Uridine,a cell division factor in pea roots

Nodulation (root nodule formation) in legume roots is initiated by the induction of cell divisions and formation of root nodule primordia in the plant root cortex, usually in front of the protoxylem ridges of the central root cylinder. We isolated a factor from the central cylinder (stele) of pea roots which enhances hormone-induced cell proliferation in root cortex explants at positions similar to those of nodule primordia. The factor was identified as uridine. Uridine may act as a morphogen in plant roots at picomolar concentrations.     

Obesity and Malnutrition in a Shantytown Population in the City of São Paulo,Brazil

To investigate the prevalence of obesity and malnutrition in the poor Brazilian population we conducted a survey on the socioeconomic and nutritional status of 535 families (comprising 2 411 individuals) living in shanty towns in the city of São Paulo. There was a 30% prevalence of malnutrition in the children, with chronic malnutrition as the most predominant problem. The prevalence of obesity was 6.4% in boys and 8.7% in girls. Overweight and obesity associated with stunting was found in 5.8% of boys and 6.8% girls. Adolescents showed a higher prevalence of malnutrition when weight-for-age distribution was used (boys 46.4%, girls 40.2%), but a right deviation in the distribution was observed with an increase in obesity and a decrease of malnutrition was observed (obesity was 21% in girls and 8.8% in boys; malnutrition was 15.5% in boys and 12.6% in girls) when the weight-for-height adjustment was made. Stunting was the most predominant type of malnutrition in both sexes. Obesity associated with stunting was more common than obesity without stunting, both in younger children and adolescents. Adults had a higher prevalence of obesity than malnutrition according to both the Metropolitan Life Insurance tables (1.7% of undernutrition, 16.7% of overweight, and 14.1% of obesity) and Body Mass Index (8.5% of undernutrition, 21.9% of overweight, and 14.6% of obesity). There was an increase in the percentage of obese children when at least one adult in the family was obese and an increased percentage of malnourished children when undernourished adults were present in the family. Obesity among the adults of the family decreased the occurrence of malnutrition among the children. In 9% of families there was a coexistence of obesity in the adults and malnutrition in the children. These results demonstrate a coexistence of malnutrition and obesity in poor urban Brazilian communities.     

Phosphorylation of the Casein Kinase II Domain of B-50 (GAP-43) in Rat Cortical Growth Cones

Abstract: Growth-associated phosphoprotein B-50 is a neural protein kinase C (PKC) substrate enriched in nerve growth cones that has been implicated in growth cone plasticity. Here we investigated whether B-50 is a physiological substrate for casein kinase II (CKII) in purified rat cortical growth cone preparations. Using site-specific proteolysis and known modulators of PKC, in combination with immunoprecipitation, mass spectrometry, and phosphoamino acid analysis, we demonstrate that endogenous growth cone B-50 is phosphorylated at multiple sites, on both serine and threonine residues. Consistent with previous reports, stimulation of PKC activity increased the phosphorylation of only those proteolytic fragments containing Ser⁴¹. Under basal conditions, however, phosphorylation was predominantly associated with fragments not containing Ser⁴¹. Mass spectrometry of tryptic digests of B-50, which had been immunoprecipitated from untreated growth cones, revealed that in situ phosphorylation occurs within peptides B-50_181–198 and B-50_82–98. These peptides contain the major and minor in vitro CKII phosphosites, respectively. In addition, cyanogen bromide digestion of immunoprecipitated chick B-50 generated a 4-kDa C-terminal B-50 phosphopeptide, confirming that phosphorylation of the CKII domain occurs across evolutionary diverse species. We conclude that B-50 in growth cones is not only a substrate for PKC, but also for CKII.     

New foliicolous lichens from New Zealand 3

Podotara gen. nov. (incertae sedis) is described. It is based on the foliicolous taxonP. pilophoriformis sp.nov., which is described and illustrated. Characteristic of the new genus and species are globose and stipitate apothecia and 3-septate, hyaline, thin-walled ascospores with annular thickening of the septa.Scoliciosporum lividum sp. nov. (Scoliciosporaceae) differs from all species of its genus known to date by its leprose grey thallus, its livid, globose, dull apothecia, and its 3-septate ascosoores spirally contorted in the ascus.     

Assessment of inhibition kinetics of the growth of strain P5 on pentachlorophenol under steady-state conditions in a nutristat

A bacterium degrading pentachlorophenol (PCP) as the only source of carbon and energy was grown in a nutristat, i.e., a continuous culture with on-line measurement and control of the substrate concentration. We improved the PCP nutristat by incorporation of a personal computer with a proportional integral derivative (PID) algorithm for controlling the medium feed pump. The controlled value deviated from the average (set-point) value by 1% maximally. In the PCP nutristat (30°C), the steadystate dilution rate, and hence, specific growth rate, showed a maximum value of 0.142±0.004 h^-1 at set-point PCP concentrations between 37 and 168 M. At PCP concentrations above 168 M, the steady-state growth rate decreased because of inhibition. The growth yield coefficient was not seriously affected by the PCP concentration, suggesting that uncoupling was not the inhibitory mechanism. It was concluded that the PCP nutristat is very useful for establishing steady-state conditions that maintain growth-inhibitory PCP concentrations and high cell concentrations, conditions for which the chemostat is not suitable.Abbreviations MCA Metabolic control analysis - NTA Nitrilotriacetic acid - PCP Pentachlorophenol - PID Proportional integral derivative     

