Effetti del cobalto,nichel e cromo sulla germinazione di Alyssum,durante il ciclo di postmaturazione e invecchiamento

Abstract

Cobalt, nickel and chromium effects on germination of Alyssum, during after-ripening and aging. Cobalt (2.5–20mM), nickel (2.5–25mM) and chromium (0.2-1mM) effects have been studied in seeds of three species of Alyssum, of which two, A. bertolonii and A. argenteum, endemics to serpentine soils and one, A. nebrodense, closely related to the others but endemic to the Madonie (Sicily) where it grows on limestone.

The results, expressed as percentage of germinated seeds (p), Kotowski coefficient of velocity (v) and germination average ratio (pτ), have been subjected to statistical analysis.

The effect per mM is generally inhibitory, not significant for cobalt, weak but significant for nickel, more marked and significant for chromium, linearly correlated to concentration. The inhibition reaches its highest level when seeds are ripe, being significantly related to the control germination capacity (pτ).

The values of the specific inhibition (effect/mM modditppτ) for all the three species are higher for chromium, which is more toxic than nickel, and increase from A. nebrodense to A. argenteum, showing that of the two serpentine endemics A. bertolonii is more tolerant both to nickel and chromium.     

The long and the short cycle. Alternative intracellular routes for trafficking of G-protein-coupled receptors

The C terminus of the human V2 vasopressin receptor contains multiple phosphorylation sites including a cluster of amino acids that when phosphorylated prevents the return of the internalized receptor to the cell surface. To identify the step where the recycling process was interrupted, the trafficking of the V2 receptor was compared with that of the recycling V1a receptor after exposure to ligand. Initially, both receptors internalized in small peripheral endosomes, but a physical separation of their endocytic pathways was subsequently detected. The V1a receptor remained evenly distributed throughout the cytosol, whereas the V2 receptor accumulated in a large aggregation of vesicles in the proximity of the nucleus where it colocalized with the transferrin receptor and Rab11, a small GTP-binding protein that is concentrated in the perinuclear recycling compartment; only marginal colocalization of Rab11 with the V1a receptor was observed. Thus, the V2 receptor was sequestered in the perinuclear recycling compartment. Targeting to the perinuclear recycling compartment was determined by the receptor subtype and not by the inability to recycle, since the mutation S363A in the phosphorylation-dependent retention signal generated a V2 receptor that was recycled via the same compartment. The perinuclear recycling compartment was enriched in beta-arrestin after internalization of either wild type V2 receptor or its recycling mutant, indicating that long term interaction between the receptors and arrestin was not responsible for the intracellular retention. Thus, the fully phosphorylated retention domain overrides the natural tendency of the V2 receptor to recycle and, by preventing its exit from the perinuclear recycling compartment, interrupts its transit via the "long cycle." The data suggest that the inactivation of the domain, possibly by dephosphorylation, triggers the return of the receptor from the perinuclear compartment to the plasma membrane.     

Transglutaminase participates in the blockade of neurotransmitter release by tetanus toxin: evidence for a novel biological function

The aim of this study was to collect evidences on the role of transglutaminase (TG, E.C.2.3.2.13) in the antineoplastic properties exerted by nimesulide (NMS), a non-steroidal anti-inflammatory drug, on murine B16-F10 melanoma cells. Treatment of melanoma cells with nimesulide produces a considerable reduction of cell proliferation, paralleled by a remarkable decrease of the intracellular concentration of polyamines spermidine and spermine. NMS treatment induces cancer cell differentiation, likely through the observed enhancement of TG and tyrosinase activities and increase of melanin production, well known markers of melanocyte differentiation. The overall results highlight the possibility that nimesulide acts as antineoplastic agent likely through the induction of intracellular TG activity.     

An intracellular role for the C1q-globular domain

The C1q-globular domain (gC1qD) is a highly conserved oligomerization motif that distinguishes a superfamily of proteins, which includes circulating factors like C1q (the first component of the complement cascade) and adiponectin. The compact structure resulting from gC1qD trimerization is well known for its versatility in establishing highly specific interactions with different ligands. Among the many binding targets are a large number of extracellular membrane-associated receptors involved in cell development, apoptosis, and immunological processes. Interestingly, proteins interacting with the prototypical globular domain of C1q have been described also inside the cell where they were shown to recognize signal transducers such as G protein coupled receptors and their downstream effectors. Afterward, it was shown that variants of the gC1qD have been adopted by intracellular proteins involved in signal transduction. This review summarizes the evidence supporting the presence of the gC1qD inside the cell and explores the possibility that the domain might play novel signaling functions in this context, such as determining highly specific protein-protein interactions aimed to organize signaling complexes on the cytosolic side of cellular membranes.     

The melanosomal/lysosomal protein OA1 has properties of a G protein‐coupled receptor

The protein product of the ocular albinism type 1 gene, named OA1, is a pigment cell‐specific integral membrane glycoprotein, localized to melanosomes and lysosomes and possibly implicated in melanosome biogenesis. Although its function remains unknown, we previously showed that OA1 shares structural similarities with G protein‐coupled receptors (GPCRs). To ascertain the molecular function of OA1 and in particular its nature as a GPCR, we adopted a heterologous expression strategy commonly exploited to demonstrate GPCR‐mediated signaling in mammalian cells. Here we show that when expressed in COS7 cells OA1 displays a considerable and spontaneous capacity to activate heterotrimeric G proteins and the associated signaling cascade. In contrast, OA1 mutants carrying either a missense mutation or a small deletion in the third cytosolic loop lack this ability. Furthermore, OA1 is phosphorylated and interacts with arrestins, well‐established multifunctional adaptors of conformationally active GPCRs. In fact, OA1 colocalizes and coprecipitates with arrestins, which downregulate the signaling of OA1 by specifically reducing its expression levels. These findings indicate that heterologously expressed OA1 exhibits two fundamental properties of GPCRs, being capable to activate heterotrimeric G proteins and to functionally associate with arrestins, and provide proof of principle that OA1 can actually function as a canonical GPCR in mammalian cells.     

