Spider silkglands contain a tissue-specific alanine tRNA that accumulates in vitro in response to the stimulus for silk protein synthesis

The large ampullate glands of the orb-web spider, Nephila clavipes provide massive amounts of fibroin throughout the lifetime of the adult female. We have developed methods to culture the glands and manipulate their biosynthetic activity. This has allowed us to monitor a series of molecular events that precede silk production in glands excised from appropriately stimulated animals. In this paper, we demonstrate that prior to the transient dramatic production of fibroin, such glands accumulate large amounts of tRNAs cognate to the abundant amino acids in spider silk. One of these, alanine tRNA, appears to consist of two isoaccepting forms--one constitutive, and the other silkgland specific. Moreover, the silkgland-specific form appears to accumulate preferentially in response to stimulation. This phenomenon of tissue-specific tRNA production appears similar to that found in the silkglands of Bombyx mori, but the spider system has the unique property of permitting manipulation in vitro. Thus, it provides an unusual opportunity to study the mechanism of regulated tRNA synthesis.     

The effects of commonly used anaesthetics on colour measurements across body regions in the poeciliid fish,Girardinus metallicus

The effects of common anaesthetics on the hue, saturation and brightness measurements of the poeciliid fish Girardinus metallicus were investigated in two experiments. For both experiments the coloration of four body regions was measured from digital images of the same males obtained under three conditions: (1) control (in a water-filled chamber); (2) anaesthetised with MS-222; and (3) anaesthetised with eugenol (clove oil). In experiment 1 anaesthetised fish were photographed out of water. In experiment 2 all photographs were taken in a water-filled chamber. Anaesthetics altered coloration in both experiments. In the more methodologically consistent experiment 2 we found significantly different hue, increased saturation and decreased brightness in anaesthetic v. control conditions, consistent with darkening caused by the anaesthetics. The body regions differed in coloration consistent with countershading but did not differentially change in response to anaesthesia. These findings suggest that photographing fish in a water-filled chamber without anaesthetic is preferable for obtaining digital images for colour analysis and that multiple body regions of fish should be measured when assessing coloration patterns meaningful in behavioural contexts, to account for the gradients caused by countershading. We are encouraged that some researchers employ such methods already and caution against using anaesthetics except when absolutely necessary for immobilisation.     

Characterization of 10 trinucleotide microsatellite loci in the Critically Endangered Pyrenean yam Borderea chouardii (Dioscoreaceae)

The low levels of allozymic variability found in the Critically Endangered Borderea chouardii prompted us to develop microsatellite markers to assess the genetic variability and population structure for the adequate conservation management of this species. A (CTT)_n‐enriched partial genomic library was constructed. Ten polymorphic microsatellite loci were isolated from it, rendering 51 alleles in 47 individuals analysed. The allelic pattern observed for all of the loci with more than two alleles suggests that B. chouardii is tetraploid.     

Solution-phase combinatorial synthesis and evaluation of piperazine-2,5-dione derivatives

An efficient one-pot synthesis of a 61-membered combinatorial chemistry library of piperazine-2,5-diones was accomplished. Results of combinatorial synthesis, purification, analysis, and biological evaluation are described.     

Controls on mass loss and nitrogen dynamics of oak leaf litter along an urban-rural land-use gradient

Using reciprocal leaf litter transplants, we investigated the effects of contrasting environments (urban vs. rural) and intraspecific variations in oak leaf litter quality on mass loss rates and nitrogen (N) dynamics along an urban-rural gradient in the New York City metropolitan area. Differences in earthworm abundances and temperature had previously been documented in the stands along this gradient. Red oak leaf litter was collected and returned to its original source stand as native litter to measure decay rates along the gradient. To separate site effects from litter quality effects on decay, reciprocal transplants of litter were also made between stands at the extremes of the environmental gradient (urban and rural stands). Land-use had no effect on mass loss and N dynamics of native litter by the end of the 22-month incubation period. The lack of differences in native litter suggests the factors affecting decay were similar across the stands in this study. However, in the transplant study both environment and litter type strongly affected decay of oak leaf litter. On average urban and rural litter decomposed faster over the incubation period in urban than in rural stands (P=0.016 and P=0.001, respectively, repeated measures ANOVA). Differences in mass loss between urban and rural stands resulted in rural environments having less mass remaining than urban environments at the end of the incubation period (25.6 and 46.2% for urban and rural sites, respectively). Likewise, less N remained in leaf residue in urban sites (71.3%) compared to that in rural sites (115.1%). Litter type also affected mass loss rates during the 22-month incubation period. On average rural litter mass loss rates were faster than urban litter rates in both urban and rural stands (P=0.030 and P=0.026, respectively, repeated measures ANOVA). By the end of the incubation period, rural litter exhibited 43 and 20% greater mass loss and retained 44 and 5% less N than urban litter decomposing in the same urban and rural sites, respectively. These results suggest that different factors were controlling mass loss and N release rates along this urban-rural gradient. In urban stands, exotic earthworms and warmer temperatures may be compensating for what would otherwise be slowly decaying leaf litter because of its lower quality. Likewise, the lower quality litter produced in the urban stands may be decreasing the net release of N from litter despite higher temperatures and earthworm activity. Even though native litter decay rates were similar, the differential importance of the factors affecting decay along this gradient could alter the response of these forests to disturbance and variations in climate.     

