Cryogenic preservation of embryos of Prochilodus lineatus (Valenciennes, 1836) (Characiforme; Prochilodontidae)

While the freezing techniques of mammal embryos have been providing promising results, the cryopreservation of teleostean eggs and embryos have remained unsuccessful up to now. Therefore, this work aimed to develop a procedure of cryogenic preservation of embryos of Prochilodus lineatus and to observe, at both structural and ultrastructural levels, the morphological alterations that took place after the application of freezing/thawing techniques. The embryos at the morula stage could not tolerate exposure to the cryoprotectants ethylene glycol monomethyl ether, propylene glycol monomethyl ether, methanol, dimethyl sulphoxide and propylene glycol, presenting 100% of mortality. Embryos at the 4- to 6-somites stage tolerated exposure to propylene glycol and dimethyl sulphoxide, and the results revealed no significant differences (alpha = 0.05) regarding survival from both treatments. None of the freezing, thawing and hydration protocols was effective on preserving embryo viability. The ultrastructural analyses of frozen and thawed embryos showed that cells from ectoderm, somites, notochord and endoderm were structurally intact, with well preserved nuclei and mitochondria. The yolk globules were able to tolerate the freezing process, but the yolk syncytial layer was unorganized, displaying an electron-dense and compacted appearance, collapsed reticules, nuclei with modified chromatin and ruptures on the plasmatic membrane at the contact zone with endoderm. It might be concluded that the procedures tested for freezing were unable to avoid the formation of intracellular ice crystals, leading to drastic morphological modifications and making P. lineatus embryos unviable.

The effect of changes in the bulk dielectric constant on the DNA torsional properties

The effect of changes in the bulk dielectric constant on the DNA torsional properties was evaluated from plasmid circularization reactions. In these reactions, pUC18 previously linearized by EcoRI digestion was recircularized with T4 DNA ligase. The bulk dielectric constant of the reaction medium was decreased by the addition of different concentrations of neutral solutes: ethylene glycol, glycerol, sorbitol, and sucrose, or increased by the addition of glycine. The topoisomers generated by the ligase reaction were resolved by agarose-gel electrophoresis. The DNA twist energy parameter (K), which is an apparent torsional constant, was determined by linearization of the Gaussian topoisomers' distribution. It was observed that the twist energy parameter for the given solutes is almost linearly dependent on the bulk dielectric constant. In the reaction buffer, the twist energy parameter was determined to be 1100 +/- 100. By decreasing the dielectric constant to 74 with the addition of sorbitol, the value of the parameter reaches K = 900 +/- 100, whereas the addition of ethylene glycol leads to kappa = 400 +/- 50. Upon addition of glycine, which resulted in a dielectric constant equal to 91, the value of the twist energy parameter increased to K 1750 +/- 100. (c) 2007 Wiley Periodicals.

A micelle nucleation model for the interaction of dodecyl sulphate with Lys49-phospholipases A(2)

Bothropstoxin-I (BthTx-I) is a Lys49-PLA(2) from the venom of Bothrops jararacussu that lacks detectable catalytic activity, yet causes rapid Ca2+-independent membrane damage. With the aim of understanding the interaction between BthTx-I and amphiphilic molecules, we have studied the interaction of sodium dodecyl sulphate (SDS) with the protein. Circular dichroism and attenuated total reflection Fourier-transform infrared spectra of BthTx-I reveal changes in the alpha-helical organization of the protein at an SDS/BthTx-I molar ratio of 20-25. At SDS/BthTx-I ratios of 40-45 the alpha-helices return to a native-like conformation, although fluorescence emission anisotropy measurements of 2-amino-N-hexadecyl-benzamide (AHBA) demonstrate that the total SDS is below the critical micelle concentration when this transition occurs. These results may be interpreted as the result of SDS accumulation by the BthTx-I homodimer and the formation of a pre-micelle SDS/BthTx-I complex, which may subsequently be released from the protein surface as a free micelle. Similar changes in the alpha-helical organization of BthTx-I were observed in the presence of dipalmitoylphosphatidylcholine liposomes, suggesting that protein structure transitions coupled to organization changes of bound amphiphiles may play a role in the Ca2+-independent membrane damage by Lys49-PLA(2)s. (c) 2006 Elsevier B.V. All rights reserved.

Interfacial concentrations of chloride and bromide in zwitterionic micelles with opposite dipoles: Experimental determination by chemical trapping and a theoretical description

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

O eletrólito suporte e suas múltiplas funções em processos de eletrodo

The aim of this work is to present the principal properties and applications of supporting electrolytes (SE) to students, teachers and researchers interested in electrode processes. Different aspects are discussed including the importance of SE in maintaining constant the activity coefficients and the diffusion coefficients and reducing the transport number of electroactive species. Its effect on the electrochemical kinetic parameters is also presented.

