Search for B-S(0) -> mu(+)mu(-) and B-0 -> mu(+)mu(-) decays

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Production of clavulanic acid and cephamycin C by Streptomyces clavuligerus under different fed-batch conditions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação do capim-elefante (Pennisetum purpureum Schum.) cultivar taiwan A-148, ensilado com diferentes técnicas de redução de umidade

O objetivo deste trabalho foi avaliar o efeito de diferentes técnicas de redução de umidade sobre a composição química, digestibilidade in vitro da matéria seca (DIVMS) e degradação ruminal da matéria seca da silagem de capim-elefante (Pennisetum purpureum Schum.) cv. Taiwan A-148. Os tratamentos (A, Controle; B, 20,0; C, 30,0; D, 40,0% de sabugo de milho; E, emurchecimento por 12 horas; F, emurchecimento por 24 horas; e G, esmagamento + emurchecimento por 24 horas) foram distribuídos em delineamento inteiramente casualizado com três repetições. Os tratamentos da silagem B e E não foram eficientes em reduzir a umidade excessiva da forragem. O emurchecimento por 24 horas aumentou o teor de MS, sem influir no teor de PB e na DIVMS. Os tratamentos C e D favoreceram o desenvolvimento da população de clostrídeos, aumentaram a concentração de N-NH3 e reduziram a concentração de ácido lático e a DIVMS das silagens. O tratamento G aumentou o teor de MS e reduziu a concentração de N-NH3 e DIVMS. A adiçao de sabugo de milho (tratamentos B, C e D) reduziu a degradação ruminal da MS das silagens. O sabugo de milho reduziu a umidade, mas apresentou efeitos negativos na qualidade da silagem, enquanto o esmagamento e/ou emurchecimento foram procedimentos que mostraram bons resultados em relação à conservação do material ensilado.

Responses of chlorophyll fluorescence parameters of the facultative halophyte and C-3-CAM intermediate species Mesembryanthemum crystallinum to salinity and high irradiance stress

Mesembryanthemum crystallinum L. (Aizoaceae) is a facultative annual halophyte and a C-3-photosynthesis/crassulacean acid metabolism intermediate species currently used as a model plant in stress physiology. Both salinity and high light irradiance stress are known to induce CAM in this species. The present study was performed to provide a diagnosis of alterations at the photosystem 11 level during salinity and irradiance stress. Plants were subjected for up to 13 days to either 0.4M NaCl salinity or high irradiance of 1000 mu mol m(-2) s(-1), as well as to both stress factors combined (LLSA = low light plus salt; HLCO = high light of 1000 mu mol m(-2)s(-1), no salt; HLSA = high light plus salt). A control of LLCO = low light of 200 mu mol m(-2) s(-1), no salt was used. Parameters of chlorophyll a fluorescence of photosystem 11 (PSII) were measured with a pulse amplitude modulated fluorometer. HLCO and LLSA conditions induced a weak degree of CAM with day/night changes of malate levels (Delta malate) of similar to 12 mM in the course of the experiment, while HLSA induced stronger CAM of Delta malate similar to 20mM. Effective quantum yield of PSII, Delta F/F'(m), was only slightly affected by LLSA, somewhat reduced during the course of the experiment by HLCO and clearly reduced by HLSA. Potential quantum efficiency of PSII, F-v/F-m, at predawn times was not affected by any of the conditions, always remaining at >= 0.8, showing that there was no acute photoinhibition. During the course of the days HL alone (HLCO) also did not elicit photoinhibition; salt alone (LLSA) caused acute photoinhibition which was amplified by the combination of the two stresses (HLSA). Non-photochemical, NPQ, quenching remained low (< 0.5) under LLCO, LLSA and HLCO and increased during the course of the experiment under HLSA to 1-2. Maximum apparent photosynthetic electron transport rates, ETRmax, declined during the daily courses and were reduced by LLSA and to a similar extent by HLSA. It is concluded that A crystallinum expresses effective stress tolerance mechanisms but photosynthetic capacity is reduced by the synergistic effects of salinity and tight irradiance stress combined. (c) 2006 Elsevier GmbH. All rights reserved.

DUCTAL CARCINOMA IN-SITU OF THE BREAST - HISTOLOGIC CATEGORIZATION AND ITS RELATIONSHIP TO PLOIDY AND IMMUNOHISTOCHEMICAL EXPRESSION OF HORMONE RECEPTORS, P53, AND C-ERBB-2 PROTEIN

