Supercharging accelerates T-tubule membrane potential changes in voltage clamped frog skeletal muscle fibers.

In voltage-clamp studies of single frog skeletal muscle fibers stained with the potentiometric indicator 1-(3-sulfonatopropyl)-4-[beta[2-(di-n-octylamino)-6-naphthyl] vinyl]pyridinium betaine (di-8 ANEPPS), fluorescence transients were recorded in response to both supercharging and step command pulses. Several illumination paradigms were utilized to study global and localized regions of the transverse tubule system (T-system). The rising phases of transients obtained from global illumination regions showed distinct accelerations when supercharging pulses were applied (95% of steady-state fluorescence achieved in 1.5 ms with supercharging pulses versus 14.6 ms with step pulses). When local transients were recorded at the edge of the muscle fiber, their kinetics resembled those of the applied waveform, but a similar relationship was not observed in transients from regions near the edge chosen to minimize the surface membrane contribution. We developed a model of the T-system capable of simulating membrane potential changes as a function of time and distance along the T-system cable and the associated fluorescence changes in regions corresponding to the experimental illumination strategies. A critical parameter was the access resistance term, for which values of 110-150 Omega.cm2 were adequate to fit the data. The results suggest that the primary mechanism through which supercharging pulses boost the kinetics of T-system voltage changes most likely involves their compensating the voltage attenuation across the access resistance at the mouth of the T-tubule.     

Phylogeny of the genus trichoderma based on sequence analysis of the internal transcribed spacer region 1 of the rDNA cluster

Sequences of the internal transcribed spacer region 1 (ITS1) of the ribosomal DNA were used to determine the phylogenetic relationships of species of Trichoderma sect. Pachybasium. To this end, 85 strains-including all the available ex-type strains-were analyzed. Parsimony analysis demonstrated that the section is nonmonophyletic, distributing the 85 strains among three main groups that were supported by bootstrap values. Group A comprises two clades (A1 and A2), with A1 including T. polysporum, T. piluliferum, and T. minutisporum, while A2 included T. hamatum, T. pubescens, and T. strigosum in addition to species previously included in sect. Trichoderma (i.e., T. viride, T. atroviride, and T. koningii). The ex-type strain of T. fasciculatum formed a separate branch basal to clade A. Clade B contained the sect. Pachybasium members T. harzianum, T. fertile, T. croceum, T. longipile, T. strictipile, T. tomentosum, T. oblongisporum, T. flavofuscum, T. spirale, and the anamorphs of Hypocrea semiorbis and H. cf. gelatinosa. Sequence differences among clades A1, A2, and B were in the same order of magnitude as between each of them and T. longibrachiatum, which was used as an outgroup in these analyses. Sequence differences within clades A1, A2, and B were considerably smaller: in some cases (i.e., T. virens and T. flavofuscum; T. strictipile and H. cf. gelatinosa), the ITS1-sequences were identical, suggesting conspecifity. In other cases (e.g., T. crassum and T. longipile; T. harzianum, T. inhamatum, T. croceum, T. fertile, and H. semiorbis; T. hamatum and T. pubescens; and T. viride, T. atroviride, and T. koningii) differences were in the range of 1-3 nt only, suggesting a very close phylogenetic relationship. The sequence of a previously described aggressive mushroom competitor group of T. harzianum strains (Th2) was strikingly different from that of the ex-type strain of T. harzianum and closely related species and is likely to be a separate species. Copyright 1998 Academic Press.     

Light quality affects morphology and polysaccharide yield and composition of Gelidium sesquipedale (Rhodophyceae)

Morphology and polysaccharide characterization of Gelidium sesquipedale (Clem.) Bornet et Thuret were studied in cultures grown under various light qualities. White light (WL), blue light (BL) and red light (RL) (all at photon fluence rate of 40 μmol m-2 s-1) were used for the study of morphological characteristics, and in addition yellow light (YL) for polysaccharide characterization. RL and BL induced a proliferating growth, which resulted in bushy plants under RL. Cortical cells of BL-grown plants were smaller and presented a higher density per unit area, whereas those of WL- and RL-grown alga were larger. Medullary cells followed the inverse pattern. Light quality also affected polysaccharide yield and composition, with the yield being higher under BL, RL or YL than WL. Most of the polysaccharide was extracted in distilled water at 100 °C, while a low amount was solubilized at 22 °C and 120 °C. Extracts from BL-grown alga presented the highest galactan content. The starch concentration was lower in extracts from RL-, BL- and YL-cultivated alga than in those from the initial plants. The degree of substitution with methoxyl groups and precursor was very low in all the agar fractions, but fractions extracted from BL- and WL-grown alga were more substituted by precursor. The highest sulfate content was reached under BL (about 9% w/w) and the highest 2-O-methyl-3,6-anhydro-L-galactose and 6-O-methyl-D-galactose content were found in extracts from alga grown under YL. This revised version was published online in June 2006 with corrections to the Cover Date.     

