Accumulation of the labdane diterpene Marrubiin in glandular trichome cells along the ontogeny of Marrubium vulgare plants

The content of the diterpene Marrubiin was assessed by GC-FID in leaves of Marrubium vulgare plants along their ontogeny. Maximum accumulation occurred just before flowering time and in fully expanded leaves. After feeding the plants with radio labeled [³H]-geranyl geranyl diphosphate, up to 70% of the radioactivity was recovered in HPLC-Rt coincidental with authentic Marrubiin, which was also characterized by GC-EIMS, thus confirming that the biosynthesis of Marrubiin proceeds through the 1-deoxy-D-xylulose-5-phosphate pathway. The major accumulation of radioactivity occurred in glandular trichome cells, and the product remained stable throughout.     

Dormancy in peach (Prunus persica L.) flower buds

Morphological studies were carried out with peach flower buds collected monthly in 1989 and 1990, from two months before leaf fall (7 March) until two to three weeks before bloom (7/8 August). Chilled (2–4°C for 30 days) and unchilled buds were exposed to 20 to 25°C, 100% RH and continuous light. Gibberellin A₃ (3 ng or 30 ng) was applied to some of the non-chilled cuttings at three days intervals. Then, 12, 19, and 26 days after they were planted, the buds were sampled and processed for histological studies. Cultured flower buds (chilled or unchilled) had accelerated anther and gynoecium morphogenesis after 12 days under controlled conditions, compared to buds processed immediately after collection from the field. Chilling treatment augmented the bud culture effect, while Gibberellin A₃ applications to the excised buds retarded bud morphogenesis to a stage comparable to that of buds collected directly from the field. This, suggests that the comparatively high levels of Gibberellin A_1/3 we previously found in mid winter [15, 18] could be at least one of the factors that controls floral bud dormancy by retarding anther and gynoecium development.     

Phytoecological importance,mutual redundancy and phytological threshold values of certain climatic factors

A combination of methods (intensity of indication, floristic and mesological redundancy analysis, beta-diversity analysis, principal components analysis and Wildi's interactive ranking procedure) were used to evaluate redundancy and relative phytoecological importance among 80 climatic variables in Galicia (N.W. Spain). The information they contained was found to be adequately summarized by just 3 factors thought to play a major role in regulating the distribution of the species considered in the study area and similar areas: Baudiere's QE index, mean minimum temperature in the coldest month and mean temperature range in the coldest month. For these three factors, phytoclimatic thresholds were determined by examining beta-diversity and were used to define phytoclimatic zone types.

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Resumen Se valora la redundancia e importancia fitoecológica relative de 80 variables climáticas en Galicia (N.O. de España) empleando una combinacion de diferentes metodos (intensidad indicadora, análisis de la redundancia florística y mesológica, análisis de beta-diversidad, análisis de componentes principales y el método de ordenación interactiva de Wildi). La información contenida en esta variables es adecuadamente resumida por 3 factores que juegan un papel predominante en regular la distribución de las especies consideradas en el area de estudio y areas similares: el índice QE de Baudiere, la temperatura media de las mínimas del mes más frío y la oscilación térmica del mes más frío. Para estos factores el análisis de la beta-diversidad permitió determinar los umbrales fitoclimáticos utilizados en la definición y cartografía de las zonas fitoclimáticas.

     

Allosteric effects of monoclonal antibodies on human growth hormone

We have previously shown that a monoclonal antibody (MAb) recognizing the human growth hormone (hGH) antigenic domain left exposed after binding to lactogenic receptors enhanced hGH binding probably through allosteric effects on the hormone binding site. Since receptors displaying different specificities would not recognize exactly the same hGH region, we explored whether some of our MAb could affect hGH binding to somatogenic receptors from rabbit liver and to human liver hGH-specific receptors.The effect of MAbAE5, AC8 and F11 on hGH binding was measured by determining the formation of¹²⁵I-MAb:hGH:receptor complexes using two different experimental approaches. Results from procedure A, which involved the previous binding of the hormone to microsomes before adding¹²⁵I-MAb, indicated that the hGH domain defined by epitopes AE5, AC8 and F11 is uncovered in the various hormone:receptor complexes.Procedure B was devised to reveal any alteration in the hGH molecule induced by the MAb. In this case preformed¹²⁵I-MAb:hGH complexes were added to microsomes. Data showed that¹²⁵I-MAb AE5:hGH complexes bound better to the various receptors than¹²⁵I-MAb AE5 to hGH:receptor complexes. On the contrary, hGH previously bound to¹²⁵I-MAb AC8 or¹²⁵I-MAb F11 was less recognized by the receptors than the free hormone. Furthermore, binding of MAb AE5 or MAb F11 to hGH 20 K (a natural hGH variant lacking residues 32–46) also enhanced its affinity to the various receptors whereas MAb AC8 did not inhibit hGH 20 K binding.Results indicated that MAb recognizing the hGH antigenic area that remains unmasked after binding to different membrane-bound receptors are able to affect hormone binding site. MAb would induce either positive or negative allosteric changes in the hormone region involved in its binding to lactogenic, somatogenic and hGH-specific receptors.     

