In Vivo Substitution of Choline for Sodium Evokes a Selective Osmoinsensitive Increase of Extracellular Taurine in the Rat Hippocampus

Recent investigations have demonstrated that taurine and phosphoethanolamine (PEA) are the amino acids most sensitive to microdialysis-perfusion with reduced concentrations of NaCl. The aim of the present work was to assess the importance of Na+ deficiency in evoking this response. Further, the previously described selectivity of replacement of Cl- with acetate with respect to amino acid release was reinvestigated. The hippocampus of urethane-anesthetized rats was dialyzed with Krebs-Ringer bicarbonate buffer, and amino acid concentrations of the perfusate were determined. Choline chloride was then stepwise substituted for NaCl, and, in some cases, mannitol (122 mM) was included in low sodium-containing media. In other experiments, NaCl was replaced with sodium acetate. The dialysate levels of taurine increased selectively in response to Na+ substitution. The elevation of taurine was linearly related to the increase in choline chloride, and maximal levels amounted to 335% of basal levels. The increase in extracellular taurine was not inhibited by perfusion with medium made hyperosmotic with mannitol. Replacement of Cl- with acetate stimulated the release of taurine to 652% of resting levels. In addition, PEA levels increased to 250% of control concentration. Other amino acids were unaffected by Cl- substitution. The results show that taurine transport is considerably more sensitive to Na+ depletion than glutamate transport, which also is known to be Na+ dependent. The taurine increase evoked by low Na+ is not caused by cellular swelling as it was unaffected by hyperosmolar medium. Finally, substitution of acetate for Cl- causes a specific elevation of extracellular taurine and PEA, possibly as a result of cytotoxic edema.     

Simulation of the dynamics in the Baker's yeast process

Simulation of the dynamics in a fed batch process for production of Baker's yeast is discussed and applied. Experimental evidences are presented for a model of the energy metabolism. The model involves the concept of a maximum respiratory capacity of the cell. If the sugar concentration is increased above a critical value, corresponding to a critical rate of glycolysis and a maximum rate of respiration, then all additional sugar consumed at higher sugar concentrations is converted into ethanol.In a fed batch process with constant sugar feed the sugar concentration declines slowly. If ethanol is present when the sugar concentration declines below the critical value of 110 mg/dm³ fructose +glucose the metabolism switches rapidly into combined oxidation of sugar and ethanol. Thus, no diauxic growth is involved under process conditions. The rate of ethanol consumption is determined by the free capacity of respiration under these conditions. The invertase activity of the cells was found to be so high that mainly fructose and glucose were present in the medium, typically in the concentration range around 100 mg/dm³. These components are consumed at the same rate but with fructose at a higher concentration, indicating a higher K _s for fructose consumption.The model was used in simulation experiments to demonstrate the dynamics of the Baker's yeast process and the influence of different process conditions.List of Symbols DOT % air sat dissolved oxygen tension - F dm³/h rate of inlet medium flow - H kg/(dm³ % air sat.) oxygen solubility - K kg/m³ saturation constant specified by index - K _L a 1/h volumetric oxygen transfer coefficient - m g/(g · h) maintenance coefficient specified by index - P kg/(m³ · h) mean productivity of biomass in the process - q g/(g · h) specific consumption or production rate - S kg/m³ concentration of sugar in reactor - S ₀ kg/m³ concentration of inlet medium sugar medium t h process time - V dm³ medium volume - X kg/m³ concentration of biomass - Y g/g yield coefficient specified by index - 1/h specific growth rate Index aa anaerobic condition - c critical value - e ethanol - ec ethanol consumption - ep ethanol production - max maximum value - o oxygen - oe oxygen for growth on ethanol - os oxygen for growth on sugar - s sugar - x biomass     

Expression of two heterologous promoters, Agrobacterium rhizogenes rolC and cauliflower mosaic virus 35S, in the stem of transgenic hybrid aspen plants during the annual cycle of growth and dormancy

