and a β‐amylase were investigated. The mechanical properties of bread were followed by uniaxial compression measurements. The microstructure was investigated by light microscopy, and starch transformations were assessed by differential scanning calorimetry (DSC) and wide‐angle X‐ray powder diffraction. Firming of bread crumb and crystallization of starch are not necessarily in agreement in systems with added amylases. Reorganization of both starch fractions, amylopectin and amylose, and the increase of starch network rigidity due to increase of polymer order are important during aging. Starch‐degrading enzymes act by decreasing the structural strength of the starch phase; for instance, by preventing the recrystallization of amylopectin or by reducing the connectivity between crystalline starch phases. On the other hand, starch‐degrading enzymes may also promote the formation of a partly crystalline amylose network and, by this, contribute to a kinetic stabilization of the starch network. Based on the results, a model for bread staling is proposed, taking into account the biphasic nature of starch and the changes in both the amylose and amylopectin fraction.     

Nitrate reductase and nitrite reductase as targets to study gene silencing phenomena in transgenic plants

Nitrate assimilation is a fundamental function in any plant including those that can fix atmospheric nitrogen in symbiosis with soil bacteria. In recent years, attempts have been made to understand the biological significance of the complex regulation of this pathway using genetic engineering techniques. Transgenic plants that either over- or under-express genes of the nitrate assimilation pathway were created in order to determine whether such directed changes affect the regulation of the metabolism. Apart from interesting physiological results, unexpected gene silencing phenomena have been observed resulting from the introduction of five different transgenes derived from either the tobacco Nia or Nii genes encoding nitrate reductase and nitrite reductase, respectively. In this review, each of these five silencing phenomena is described, with the emphasis on the advantages provided by the use of both Nia and Nii genes to analyze the molecular and genetic basis of gene silencing in transgenic plants.     

Dietary protein/carbohydrate ratio in low‐lipid diets for Senegalese sole (Solea senegalensis,Kaup 1858) juveniles. Influence on growth performance,nutrient utilization and flesh quality

Four isoenergetic (21 kJ/g dry matter, DM) and isolipidic (65 g/kg DM) diets containing different crude protein/total carbohydrate (CHO) ratios: 60/26, 56/30, 52/34 and 48/38, were tested in 22 g Senegalese sole for 104 days. Apparent digestibility coefficients (ADC) were not affected by the treatments, and all groups presented extremely low starch ADC values (22.8%–36.5%). Replacement of dietary protein by CHO did not affect daily growth index (0.9–1.0), but significantly increased voluntary feed intake of fish. Regression analyses demonstrated that digestible protein content, rather than digestible energy, was the main dietary factor influencing such feeding activity (R² = .952). A significantly increased feed conversion ratio was observed in sole fed increasing CHO contents. The dietary protein/CHO ratio did not influence whole‐body composition. Sole fed the 48/38 diet showed the lowest efficiency in terms of N and energy utilization. PUFA were the most represented fatty acid fraction in fillet, regardless of the dietary protein/CHO ratio, mainly due to the high content of DHA. Senegalese sole increase feed intake under low dietary protein/CHO ratios to ensure an adequate N intake. Such compensatory mechanism seems to be triggered to satisfy a specific protein metabolic requirement for energy purposes as tissue accretion remained unchanged.