Cyclic AMP as the second hormonal messenger in the in vitro folliculogenesis of the foetal thyroid gland of the rat

Thyroid-stimulating hormone, the catecholamine isoproterenol, and prostaglandins E1 and E2, all substances known to increase cAMP concentration in thyroid tissue, accelerate the formation of follicular cavities in explanted thyroid of 15-day-old rat foetuses. Dibutyryl-cAMP added to the medium, but not sodium fluoride, also stimulates the folliculogenesis. Since fluoride stimulates membrane adenylate cyclase but does not increase the intracellular cAMP level, these results show that cAMP is involved as a second messenger in the activation of foetal thyroid morphogenesis induced by hormones. They indicate also that the thyroid gland of the foetal rat is capable of responding to hormonal stimulation as early as the 15th day of pregnancy; this implies that on day 15, the foetal thyroid possesses receptors not only for the thyroid-stimulating hormone, but also for catecholamines and prostaglandins.     

Interactive signalling by phytochromes and cryptochromes generates de-etiolation homeostasis in Arabidopsis thaliana

Single, double, triple and quadruple mutants of phyA, phyB, cry1 and cry2 were exposed to different sunlight irradiances and photoperiods to investigate the roll played by phytochrome A, phytochrome B, cryptochrome 1 and cryptochrome 2 during de-etiolation of Arabidopsis thaliana seedlings under natural radiation. Even the quadruple mutant retained some hypocotyl-growth inhibition by sunlight. Hypocotyl length was strongly affected by interactions among photoreceptors. Double phyA phyB, phyA cry1, and cry1 cry2 mutants were taller than expected from the additive action of single mutations. Some of these redundant interactions required the presence of phytochromes A and/or B. Interactions among photoreceptors resulted in a 44% reduction of the response to irradiance and a 70% reduction of the response to photoperiod. The complex network of interactions among photoreceptors is proposed to buffer de-etiolation against changes in irradiance and photoperiod, i.e light fluctuations not related to the positions of the shoot above or below soil level     

Hemocyanin of the chiton Acanthopleura granulata

The subunit structure and solution conformation of the hemocyanin of the chiton Acanthopleura granulata were investigated by light-scattering, ultracentrifugation, viscosity, absorbance, and circular dichroism methods. The molecular weight, determined by light scattering at pH 7.4 in the presence of 0.05 M Mg2+ and 0.01 M Ca2+, was (4.2 +/- 0.3) X 10(6), while those of dissociated subunits in the presence of 8.0 M urea (at pH 7.4) and at pH 10.7 were found to be 4.57 X 10(5) and 4.58 X 10(5), respectively. Circular dichroism and absorbance measurements at 222 and 346 nm indicate only minor changes in the conformation of the folded domains of the hemocyanin subunits in these dissociating solvents. As with the hemocyanins of the snails Busycon canaliculatum, Lunatia heros, and Littorina littorea, exposure to 4.0-6.0 M guanidinium chloride (GdmCl) is found to produce unfolding of the domains, resulting in much more pronounced spectral changes and a further drop in molecular weight. A Mw of 3.2 X 10(5) was obtained with Acanthopleura hemocyanin in 6.0 M GdmCl, suggesting hidden breaks in the polypeptide chains analogous to those observed with the gastropodan hemocyanins. Both urea and pH dissociation showed gradual declines in the molecular weights, consistent with a decamer-dimer-monomer scheme of subunit dissociation. The bell-shaped molecular weight profiles obtained in the pH region from 5 to 11 can be accounted for by assuming two proton-linked groups per dimer, characterized by apparent pK values of 5.5 and 9.5, and the further involvement of five to eight acidic and five to eight basic groups per monomer, having apparent pK values of 5.0 and 10.2.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of the 3''-leaving group on turnover of cephem antibiotics by a class C beta-lactamase.

1. We have previously shown that the capacity for specific binding of human 125I-labelled low-density lipoprotein (LDL) to rat hepatocytes increases with time in culture [Salter, Bugaut, Saxton, Fisher & Brindley (1987) Biochem. J. 247, 79-84]. 2. In the present study we show that this up-regulation is accompanied by a rise in the cholesterol ester content of the cells. 3. Inhibition of cholesterol esterification with the drug 58-035 (Sandoz) significantly decreases the time-dependent 'up-regulation' of LDL receptors. 4. Incubation of hepatocytes with LDL itself has little effect on subsequent LDL binding. However, when cholesterol esterification is inhibited, incubation with LDL decreases binding below that attained with the drug alone. 5. Inhibition of cholesterol synthesis with Lovastatin significantly increases LDL binding and antagonizes the effect of 58-035. 6. We conclude that in hepatocytes the rate of cellular cholesterol esterification can become the major determinant of LDL-receptor activity.     