EFFECT OF ROCKY INTERTIDAL AMPHIPODS ON ALGAL RECRUITMENT: A FIELD STUDY1

Evidence about the potential of mobile marine invertebrates to act as algal spore dispersal agents is presently circumstantial. Using a field correlational and experimental protocol, our study tested the hypothesis that amphipods can increase the spore recruitment of the red alga Iridaea laminarioides Bory. Iridaea laminarioides spore recruitment onto glass slides was measured at a site with high amphipod abundance and a site with low density of amphipods. To evaluate the effect of an Ulva canopy on recruitment, replicated glass slides with and without a surrounding Ulva canopy were installed at both sites. The number of I. laminarioides spores recruited on the glass slides was four to eight times higher at the high amphipod abundance site than at the low density site. However, the presence of an Ulva canopy covering the glass slides did not significantly increase the recruitment of I. laminarioides. Because the abundance of I. laminarioides, the proportion of cystocarpic plants, and the percentage of open cystocarps only differed slightly between the low and high abundance amphipod sites, we suggest that the variation in recruitment between the sites is due to the differences in amphipod abundance (and their movements) and not to differences in spore production. Moreover, the presence of I. laminarioides cystocarps showing amphipod grazing scars was significantly higher at the high amphipod density site than at the low density site.     

Études histologiques et ultrastructurales des larves dePandemis heparana [Lep. Tortricidae] au cours des stades avancés d'une baculovirose due a un nouveau virus inducteur de diapause

Résumé Une polyédrose nucléaire de la tordeuse de la pelure,Pandemis heparana Denis & Schiffermuller (Lep.: Tortricidae) due à un baculovirus isolé dans un verger de pommiers en France est étudiée. Le baculovirus, uniquement actif sur les larves nouvelles nées, induit un arrêt de développement des larves au stade L3 et une mortalité ultérieue des larves aux stades L5 et L6. L'étude en microscopie optique et en microscopie électronique des larves L5 et L6 présentant des sympt?mes très prononcés de la viroise (début de paralysie et couleur blanchatre des segments abdominaux) montre que c'est essentiellement le tissu adipeux et accessoirement certains hémocytes et quelques cellules de l'hypoderme qui sont le siège de la multiplication virale. La morphogénèse virale dans les cellules du tissu adipeux est décrite en détail. Certains aspects de l'étude ultrastructurale liés à la forme en massue des virions, au processus de cristallisation des polyèdres et à l'accumulation de masses fibrillaires viro-induites sont discutés.

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Summary A nuclear polyhedrosis virus infecting the torticidPandemis heparana Denis & Schiffermuller, collected in an apple-orchard in France was isolated and studied. The baculovirus is active only in the newly hatched larvae. It induces a delayed arrest of development in the L3 larvae and subsequently the death of L5 and L6 larvae. Study by light and electron microscopy of L5 and L6 larvae exhibiting very marked symptoms of the virus disease (early stages of paralysis and whitish colour of abdominal segments) shows that the site of multiplication of the virus is mainly located in the fat body cells and occasionally in some hemocytes or hypoderm cells. Virus morphogenesis within the fat body cells is described in detail. Some aspects of the ultrastructure such as clubbed virions, process of crystallization of polyhedral bodies and clumps of fibrous material induced by the virus, are discussed.

     

Copper, copper mine tailings and their effect on marine algae in Northern Chile

Results are presented of a long-term research programme on the effect of copper contamination on biota in Chilean coastal waters. In spite of the magnitude of the copper mining tailings that affected Caleta Palito and surroundings in northern Chile, the effects on the intertidal assemblages remain restricted to a small geographic area. Even within the affected area, the effects are not homogeneous and there is evidence of active recovery in biological diversity in recent few years. Experimental evidence suggests that the current low algal diversity and abundance is strongly influenced by herbivory, although chronic effects of the discharges cannot be ruled out. Cellular changes in Enteromorpha compressa from the impacted area were characterised by abnormal granules in the cytoplasm, though these granules did not contain detectable levels of copper or other heavy metals. This revised version was published online in June 2006 with corrections to the Cover Date.     

Nonglucosylated oligosaccharides are transferred to protein in MI8-5 Chinese hamster ovary cells

A CHO mutant MI8-5 was found to synthesize Man9-GlcNAc2-P-P-dolichol rather than Glc3Man9GlcNAc2-P-P-dolichol as the oligosaccharide-lipid intermediate in N-glycosylation of proteins. MI8-5 cells were incubated with labeled mevalonate, and the prenol was found to be dolichol. The mannose-labeled oligosaccharide released from oligosaccharide-lipid of MI8-5 cells was analyzed by HPLC and alpha-mannosidase treatment, and the data were consistent with a structure of Man9GlcNAc2. In addition, MI8-5 cells did not incorporate radioactivity into oligosaccharide- lipid during an incubation with tritiated galactose, again consistent with MI8-5 cells synthesizing an unglucosylated oligosaccharide-lipid. MI8-5 cells had parental levels of glucosylphosphoryldolichol synthase activity. However, in two different assays, MI8-5 cells lacked dolichol- P-Glc:Man9GlcNAc2-P-P-dolichol glucosyltransferase activity. MI8-5 cells were found to synthesize glucosylated oligosaccharide after they were transfected with Saccharomyces cerevisiae ALG 6, the gene for dolichol-P-Glc:Man9GlcNAc2-P-P-dolichol glucosyltransferase. MI8-5 cells were found to incorporate mannose into protein 2-fold slower than parental cells and to approximately a 2-fold lesser extent.