Background. Ductal carcinoma in situ (DCIS) of the breast has been diagnosed increasingly since the advent of mammography. However, the natural history of these lesions remains uncertain. Ductal carcinoma in situ of the breast does not represent a single entity but a heterogeneous group with histologic and clinical differences. The histologic subtype of DCIS seems to have an influence on its biologic behavior, but there are few studies correlating subtype with biologic markers.Methods. The authors studied a consecutive series of 40 cases of DCIS and after its histologic categorization verified its relationship with ploidy using image analysis and analyzing estrogen receptor (ER), progesterone receptor (PR), p53 and c-erbB-2 expression using immunohistochemistry.Results. The three groups proposed according to the grade of malignancy were correlated significantly with some of the additional parameters studied, including aneuploidy and c-erB-2 expression. Aneuploidy was detected in 77.5% of cases of DCIS mainly in high and intermediate grade subtypes (100% and 80% vs. 35.7% in low grade) whereas immunoreactivity for c-erbB-2 was detected in 45% of cases of DCIS mainly in the high grade group. Expression of ER and PR were observed frequently in this study (63.9% and 65.7% respectively), but without correlation with the histologic subtype of DCIS, although we found a somewhat significant association between high grade DCIS and lack of ER. p53 protein expression was detected in 36.8% of these cases, but no relationship between this expression and histologic subtype or grading of DCIS was found.Conclusions. These results provide further evidence for the morphologic and biologic heterogeneity of DCIS. Besides histologic classification and nuclear grading, some biologic markers such as aneuploidy and c-erbB-2 expression constitute additional criteria of high grade of malignancy.

Structure, genomic DNA typing, and kinetic characterization of the D allozyme of placental alkaline phosphatase (PLAP/ALPP)

The D allozyme of placental alkaline phosphatase (PLAP) displays enzymatic properties at variance with those of the common PLAP allozymes. We have deduced the amino acid sequence of the PLAP D allele by PCR cloning of its gene, ALPP Two coding substitutions were found in comparison With the cDNA of the common PLAP F allele, i.e., 692C>G and 1352A>G, which translate into a P209R and E429G substitution. A single nucleotide primer extension (SNuPE) assay was developed using PCR primers that enable the amplification of a 1.9 kb PLAP fragment. Extension primers were then used on this PCR fragment to detect the 692C>G and 1352A>G substitution. The SNuPE assay on these two nucleotide substitutions enabled us to distinguish the PLAP F and D alleles from the PLAP S/I alleles. Functional studies on the D allozyme were made possible by constructing and expressing a PLAP D cDNA, i.e., [Arg209, Gly429] PLAP, into wildtype Chinese hamster ovary cells. We determined the k(cat) and K-m, of the PLAP S, F. and D allozymes using the non,physiological substrate p-nitrophenylphosphate at an optimal pH (9.8) as well as two physiological substrates, i.e., pyridoxal-5'-phosphate and inorganic pyrophosphate at physiological pH (7.5). We found that the biochemical properties of the D allozyme of PLAP are significantly different from those of the common PLAP allozymes. These biochemical findings suggest that a suboptimal enzymatic function by the PLAP D allozyme may be the basis for the apparent negative selective pressure of the PLAP D allele. The development of the SNuPE assay will enable us to test the hypothesis that the PLAP D allele is subjected to intrauterine selection by examining genomic DNA from statistically informative population samples. Hum Mutat 19:258-267, 2002. (C) 2002 Wiley-Liss, Inc.

Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity chromatography

Cathepsin D, a lysosomal aspartic protease, has been purified from porcine liver using a combination of pepstatin-A agarose and Affi-Gel Blue affinity chromatography, followed by size-exclusion chromatography. The purified protein consists of two polypeptide chains of 15 and 30 kDa, and has an isoelectric point of 6.8. Porcine liver cathepsin D has maximum activity at pH 2.5-3.0 as determined by its activity against hemoglobin, with a K-cat of 14.3 s(-1) and a k(cat)/K-M of 2.70 x 10(6) s(-1) M-1 as determined by the hydrolysis of a fluorogenic peptide substrate.

Water regulation of actinomycin-D binding to DNA: the interplay among drug affinity, DNA long-range conformation, and hydration

