Colleters in Caryocar brasiliense (Caryocaraceae) ontogenesis, ultrastructure and secretion

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

First gonadal maturation of Pinirampus pirinampu (Siluriformes: Pimelodidae) in the Pantanal, Mato Grosso do Sul State, Brazil

Informações sobre o ciclo reprodutivo de espécies exploradas pela pesca são imprescindíveis para o seu manejo adequado. O tamanho de início da atividade reprodutiva do Pinirampus pirinampu (Spix, 1829) foi determinado. Os exemplares foram capturados em setembro, outubro e dezembro de 1997 e fevereiro e março de 1998. O estádio de desenvolvimento gonadal foi identificado macroscopicamente. O índice gonadossomático médio (IGS), calculado mensalmente, foi utilizado como indicador da época de desova. Para determinação do comprimento da primeira maturação gonadal agruparam-se, separadamente, machos e fêmeas por classes de comprimento em imaturos e adultos. Os resultados referentes aos indivíduos adultos foram lançados em gráficos e a mediana correspondeu à estimativa do comprimento no qual 50% dos indivíduos atingem a maturidade (L50). Foi também determinado o L100, estimativa do comprimento em que todos os indivíduos estão aptos à reprodução. Machos e fêmeas em processo de maturação gonadal foram encontrados a partir de outubro, com maior freqüência em fevereiro, e, somente a partir deste mês, foram encontrados indivíduos com gônadas esvaziadas. O índice gonadossomático mostrou que a partir de setembro inicia-se o processo de desenvolvimento gonadal, com seu valor máximo em fevereiro. O L50 para fêmeas foi 574 mm e para machos foi 536 mm. O L100 para fêmeas foi 590 mm e para os machos, 580 mm.

In vitro anti-Mycobacterium tuberculosis activity of some Brazilian Cerrado plants

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

Phenomenology of infrared finite gluon propagator and coupling constant

We report on some recent solutions of the Dyson-Schwinger equations for the infrared behavior of the gluon propagator and coupling constant, discussing their differences and proposing that these different behaviors can be tested through hadronic phenomenology. We discuss which kind of phenomenological tests can be applied to the gluon propagator and coupling constant, how sensitive they are to the infrared region of momenta and what specific solution is preferred by the experimental data.

B-chromosomes in two Brazilian populations of Dendropsophus nanus (Anura, Hylidae)

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

Co-infecção HIV/HCV em pacientes de Botucatu e região

Devido à similaridade nas rotas de transmissão, a co-infecção HIV/HCV é freqüente, afetando em média 30 a 50% dos portadores de HIV. O presente estudo visou avaliar uma possível associação entre os subtipos do HIV e genótipos do HCV em pacientes co-infectados, com base na análise das freqüências em pacientes mono e co-infectados. Para determinação da freqüência dos subtipos HIV e genótipos HCV, foram analisados respectivamente 124 e 496 pacientes mono-infectados. O estudo da co-infecção foi realizado num grupo de 150 pacientes HIV positivos e esteve presente em 22 (14,7%) dos pacientes. A freqüência dos subtipos do HIV-1 em mono-infectados foi: subtipo B (85,5%), subtipo F (12,9%) e recombinante B/F (1,6%), enquanto nos genótipos HCV foi: 1a (25%), 1b (29,4%), 1a/1b (3,6%), 3a (35%), 2 (1,8%) e 5 (0,4%). Nos co-infectados o padrão de distribuição dos subtipos HIV-1 é semelhante aos mono-infectados, ou seja, subtipo B (85,0%), seguido do subtipo F (15,0%). A distribuição de freqüência de genótipos HCV nos co-infectados foi: 1a (36,3%), 1b (27,3%), 1a/1b (9,1%) e 3a (27,3%) mostrando um aumento de 10% na freqüência do genótipo 1, queda de 7,7% no genótipo 3 e ausência de outros genótipos. A análise estatística de associação entre os subtipos HIV e genótipos HCV (Goodman) mostrou que no genótipo 1 (HCV) ocorreu predominância do subtipo B, enquanto no genótipo 3 (HCV) a distribuição dos subtipos B e F (HIV-1) foi casual. Isto aponta para a necessidade de mais estudos desse grupo e um maior valor amostral.

