Immunocytochemical localization of nitrite reductase in green algae

The distribution of nitrite reductase (EC 1.7.7.1) in the green algae Chlamydomonas reinhardtii, Monoraphidium braunii, Chlorella fusca, and Scenedesmus obliquus was studied by immunoelectron microscopy. The labeling of ultrathin cryosections was performed with anti-nitrite reductase antibodies followed by gold-labeled goat anti-rabbit antibodies. In C. reinhardtii sections, gold label was mainly associated with the pyrenoid, tonoplast, and plasmalemma. Significant labeling was also detected in the thylakoid region. In all other organisms, label density was lower but distributed in the same locations, except that the plasmalemma of S. obliquus was not significantly labeled. From estimates of the relative volume of different cell regions, we found that approximately 80% of the total enzyme is located in the chloroplastic region (thylakoids plus pyrenoid) of C. reinhardtii, M. braunii, and C. fusca, and 97% in the case of S. obliquus.     

Multimodal magnetic resonance imaging of the central nervous system

The physiological and biochemical properties of the diseased brain that can be explored with magnetic resonance imaging (MRI) are increasing. Progress in MR-based technology affords a large panel of MRI sequences that explore different phenomena and, thus, provide complementary informations. The diagnostic accuracy of MRI is improved by the combination of all MR modalities. However, this abundance of data requires an efficient multiparametric analysis to fully achieve the goal of the multimodal strategy. We will discuss the potential impact of this advanced MRI analysis in the clinical management and the therapeutical strategies of the most common brain pathologies (intracranial tumors, multiple sclerosis, stroke, epilepsy and dementia). This non-invasive approach is of utmost importance since it already improves the diagnosis and the therapeutic choice in the management of several central nervous system diseases.     

Ultrastructural study and ontogenesis of the appendages and related musculature of Paraspadella (Chaetognatha)

A lineage of benthic chaetognaths has developed limb-like appendages on the caudal part of the body, resulting from a local modification of the lateral fins, which are folds of the epidermis and have a role in balance when swimming. The most complex are those of Paraspadella gotoi which are used as props with the tip of the tail, allowing an elaborated mating behaviour comprising different movements: complete erection of the body, swings and jumps, astonishing for so simple-bodied animals. In the tail, the epidermis and the connective tissue, together with the longitudinal musculature, are involved in this innovation. All the components of the fins, i.e. connective tissue, fin rays and multilayered epidermic cells are conserved, but their function has changed. The movements of appendages are adjusted by one pair of small appendicular muscles localised in the body wall, while posture movements of the body are allowed by four longitudinal bundles of raising muscle. These two new muscles have successively appeared in the evolutive series previously described in Paraspadella. They have definitely arisen from the secondary muscle: the two lateral bundles for the former, and the two dorsal and two ventral ones for the latter. All are supercontracting muscles, a muscle kind also observed in the other benthic genus Spadella, but unknown in planktonic and benthoplanktonic chaetognaths.     

Immunological evidence for the presence of plant homologues of the actin- related protein Arp3 in tobacco and maize: subcellular localization to actin-enriched pit fields and emerging root hairs

Summary. The actin-nucleating and -organizing Arp2/3 protein complex is well known to be conserved throughout the eukaryotic kingdom. For higher plants, however, only limited evidence is available for the presence of the Arp2/3 complex so far. Using heterologous antibodies against the Dictyostelium discoideum and Schizosaccharomyces pombe proteins and a bovine peptide, we found immunological evidence for the presence of Arp3 homologues in plants. First, proteins with a molecular mass of about 47–50 kDa were clearly recognized in extracts of both a dicotyledonous plant (tobacco) and a monocotyledonous plant (maize) in immunoblots with the anti-Arp3 antibodies. Second, immunolocalization with these Arp3 antibodies was performed on different plant cells, selected for their diverse actin organizations and functions. On isolated plasma membrane ghosts derived from tobacco leaf protoplasts, a putative Arp3 was localized along cortical actin filaments. In the inner cortex of maize roots, Arp3 was localized to actin-rich plasmodesmata and pit fields and to multivesicular bodies in the cytoplasm. During root hair formation, distinct site-specific localization was found at the protruding apical plasma membrane portions of these tip-growing cells.Correspondence and reprints: Department of Biology, Universitaire Instelling Antwerpen, Universiteitsplein 1, 2610 Wilrijk, Belgium.Received January 3, 2003; accepted February 7, 2003; published online August 26, 2003     

Mathematical analysis of competition between sensory ganglion cells for neurotrophic factor in the skin

A model is presented of competition between sensory axons for trophic molecules (e.g. a neurotrophin such as NGF), produced in a region of skin small enough to permit their free diffusion throughout it; e.g., a touch dome, or a vibrissal follicle hair sinus. The variables specified are the number of high affinity trophic factor receptors per axon terminal and the concentration of trophic factor in the extracellular space. Previous models of this class predicted the loss of all the axons innervating the region except the one requiring least trophic factor for its maintenance, even with high rates of trophic factor production. In the present model, we have imposed upper limits to axonal growth, thereby introducing new equilibria, and we show by a global analysis using LaSalle's theorem, and also by local analysis, that several axons can then coexist if the rate of production of trophic molecules is sufficiently high.     