Reações ácido-base: conceito, representação e generalização a partir das energias envolvidas nas transformações

Undergraduate students on the first year of Chemistry Courses are unfamiliar with the representation of acid-base reactions using the ionic equation H+ + OH- → H2O. A chemistry class was proposed about acid-base reactions using theory and experimental evaluation of neutralization heat to discuss the energy involved when water is formed from H+ and OH- ions. The experiment is suggested using different strong acids and strong base pairs. The presentation of the theme within a chemistry class for high school teachers increased the number of individuals that saw the acid-base reaction from this perspective.

Rendimentos dos cortes e não-componentes das carcaças de cordeiros terminados com dietas contendo diferentes fontes de óleo vegetal

Objetivou-se avaliar os rendimentos dos cortes e dos não-componentes das carcaças de cordeiros Santa Inês puros e ½ Dorset ½ Santa Inês, alimentados com dietas contendo diferentes fontes de óleo vegetal (óleo de soja, óleo de canola e óleo de linhaça) e uma dieta controle (sem adição de óleo vegetal). Após o abate, foram coletados sangue, pele, aparelho gastrintestinal cheio (esôfago + estômagos + intestinos delgado e grosso com seus conteúdos), aparelho gastrintestinal vazio (esôfago + estômagos + intestinos delgado e grosso, previamente esvaziados e limpos), aparelho reprodutor + bexiga, baço, fígado, coração, aparelho respiratório, rins com gordura perirrenal, cabeça, patas e cauda, que foram pesados para determinação do rendimento em relação ao peso vivo ao abate. Após resfriamento por 24 horas em câmara fria, pesou-se a carcaça e, posteriormente, dividiu-se longitudinalmente, sendo a metade esquerda seccionada em sete regiões anatômicas: perna, lombo, paleta, costelas flutuantes, costelas verdadeiras, baixos e pescoço. O estudo dos não-componentes da carcaça destacou a representabilidade dos pesos da pele (8,74%) e do conteúdo gastrintestinal (10,65%) na determinação do rendimento. As porcentagens dos cortes não apresentaram diferenças (p>0,05) em relação às dietas e grupos genéticos estudados.

Fontes de óleo vegetal na dieta de cordeiros em confinamento

Objetivou-se avaliar a ingestão de matéria seca, o ganho de peso diário, a conversão alimentar, o peso vivo ao abate e o período de confinamento em cordeiros Santa Inês puros e ½Dorset ½Santa Inês, alimentados com dietas isoenergéticas (76,59% de NDT) e isoprotéicas (17,48% de PB) contendo diferentes fontes de óleo vegetal (óleos de soja, canola e linhaça) e uma dieta controle (sem inclusão de óleo vegetal). A relação volumoso:concentrado foi de 30:70 e utilizou-se feno de aveia como volumoso. Realizou-se também um ensaio de digestibilidade, utilizando quatro cordeiros não-castrados, distribuídos em delineamento quadrado latino, avaliando-se ingestão, excreção fecal e digestibilidade total dos nutrientes das rações. A ingestão de matéria seca, expressa em porcentagem do peso vivo, foi menor nos cordeiros que receberam dieta contendo óleo de canola que naqueles que receberam dieta controle. Porém, todas as rações proporcionaram ganhos de peso e conversão alimentar satisfatórios. Os valores de digestibilidade total da matéria seca (76,02%) e matéria orgânica (76,82%) da dieta controle foram superiores aos da dieta contendo óleo de linhaça (72,11% e 72,97%, respectivamente), embora não tenham diferido das dietas contendo óleos de soja (72,94 e 73,71%) e canola (73,45 e 74,25%). A digestibilidade do extrato etéreo foi menor na dieta controle (84,02%), enquanto as demais dietas apresentaram valor médio de 91,98%. Os óleos vegetais reduziram a digestibilidade da matéria seca e da matéria orgânica, não afetando a ingestão e digestão dos demais nutrientes.

