Influence of calcium oxide level and time of exposure to sugarcane on in vitro and in situ digestive kinetics

These experiments were carried out to evaluate, using in vitro and in situ techniques, the effects of three inclusion levels of calcium oxide (0, 5, and 10 g/kg of sugarcane fresh matter) and four exposure times (0, 24, 48, and 72 h) of sugarcane to calcium oxide on the chemical composition and digestive kinetic parameters of sugarcane. The treatments were arranged in a 3 by 4 factorial design. Freshly-cut sugarcane (whole plant) was treated with calcium oxide and separated into 12 piles inside a barn to prevent direct exposure to sunlight, rain, and wind. Every day, before and after animal feeding, the calcium oxide was proportionally hand-mixed with approximately 150 kg of freshly-cut sugarcane to make up the dietary treatments. The lowest (Ti) and greatest (Ts) temperature and pH of the treated sugarcane piles were measured immediately before and after sampling, respectively. The ether extract (EE) and DM were not affected (P>0.05) by either exposure time or inclusion level. However, CP increased linearly (P=0.01) and OM decreased linearly (P<0.0001) as the exposure time and calcium oxide inclusion level increased. Interactions between inclusion level and exposure time on DM, OM, CP, EE, Ti, and Ts were not observed. However, significant interactions were detected for non-fibre carbohydrate (NFC), neutral detergent fibre (aNDF), and pH. A quadratic effect of exposure time on the Ti and Ts was observed (P=0.001 and P=0.001, respectively). The maximum temperature was reached with approximately 51 h of exposure time. Calcium oxide positively affected the insoluble potentially digestible fraction of sugarcane DM and aNDF (P=0.001 and P=0.001, respectively), and the indigestible fraction of sugarcane aNDF (P=0.001). Interactions between inclusion level and exposure time on soluble and indigestible fractions of sugarcane DM (P=0.0001 and P=0.01, respectively) were found. However, no interactions (P>0.27) were found between inclusion level and exposure time on aNDF digestive kinetic parameters. The fractional digestion rate (kd) of sugarcane DM and aNDF was not influenced by treatments (P>0.05). The mean values of kd for sugarcane DM and aNDF were 0.0235 and 0.0215/h, respectively. The gas production kinetics parameters were not affected (P>0.05) by treatments. In conclusion, the inclusion of calcium oxide improved the in situ potentially digestible fraction of sugarcane DM and aNDF; however, it did not influence the fractional digestion rate. No effects were observed on the in vitro digestive kinetic parameters.     

Screening of some plants used in the Brazilian folk medicine for the treatment of infectious diseases

Extracts of 13 Brazilian medicinal plants were screened for their antimicrobial activity against bacteria and yeasts. Of these, 10 plant extracts showed varied levels of antibacterial activity. Piper regnellii presented a good activity against Staphylococus aureus and Bacillus subtilis, a moderate activity on Pseudomonas aeruginosa, and a weak activity against Escherichia coli. Punica granatum showed good activity on S. aureus and was inactive against the other standard strains. Eugenia uniflora presented moderate activity on both S. aureus and E. coli. Psidium guajava,Tanacetum vulgare, Arctium lappa, Mikania glomerata, Sambucus canadensis, Plantago major and Erythrina speciosa presented some degree of antibacterial activity. Spilanthes acmella, Lippia alba, and Achillea millefolium were considered inactive. Five of the plant extracts presented compounds with Rf values similar to the antibacterial compounds visible on bioautogram. Of these, three plants belong to the Asteraceae family. This may mean that the same compounds are responsible for the antibacterial activity in these plants. Anticandidal activity was detected in nine plant extracts (P. guajava, E. uniflora, P. granatum, A. lappa, T. vulgare, M. glomerata, L. alba, P. regnellii, and P. major). The results might explain the ethnobotanical use of the studied species for the treatment of various infectious diseases.     

A GPU-Based Implementation of the Firefly Algorithm for Variable Selection in Multivariate Calibration Problems

Several variable selection algorithms in multivariate calibration can be accelerated using Graphics Processing Units (GPU). Among these algorithms, the Firefly Algorithm (FA) is a recent proposed metaheuristic that may be used for variable selection. This paper presents a GPU-based FA (FA-MLR) with multiobjective formulation for variable selection in multivariate calibration problems and compares it with some traditional sequential algorithms in the literature. The advantage of the proposed implementation is demonstrated in an example involving a relatively large number of variables. The results showed that the FA-MLR, in comparison with the traditional algorithms is a more suitable choice and a relevant contribution for the variable selection problem. Additionally, the results also demonstrated that the FA-MLR performed in a GPU can be five times faster than its sequential implementation.     

Susceptibility of guava genotypes to natural infestation by Anastrepha spp. (Diptera: Tephritidae) in the municipality of Monte Alegre do Sul, state of São Paulo, Brazil

The infesting species and their infestation indices of fruit flies were determined for eleven guava genotypes (Psidium guajava L.). From March to April 2000, ten mature fruits of each genotype were harvested at weekly intervals from insecticide unsprayed trees located in the municipality of Monte Alegre do Sul, SP, Brazil. Fruits were brought to the laboratory, weighed and placed in individual plastic cups containing sand at the bottom to obtain the tephritid pupae. About 95% of guavas produced fruit fly puparia. Of the 682 Anastrepha females recovered, four species were identified: A. fraterculus (Wied.) (86.5%), A. obliqua (Macquart) (10.8%), A. bistrigata Bezzi (1.8%) and A. sororcula Zucchi (0.9%). Three species of parasitoids Braconidae (Opiinae) were recovered: Doryctobracon areolatus (Szépligeti), Doryctobracon brasiliensis (Szépligeti) and Utetes anastrephae (Viereck). The genotypes differ in level of infestation depend on the collecting time. The genotypes 'L2P4 Vermelha', 'Ruby Suppreme' and 'Webber Suppreme' showed the lowest susceptibility to tephritids in terms of puparia per fruit. The variability of infestation among the guava genotypes and the reasons for increasing fruit fly infestations along the time were discussed.     