Production of recombinant proteins in Chinese Hamster Ovary cells overexpressing the subtilisin-like proprotein converting enzyme furin

The proprotein processing enzyme furin is the mammalian prototype of a novel family of subtilisin-like serine endoproteases which possess cleavage specificity for sites involving multiple basic amino acid residues and are involved in the processing of precursor proteins of a variety of regulatory peptides and proteins. One of the limiting steps in the engineering of mammalian cells designed for the overproduction of secreted proteins is the endoproteolytic cleavage of the precursor molecule to its mature biologically active form. The extremely low level of endogenous furin is likely the reason why cells are not able to fully mature overexpressed precursor proteins to their mature form. Here, we report a CHO-derived cell line genetically engineered for the production of high levels of recombinant proteins that need such endoproteolytic maturation. First, the human furin cDNA under the control of the cytomegalovirus early promoter and enhancer was introduced and overexpressed in a DHFR-deficient CHO cell line. A permanent cell line CHO-D3-FUR was established that expressed biologically active furin. Subsequently, to demonstrate the capacity of CHO-D3-FUR cells to produce recombinant proteins in a fully matured form, two derivative cell lines were established that overexpressed the von Willebrand factor (vWF) and transforming growth factor 1 (TGF1); CHO-D3-vWF and CHO-D3-TGF1, respectively. Both derivative cell lines were able to produce relatively high levels of recombinant protein in a fully matured and biologically active form. Our results illustrate the potential of the CHO-D3-FUR cell line in the production of recombinant secretory proteins that need endoproteolytic activation at the consensus furin cleavage sequence Arg-X-Lys/Arg-Arg.Abbreviations ATCC American Type Culture Collection - CHO Chinese hamster ovary - CMV cytomegalovirus - DHFR dihydrofolate reductase - DMEM Dulbecco's modified Eagle's medium - DT diphtheria toxin - EDTA ethylenediaminetetra-acetate - ECL enhanced chemiluminescence - ER endoplasmic reticulum - FITC fluorescein isothiocyanate - FUR FES upstream region - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - MEM minimal essential medium - MTX methotrexate - PAGE polyacrylamide gel electrophoresis - SDS sodium dodecyl sulfate - TCA trichloroacetic acid - TFG transforming growth factor - vWF von Willebrand Factor     

Physiological responses and central nervous projections of antennal olfactory receptor neurons in the adult desert locust, Schistocerca gregaria (Orthoptera: Acrididae)

Olfactory receptor neurons present in two morphological sensillum types on the male Schistocerca gregaria antenna were for the first time investigated physiologically when stimulated with behaviourally relevant odours. Neurons present in trichoid/basiconic sensilla showed clear excitatory responses to compounds present in the male-produced aggregation pheromone and also to a plant produced compound. Sensilla could be categorised physiologically according to the responses of their receptor neurons to the tested stimuli. Also receptor neurons present in sensilla coeloconica responded to aggregation pheromone components, but always in an inhibitory fashion. These neurons could, however, be excited by a plant produced compound and by some acids present in the nymphal odour. The antennal lobe of the male S. gregaria was observed to contain about 1000 very small glomerular structures. Single receptor neurons were stained from the antenna to the antennal lobe using a cobalt lysine technique. These stainings revealed a multi glomerular axonal branching pattern of antennal receptor neurons.Abbreviations AN antennal nerve - AL antennal lobe - RN receptor neuron     

Suppression of floral induction inBryophyllum daigremontianum by a growth retardant

Summary Flower formation in the long-short-day plantBryophyllum daigremontianum induced by the shift from long to short day is fully suppressed by applying the growth retardant CCC [(2-chloroethyl)-trimethylammonium chloride]via the roots during short day treatment at a level of at least 1 g per plant. At the same time CCC applications strongly reduce the internode length, but the rate of leaf formation does not seem to be affected. CCC also causes the production of more anthocyanin in the leaves.Gibberellin A₃ appliedvia the apices and youngest leaves at a dose of 1.5 g per plant, completely overcomes the inhibition of flower formation caused by CCC. Ten times higher quantities are needed for a complete reversal of the reduction in stem elongation.The growth retardant CCC seems to keep the level of physiologically active GA below that which is normally required for floral initiation and stem elongation. Thus, the results are in agreement with the hypothesis that a high GA level is necessary for the production of floral stimulus inBryophyllum daigremontianum.With 2 Figures in the TextThis work was supported by the National Science Foundation, grants G-16408 and G-17483.     

Modification of the free sulphydryl groups of bovine serum albumin to probe conformational transitions in the neutral region

The free SH group in bovine serum albumin has been modified by covalent coupling with 2-chloromercuri-4-nitrophenol and 2-chloromercuri-2,4-dinitrophenol. The ionization of the phenolic OH group of the former label when bound to albumin can be followed spectrophotometrically. The pK of this group was influenced by the presence of Ca²⁺. This is not a direct effect but proceeds via an effect of Ca²⁺ on the protein conformation. Similar results were obtained by following the c.d. signal of this label. This conformational change seems to be different from the one which can be detected by measuring the induced c.d. of a non-covalently bound ligand like diazepam.