Caracterização morfológica dos estádios de desenvolvimento do aparelho reprodutor masculino da rã-touro, Rana catesbeiana, no sistema anfigranja de criação intensiva

The objective of this work was to caracterize the morphological characterization of the development stages of male reproduction apparel of bullfrog Rana catesbeiana from intensive Anfigranja system. During the period of one year, 657 bullfrog males were removed from regrowing and reproduction sections in Anfigranja system to study basic aspects of the reproduction of this specie. The collections were realized monthly and, in that occasion, the body and testicles weights for each animal were recorded and their macroscopic gonadal characteristics, registred. Testicles from 274 males were removed and histologically analyzed. From the macro and microscopic gonadal characteristics, analysis set of the testicles, five stages of gonadal maturation for the male of Rana catesbeiana were determined: juvenile, beginning of maturation, intermediate maturation, advanced maturation and spent.

pH at the micellar interface: Synthesis of pH probes derived from salicylic acid, acid-base dissociation in sodium dodecyl sulfate micelles, and Poisson-Boltzmann simulation

The study of the H+ concentration at the micellar interface is a convenient system for modeling the distribution of H+ at interfaces. We have synthesized salicylic acid derivatives to analyze the proton dissociation of both the carboxylic and phenol groups of' the probes, determining spectrophotometrically the apparent pK(a)'s (pK(ap)) in sodium dodecyl Sulfate, SDS, micelles with and without added salt. The synthesized probes were 2-hydroxy-5-(2-trimethylammoniumacetyl)benzoate; 2-hydroxy-5-(2-dimethylhexadecylammoniumacetyl)benzoate- 2-hydroxy-5-(2-dimethylhexadecylammoniumhexanoyl)benzoate-, 2-hydroxy-5-(2-diniethylhexadecylammoniumundecanoyl)betizoate; 2-hydroxy-5-acetylbenzoic acids and 2-hydroxy-5-dodecanoylbenzoic acid. Upon incorporation into SDS micelles the pK(ap)'s of both carboxylic and phenol groups increased by ca. 3 pH units and NaCl addition caused a decrease in the probe-incorporated pKap. The experimental results were fitted with a cell model Poisson-Boltzmann (P-B) equation taking in consideration the effect of salt on the aggregation number of SDS and using the distance of' the dissociating group as a parameter. The conformations of the probes were analyzed theoretically using two dielectric constants, e.g., 2 and 78. Both the P-B analysis and conformation calculations can be interpreted by assuming that the acid groups dissociate very close to, or at, the interface. Our results are consistent with the assumption that the intrinsic pK(a)'s of both carboxylic and phenol groups of the salicylic acid probes used here can be taken as those in water. Using this assumption the micellar and salt effects on the pKap's of the (trialkylammonium)benzoate probes were described accurately using a cell model P-B analysis. (c) 2005 Elsevier B.V. All rights reserved.

Planar membranes interaction

The system of two parallel planar, arbitrarily charged surfaces immersed in a solution containing only one ionic species, the counterions, is completely analyzed under a mean field Poisson-Boltzmann approach. Results for the pressure, reduced potential, and counterionic concentration are graphically displayed for two dissociating membranes and for a dissociating and an adsorbing membrane. The results indicate that the system of two planar parallel dissociating membranes acts as a buffer for pressure values and for counterionic concentration values in regions interior to and far from the membranes. The results are related to properties of planar or quasiplanar structures in biological cells.

Uroflow studies in children: Correlation with age, weight and body surface area.

Peak flow rate data (Q(max)) are important for diagnosis of lower urinary tract obstruction. However, the lack of uroflowmetry studies in children, makes their interpretation difficult. With this aim, we studied 167 boys classified in four groups according to their age (G1 : n = 48, 6-7 years; G2 : n = 43, 8-9 years; G3 : n = 37, 10-11 years; G4: n = 39, 12-14 years). We studied in all children, weight, height, body surface area, peak flow rate (Q(max) and the correspondant urinary volume (Vol). Means and standard deviations of Q(max) (ml/sec) were : 15 +/- 5 (G1), 15 +/- 5 (G2), 17 +/- 5 (G3) and 22 +/- 7 (G4) respectively. Corresponding urinary volumes (Vol) (mean standard deviation - in ml) were : G1 = 123 +/- 75; G2 = 122 +/- 79; G3 = 158 +/- 96 and G4 = 162 +/- 101. We found a significant correlation (p < 0,01) between Q(max) and Vol in groups G2, G3 and G4; and between Q(max) and height in groups G1 and G4.The authors demonstrated a positive correlation between maximum flow and voided volume, and an increase of Q(max) with age.

Estimativa do Tamanho da Primeira Maturação Sexual da Rã - Touro, Rana catesbeiana, no Sistema Anfigranja de Criação Intensiva

The objective of this research was to study the basic aspects of bullfrog reproduction in Anfigranja system from June 1989 to July 1990. For the 1493 animals involved in this experiment (657 males and 836 females) biometrical data were obtained (weight, length and tympanum diameter) and developmental stages of gonads were determined. Female sizes at first maturation were 10,33 cm and 109,78 g and for males it was around 8,09 cm; all males above 45 g were already in the maturation process. The relationship between tympanum diameter and body length (RTC) was considered a good index for male maturation.