SHP-1 negatively regulates neuronal survival by functioning as a TrkA phosphatase

Nerve growth factor (NGF) mediates the survival and differentiation of neurons by stimulating the tyrosine kinase activity of the TrkA/NGF receptor. Here, we identify SHP-1 as a phosphotyrosine phosphatase that negatively regulates TrkA. SHP-1 formed complexes with TrkA at Y490, and dephosphorylated it at Y674/675. Expression of SHP-1 in sympathetic neurons induced apoptosis and TrkA dephosphorylation. Conversely, inhibition of endogenous SHP-1 with a dominant-inhibitory mutant stimulated basal tyrosine phosphorylation of TrkA, thereby promoting NGF-independent survival and causing sustained and elevated TrkA activation in the presence of NGF. Mice lacking SHP-1 had increased numbers of sympathetic neurons during the period of naturally occurring neuronal cell death, and when cultured, these neurons survived better than wild-type neurons in the absence of NGF. These data indicate that SHP-1 can function as a TrkA phosphatase, controlling both the basal and NGF-regulated level of TrkA activity in neurons, and suggest that SHP-1 regulates neuron number during the developmental cell death period by directly regulating TrkA activity.     

EFFECTS OF HUMAN ACTIVITIES ON STRUCTURE AND COMPOSITION OF WOODY SPECIES OF THE NOKREK BIOSPHERE RESERVE OF MEGHALAYA,NORTHEAST INDIA

Aims Our study was conducted in the Nokrek Biosphere Reserve (NBR) in the Garo hills districts of Meghalaya, Northeast India. Our aim was to assess the effects of human activities on plant diversity,population structure and regeneration.Methods We selected a representative 1.2 hm2 stand in both the core and buffer zones of NBR. Structure and composition were determined by randomly sampling square quadrats, population structure was assessed by determining age structure, and regeneration was assessed by measuring densities of seedling, sapling and adult trees.Important findings More woody species were recorded from the core zone than the buffer zone (87 vs. 81 species), and there were a large number of tropical, temperate, and Sino-Himalayan, Burma-Malaysian and Malayan elements, primitive families and primitive genera. The trees were distributed in three distinct strata,canopy, subcanopy and sapling. Subcanopy and sapling layers had the highest species richness (81％ -88％ ). Lauraceae and Euphorbiaceae were the dominant families in terms of the number of species, and a large number of families were represented by single species. Most woody species (57 ％ - 79 ％ ) were contagiously distributed and had low frequency ( ＜ 20％ ). Although stand density was high in the buffer zone, its basal area was low compared to the stand in the core zone. Low similarity and high β-diversity indicate marked differences in species composition of the stands. Shannon diversity index was high in both the stands, while Simpson dominance index was low. The diameter-class distribution for dominant species revealed that the most had a large number of young individuals in their populations. Preponderance of tree seedlings, followed by a steep decline in population density of saplings and adult trees, indicated that the seedling to sapling stage was the most critical in the life cycle of the tree populations. Most species (42 ％ - 48 ％ ) had no regeneration,25 ％ - 35 ％ had good/fair regeneration, and the rest had poor regeneration or reoccurred as immigrants.     

Lipid Auxotrophy and the Effect on Lipid Composition of Entamoeba invadens*†

SYNOPSIS. A medium for the axenic cultivation of Entamoeba invadens has been developed. Serum, an essential constituent of conventional media, has been replaced by a mixture of albumin, unsaturated fatty acids, Tween, and cholesterol to control the lipid composition of the medium. Entamoeba invadens requires both cholesterol and unsaturated fatty acids for growth. The fatty acid composition of the phospholipids of the ameba reflects that of the medium to a great extent, especially with regard to the unsaturated fatty acids. The amount of membrane bounded cholesterol depends on the cholesterol concentration in the medium.     