Environmental vibrio phage–bacteria interaction networks reflect the genetic structure of host populations

Phages depend on their bacterial hosts to replicate. The habitat, density and genetic diversity of host populations are therefore key factors in phage ecology, but our ability to explore their biology depends on the isolation of a diverse and representative collection of phages from different sources. Here, we compared two populations of marine bacterial hosts and their phages collected during a time series sampling program in an oyster farm. The population of Vibrio crassostreae, a species associated specifically to oysters, was genetically structured into clades of near clonal strains, leading to the isolation of closely related phages forming large modules in phage–bacterial infection networks. For Vibrio chagasii, which blooms in the water column, a lower number of closely related hosts and a higher diversity of isolated phages resulted in small modules in the phage–bacterial infection network. Over time, phage load was correlated with V. chagasii abundance, indicating a role of host blooms in driving phage abundance. Genetic experiments further demonstrated that these phage blooms can generate epigenetic and genetic variability that can counteract host defence systems. These results highlight the importance of considering both the environmental dynamics and the genetic structure of the host when interpreting phage–bacteria networks.     

Assessment of capybara (Hydrochoerus hydrochaeris) populations in the wetlands of Corrientes,Argentina

Ten sampling sites were selected to represent six distinct habitat types used by capybaras (clean lagoons, dirty lagoons, cutwaters, fens and marshes, gallery forests, and erosion ditches). The sites were sampled during winter (July and August); densities were expressed as number of capybaras per linear km of shoreline (C/LKS). The sites were classified as protected from poachers (P), under light hunting pressure (LHP), and under heavy hunting pressure (HHP). Clean protected (P) lagoons had three times as many capybaras as LHP ones (30.7 and 10.9 C/LKS, respectively), and thirty times those under HHP (1.0 C/LKS). Protected marshes and dirty lagoons had even higher capybara densities (52.5 and 50.0 C/LKS, respectively). Gallery forests under LHP had low densities (6.3 C/LKS), and protected cutwaters intermediate densities (27.5 C/LKS). Erosion ditches had exceptionally high densities (900 C/LKS), probably because cattle were fenced out, reducing forage competition. These densities, when converted to the standard unit area measurement (individuals/ha), were similar to those obtained by other researchers in the Brazilian Pantanal, and somewhat smaller than those in the Venezuelan Llanos. Mean number of capybaras per group remained relatively constant in all habitats (averages ranged between 9.2 and 11.8 individuals/group) but its coefficient of variation was much higher in LHP sites, probably because social structure was altered severely by hunting. The overall ratio of young to adults and juveniles was 1:7.4. In one of the sites, 13 of 34 groups (38.2%) were with young (average of 17 capybaras per group, 4.7 of which were young), confirming that this species can reproduce all year long.Requests for reprints should be sent to: Dr. J. Rabinovich.     

Effect of ovarian steroid hormones on superficial activator calcium in the rabbit uterus.

Effect of oestradiol (E2) and progesterone (P) on activator calcium binding was studied in the rabbit uterus. Superficial Ca-binding of isolated uterine strips was characterized by determining the rate of loss of isometric tension evoked by low (2.4 V/cm) field strength electrical stimulation in Ca-free Krebs solution. Intramuscular injection of 10 mg P increased superficial Ca-binding significantly in the postpartum and E2-treated virgin uterus with a latency period of 8--12 hours. Bilateral ovariectomy on the 25th day of pregnancy decreased superficial Ca-binding progressively, which could be avoided by P-substitution. 72 hours after ovariectomy P-treatment failed to increase Ca-binding. Local application of cycloheximide increased Ca-binding in the E2-treated virgin uterus. The results suggest that a high P-level plays an important role in the induction and maintenance of a strong binding of superficial activator calcium in the rabbit uterus. Progressive disappearance of the strong binding near term and after bilateral ovariectomy correlates well with P-withdrawl in this species.     