Improvement of acetate production from lactose by growing Clostridium thermolacticum in mixed batch culture

AIMS: The objective of this study was to increase the acetate production by Clostridium thermolacticum growing on lactose, available as a renewable resource in the milk and whey permeate from the cheese industry. METHODS AND RESULTS: Experiments for increased acetate productivity by thermophilic anaerobes grown on lactose were carried out in batch cultures. Lactose at concentration of 30 mmol l(-1) (10 g l(-1)) was completely degraded by Cl. thermolacticum and growth rate was maximal. High concentrations of by-products, ethanol, lactate, hydrogen and carbon dioxide were generated. By using an efficient hydrogenotroph, Methanothermobacter thermoautotrophicus, in a defined thermophilic anaerobic consortium (58 degrees C) with Cl. thermolacticum and the acetogenic Moorella thermoautotrophica, the hydrogen partial pressure was dramatically lowered. As a consequence, by-products concentrations were significantly reduced and acetate production was increased. CONCLUSION: Through efficient in situ hydrogen scavenging in the consortium, the metabolic pattern was modified in favour of acetate production, at the expense of reduced by-products like ethanol. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of this thermophilic anaerobic consortium opens new opportunities for the efficient valorization of lactose, the main waste from the cheese industry, and production of calcium-magnesium acetate, an environmentally friendly road de-icer.     

Slight respiratory irritation but not inflammation in mice exposed to (1-->3)-beta-D-glucan aerosols

Airway irritation effects after single and repeated inhalation exposures to aerosols of beta-glucan (grifolan) were investigated in mice. In addition, the effects on serum total immunoglobulin E (IgE) production and histopathological inflammation in the respiratory tract were studied. The beta-glucan aerosols provoked slight sensory irritation in the airways, but the response was not concentration dependent at the levels studied. Slight pulmonary irritation was observed after repeated exposures. No effect was found on the serum total IgE levels, and no signs of inflammation were seen in the airways 6 h after the final exposure. The results suggest that, irrespective of previous fungal sensitization of the animals, inhaled beta-glucan may cause symptoms of respiratory tract irritation but without apparent inflammation. Respiratory tract irritation reported after inhalation of fungi may not be entirely attributed to beta-glucan.     

The complex relationship between soluble and insoluble tau in tauopathies revealed by efficient dephosphorylation and specific antibodies

Phosphorylated tau is deposited as insoluble inclusion bodies in the tauopathies. We have used a new efficient method to dephosphorylate tau extracted from control and tauopathy brain. In some tauopathies, including Alzheimer's disease and progressive supranuclear palsy, the pattern of insoluble tau isoforms reflected that of soluble tau. In contrast, in corticobasal degeneration, Pick's disease, and some forms of fronto-temporal dementia, specific tau isoforms were selectively sequestered into insoluble inclusion-forming tau. Therefore the overall expression of individual tau isoforms does not predict which tau isoforms are deposited in all tauopathies and different mechanisms must operate that result in the deposition of specific tau isoforms.     

Ca-dependence of diastolic properties of cardiac sarcomeres: involvement of titin

The stiffness of the sarcomeres was studied during the diastolic interval of 18 stimulated (0.5 Hz) cardiac trabeculae of rat (pH 7.4; temperature = 25°C). Sarcomere length (SL) and force (F) were measured using, respectively, laser diffraction techniques (resolution: 4 nm) and a silicon strain gauge (resolution: 0.63 μN). Sinusoidal perturbations (frequency = 500 Hz) were imposed to the length of the preparation. The stiffness was evaluated from the corresponding F and SL sinusoids by analysis of both signals together either in the time domain or in the frequency domain. A short burst (duration = 30 ms) of sinusoidal perturbations was repeated at 5 predetermined times during diastole providing 5 measurements of stiffness during the time interval separating two twitches. These measurements revealed that stiffness increases by 30% during diastole, while a simultaneous expansion of the sarcomeres (amplitude = 10-60 nm) was detected. Measurements of the fluorescence of fura-2 under the same conditions revealed a continuous exponential decline of [Ca²⁺]_i from 210 to 90 nM (constant of time 300 ms) during diastole. In order to test the possibility that the increase of sarcomere stiffness and the decline of [Ca²⁺]_i were coupled during diastole of intact trabeculae, we studied the effect of different free Ca²⁺-concentrations ([Ca²⁺]) between 1 and 430 nM on sarcomere stiffness in rat cardiac trabeculae skinned by saponin (n = 17). Stiffness was studied using 500 Hz sinusoidal perturbations of muscle length (ML). We found that, below 70 nM, the stiffness was independent of [Ca²⁺]; between 70 and 200 nM, the stiffness declined with increase of [Ca²⁺]; above 200 nM, the stiffness increased steeply with [Ca²⁺]. The data fitted accurately to the sum of two sigmoids (Hill functions): (1) at [Ca²⁺] < 200 nM the stiffness decreased with [Ca²⁺] (EC₅₀ = 160 ± 13 nM; n = −2.6±0.7) and (2) at [Ca²⁺] > 200 nM, stiffness increased with [Ca²⁺] (EC₅₀ = 3.4±0.3 μM; n = 2.1±0.2) due to attachment of cross-bridges. From these results, it was possible to reproduce accurately the time course of diastolic stiffness observed in intact trabeculae and to predict the effect on stiffness of a spontaneous elevation of the diastolic [Ca²⁺]. Identical stiffness measurements were performed in 4 skinned preparations exposed to a cloned fragment of titin (Ti I-II) which has been shown to exhibit a strong interaction with F-actin in vitro. It was anticipated that Ti I-II would compete with endogenous titin for the same binding site on actin in the I-band. Below 200 nM, Ti I-II (2 μM) eliminated the Ca²⁺-dependence of stiffness. These results are consistent with the hypothesis that the Ca²⁺-sensitivity of the sarcomeres at [Ca²⁺] < 200 nM, i.e. where the myocytes in intact muscle operate during diastole, involves an association between titin molecules and the thin filament.