Taxonomic review of the “Bulimes”, terrestrial gastropods from the continental Eocene of the Hamada de Méridja (northwestern Sahara,Algeria) (Mollusca: Stylommatophora: Strophocheilidae?), with a discussion of the genera of the family Vidaliellidae

Terrestrial gastropods occur in many North African localities in Eocene continental deposits. Here we analyse the faunal assemblage from the Hamada de Méridja Formation in southwestern Algeria, dated as Early to Middle Eocene on the basis of charophytes. The assemblage consists of three closely related species that to date have been classified either in the extant Madagascan genus Leucotaenius v. Martens, 1860, or in the SW European Eocene genera Romanella Jodot, 1957 and Vicentinia Jodot, 1957. This is rejected for shell morphological and phylogeographical reasons, and a new classification as Maghrebiola gen. nov. is proposed. Maghrebiola is tentatively placed in the South American family Strophocheilidae, as species from the Early Eocene Itaboraí Basin of Brazil, currently placed in the genus Eoborus Klappenbach and Olazarri, 1970 in the family Strophocheilidae, superfamily Acavoidea, have a very similar shell habitus. This record possibly extends the known geographical range of the Strophocheilidae into the African continent during the Eocene. Immigration of this stock into North Africa during the Cretaceous via a still existing plate connection is assumed. An attribution of Maghrebiola to the African family Achatinidae is unlikely for shell morphological reasons despite certain habitus similarities, although the Priabonian genera Arabicolaria and Pacaudiella from Oman most likely belong into this family, and not to the Vidaliellidae as originally proposed. Possible causes for the very low diversity of the assemblage are mainly unfavourable living conditions, i.e. a relatively dry climate resulting in sparse vegetation and only occasional presence of water bodies, which may have had increased salinities, accounting for the lack of freshwater mollusks. The absence of any competing large gastropods may possibly have facilitated high intraspecific variability leading to sympatric occurrence of three closely related species, due to the animals occupying a wide range of available ecological niches. As the species discussed here have also been attributed to the genera Romanella and Vicentinia in the Vidaliellidae, we provide an appendix with annotated characterisations of most genera of the Vidaliellidae and list the nominal species assigned to them. This family is tentatively placed in the South American superfamily Orthalicoidea; its stock would have similarly immigrated from South America, but have successfully colonized mainly SW Europe, with only one Eocene species [Romanella kantarensis (Jodot, 1936)] recognized in Algeria.     

LaPSvS1, a (1-->3)-beta-galactan sulfate and its effect on angiogenesis in vivo and in vitro

LaPSvS1, a highly sulfated branched (1-->3)-beta-galactan was prepared from the arabino-galactan from Larix decidua Miller by partial hydrolysis and subsequent sulfation with SO(3)-pyridine in DMF. The molecular weight was analyzed by GPC and the sulfate content was determined by ion chromatography. LaPSvS1 exhibited good antiangiogenic and antiinflammatory effects in two different modifications of the known CAM-assay. In vitro results obtained in the FGF-2-trypsin-assay and in fluorospectrometric experiments revealed that LaPSvS1 interacts with the fibroblast growth factor 2 system. This interaction is correlated with the in vivo effect of LaPSvS1 on FGF-2 induced angiogenesis.     

Nimodipine confers clinical improvement in two models of experimental autoimmune encephalomyelitis

Multiple sclerosis is characterised by inflammatory neurodegeneration, with axonal injury and neuronal cell death occurring in parallel to demyelination. Regarding the molecular mechanisms responsible for demyelination and axonopathy, energy failure, aberrant expression of ion channels and excitotoxicity have been suggested to lead to Ca²⁺ overload and subsequent activation of calcium‐dependent damage pathways. Thus, the inhibition of Ca²⁺ influx by pharmacological modulation of Ca²⁺ channels may represent a novel neuroprotective strategy in the treatment of secondary axonopathy. We therefore investigated the effects of the L‐type voltage‐gated calcium channel blocker nimodipine in two different models of mouse experimental autoimmune encephalomyelitis (EAE ), an established experimental paradigm for multiple sclerosis. We show that preventive application of nimodipine (10 mg/kg per day) starting on the day of induction had ameliorating effects on EAE in SJL /J mice immunised with encephalitic myelin peptide PLP _139–151, specifically in late‐stage disease. Furthermore, supporting these data, administration of nimodipine to MOG _35–55‐immunised C57BL /6 mice starting at the peak of pre‐established disease, also led to a significant decrease in disease score, indicating a protective effect on secondary CNS damage. Histological analysis confirmed that nimodipine attenuated demyelination, axonal loss and pathological axonal β‐amyloid precursor protein accumulation in the cerebellum and spinal cord in the chronic phase of disease. Of note, we observed no effects of nimodipine on the peripheral immune response in EAE mice with regard to distribution, antigen‐specific proliferation or activation patterns of lymphocytes. Taken together, our data suggest a CNS ‐specific effect of L‐type voltage‐gated calcium channel blockade to inflammation‐induced neurodegeneration.

