Involvement of nitric oxide synthase in the mechanism of histamine-induced inhibition of Leydig cell steroidogenesis via histamine receptor subtypes in Sprague-Dawley rats

This study was conducted to shed light on the so far unexplored intracellular mechanisms underlying negative modulation of Leydig cell steroidogenesis by histamine (HA). Using the MA-10 cell line and highly purified rat Leydig cells as experimental models, we examined the effect of the amine on biochemical steps known to be modulated by HA or involved in LH/hCG action. In agreement with previous findings, HA at 10 microM showed a potent inhibitory effect on hCG-stimulated steroid synthesis, regardless of the gonadotropin concentration used. Moreover, HA decreased not only LH/hCG-induced cAMP production but also steroid synthesis stimulated by the permeable cAMP analog dibutyryl cAMP (db-cAMP). Considering the post-cAMP sites of HA action, it is shown herein that HA markedly inhibited db-cAMP-stimulated steroidogenic acute regulatory (STAR) protein expression, as well as steps catalyzed by P450-dependent enzymes, mainly the conversion of cholesterol to pregnenolone by cholesterol side-chain cleavage enzyme (CYP11A). The antisteroidogenic action of HA was blocked by addition of the phospholipase C (PLC) inhibitor U73122, and HA significantly augmented inositol triphosphate (IP3) production, suggesting a major role for the PLC/IP3 pathway in HA-induced inhibition of Leydig cell function. Finally, HA increased nitric oxide synthase (NOS) activity, and the NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) markedly attenuated the effect of the amine on steroid synthesis. On the basis of our findings, HA antagonizes the gonadotropin action in Leydig cells at steps before and after cAMP formation. NOS activation is the main intracellular mechanism by which HA exerts its antisteroidogenic effects.     

Comparison of the muscle activation pattern for the vastus lateralis before and after an 8-week resistance training program

The purpose of this investigation was to use wavelet analyses and pattern classification techniques to examine potential changes in the joint time–frequency distribution of surface electromyographic (EMG) signals due to an 8-week resistance training program. Thirteen untrained men (mean ± S.D. age = 22.2 ± 4.0 yrs) performed eight separate submaximal isometric muscle actions of the dominant leg extensors in 10% increments from 10 to 80% of the maximum voluntary contraction (MVC). During each muscle action, a monopolar surface EMG signal was detected from the vastus lateralis. All signals were then analyzed with a wavelet analysis designed specifically for EMG signals, and the resulting intensity patterns were classified using pattern classification techniques into their respective pre-training versus post-training categories. The results showed accuracy rates (% of correctly classified patterns) that ranged from approximately 62 to 92%, but these rates did not change consistently with increases in force. In addition, for five of the eight submaximal force levels, the classification was considered to be significantly different from random. Thus, although the differences between the pre- and post-training EMG intensity patterns were not always consistent, our findings did suggest that there is the potential for wavelet analyses and pattern classification techniques to be used to examine neuromuscular adaptations during resistance training.     

Differentially-expressed genes in Candida albicans exposed to ε-poly-l-lysine

Candida albicans is an opportunistic human pathogen whose disinfection is a challenge. ε-Poly-l-lysine (ε-PL), an antagonistic agent, can disrupt cell membranes and inhibit the growth of C. albicans. Genes that were differentially-expressed in response to ε-PL were isolated from C. albicans and identified by suppression subtractive hybridization. Ten subtracted clones, that share >98 % homology with known genes of C. albicans, were isolated. Among these, four genes encoded cell wall-associated proteins. Real-time quantitative PCR and northern blot hybridization suggest that these genes are involved in the response to ε-PL. These findings will help to determine the mechanism of the antimicrobial activity of ε-PL against C. albicans.     