Transformation of potato (Solanum tuberosum) cv. Mantiqueira using Agrobacterium tumefaciens and evaluation of herbicide resistance

Summary In search of establishing a system for genetic transformation of Brazilian potato cultivars, Agrobacterium tumefaciens carrying the plasmid pGV1040, was used to transform leaf discs of three cultivars of local importance, i.e., Aracy, Baronesa and Mantiqueira. This plasmid contains marker genes for resistance to kanamycin and phosphinothricin plus the gene for the enzyme -glucuronidase. A two step regeneration/selection procedure produced shoots of potato cultivar Mantiqueira with in vitro resistance to kanamycin and to phosphinothricin. After transfer to the greenhouse, the potentially transgenic plants, sprayed with the herbicide Finale^® (20% a.i.; Hoechst^®) remained green as compared to control clones that died immediately afterwards. Southern blot analysis and histochemical and fluorimetric assay for -glucuronidase indicated that the gene coding for the enzyme was integrated in the potato genome and could be expressed in potato tissues. No success was obtained for transformation of cultivars Aracy and Baronesa using this procedure.Abbreviations NAA Naphthalene Acetic Acid - BAP Benzyl-aminopurine - GA₃ Gibberellic Acid - PPT Phosphinothricin - PAT Phosphinothricin Acetyl Transferase     

Third molar agenesis in a trihybrid Brazilian population

The frequency of missing third molars is about eight per cent per quadrant in a sample of 490 males of the mixed White/Negro/Indian population of Natal, Brazil. Absence of the four third molars appears in two per cent of the individuals of the sample. The degree of asymmetry for the trait was not marked. No differences were observed when the persons were separated on morphological grounds as Whites or Negroids suggesting that these segregants may be genetically homogeneous in relation to this and other characteristics.     

Strategies for ex situ conservation of Centaurea cineraria subsp. circae (Asteraceae), an endemic plant from Lazio (Italy)

Centaurea cineraria subsp. circae is an endemic plant with a distribution area limited to Circeo mountain (Lazio, Italy), whose population was estimated in a very low number of individuals. The aim of this work was to investigate ex situ conservation strategies such as achene collection and in vitro plant propagation, which will permit to carry out restoration programmes. The test carried out on the achenes demonstrated that only 5.5% of them were morphologically healthy. Seed germination tests showed that seeds do not display dormancy and that germination does not require pre-treatments. The higher germination rate (67.5%) was observed under a photoperiod of 12/12 h (light/dark) and temperature regime +20/+10°C. The in vitro studies demonstrated that micropropagation, acclimatization and the transfer outdoors of C. cineraria subsp. circae are not particularly difficult: 74% of shoot explants in a Murashige and Skoog (MS) medium added with 0.5 mg/l benzylaminopurine and 2 mg/l kinetin formed multiple shoots; 100% of shoots rooted in the MS medium added with 0.5 mg/l indole-3-butyric acid and over 90% survived the acclimatization phase. After been transferred outdoors, the totality of in vitro-propagated plants bloomed and appeared morphologically indistinguishable from wild plants. Preliminary chemical analyses showed a similar profile for in vitro-propagated and wild plants.     

Removal of proteases from <Emphasis Type="Italic">Clostridium perfringens</Emphasis> fermented broth by aqueous two-phase systems (PEG/citrate)

In order to reduce the toxicity of Clostridium perfringens fermentation broths used in vaccine preparation, we developed two-phase aqueous systems for removal of toxin-activating proteases. Removal of the proteases inhibits the conversion of protoxin to active toxin. In order to establish the conditions under which the phase separation occurs, binodal curves, formed by poly(ethylene glycol) (PEG) and sodium citrate, were investigated at different values of pH and PEG molar mass. A 2⁴-experimental design was used to evaluate the influence of PEG molar mass and concentration, citrate concentration and pH on protease partition coefficient, removal factor and protease removal yield. It has been found that simultaneous increase in PEG molar mass and decrease in citrate concentration remarkably improved the removal factor, whereas the protease removal yield showed an opposite trend. The best conditions for the system under consideration (removal factor of 2.69 and yield of 116%) were obtained at pH 8.0 using PEG molar mass of 8000 g mol⁻¹ and concentrations of PEG and citrate of 24 and 15%, respectively.     

Retinol and retinoic acid increase MMP-2 activity by different pathways in cultured Sertoli cells

Diseases such as atherosclerosis, arthritis and cancer have been related with imbalance in ROS production and failures in regulation of the MMPs. Authors suggested a relationship between MPP activity and ROS. Our research group has demonstrated that retinol 7µM induced changes in Sertoli cell metabolism linking retinol treatment and oxidative stress. We verified MMP activity in Sertoli cells treated with vitamin A using gelatin zymography. We found that retinol (7µM) and retinoic acid (1nM) induced MMP-2 activity in Sertoli cells. Antioxidants reversed retinol-induced but not retinoic acid-induced MMP-2 activity. Moreover, retinol but not retinoic acid increased ROS production quantified by DCFH-DA oxidation. We found that retinol and retinoic acid induced ERK1/2 phosphorylation, but only retinol-increased MMP-2 activity was inhibited by UO126, an ERK1/2 phosphorylation inhibitor. Our findings suggested that retinol-induced MMP-2 activity, but not retinoic acid-induced MMP-2 activity, was related to ERK1/2 phosphorylation and ROS production.