Differential distribution of functional alpha(1)-adrenergic receptor subtypes along the rat tail artery

The rat tail artery has been used for the study of vasoconstriction mediated by alpha(1A)-adrenoceptors (ARs). However, rings from proximal segments of the tail artery (within the initial 4 cm, PRTA) were at least 3- fold more sensitive to methoxamine and phenylephrine (n = 6 - 12; p < 0.05) than rings from distal parts (between the sixth and 10th cm, DRTA). Interestingly, the imidazolines N-[ 5-( 4,5- dihydro- 1H- imidazol-2-yl)-2-hydroxy-5,6,7,8- tetrahydronaphthalen- 1- yl] methanesulfonamide hydrobromide (A-61603) and oxymetazoline, which activate selectively alpha(1A)- ARs, were equipotent in PRTA and DRTA (n = 4 - 12), whereas buspirone, which activates selectively alpha(1D)-AR, was approximate to 70-fold more potent in PRTA than in DRTA (n = 8; p < 0.05). The selective alpha(1D)-AR antagonist 8-[2-[4-(methoxyphenyl)-1-piperazinyl] ethyl]-8-azaspiro[4.5] decane-7,9-dione dihydrochloride (BMY- 7378) was approximate to 70- fold more potent against the contractions induced by phenylephrine in PRTA (pK(B) of approximate to 8.45; n = 6) than in DRTA (pK B of approximate to 6.58; n = 6), although the antagonism was complex in PRTA. 5-Methylurapidil, a selective alpha(1A)-antagonist, was equipotent in PRTA and DRTA (pK(B) of approximate to 8.4), but the Schild slope in DRTA was 0.73 +/- 0.05 ( n = 5). The noncompetitive alpha(1B)-antagonist conotoxin rho-TIA reduced the maximal contraction induced by phenylephrine in DRTA, but not in PRTA. These results indicate a predominant role for alpha(1A)-ARs in the contractions of both PRTA and DRTA but with significant coparticipations of alpha(1D)-ARs in PRTA and alpha(1B)-ARs in DRTA. Semiquantitative reverse transcription-polymerase chain reaction revealed that mRNA encoding alpha(1A)- and alpha(1B)-ARs are similarly distributed in PRTA and DRTA, whereas mRNA for alpha(1D)-ARs is twice more abundant in PRTA. Therefore, alpha(1)-ARs subtypes are differentially distributed along the tail artery. It is important to consider the segment from which the tissue preparation is taken to avoid misinterpretations on receptor mechanisms and drug selectivities. antagonism was complex in PRTA. 5- Methylurapidil, a selective alpha(1A)-antagonist, was equipotent in PRTA and DRTA (pK(B) of approximate to 8.4), but the Schild slope in DRTA was 0.73 +/- 0.05 ( n = 5). The noncompetitive alpha(1B)-antagonist conotoxin rho-TIA reduced the maximal contraction induced by phenylephrine in DRTA, but not in PRTA. These results indicate a predominant role for alpha(1A)-ARs in the contractions of both PRTA and DRTA but with significant coparticipations of alpha(1D)-ARs in PRTA and alpha(1B)-ARs in DRTA. Semiquantitative reverse transcription-polymerase chain reaction revealed that mRNA encoding alpha(1A)- and alpha(1B)- ARs are similarly distributed in PRTA and DRTA, whereas mRNA for alpha(1D)-ARs is twice more abundant in PRTA. Therefore, alpha(1)-ARs subtypes are differentially distributed along the tail artery. It is important to consider the segment from which the tissue preparation is taken to avoid misinterpretations on receptor mechanisms and drug selectivities.

The N terminus of the human alpha(1D)-adrenergic receptor prevents cell surface expression