GIP,a G-protein-coupled receptor interacting protein

A novel protein was cloned while screening for partners interacting with the second intracellular loop of the V2 vasopressin receptor (V2R). The protein was named GIP as in G-protein-coupled receptor Interacting Protein; the corresponding gene was located on the 17th chromosome where three exons encode for a 379-amino-acid protein.GIP subcellular localization was studied by immunocytochemistry and also using a biotinylating agent. The protein was found to be localized, at least in part, on the plasma membrane, probably in the form of a trimer.The results indicated that GIP is a transmembrane protein and the most part of the molecule is intracellular. Sequence homology inferred that GIP cytosolic domain is folded as a collagen-like helix followed by a globular domain.The interaction of the globular domain with the V2R was confirmed by pull-down experiments indicating that this structural motif can also interact with cytosolic proteins.     

L'ipotesi dell'effetto del campo geomagnetico e la variabilità tra le ripetizioni in prove di germinazione ed accrescimento in Triticum

Abstract

The geomagnetic field effect hypothesis and the variability among replications in germination and growth tests in Triticum. – During previous eperiments on germination and growth of Triticum, we have often found many cases of heteroscedasticity and of statistically significant differences among contemporary replications (i.e. germinators). In such experiments all the caryopses were oriented in the same way inside the germinators, whilst the germinators were randomly set inside the thermostats: therefore the seeds of different germinators might result differently oriented. Some researchers report that seeds differently oriented according to the lines of force of the geomagnetic field, may respond with different germination, growth rates and growth directions of shoots and roots (magnetotropism). The present investigation was designed to ascertain if these effects might be responsible of the variability among the contemporary replications. The different caryopses orientation within laboratory geomagnetic field, does not give evidence of any effect on germination, growth rate and direction. The caryopses show no germination sensibility neither related to different age, light and temperature conditions, neither to different cultivars, neither to varying orientation from the geomagnetic North every 15° and to varying time of ? preorientation ? in dry conditions. Differences between the variability of randomly and North oriented subsamples are not observed. No statistically significant differences are observed among shoots and roots lenghts of seedlings from differently oriented caryopses neither in different environmental conditions, nor as function of the orientation (24 angles of 15°), nor after different preorientations, nor after repeating the test on 608 subsamples (12.000 seedlings). Therefore the variability among contemporary replications is not imputable to the geomagnetic field. The variability and the distributions structure analysis suggest that both the heteroscedasticity and the significant differences among the replications may be due to an interaction among the caryopses inside the germinators through the substrate. This is probably related to the cultivar characteristics. We emphasize that an accurate evaluation of the variability is particularly necessary in germination and growth tests.     

Osservazioni Preliminari Sulla Struttura Submicroscopica dei Granuli di Amido dell'endosperma Maturo di Triticum Durum Desf

Abstract

Preliminary observations on the submicroscopic structure of the endospermic starch granules in TRITICUM DURUM desf. (The electronic transparency of the starch granules). — The starch granules of Triticum durum Desf. kernels have been examined by electron microscopy.

The high degree of transparency of the granules appears to depend upon the regular disposition of the structural units, while their opacity probably is in relation with their orderless arrangement. The opacity of amorphous portions of the layers, of the median fissure and of the black bands seems to be in relation with the amorphous condition of such formations.

The black bands and the bubbles which have been considered respectively as protein layers and as the result of enzymes action, are only artifacts. The bubbles are interpreted as pyrodestrines.     

G protein-coupled receptor oligomerization provides the framework for signal discrimination

The idea that G protein-coupled receptors (GPCRs) may undergo homo- or hetero-oligomerization, although highly controversial up to a few years ago, has recently gained wide acceptance. The recognition that GPCRs may exhibit either dimeric or oligomeric structures is based upon a large body of biochemical and biophysical evidence. While much effort has been spent to demonstrate the mechanism(s) by which GPCRs interact with each other, the physiological relevance of this phenomenon remains rather elusive. GPCR oligomerization has been proposed to play a role in receptor ontogeny by either chaperoning protein folding or controlling trafficking to the cell surface. However, the acquisition of these roles does not rule out the possibility that oligomeric receptors may have additional functions, once they are brought to the cell surface. Herein, we propose that protein-protein as well as protein-lipid interactions may provide the structural basis for organizing distinct cell compartments along the plasma membrane where different extracellular signals may be perceived and discriminated.     

Serum/Plasma depletion with chicken immunoglobulin Y antibodies for proteomic analysis from multiple Mammalian species.

Plasma from different species is the most accessible and valuable source for biomarker discovery in clinical and animal samples. However, due to the high abundance of some proteins such as albumin and immunoglobulins, low-abundant proteins are often undetectable in proteomic analysis of plasma. We have established a plasma depletion scheme using chicken antibodies against various abundant proteins. This immunoaffinity purification procedure is able to deplete albumin across multiple species. The high binding capacity and specificity of the chicken antibody enables the efficient capture of its ligand from microliter volumes of plasma sample. The resulting two-dimensional gel analyses of the depleted and captured samples show significant enhancement of the low-abundant proteins and specific capture of the abundant ligand. By utilizing this sample preparation scheme, it is now possible to analyze the plasma proteome from multiple species in a potentially rapid and large-scale capacity for biomarker discovery, drug target discovery, and toxicology studies.