We monitored, for the first time, the activity of two model heterologous promoters, the Agrobacterium rhizogenes rolC and the cauliflower mosaic virus (CaMV) 35S, throughout the annual cycle of growth and dormancy in a perennial species, hybrid aspen. Each promoter was fused to the uidA -glucuronidase (GUS) reporter gene and the constructs were introduced into the hybrid aspen genome by Agrobacterium-mediated transformation. Both wildtype and transgenic plants were cultivated under different regimes of photoperiod and temperature to induce passage through one growth-dormancy-reactivation cycle, and at intervals GUS staining was assessed in stem sections. In rolC::uidA transformants, GUS activity in rapidly growing current-year shoots was not only tissue-specific, being localized to the phloem, but also cell-specific at the shoot base, where it was present only in the companion cells. However, during the onset of dormancy induced by short photoperiod, GUS activity shifted laterally from the phloem to include the cortex and pith. After subsequent exposure to chilling temperatures to induce the transition between the dormancy stages of rest and quiescence, GUS activity almost disappeared from all stem tissues, but regained its original phloem specificity and intensity after the shoots were reactivated by exposing them to long photoperiod and high temperatures. In contrast, GUS activity in the stem of 35S::uidA transformants was strong in all tissues except for the vascular cambium and xylem, and did not vary in intensity during the growth-dormancy-reactivation cycle. The lateral shift and increased intensity of GUS activity in the stem of rolC::uidA transformants during dormancy induction was shown to be associated with the accumulation of starch, and to be mimicked by incubating stem sections in sucrose, as well as glucose and fructose, but not sorbitol, prior to the GUS assay. Our results demonstrate that the activities of the rolC and 35S promoters varied in very different, unpredictable ways during the annual cycle of growth and dormancy in a perennial species, and indicate that the spatial and temporal variation in rolC promoter activity that we observed in the stem of transgenic hybrid aspen plants is attributable to cellular and seasonal changes in sucrose content.     

Distribution of PACAP (pituitary adenylate cyclase-activating polypeptide)-like and helospectin-like peptides in the teleost gut

Pituitary adenylate cyclase-activating polypeptide (PACAP) and helospectin are two vasoactive intestinal polypeptide (VIP)-related neuropeptides that have recently been demonstrated in the mammalian gut; the aim of this study was to reveal their occurrence and localisation in the gastrointestinal tract, swimbladder, urinary bladder and the vagal innervation of the gut of teleosts, using immunohistochemical methods on whole-mounts and sections of these tissues from the Atlantic cod, Gadus morhua and the rainbow trout, Oncorhynchus mykiss. Both PACAP-like and helospectin-like peptides were present in the gut wall of the two species. Immunoreactive nerve fibres were found in all layers but were most frequent in the myenteric plexus and along the circular muscle fibres. Immunoreactivity was also demonstrated in nerves innervating the swimbladder wall, the urinary bladder and blood vessels to the gut. Immunoreactive nerve cell bodies were found in the myenteric plexus of the gut and in the muscularis mucosae of the swimbladder. In the vagus nerve, non-immunoreactive nerve cells were surrounded by PACAP-immunoreactive fibres. Double staining revealed the coexistence of PACAP-like and helospectin-like peptides with VIP in all visualized nerve fibres and in some endocrine cells. It is concluded that PACAP-like and helospectin-like peptides coexist with VIP in nerves innervating the gut of two teleost species. The distribution suggests that both PACAP and helospectin, like VIP, are involved in the control of gut motility and secretion.     

Factors controlling the population dynamics of the clonal helophyte Ranunculus lingua

Abstract. Two marginal and two central populations of the pseudo-annual aquatic plant Ranunculus lingua were studied over four years. The main purpose was to quantify potentially influential abiotic and biotic factors and to derive predictions about life-history differences between the populations. Variation in abundance and height of R. lingua ramets at different depths were related to water-level fluctuations, to abundance of other helophyes (emergent macrophytes), and to the occurrence of invertebrate grazing and fungal pathogens. Clear differences between marginal and central populations were shown in the depth distribution of ramet numbers and ramet heights, as well as in the dynamic patterns, where marginal populations had a higher flux of ramets. These patterns and regression analyses indicated that abiotic factors have a greater influence in marginal populations, whereas biotic factors are more important in central populations. It is suggested that marginal habitats for R. lingua would favour life-histories with a high reproductive capacity, whereas a large size of ramet would be the most important life-history feature in central habitats. This was supported by the fact that ramets in marginal populations, in spite of their smaller size, produced higher number of rhizomes than ramets in central populations. Variation in regional abundance was finally related to differences in demographic processes and dispersal potential between the populations.     

A new species of Croton (Euphorbiaceae) from Somalia

Croton crispatus , sp. nov., from Acacia-Commiphora bushland on sand in central Somalia, is described and illustrated.     

Acacia ochracea (Leguminosae-Mimosoideae), a new species from Somalia

Acacia ochracea , a new species in the A. Senegal complex, is described and illustrated. It is a common and conspicuous tree over large areas in the western Bay Region and in the Gedo Region in SW Somalia.     