Calcium distribution and exchange in the pregnant and post partum rabbit uterus

Calcium content and distribution of the 25-day pregnant (PR) and post partum (PP) rabbit uterus was studied by atomic absorption spectrophotometry and 45Ca determination. Total Ca content [2.28 +/- 0.28 (PR) and 2.19 +/- 0.12 (PP) mM/kg wet wt] extracellular [1.21 +/- 0.09 (PR) and 1.25 +/- 0.11 (PP) mM/kg wet wt] cellular [1.07 +/- 0.08 (PR) and 0.94 +/- 0.09 (PP) mM/kg wet et], total exchangeable [1.86 +/- 0.11 (PR) and 1.84 +/- 0.09 (PR) mM/kg wet wt] and inexchangeable [0.43 +/- 0.05 (PR) and 0.35 +/- 0.04 (PP) mM/kg wet wt] Ca fractions were identical in the two extreme endocrinological conditions. In contrast compartment size and rate constant of different exchangeable Ca fractions determined by kinetic analysis of 45Ca desaturation "urves (curve-peeling tecnique and computer method), revealed significant differences between PR and PP uteri. Two exchangeable phases could be identified in both endocrinological states. The rate constants of both phases of efflux were significantly higher in the PP (alpha 1 = 0.173 +/- 0.02 min-1; alpha 2 = 0.023 +/- 0.001 min-1) than in the PR uterus (alpha 1 = 0.099 +/- 0.01 min-1; alpha 2 = 0.018 +/- 0.01 min-1). Compartment size of phase 1 (fast component) was significantly higher in the PR (1.13 +/- 0.1 mM/kg wet wt) than in the PP uterus (0.77 +/- 0.06 mM/kg wet wt). In contrast, compartment size of phase 2 (slow component) was significantly smaller in PR than in PP uterine strips (0.74 +/- 0.06 and 1.08 +/- 0.11 mM/kg wet wt). The last portion of desaturation curves represents efflux from one homogenous compartment. The present results suggest that endocrinological control of the rabbit myometrium is linked to the regulation of the binding of a superficial exchangeable Ca fraction.     

Possible role of ionic gradients in the apical growth of <Emphasis Type="Italic">Neurospora crassa</Emphasis>

The effects of the Ca²⁺/H⁺ exchanger A23187 and the K⁺/H⁺ exchanger nigericin on the growth of Neurospora crassa were analyzed. Both ionophores had the same effects on the fungus. They both inhibited growth in liquid media, apical extension being more affected than protein synthesis. A sudden challenge to either ionophore on solid media rapidly stopped hyphal extension. Additionally, both ionophores induced profuse mycelium branching and upward hyphal growth. Hyphae growing on nigericin-containing media also burst at the apex. Both ionophores caused a rapid inhibition in the apically-occurring synthesis of structural wall polysaccharides, but they did not affect mitochondrial energy conservation. With the use of DiBAC, a membrane-potential sensitive fluorophore, it was excluded that their effects were due to depletion of the plasma membrane potential. Considering that both ionophores exchange H⁺ for different metallic ions, we concluded that their effect was due to dissipation of a proton gradient, which is directly or indirectly involved in the apical growth of the fungus. This revised version was published online in June 2006 with corrections to the Cover Date.     

Rapid up-regulation of alpha4 integrin-mediated leukocyte adhesion by transforming growth factor-beta1

The alpha4 integrins (alpha4beta1 and alpha4beta7) are cell surface heterodimers expressed mostly on leukocytes that mediate cell-cell and cell-extracellular matrix adhesion. A characteristic feature of alpha4 integrins is that their adhesive activity can be subjected to rapid modulation during the process of cell migration. Herein, we show that transforming growth factor-beta1 (TGF-beta1) rapidly (0.5-5 min) and transiently up-regulated alpha4 integrin-dependent adhesion of different human leukocyte cell lines and human peripheral blood lymphocytes (PBLs) to their ligands vascular cell adhesion molecule-1 (VCAM-1) and connecting segment-1/fibronectin. In addition, TGF-beta1 enhanced the alpha4 integrin-mediated adhesion of PBLs to tumor necrosis factor-alpha-treated human umbilical vein endothelial cells, indicating the stimulation of alpha4beta1/VCAM-1 interaction. Although TGF-beta1 rapidly activated the small GTPase RhoA and the p38 mitogen-activated protein kinase, enhanced adhesion did not require activation of both signaling molecules. Instead, polymerization of actin cytoskeleton triggered by TGF-beta1 was necessary for alpha4 integrin-dependent up-regulated adhesion, and elevation of intracellular cAMP opposed this up-regulation. Moreover, TGF-beta1 further increased cell adhesion mediated by alpha4 integrins in response to the chemokine stromal cell-derived factor-1alpha. These data suggest that TGF-beta1 can potentially contribute to cell migration by dynamically regulating cell adhesion mediated by alpha4 integrins.