We previously reported that truncation of the N-terminal 79 amino acids of alpha(1D)-adrenoceptors (Delta(1-79)alpha(1D)-ARs) greatly increases binding site density. In this study, we determined whether this effect was associated with changes in alpha(1D)-AR subcellular localization. Confocal imaging of green fluorescent protein (GFP)-tagged receptors and sucrose density gradient fractionation suggested that full-length alpha(1D)-ARs were found primarily in intracellular compartments, whereas Delta(1-79)alpha(1D)-ARs were translocated to the plasma membrane. This resulted in a 3- to 4-fold increase in intrinsic activity for stimulation of inositol phosphate formation by norepinephrine. We determined whether this effect was transplantable by creating N-terminal chimeras of alpha(1)-ARs containing the body of one subtype and the N terminus of another (alpha(1A) NT-D, alpha(1B) NT-D, alpha(1D) NT-A, and alpha(1D)NT-B). When expressed in human embryonic kidney 293 cells, radioligand binding revealed that binding densities of alpha(1A)- or alpha(1B)-ARs containing the alpha(1D)-N terminus decreased by 86 to 93%, whereas substitution of alpha(1A)- or alpha(1B)-N termini increased alpha(1D)-AR binding site density by 2- to 3-fold. Confocal microscopy showed that GFP-tagged alpha(1D)NT-B-ARs were found only on the cell surface, whereas GFP-tagged alpha(1B)NT-D-ARs were completely intracellular. Radioligand binding and confocal imaging of GFP-tagged alpha(1D)- and Delta(1-79)alpha(1D)-ARs expressed in rat aortic smooth muscle cells produced similar results, suggesting these effects are generalizable to cell types that endogenously express alpha(1D)-ARs. These findings demonstrate that the N-terminal region of alpha(1D)-ARs contain a transplantable signal that is critical for regulating formation of functional bindings, through regulating cellular localization.

Differential antagonism by conotoxin p-TIA of contractions mediated by distinct alpha(1)-adrenoceptor subtypes in rat vas deferens, spleen and aorta

The ability of the conotoxin p-TIA, a 19-amino acid peptide isolated from the marine snail Conus tulipa, to antagonize contractions induced by noradrenaline through activation of alpha(1A)-adrenoceptors in rat vas deferens, alpha(1B)-adrenoceptors in rat spleen and alpha(ID)-adrenoceptors in rat aorta, and to inhibit the binding of [I-125]HEAT (2-[[beta-(4-hydroxyphenyl)ethyl]aminomethyl]-1-tetralone) to membranes of human embryonic kidney (HEK) 293 cells expressing each of the recombinant rat alpha(1)-adrenoceptors was investigated. p-TIA (100 nM to 1 muM) antagonized the contractions of vas deferens and aorta in response to noradrenaline without affecting maximal effects and with similar potencies (pA(2)similar to7.2, n=4). This suggests that p-TIA is a competitive antagonist of alpha(1A)- and alpha(1D)-adrenoceptors with no selectivity between these subtypes. Incubation of p-TIA (30 to 300 nM) with rat spleen caused a significant reduction of the maximal response to noradrenaline, suggesting that p-TIA is a non-competitive antagonist at alpha(1B)-adrenoceptors. After receptor inactivation with phenoxybenzamine, the potency of p-TIA in inhibiting contractions was examined with similar occupancies (similar to25%) at each subtype. Its potency (pIC(50)) was 12 times higher in spleen (8.3 +/- 0.1, n=4) than in vas deferens (7.2 +/- 0.1, n=4) or aorta (7.2 0.1, n=4). In radioligand binding assays, p-TIA decreased the number of binding sites (B,,,,,,) in membranes from HEK293 cells expressing the rat alpha(1B)-adrenoceptors without affecting affinity (K-D), In contrast, in HEK293 cells expressing rat alpha(1A)- or alpha(1D)-adrenoceptors, p-TTA decreased the KD without affecting the B-max. It is concluded that p-TIA will be useful for distinguishing the role of particular alpha(1)-adrenoceptor subtypes in native tissues. (C) 2004 Elsevier B.V. All rights reserved.

Effects of subtypes alpha- and beta-adrenoceptors of the lateral hypothalamus on the water and sodium intake induced by angiotensin II injected into the subfornical organ

The present experiments were conducted to investigate the role of the alpha (1A)-, alpha (1B), beta (1),- and beta (2)-adrenoceptors of the lateral hypothalamus (LH) on the water and salt intake responses elicited by subfornical organ (SFO) injection of angiotensin II (ANG II) in rats. 5-methylurapidil (an alpha (1A)-adrenergic antagonist), cyclazosin (an alpha (1B)-adrenergic antagonist) and ICI-118,551 (a beta (2)-adrenergic antagonist) injected into the LH produced a dose-dependent reduction, whereas efaroxan (an alpha (2)-antagonist) increased the water intake induced by administration of ANG II into the SFO. These data show that injection of 5-methylurapidil into the LH prior to ANG II into the SFO increased the water and sodium intake induced by the injection of ANG II. The present data also show that atenolol (a beta (1)-adrenergic antagonist), ICI-118,551, cyclazosin, or efaroxan injected into the LH reduced in a dose-dependent manner the water and sodium intake to angiotensinergic activation of SFO. Thus, the alpha (1)- and beta -adrenoceptors of the LH are possibly involved with central mechanisms dependent on ANG II and SFO that control water and sodium intake. (C) 2000 Elsevier B.V. B.V. All rights reserved.