A small-scale analysis of the spatial structure of a Posidonia oceanica meadow off the Island of Ischia (Gulf of Naples,Italy): Relationship with the seafloor morphology

Posidonia oceanica L. (Delile), an endemic species of the Mediterranean, forms extensive meadows which are continuously endangered by anthropic impacts. The availability of up-to-date information about interannual changes in shoot density of meadows and the knowledge of its expansion capabilities are crucial elements for the development of effective protection plans. Conversely, spatial ecology is becoming an increasingly important component of resource management, and the use of quantitative data for constructing prognosis maps of the dynamics of ecosystem degradation and restoration by nonlinear simulation methods is a topical field of landscape ecology. Unfortunately, little is known on spatial patterns of shoot density of P. oceanica on a small scale, despite their increasing use as indicators of the status and/or trends of meadows. The spatial structure of a continuous P. oceanica meadow, extending from 1 to 33 m depth in Lacco Ameno (Gulf of Naples, Italy), is investigated here by the “kriging” technique, a method widely used for geostatistical purposes. The analysis detected peculiar spatial patterns of shoot density and facilitated a small-scale (square meters) model of the distribution of P. oceanica. The highest shoot densities were found at the shallow stand (430 shoots m⁻², on the average, with a peak of 1000 shoots m⁻² in a relatively small area, at 1 m depth) and the lowest at the deep stand (average density <300 shoots m⁻² below 15 m depth). A high degree of patchiness was found in the shallow stand, down to 10 m depth. Nestlike patterns with a rounded shape, characterized by shoot density radially decreasing from the center, were demonstrated mainly in the shallow stand. An opposite trend was detected in one case, where the main nestlike pattern was characterized by a decrease of density towards the center. Nestlike patterns may be generated by the confluence and overlap of stolons expanding from proximal areas. The comparison of density and depth models indicated that the shape of the seafloor influences the density structure of meadows and the growth patterns of P. oceanica.     

Curcuma longa L. as a Therapeutic Agent in Intestinal Motility Disorders. 2: Safety Profile in Mouse

Background

Curcuma extract exerts a myorelaxant effect on the mouse intestine. In view of a possible use of curcuma extract in motor functional disorders of the gastrointestinal tract, a safety profile study has been carried out in the mouse.

Methods

Thirty mice were used to study the in vitro effect of curcuma on gallbladder, bladder, aorta and trachea smooth muscular layers and hearth inotropic and chronotropic activity. The myorelaxant effect on the intestine was also thoroughly investigated. Moreover, curcuma extract (200 mg/Kg/day) was orally administered to twenty mice over 28 days and serum liver and lipids parameters were evaluated. Serum, bile and liver bile acids qualitative and quantitative composition was were also studied.

Results

In the intestine, curcuma extract appeared as a not competitive inhibitor through cholinergic, histaminergic and serotoninergic receptors and showed spasmolytic effect on K⁺ induced contraction at the level of L type calcium channels. No side effect was observed on bladder, aorta, trachea and heart when we used a dose that is effective on the intestine. An increase in gallbladder tone and contraction was observed. Serum liver and lipids parameters were normal, while a slight increase in serum and liver bile acids concentration and a decrease in bile were observed.

Conclusions

Although these data are consistent with the safety of curcuma extract as far as its effect on the smooth muscular layers of different organs and on the heart, the mild cholestatic effect observed in absence of alteration of liver function tests must be further evaluated and the effective dose with minimal side effects considered.     

Genetic diversity among endangered Uruguayan populations of Butia Becc. species based on ISSR

The genus Butia Becc. (Arecaceae) comprises 18 species distributed exclusively in South America, four of them in Uruguay: Butia lallemantii, B. paraguayensis, B. yatay and B. capitata. All species show serious regeneration problems which jeopardise their continuity in the near future. Butia lallemantii and B. paraguayensis are in the most critical situation, with restricted distributions (fragmented and reduced by cattle grazing and forestation) and populations with few individuals. Taxonomy in the genus is complex, and species delimitation remains unclear. The aim of this study was to examine the genetic variability of natural populations of B. paraguayensis, B. lallemantii and B. yatay, based on inter?Csimple sequence repeat markers, in view of long term conservation plans and to assist the taxonomic resolution of these closely related species. Twenty individuals from each population were sampled including four populations of B. lallemantii, three of B. yatay and the only B. paraguayensis population reported for Uruguay. Syagrus romanzoffiana was used as outgroup. Five primers were selected based on amplification profiles. Relationships among species were evaluated by constructing dendrograms and principal coordinates analysis. Genetic distance analyses indicate the existence of low variability among Butia species. Variability within populations was high, possibly due to gene flow, past hybridisation or life history traits. This variability provides great potential for recovery. Results do not allow us to suggest changes to the present taxonomic status of these species. We propose management recommendations involving restoration of recruitment and augmentation via stocks from within populations.     

Highly sensitive detection of Staphylococcus aureus directly from patient blood

BackgroundRapid detection of bloodstream infections (BSIs) can be lifesaving. We investigated the sample processing and assay parameters necessary for highly-sensitive detection of bloodstream bacteria, using Staphylococcus aureus as a model pathogen and an automated fluidic sample processing – polymerase chain reaction (PCR) platform as a model diagnostic system.ConclusionsWe have demonstrated a highly sensitive two-hour assay for detection of sepsis causing bacteria like S. aureus directly in 1 ml of whole blood, without the need for blood culture.