Glutathione status and antioxidant enzymes in a crocodilian species from the swamps of the Brazilian Pantanal

In a previous study oxidative damage markers - lipid peroxidation and protein oxidation - were determined in organs of wild Caiman yacare captured in winter-2001 and summer-2002 at various developmental stages. An increase in oxidative damage occurred in the hatchling-juvenile transition (but not in the juvenile-adult transition) and winter-summer transition (in juveniles), suggesting that oxidative stress is associated with development and season. Herein the effect of development and season on glutathione (GSH) metabolism and the effect of development on the activity of antioxidant enzymes (catalase, glutathione peroxidase, glutathione reductase and glutathione S-transferase) and glucose 6-phosphate dehydrogenase were analyzed. The ratio GSSG:GSH-eq increased in lung, liver, kidney and brain by 1.8- to 4-fold in the embryo/hatchling to juvenile transition. No changes occurred in juvenile-adult transition. GSSG:GSH-eq across seasons was significantly elevated in summer. Total-glutathione content was mostly stable in various organs; in liver it increased in the embryo-juvenile transition. Enzyme activities were only determined in summer-animals (embryos, hatchlings and juveniles). For most antioxidant enzymes, activities increased from embryo/hatchling to juvenile in liver and Kidney. In lung, there was an inverse trend for enzyme activities and total glutathione content. Thus, increased metabolic rates during early caiman growth - in embryo-juvenile transition - appears to be related to redox imbalance as suggested by increased GSSG:GSH-eq and activation of antioxidant defenses. Differences in oxidative stress across seasons were related with summer-winter nocturnal temperatures. These results, as a whole, were interpreted in the context of ecological biochemistry.     

Electromyographic evaluation of masseter and anterior temporalis muscles in rest position of edentulous patients with temporomandibular disorders, before and after using complete dentures with sliding plates

Objectives: This study was performed with the purpose of investigating electromyographic (EMG) activity of the anterior temporalis and masseter muscles in edentulous individuals with temporomandibular disorder (TMD), before and after using sliding plates on complete dentures in the mandibular rest position. Background: Edentulous patients may present TMD, which is characterised by pain in temporomandibular joints, masticatory and neck muscles, uncoordinated and limited mandible movements, joint sounds and an altered occlusal relationship. It is imperative to offer treatment in order to re‐establish stomatognathic system structures before submitting the individual to any definitive restorative treatment. Materials and methods: The patients were edentulous for at least 10 years. EMG recordings were made before the insertion of the dentures (0 months) and also after using the sliding plates at the fourth month, 9th month and 12th month, using computerised electromyography K6‐I/ EMG Light Channel Surface. EMG evaluations of the muscles were performed under the following clinical conditions: rest position with dentures (R1), rest position without dentures (R2), rest position with dentures post‐activity (chewing) (R3), rest position without dentures post‐activity (chewing) (R4). Results: All patients obtained remission of muscular fatigue and reduced pain in stomatognathic system structures. Temporalis muscle showed significant increase in EMG activity compared with initial values (p < 0.01). Masseter muscles showed significantly lower mean values (p < 0.01) compared with initial values. Conclusion: The sliding plates allowed the process of neuromuscular deprogramming, contributing to muscular balance of the masticatory system, and are therefore indicated to be used before the fabrication of definitive complete dentures in patients with TMD.