New or noteworthy species of Leguminosae in NE tropical Africa

The following new species are described: Chamaecrista dunensis (Somalia), Acacia densispina (Somalia), A. flagellaris (Somalia), Tephrosia karkarensis (Somalia), T. scopulata (Somalia), Indigofera ancistrocarpa (Kenya), I. curvirostrata (Ethiopia), I. hiranensis (Somalia), I. karinensis (Somalia), I. kucharii (Somalia), I. nugalensis (Somalia), I. pellucida (Kenya, Somalia), I. sabulosa (Ethiopia, Somalia), I. sessilis (Somalia), I. ienuirostris (Somalia), Rhynchosia megalocalyx (Somalia), Crotalaria allophylla (Somalia), C. intricata (Somalia), C. warfae (Somalia), and Coronilla somalensis (Somalia). 2n = 22 is reported in Tephrosia karkarensis. Cassia somalensis, Acacia puccioniana, Sophora somalensis , and Crotalaria sennii are reduced to synonyms of respectively Senna hookeriana, Dichrostachys kirkii, Millettia usaramensis and Crotalaria boranica subsp. trichocarpa . Thirty-seven taxa are reported from Somalia for the first time, including Ptycholobium biflorum , known previously only from Southern Africa. Crotalaria pieropoda , known previously only from Socotra, is reported from Somalia and Oman. Neptunia oleracea and Vatovaea pseudolablab are new generic records for respectively Ethiopia and Oman. The fruits of Indigofera boranica are described. Vigna somaliensis and an allied species are discussed. Rhynchosia gansole is lectotypified and its flowers and fruits are described.     

Physiological effects of translation initiation factor IF3 and ribosomal protein L20 limitation inEscherichia coli

To investigate the physiological roles of translation initiation factor IF3 and ribosomal protein L20 inEscherichia coli, theinfC, rpmI andrpIT genes encoding IF3, L35 and L20, respectively, were placed under the control oflac promoter/operator sequences. Thus, their expression is dependent upon the amount of inducer isopropyl thiogalactoside (IPTG) in the medium. Lysogenic strains were constructed with recombinant lambda phages that express eitherrpmI andrplT orinfC andrpmI in trans, thereby allowing depletion of only IF3 or L20 at low IPTG concentrations. At low IPTG concentrations in the IF3-limited strain, the cellular concentration of IF3, but not L20, decreases and the growth rate slows. Furthermore, ribosomes run off polysomes, indicating that IF3 functions during the initiation phase of protein synthesis in vivo. During slow growth, the ratio of RNA to protein increases rather than decreases as occurs with control strains, indicating that IF3 limitation disrupts feedback inhibition of rRNA synthesis. As IF3 levels drop, expression from an AUU-infC-lacZ fusion increases, whereas expression decreases from an AUG-infC-lacZ fusion, thereby confirming the model of autogenous regulation ofinfC. The effects of L20 limitation are similar; cells grown in low concentrations of IPTG exhibited a decrease in the rate of growth, a decrease in cellular L20 concentration, no change in IF3 concentration, and a small increase in the ratio of RNA to protein. In addition, a decrease in 50S subunits and the appearance of an aberrant ribosome peak at approximately 41–43S is seen. Previous studies have shown that the L20 protein negatively controls its own gene expression. Reduction of the cellular concentration of L20 derepresses the expression of anrplT-lacZ gene fusion, thus confirming autogenous regulation by L20.     

β-Sheet Models for the Ordered Filamentous Structure Formed by a Peptide That Enhances the Action of Insulin

Certain peptides with sequences related to part of the major histocompatibility complex class I antigen enhance the action of insulin. These peptides also aggregate into fibrous structures that seem to be related to their biological activity. In the current study, the 17-residue peptide with amino acid sequence Gly-Asn-Glu-Gln-Ser-Phe-Arg-Val-Asp-Leu-Arg-Thr-Leu-Leu-Arg-Tyr-Ala is used as a representative example of these bioactive molecules. As seen by electron microscopy, the peptide associates into gently twisted ribbons, 50 Å thick, in which the amount of twist decreases as the ribbons become wider. X-ray diffraction analysis suggests that the peptides are arranged as in an antiparallel β-sheet extending essentially endlessly along the fiber axis. The amino acid sequence of the peptide is such that one side of the β-sheet is predominantly polar while the opposite side is nonpolar. This allows the β-sheets to form multilayers with alternating hydrophobic and hydrophilic interfaces. The length of the extended peptide (≈54 Å) determines the thickness of the ribbon and the tendency of individual β-sheets to twist accounts for the twisting of the ribbons. An alternative model is also discussed, again based on antiparallel β-sheets, but with adjacent sheets interdigitated in a “side-by-side” fashion rather than forming stacked layers. Comparable inactive peptides such as Gly-Asn-Glu-Gln-Ser- A_l_a_-Arg-Val-Asp-Leu-Arg-Thr-Leu-Leu-Arg-Tyr-T_y_r_ (changed amino acids underlined) do not form ordered filamentous structures.