Role of calcium during biosynthesis,secretion and organization of cell-wall polysaccharides

Abstract

The role of calcium during the synthesis, secretion and molecular organization of the primary cell-wall polysaccharides is the topic of this review. With the exception of callose synthase, the in vitro activity of all polysaccharide synthases is not controlled by Ca²⁺ ions. However, changes in the intracellular Ca²⁺ level could control the rate of exocytotic fusion of the secretory vesicles containing cell-wall matrix polysaccharides. In particular, the ability of Ca²⁺ to regulate the fusion of secretory vesicles with the plasma membrane is due to a class of Ca²⁺-dependent phospholipid-binding proteins known as annexins. The ionic interactions between calcium and the negatively charged homogalacturonan domains of the pectins are important not only for the mechanical properties of the wall but also for the gel-properties of these complex biopolymers.     

Pentylenetetrazol-Induced Epileptiform Activity Affects Basal Synaptic Transmission and Short-Term Plasticity in Monosynaptic Connections

Epileptic activity is generally induced in experimental models by local application of epileptogenic drugs, including pentylenetetrazol (PTZ), widely used on both vertebrate and invertebrate neurons. Despite the high prevalence of this neurological disorder and the extensive research on it, the cellular and molecular mechanisms underlying epileptogenesis still remain unclear. In this work, we examined PTZ-induced neuronal changes in Helix monosynaptic circuits formed in vitro, as a simpler experimental model to investigate the effects of epileptiform activity on both basal release and post-tetanic potentiation (PTP), a form of short-term plasticity. We observed a significant enhancement of basal synaptic strength, with kinetics resembling those of previously described use-dependent forms of plasticity, determined by changes in estimated quantal parameters, such as the readily releasable pool and the release probability. Moreover, these neurons exhibited a strong reduction in PTP expression and in its decay time constant, suggesting an impairment in the dynamic reorganization of synaptic vesicle pools following prolonged stimulation of synaptic transmission. In order to explain this imbalance, we determined whether epileptic activity is related to the phosphorylation level of synapsin, which is known to modulate synaptic plasticity. Using western blot and immunocytochemical staining we found a PTZ-dependent increase in synapsin phosphorylation at both PKA/CaMKI/IV and MAPK/Erk sites, both of which are important for modulating synaptic plasticity. Taken together, our findings suggest that prolonged epileptiform activity leads to an increase in the synapsin phosphorylation status, thereby contributing to an alteration of synaptic strength in both basal condition and tetanus-induced potentiation.     

The Footprint of Continental-Scale Ocean Currents on the Biogeography of Seaweeds

Explaining spatial patterns of biological organisation remains a central challenge for biogeographic studies. In marine systems, large-scale ocean currents can modify broad-scale biological patterns by simultaneously connecting environmental (e.g. temperature, salinity and nutrients) and biological (e.g. amounts and types of dispersed propagules) properties of adjacent and distant regions. For example, steep environmental gradients and highly variable, disrupted flow should lead to heterogeneity in regional communities and high species turnover. In this study, we investigated the possible imprint of the Leeuwin (LC) and East Australia (EAC) Currents on seaweed communities across ~7,000 km of coastline in temperate Australia. These currents flow poleward along the west and east coasts of Australia, respectively, but have markedly different characteristics. We tested the hypothesis that, regional seaweed communities show serial change in the direction of current flow and that, because the LC is characterised by a weaker temperature gradient and more un-interrupted along-shore flow compared to the EAC, then coasts influenced by the LC have less variable seaweed communities and lower species turnover across regions than the EAC. This hypothesis was supported. We suggest that this pattern is likely caused by a combination of seaweed temperature tolerances and current-driven dispersal. In conclusion, our findings support the idea that the characteristics of continental-scale currents can influence regional community organisation, and that the coupling of ocean currents and marine biological structure is a general feature that transcends taxa and spatial scales.     

Hidden diversity in high-latitude Southern Hemisphere environments: Reinstatement of the genus Rama and description of Vandenhoekia gen. nov. (Cladophoraceae,Ulvophyceae, Chlorophyta), two highly variable genera

The continental coasts and remote islands in the high-latitude Southern Hemisphere, including the subantarctic region, are characterized by many endemic species, high abundance of taxa, and intermediate levels of biodiversity. The macroalgal flora of these locations has received relatively little attention. Filamentous green algae are prolific in the intertidal of southern islands, but the taxonomy, distribution, and evolutionary history of these taxa are yet to be fully explored, mostly due to the difficulty of access to some of these locations. In this study, we examined specimens of the order Cladophorales from various locations in the high-latitude Southern Hemisphere including the subantarctic (the Auckland Islands, Bounty Islands, Campbell Island, Macquarie Island, and Kerguelen Islands), as well as mainland New Zealand, the Chatham Islands, Chile, and Tasmania. The analyses of the rDNA sequences of the samples revealed the existence of two new clades in a phylogeny of the Cladophoraceae. One of these clades is described as the novel genus Vandenhoekia gen. nov., which contains three species that are branched or unbranched. The amended genus Rama is reinstated to accommodate the other clade, and contains four species, including the Northern Hemisphere “Cladophora rupestris.” In Rama both branched and unbranched morphologies are found. It is remarkable that gross morphology is not a predictor for generic affiliations in these algae. This study illustrates that much can still be learned about diversity in the Cladophorales and highlights the importance of new collections, especially in novel locations.     