     

Chemotaxis and nod Gene Activity of Bradyrhizobium japonicum in Response to Hydroxycinnamic Acids and Isoflavonoids

For Bradyrhizobium japonicum, the chemotactic and the nod gene-inducing effects of hydroxycinnamic acids and two of their derivatives were compared with those of isoflavonoids. Only the hydroxycinnamic acids were strong chemoattractants, while the other substances tested were chemotactically inactive. Besides the known nod gene induction by isoflavonoids, a weak nod gene induction by coniferyl alcohol, chlorogenic acid, and ferulic acid was found.     

Effects of temperature and photoperiod on flowering in Hyptis brevipes

Few tropical species have been tested for their flowering response under controlled conditions. Hyptis brevipes Poit, is an annual herb, commonly found in wet margins of streams and ponds, being considered a weed for some perennial plantations in Brazil. Under experimental glasshouse conditions, this species proved to be an obligate short-day plant. Flowering was delayed when photoperiods longer than 8 h were given, the critical photoperiod being between 12 and 13 h. When both temperature and photoperiod were controlled, at 20°C a longer protoperiod (by almost 1 h) is still inductive compared to 25 and 30°C. The number of short-day cycles required for full induction is relatively high and dependent upon temperature; at 20°C or above, 10 cycles are adequate, but at 15°C, more short-day cycles are needed. The number of inflorescences formed as well as the floral index vary according to daylength × temperature × inductive cycle number, allowing flowering to be assessed quantitatively. Long days are inhibitory to flowering, either suppressing it completely (when symmetrically intercalated among 24 inductive cycles) or preventing the floral index from increasing.     

Performance Evaluation of Kits for Bisulfite-Conversion of DNA from Tissues,Cell Lines,FFPE Tissues,Aspirates, Lavages,Effusions, Plasma,Serum, and Urine

DNA methylation analyses usually require a preceding bisulfite conversion of the DNA. The choice of an appropriate kit for a specific application should be based on the specific performance requirements with regard to the respective sample material. In this study, the performance of nine kits was evaluated: EpiTect Fast FFPE Bisulfite Kit, EpiTect Bisulfite Kit, EpiTect Fast DNA Bisulfite Kit (Qiagen), EZ DNA Methylation-Gold Kit, EZ DNA Methylation-Direct Kit, EZ DNA Methylation-Lightning Kit (Zymo Research), innuCONVERT Bisulfite All-In-One Kit, innuCONVERT Bisulfite Basic Kit, innuCONVERT Bisulfite Body Fluids Kit (Analytik Jena). The kit performance was compared with regard to DNA yield, DNA degradation, DNA purity, conversion efficiency, stability and handling using qPCR, UV, clone sequencing, HPLC, and agarose gel electrophoresis. All kits yielded highly pure DNA suitable for PCR analyses without PCR inhibition. Significantly higher yields were obtained when using the EZ DNA Methylation-Gold Kit and the innuCONVERT Bisulfite kits. Conversion efficiency ranged from 98.7% (EpiTect Bisulfite Kit) to 99.9% (EZ DNA Methylation-Direct Kit). The inappropriate conversion of methylated cytosines to thymines varied between 0.9% (innuCONVERT Bisulfite kits) and 2.7% (EZ DNA Methylation-Direct Kit). Time-to-result ranged from 131 min (innuCONVERT kits) to 402 min (EpiTect Bisulfite Kit). Hands-on-time was between 66 min (EZ DNA Methylation-Lightning Kit) and 104 min (EpiTect Fast FFPE and Fast DNA Bisulfite kits). Highest yields from formalin-fixed and paraffin-embedded (FFPE) tissue sections without prior extraction were obtained using the innuCONVERT Bisulfite All-In-One Kit while the EZ DNA Methylation-Direct Kit yielded DNA with only low PCR-amplifiability. The innuCONVERT Bisulfite All-In-One Kit exhibited the highest versatility regarding different input sample materials (extracted DNA, tissue, FFPE tissue, cell lines, urine sediment, and cellular fractions of bronchial aspirates, pleural effusions, ascites). The innuCONVERT Bisulfite Body Fluids Kit allowed for the analysis of 3 ml plasma, serum, ascites, pleural effusions and urine.     