Involvement of Ethylene Biosynthesis and Signalling in the Transition from Male to Female Flowering in the Monoecious Cucurbita pepo

It is well established that ethylene is the main hormonal regulator of sexual expression in the Cucurbitaceae family, controlling not only the sexual fate of individual floral buds, but also the female flower transition, that is, the time at which the first female flower appears and therefore the number of female flowers per plant. Although sex determination of individual flower buds is known to be controlled by specific ethylene biosynthesis ACS genes in melon and cucumber, the role of ethylene genes in the control of the transition to female flowering is still unknown. We have identified two contrasting monoecious inbred lines of Cucurbita pepo, Bolognese (Bog) and Vegetable spaghetti (Veg), which differ in female flower transition but not in flower development. In Bog, which is very sensitive to ethylene, the transition to female flowering is very early, whereas in Veg, which is much less sensitive to ethylene, the transition occurs much later. In this article we compare the production of ethylene and the expression profiles of seven genes involved in the biosynthesis, perception, and signalling of ethylene in the two contrasting lines. Bog, with earlier female flower transition, showed higher ethylene production and CpACO1 expression in the apex at an earlier stage of plant development, when Bog is already producing female flowers, but Veg has not transitioned to female flowering yet. Moreover, the expression of the ethylene receptor and CTR-like genes in the apex of Veg and Bog plants indicates that these genes negatively regulate female flower transition during the earlier stages of plant development. The earlier transition to female flowering in Bog is not only associated with a higher production of ethylene in the apex but also with a premature decline of ethylene negative regulators (receptors and CTR-like) in the apex of the plant. These results provide the basis for a model that explains the regulation of female flowering transition in monoecious cucurbits.     

The role of ethylene and brassinosteroids in the control of sex expression and flower development in <Emphasis Type="Italic">Cucurbita pepo</Emphasis>

In this paper we compare the sensitivity of different squash genotypes to ethylene and brassinosteroids by studying the effects of different ethylene and brassinosteroid treatments on the sexual expression and flower development of different C. pepo genotypes: Bolognese (Bog) and Vegetable Spaghetti (Veg), two contrasting lines for ethylene production and sensitivity, as well as Cora, a standard commercial hybrid. Results have demonstrated that ethylene has a much greater effect on sexual expression and flower development in C. pepo than brassinosteroids. Ethephon increases the number of female flowers per plant and reduces the first male phase of development, while treatments with the ethylene inhibitors AVG and STS reduce the number of female flowers per plant and expand the first male phase of development. The differential response observed between genotypes appears to be related to ethylene production and sensitivity. Bog, which produces more ethylene and is more sensitive to this hormone, responded much better to AVG and STS, reducing the number of female flowers per plant, while Veg, which is characterised by lower production of and sensitivity to ethylene, responded better to ethephon by reducing the first male phase of development and increasing the number of female flowers per plant. The differential abortion of female and male flowers in ethephon, AVG and STS treatments, as well as the occurrence of bisexual flowers in the AVG and STS treated plants of the more ethylene sensitive genotypes, demonstrate that ethylene is also involved in the development of female flowers. Female flower buds require a minimal level of ethylene not only to complete their development and maturation without a premature abortion, but also to arrest the development of stamens in the third whorl and to promote the appropriate growth of the carpels. On the contrary, the role of brassinosteroids in the sexual expression of C. pepo was not so evident. The application of brassinazole, an inhibitor of brassinosteroid biosynthesis slightly changes the production of ethylene in the three analysed genotypes, but those changes have little effect on their sexual phenotypes, and they do not alter the development of the unisexual flowers.     

Acyl coenzyme A synthetase long-chain 1 (ACSL1) gene polymorphism (rs6552828) and elite endurance athletic status: a replication study

The aim of this study was to determine the association between the rs6552828 polymorphism in acyl coenzyme A synthetase (ACSL1) and elite endurance athletic status. We studied 82 Caucasian (Spanish) World/Olympic-class endurance male athletes, and a group of sex and ethnically matched healthy young adults (controls, n=197). The analyses were replicated in a cohort of a different ethnic origin (Chinese of the Han ethnic group), composed of elite endurance athletes (runners) [cases, n=241 (128 male)] and healthy sedentary adults [controls, n=504 (267 male)]. In the Spanish cohort, genotype (P=0.591) and minor allele (A) frequencies were similar in cases and controls (P=0.978). In the Chinese cohort, genotype (P=0.973) and minor allele (G) frequencies were comparable in female endurance athletes and sedentary controls (P=0.881), whereas in males the frequency of the G allele was higher in endurance athletes (0.40) compared with their controls (0.32, P=0.040). The odds ratio (95%CI) for an elite endurance Chinese athlete to carry the G allele compared with ethnically matched controls was 1.381 (1.015-1.880) (P-value=0.04). Our findings suggest that the ACSL1 gene polymorphism rs6552828 is not associated with elite endurance athletic status in Caucasians, yet a marginal association seems to exist for the Chinese (Han) male population.     