Compartmentation of newly synthesized phosphatidylethanolamine in rat brain microsomes

Summary The compartmentation of the phosphatidylethanolamine newly synthesized in brain microsomesin vitro either by base exchange or net synthesis has been studied, using difluorodinitrobenzene as a chemical probe. The experimental results demonstrate that in rat brain microsomes the phosphatidylethanolamine molecules synthesized by base exchange and the bulk membrane lipid belong to different pools. Ca²⁺ bound to microsomes seems to be involved in the maintenance of the compartmentation of phosphatidylethanolamine. In the presence of Ca²⁺ the newly synthesized phosphatidylethanolamine molecules react with difluorodinitrobenzene as though they are organized in clusters. After biosynthesisin vivo orin vitro through the cytidine pathway, the compartmentation of the newly formed phosphatidylethanolamine appears less marked than after the synthesis through base exchange.     

Midline (dermoid) cysts of the floor of the mouth: report of 16 cases and review of surgical techniques

A retrospective review of 16 cases of midline (dermoid) cysts of the floor of the mouth is presented, evaluating the different surgical approaches. Sixteen cases of patients with a diagnosis of midline cyst of the floor of the mouth, treated at the Maxillofacial Surgery Department of the School of Medicine and Surgery of the "Federico II" University of Naples (Naples, Italy), were observed over a 10-year period, between 1988 and 1998; age, sex, localization, diagnostic technique, and type of treatment were evaluated. Male patients were more frequently affected, with a male-to-female ratio of 3:1 (12:4 cases). Patients ranged in age from 5 to 51 years (average age, 27.8 years). The preoperative assessment was made using ultrasonography in all cases but one, computed tomography in eight cases, and magnetic resonance imaging in three cases. Regarding surgical techniques used, a transcutaneous approach was adopted for median geniohyoid cysts, an extended median glossotomy technique was used for very large median genioglossal cysts, a median glossotomy technique was used for median genioglossal cysts, and a midline incision of the oral mucosa along the lingual frenulum was used for sublingual cysts. During the postoperative course, there were no complications except for modest edema in three cases. Follow-up ranged between 24 months and 12 years; no relapses or malignant changes were observed. In the authors' experience, the intraoral approach was also effective for the treatment of large lesions and led to very good cosmetic and functional results, whereas the extraoral incision was necessary only when the cysts were under the geniohyoid muscle.     

Kinematic and electromyographic analysis of the Nordic Hamstring Exercise

The Nordic Hamstring Exercise (NHE) has been introduced as a training tool to improve the efficiency of eccentric hamstring muscle contraction. The aim of this study was to perform a biomechanical analysis of the NHE. Eighteen participants (20.4 ± 1.9 years) performed two sets of five repetitions each of the NHE and maximal eccentric voluntary contraction (MEVC) of the knee flexors on an isokinetic dynamometer whilst knee angular displacement and electrical activity (EMG) of biceps femoris were measured. EMG was on average higher during the NHE (134.3% of the MEVC). During the forward fall of the NHE, the angle at which a sharp increase in downward velocity occurred varied between 47.9 and 80.5 deg, while the peak knee angular velocity (pVelocity) varied between 47.7 and 132.8 deg s^?1. A significant negative correlation was found between pVelocity and peak EMG (r = ?0.62, p < 0.01) and EMG at 45 deg (r = ?0.75, p < 0.01) expressed as a percentage of peak MEVC EMG. Some of the variables analyzed exhibited good to excellent levels of intra- and inter-session reliability. This type of analysis could be used to indirectly monitor the level of eccentric strength of the hamstring muscles while performing the NHE and potentially any training- or injury-related changes.     

Preservation of capsular material of streptococcal cells by specific lectins determined by immunoelectron microscopy

Summary We describe the use of lectins as specific stabilizing agents for the polysaccharide capsular components of two Gram-positive bacteria,Streptococcus agalactiae andStreptococcus bovis. Treatment of bacterial suspensions with wheatgerm agglutinin and concanavalin A allowed better morphological preservation as well as immunoelectron microscopic localization of a capsular component (lipoteichoic acid) by employing specific antibodies and the protein A-gold technique. Data obtained indicate that lectins are useful agents in preserving highly water-soluble capsular components during the electron microscopy procedures for both unembedded and embedded samples.     

Structure of a novel class II phospholipase D: catalytic cleft is modified by a disulphide bridge

Phospholipases D (PLDs) are principally responsible for the local and systemic effects of Loxosceles envenomation including dermonecrosis and hemolysis. Despite their clinical relevance in loxoscelism, to date, only the SMase I from Loxosceles laeta, a class I member, has been structurally characterized. The crystal structure of a class II member from Loxosceles intermedia venom has been determined at 1.7 Å resolution. Structural comparison to the class I member showed that the presence of an additional disulphide bridge which links the catalytic loop to the flexible loop significantly changes the volume and shape of the catalytic cleft. An examination of the crystal structures of PLD homologues in the presence of low molecular weight compounds at their active sites suggests the existence of a ligand-dependent rotamer conformation of the highly conserved residue Trp230 (equivalent to Trp192 in the glycerophosphodiester phosphodiesterase from Thermus thermophofilus, PDB code: 1VD6) indicating its role in substrate binding in both enzymes. Sequence and structural analyses suggest that the reduced sphingomyelinase activity observed in some class IIb PLDs is probably due to point mutations which lead to a different substrate preference.