Isolation of root hairs from seedlings of Pisum sativum. Identification of root hair specific proteins by in situ labeling

A procedure was developed which allows the large-scale isolation of root hairs from seedlings of Pisum sativum . L. cvs. Kleine Rheinländerin and Rosa Krone. The method may yield up to 50 g fresh weight of root hairs per 3.10⁴ seedlings. In a modified form considerable amounts of root hair material may be harvested, even after incubation of the roots in aqueous solutions. Thus, detailed biochemical studies on the root hair system have become feasible.
The occurrence of specific proteins in membrane fractions of P. sativum root hairs was demonstrated as follows: Incubation of root hairs in situ with 3-azidonaphthalene-2,7-disulfonate – a strongly anionic, photoactivated fluorescent marker – followed by gel electrophoresis of membrane fractions showed the presence of root-hair specific proteins which, since the system was intact, suggests that they are on the outer surface of the cells.     

A sequential analysis of the development of the synaptonemal complex in spermatocytes of the mouse by electron microscopy using hydroxyurea and agar filtration

A method is presented for sequential analysis of the development and behaviour of the Synaptonemal Complex (SC) in primary spermatocytes of male mice, using agar filtration for electron microscope grid preparation. The mice were treated with hydroxyurea (HU) to produce a gap in the spermatogenic line. The front of surviving cells behind the gap was examined day by day. The first visible parts of unpaired axial elements, with some barely recognizable paired regions were found 9 days after the last HU injection i.e. directly after the last S-phase before meiosis. During mid zygotene and late zygotene the axes of the autosomes had a fuzzy ill-defined appearance with irregular regions of apparent thickening. The axes of the XY pair could be recognized only at late zygotene. During pachytene the SCs of the autosomal pairs did not show a significant change except for a slight increase in size of the attachment points of the axial elements. On the first day of pachytene the axes of the XY pair appeared thin and long. On the second day the axes of the XY pair showed maximal pairing of about 50% of the axis of the Y chromosome. From the third to the fifth day a decrease of the paired region of the sex chromosomes was found together with an increase in thickness of the axes, which reached its maximum on the fourth day. Diplotene could be easily recognized: the autosomal axes showed a sharp, well-defined outline with thick attachment points with deltoid structure, and desynapsis was very clear. The axes of the XY pair showed variation during diplotene but on the third day of diplotene a characteristic bulging could be seen. The axes of the autosomes disappeared at this time and in most cases only the attachment points remained visible. The duration of the prophase classes of meiosis I was found to be: zygotene approximately 2 days; pachytene a little more than 5 days and diplotene approximately 3 days. Leptotene could not be traced by the method used. If it exists at all, it must be a stage of very short duration.     

Can root electrical capacitance be used to predict root mass in soil?

Background

Electrical capacitance, measured between an electrode inserted at the base of a plant and an electrode in the rooting substrate, is often linearly correlated with root mass. Electrical capacitance has often been used as an assay for root mass, and is conventionally interpreted using an electrical model in which roots behave as cylindrical capacitors wired in parallel. Recent experiments in hydroponics show that this interpretation is incorrect and a new model has been proposed. Here, the new model is tested in solid substrates.

Methods

The capacitances of compost and soil were determined as a function of water content, and the capacitances of cereal plants growing in sand or potting compost in the glasshouse, or in the field, were measured under contrasting irrigation regimes.

Key Results

Capacitances of compost and soil increased with increasing water content. At water contents approaching field capacity, compost and soil had capacitances at least an order of magnitude greater than those of plant tissues. For plants growing in solid substrates, wetting the substrate locally around the stem base was both necessary and sufficient to record maximum capacitance, which was correlated with stem cross-sectional area: capacitance of excised stem tissue equalled that of the plant in wet soil. Capacitance measured between two electrodes could be modelled as an electrical circuit in which component capacitors (plant tissue or rooting substrate) are wired in series.

Conclusions

The results were consistent with the new physical interpretation of plant capacitance. Substrate capacitance and plant capacitance combine according to standard physical laws. For plants growing in wet substrate, the capacitance measured is largely determined by the tissue between the surface of the substrate and the electrode attached to the plant. Whilst the measured capacitance can, in some circumstances, be correlated with root mass, it is not a direct assay of root mass.