Time-frequency analysis of surface electromyographic signals during fatiguing isokinetic muscle actions

The purpose of this study was to use a wavelet analysis designed specifically for electromyography (EMG) signals in combination with a trend plot to examine changes in EMG intensity patterns during maximal, fatiguing isokinetic muscle actions. Eleven men (mean ± SD age = 22.4 ± 1.1 years) and 7 women (mean ± SD age = 22.7 ± 2.1 years) performed 50 consecutive maximal concentric isokinetic muscle actions of the dominant leg extensors at a velocity of 180°·s(-1). During each muscle action, a bipolar surface EMG signal was detected from the vastus lateralis. All signals were then processed with a wavelet analysis designed specifically for EMG signals, which resulted in EMG intensity patterns. The patterns for each subject were then analyzed with a trend plot, which provided information regarding the changes that occurred because of fatigue. The results indicated that for all the 18 subjects, the EMG intensity patterns moved in a predictable manner in pattern space, but the changes to the patterns were different for each subject. These findings reflect the complex changes that occur in the EMG signal during fatigue. These changes cannot be characterized fully with a single amplitude and center frequency parameter and can be useful for athletes and coaches who need to track the fatigue status of individual muscles.     

GROWTH RATES AND ULTRASTRUCTURE OF SILICEOUS SETAE OF CHAETOCEROS GRACILIS (BACILLARIOPHYCEAE) 12

The growth of setae in post-division Chaeloceros gracilis Schütt was triphasic. Following cell separation there was an initial lag phase of about 30 min after which setae grew linearly at rates ranging from 0.20 to 0.38 μm-min^?1. The growth rate was independent of whether diatoms cultured in medium containing 200 μM Si were transferred for experimentation to media containing 10 or 200 μM Si. When developing setae had attained a length of approximately 2 μm, their growth rate slowed as they entered a clearly defined plateau phase. The amount of silicon per unit length of setae was found to be 0.02 pg Si μm^?1, and the deposition rate was estimated as 0.005 Pg Si min^?1. Transmission electron microscopy revealed an axial structure resembling a microlubule extending the length of each seta and membrane bound polyphosphate bodies postulated to be the energy source for growth and associated biomineralization processes.     

Molecular and functional characterization of CpACS27A gene reveals its involvement in monoecy instability and other associated traits in squash (Cucurbita pepo L.)

A number of Cucurbita pepo genotypes showing instable monoecy or partial andromonoecy, i.e. an incomplete conversion of female into bisexual flowers, have been detected. Given that in melon and cucumber andromonoecy is the result of reduction of ethylene production in female floral buds, caused by mutations in the ethylene biosynthesis genes CmACS7 and CsACS2; we have cloned and characterized two related C. pepo genes, CpACS27A and CpACS27B. The molecular structure of CpACS27A and its specific expression in the carpels of female flowers during earlier stages of flower development suggests that this gene is the Cucurbita ortholog of CmACS7 and CsACS2. CpACS27B is likely to be a paralogous pseudogene since it has not been found to be expressed in any of the analyzed tissues. CpACS27A was sequenced in Bolognese (Bog) and Vegetable Spaghetti (Veg), two monoecious inbred lines whose F2 was segregating for partial andromonoecy. The Bog allele of CpACS27A carried a missense mutation that resulted in a substitution of the conserved serine residue in position 176 by an alanine. Segregation analysis indicated that this mutant variant is necessary but not sufficient to confer the andromonoecious phenotype in squash. In concordance with its involvement in stamen arrest, a reduction in CpACS27A expression has been found in bisexual flower buds at earlier stages of development. This reduction in CpACS27A expression was concomitant with a downregulation of other ethylene biosynthesis and signaling genes during earlier and later stages of ovary development. The role of CpACS27A is discussed regarding the regulation of ethylene biosynthesis and signaling genes in the control of andromonoecy-associated traits, such as the delayed maturation of corolla and stigma as well as the parthenocarpic development of the fruit.