Polysiloxane-poly(propylene oxide) hybrid discs as solid phase in anti-HCV detection using a recombinant core protein

In this work, siloxane-poly(propylene oxide) discs (PPO disc) prepared using the sol-gel process were used as solid phase in enzyme-linked immunosorbent assays (ELISA) for the detection of anti-hepatitis C virus (HCV) antibodies. The HCV RNA from serum (genotype 1b) was submitted to the RT-PCR technique and subsequent amplification of the HCV core 408 pb. This fragment was cloned into expression vector pET42a and expressed in Escherichia coli as recombinant protein with glutathione S-transferase (GST). Cell cultures were grown and induced having a final concentration of 0.4 x 10(-3) mol L-1 of IPTG. After induction, the cells were harvested and the soluble fraction was analyzed using polyacrilamide gel 15% showing a band with an approximate molecular weight of 44 kDa, the expected size for this GST-fused recombinant protein. The recombinant protein was purified and continued by immunological detection using HCV-positive serum and showed no cross-reactivity with positive samples for other infectious diseases. An ELISA was established using 1.25 ng of recombinant protein per PPO disc, a dilution of 1: 10,000 and 1:40 for a peroxidase conjugate and serum, respectively, and solutions of hydrogen peroxide and 3,3',5,5'-tetra-methylbenzidine in a ratio of 1: 1. The proposed methodology was compared with the ELISA conventional polystyrene-plate procedure and the performance of the PPO discs as a matrix for immunodetection gave an easy synthesis, good performance and reproducibility for commercial application. (c) 2007 Elsevier B.V. All rights reserved.

Catecholamine effects on human melanoma cells evoked by α1-adrenoceptors

The biological effects of catecholamines in mammalian pigment cells are poorly understood. Our previous results showed the presence of α1-adrenoceptors in SK-Mel 23 human melanoma cells. The aims of this work were to (1) characterize catecholamine effects on proliferation, tyrosinase activity and expression, (2) identify the α1- adrenoceptor subtypes, and (3) verify whether chronic norepinephrine (NE) treatment modified the types and/or pharmacological characteristics of adrenoceptors present in SK-Mel 23 human melanoma cells. Cells treated with the aradrenergic agonist, phenylephrine (PHE, 10-5 or 10-4 M), for 24-72 h, exhibited decreased cell proliferation and enhanced tyrosinase activity, but unaltered tyrosinase expression as compared with the control. The proliferation and tyrosinase activity responses were inhibited by the α1-adrenergic antagonist prazosin, suggesting they were evoked by α1-adrenoceptors. The presence of actinomycin D, a transcription inhibitor, did not diminish PHE-induced effects. RT-PCR assays, followed by cloning and sequencing, demonstrated the presence of α1A- and α1B-adrenoceptor subtypes. NE-treated cells (24 or 72 h) were used in competition assays, and showed no significant change in the competition curves of α1-adrenoceptors as compared with control curves. Other adrenoceptor subtypes were not identified in these cells, and NE pretreatment did not induce their expression. In conclusion, the activation of SK-Mel 23 human melanoma α1- radrenoceptors elicit biological effects, such as proliferation decrease and tyrosinase activity increase. Desensitization or expression of other adrenoceptor subtypes after chronic NE treatment were not observed.

Design of SPR systems with dynamic compensators and output variable structure control

This paper presents necessary and sufficient conditions for the following problem: given a linear time invariant plant G(s) = N(s)D(s)-1 = C(sI - A]-1B, with m inputs, p outputs, p > m, rank(C) = p, rank(B) = rank(CB) = m, £nd a tandem dynamic controller Gc(s) = D c(s)-1Nc(s) = Cc(sI - A c)-1Bc + Dc, with p inputs and m outputs and a constant output feedback matrix Ko ε ℝm×p such that the feedback system is Strictly Positive Real (SPR). It is shown that this problem has solution if and only if all transmission zeros of the plant have negative real parts. When there exists solution, the proposed method firstly obtains Gc(s) in order to all transmission zeros of Gc(s)G(s) present negative real parts and then Ko is found as the solution of some Linear Matrix Inequalities (LMIs). Then, taking into account this result, a new LMI based design for output Variable Structure Control (VSC) of uncertain dynamic plants is presented. The method can consider the following design specifications: matched disturbances or nonlinearities of the plant, output constraints, decay rate and matched and nonmatched plant uncertainties. © 2006 IEEE.