Actiaomycin-D (actD) binds to natural DNA at two different classes of binding sites, weak and strong. The affinity for these sites is highly dependent on DNA se(sequence and solution conditions, and the interaction appears to be purely entropic driven Although the entropic character of this reaction has been attributed to the release of water molecules upon drug to DNA complex formation, the mechanism by which hydration regulates actD binding and discrimination between different classes of binding sites on natural DNA is still unknown. In this work, we investigate the role of hydration on this reaction using the osmotic stress method. We skew that the decrease of solution water activity, due to the addition of sucrose, glycerol ethylene glycol, and betaine, favors drug binding to the strong binding sites on DNA by increasing both the apparent binding affinity Delta G, and the number of DNA base pairs apparently occupied by the bound drug n(bp/actD). These binding parameters vary linearly with the logarithm of the molar fraction of water in solution log(X-w), which indicates the contribution of water binding to the energetic of the reaction. It is demonstrated that the hydration change measured upon binding increases proportionally to the apparent size of the binding site n(bp/uctD). This indicates that n(bp/actD) measured from the Scatchard plod is a measure of the size of the DNA molecule changing conformation due to ligand binding. We also find that the contribution of DNA deformation, gauged by n(bp/act) to the total free energy of binding Delta G, is given by Delta G = Delta G(local) + n(bp/actD) x delta G(DNA), where Delta G(local), = -8020 +/- 51 cal/mol of actD bound and delta G(DNa) = -24.1 +/- 1.7cal/mol of base pair at 25 degrees C. We interpret Delta G(local), as the energetic contribution due to the direct interactions of actD with the actual tetranucleotide binding site, and it n(bp/actB) X delta G(DNA) as that due to change inconformation, induced by binding, of it n(bp/actD) DNA base pairs flanking the local site. This interpretation is supported by the agreement found between the value of delta G(DNA) and the torsional free energy change measured independently. We conclude suggesting an allosteric model for ligand binding to DNA, such that the increase in binding affinity is achieved by increasing the relaxation of the unfavorable free energy of binding storage at the local site through a larger number of DNA base pairs. The new aspect on this model is that the size of the complex is not fixed but determined by solutions conditions, such as water activity, which modulate the energetic barrier to change helix conformation. These results may suggest that long-range allosteric transitions of duplex DNA are involved in the inhibition of RNA synthesis by actD, and more generally, in the regulation of transcription. (C) 2000 John Wiley & Sons, Inc.

Changes in physiological and stroking parameters during interval swims at the slope of the d-t relationship

The slope of the distance-time relationship from maximal 200 and 400 in bouts (S(200-400)) has been increasingly employed for setting training intensities in swimming. However, physiological and mechanical responses at this speed are poorly understood. Thus, this study investigated blood lactate, heart rate (HR), stroke rate (SR), stroke length (SL) and RPE responses to an interval swimming set at S(200-400) in trained swimmers. In a 50-m pool, twelve athletes (16.5 +/- 1.2 yr, 176 +/- 7 cm, 68.4 +/- 5.4 kg, and 7.8 +/- 2.5% body fat) performed maximal 200 and 400 m crawl trials for S(200-400) determination (1.28 +/- 0.05 m/s). Thereafter, swimmers were instructed to perform 5 x 400 in at this speed with 1.5 min rest between repetitions. Three athletes Could not complete the set (exhaustion at 21.0 +/- 3.1 min). For the remaining swimmers (total set duration = 32.0 +/- 1.3 min) significant increases) (p < 0.05) in blood lactate (5.7 +/- 0.8-7.9 +/- 2.4 mmol/l), SR (29.6 +/- 3.2-32.1 +/- 4.1 cycles/min), HR (169 +/- 11-181 +/- 8 bpm) and RPE (13.3 +/- 1.6-16.3 +/- 2.6) were observed through the IS. Conversely, SL decreased significantly (p < 0.05) from the first to the fifth repetition (2.48 +/- 0.22-2.31 +/- 0.24 m/cycle). These results suggest that interval swimming at S(200-400) represents an intense physiological, mechanical and perceptual stimulus that can be sustained for a prolonged period by most athletes. (C) 2008 Sports Medicine Australia. Published by Elsevier Ltd. All rights reserved.

Downregulation of nuclear factor-kappa B (NF-kappa B) pathway by silibinin in human monocytes challenged with Paracoccidioides brasiliensis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Increased Levels of NOTCH1, NF-kappa B, and Other Interconnected Transcription Factors Characterize Primitive Sets of Hematopoietic Stem Cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The Effects of N Helix Deletion and Mutant F29W on the Ca2+ Binding and Functional Properties of Chicken Skeletal Muscle Troponin C

To assess the structural and functional significance of the N helix (residues 3-13) of avian recombinant troponin C (rTnC), we have constructed NHdel, in which residues 1-11 have been deleted, both in rTnC and in the spectral probe mutant F29W (Pearlstone, J. R., Borgford, T., Chandra, M., Oikawa, K., Kay, C. M., Herzberg, O., Moult, J., Herklotz, A., Reinach, F. C., and Smillie, L.B. (1992) Biochemistry 31, 6545-6553). Comparison of the far- and near-UV CD spectra (±Ca2+) of F29W and F29W/ NHdel and titration of the Ca2+-induced ellipticity and fluorescence changes indicates that the deletion has little effect on the global fold of the molecule but reduces the Ca2+ affinity of the N domain, but not the C domain, by 1.6-1.8-fold. Comparisons of the mutants NHdel, F29W, and F29W/NHdel with rTnC have been made using several functional assays. In reconstituted troponin-tropomyosin actomyosin subfragment 1 and myofibrillar ATPase systems, both F29W and NHdel have significantly reduced Ca2+-activated enzymic activities. These effects are cumulative in the double mutant F29W/ NHdel. On the other hand, maximal isometric tension development in Ca2+-activated reconstituted skinned fibers is not affected with F29W and NHdel, although the Ca2+ sensitivity of NHdel in this system is markedly reduced. We conclude that both mutations, NHdel and F29W, are functionally deleterious, possibly affecting interactions of the N domain with troponin I and/or T.