Response of rootstocks to stem canker and the production and quality of melon under protected cultivation

Two tests were performed. In the first, resistance to Didymella bryoniae was determined for the following genotypes: the pumpkins 'Ikky', 'Agroceres', 'Kirameki' and 'Shelper', watermelon progenies 1a, 2a, 3a, 5a, 1b, 2b, 3b and 5b, and 'Gherkin' (C. anguria). The plants were inoculated with the fungus during transplanting. The evaluations of the test were performed every 15 d according to a scoring scale adopted by Dusi et al. (1994). The second test examined compatibility among the rootstocks x grafts, and their effects on production. The rootstocks, 5 pumpkins including 'Ikky', 'Agroceres', 'Kirameki', 'Shelper', six watermelon progenies 1a, 2a, 5a, 1b, 2b and 5b, and one 'Gherkin', were planted one week before planting of the grafted 'Bônus No. 2' melon. The experiments were carried out with 12 treatments, including the control ('Bônus No. 2') with 3 replications with 14 grafted plants per each replication. For the first test, the first three evaluations (at 15, 30 and 45 d after inoculation) did not show characteristic lesions of stem canker, but progeny 3b was found to be susceptible in evaluations performed at 60 and 75 d after inoculation. Progeny 3a demonstrated intermediate susceptibility, while progenies 1a, 2a, 5a, 1b, 2b and 5b, the pumpkins 'Kirameki', 'Shelper', 'Ikky' and 'Agroceres', and 'Gherkin', showed resistance to Didymella bryoniae. In the second test, watermelon progenies 1a, 5a, 1b and 2b, and the pumpkins 'Kirameki', 'Shelper', 'Ikky' and 'Agroceres' showed a level of grafting success of 100%, while results with progenies 2a and 5b, and 'Gherkin' were different in grafting success, respectively 91.67, 98.33 and 43.33%. For other fruit parameters, weight, longitudinal and transverse diameters, pulp thickness and level of total soluble solids, there were no differences among the treatments.

Influence of acid-etching and ceramic primers on the repair of a glass ceramic

The objective of this study was to evaluate the influence of different primers on the microtensile bond strength (μT BS) between a feldspathic ceramic and two composites. Forty blocks (6.0 × 6.0 × 5.0 mm 3) were prepared from Vita Mark II . After polishing, they were randomly divided into 10 groups according to the surface treatment: Group 1, hydrofluoric acid 10% (HF) + silane; Group 2, CoJet + silane; Group 3, HF + Metal/Zirconia Primer; Group 4, HF + Clearfil Primer; Group 5, HF + Alloy Primer; Group 6, HF + V-Primer; Group 7, Metal/Zirconia Primer; Group 8, Clearfil Primer; Group 9, Alloy Primer; Group 10, V-Primer. After each surface treatment, an adhesive was applied and one of two composite resins was incrementally built up. The sticks obtained from each block (bonded area: 1.0 mm2 ± 0.2 mm) were stored in distilled water at 37°C for 30 days and submitted to thermocycling (7,000 cycles; 5°C/55°C ± 1°C). The μT BS test was carried out using a universal testing machine (1.0 mm/min). Data were analyzed using ANOVA and a Tukey test (α = 0.05). The surface treatments significantly affected the results (P < 0.05); no difference was observed between the composites (P > 0.05). The bond strength means (MPa) were as follows: Group 1a = 29.6; Group 1b = 33.7; Group 2a = 28.9; Group 2b = 27.1; Group 3a = 13.8; Group 3b = 14.9; Group 4a = 18.6; Group 4b = 19.4; Group 5a = 15.3; Group 5b = 16.5; Group 6a = 11; Group 6b = 18; Groups 7a to 10b = 0. While the use of primers alone was not sufficient for adequate bond strengths to feldspathic ceramic, HF etching followed by any silane delivered higher bond strength.