Immunogenicity and cross-reactivity of 2009-2010 inactivated seasonal influenza vaccine in US adults and elderly

The campaign of 2009-2010 Northern Hemisphere seasonal vaccination was concurrent with the 2009 H1N1 pandemic. Using a hemagglutination inhibition (HAI) assay, we evaluated the immunogenicity and cross-reactivity of 2009-2010 inactivated trivalent influenza vaccine (TIV) in US adult and elderly populations. Vaccination of TIV resulted in a robust boost on the antibody response of all subjects to seasonal A/Brisbane/59/2007 (H1N1) and A/Uruguay/716/2007 (H3N2) with over 70% of recipients reaching a seroprotective titer of 40. B/Brisbane/60/2008 was the least immunogenic among the three seasonal vaccine strains with <30% of TIV recipients reaching a seroprotective titer of 40. TIV vaccination also induced a moderate boost on the pandemic specific antibody responses. Twenty-four percent of adults and 36% of elderly reached a seroprotective HAI titer of 40 or more against pandemic A/South Carolina/18/2009 (H1N1) after receiving TIV compared to 4% and 7% at the beginning of vaccination, respectively. In addition, 22% of adults and 34% of elderly showed an increase of 4-fold or more in A/South Carolina/18/2009 specific HAI titers after TIV vaccination. The pandemic specific cross-reactive antibodies strongly correlated with the post-vaccination HAI titers against the seasonal H3N2 vaccine strain in all subjects.     

Liver cytochrome P-450 system in rats with a varying susceptibility to the development of experimental alcoholism

The cytochrome P-450-dependent monooxygenase system of the liver was studied in laboratory noninbred male rats selected according to the intensity of their initial alcohol motivation and the dynamics of these parameters was followed up during 10-day alcoholisation. It was shown that in the animals inclined to the development of alcoholism the activity of the monooxygenase system (cytochrome P-450, B5; enzymes: aminopyrine N-demethylase, aniline p-hydroxylase, NADPH-cytochrome c-reductase) is higher than in the animals noninclined to the development of this disease. 10-day alcohol consumption in the free-choice situation between water and 15% ethanol solution did not change the parameters investigated. The only exception was NADPH-cytochrome c-reductase: its activity grew in both the groups of the animals by 40-75%.     

The chemical synthesis of glucosaminylphosphatidylglycerol. Comparison with a new phospholipid isolated from Bacillus megaterium

1. We describe the synthesis of a glucosamine derivative of phosphatidylglycerol having the same structure as that of the natural compound isolated from Bacillus megaterium. 2. 2-O-(3,4,6-Tri-O-acetyl-2-deoxy-2-phthalimido-d-glucopyranosyl)-3-O-benzyl-1-iodo-sn-glycerol was prepared by a Königs–Knorr condensation between 3-O-benzyl-1-toluene-p-sulphonyl-sn-glycerol and 3,4,6-tri-O-acetyl-1-bromo-2-deoxy-2-phthalimido-d-glucopyranose followed by replacement of the toluene-p-sulphonyl group with iodine. The iodide was treated with the silver salt of 2-isolauroyl-1-oleoyl-sn-glycerol 3-(monobenzyl hydrogen phosphate) to form the fully protected phosphoglycolipid. 3. Removal of benzyl protecting groups by catalytic hydrogenolysis, phthaloyl group with hydrazine and acetyl groups with pH10 buffer furnished 2-O-(2-amino-2-deoxy-d-glucopyranosyl)-1-(2-isolauroyl-1-stearoyl-sn-glycero-3-phosphoryl)-sn-glycerol. 4. The synthetic and natural compounds appeared identical when compared by chromatography and by identification of hydrolysis products from chemical and enzymic degradations.     

Analysis of pCU1 replication origins: dependence of oriS on the plasmid-encoded replication initiation protein RepA

The broad-host-range replicon of the plasmid pCU1 has three origins of vegetative replication called oriB, oriS, and oriV. In the multi-origin replicon, individual origins can distinguish among replication factors provided by the host. It has been found that during replication in Escherichia coli polA(-) host, oriS was the only active origin of a mutant pCU1 derivative bearing a mutation in the gene encoding replication initiation protein RepA. To further investigate the capacity of oriS to function in an E. coli polA(-) host we constructed a number of clones of the basic replicon of pCU1 containing oriS as the only replication origin. An oriS construct created with pUC18 could transform the polA(-) strain when RepA was supplied in trans. When the oriS region (between nucleotides 290 and 832) was ligated to an antibiotic resistance Omega fragment, the construct could be recovered as a plasmid from polA(+) strain if functional RepA was provided in trans. Our results therefore indicate that the basic replicon of pCU1, containing oriS as the sole origin, does require RepA to initiate plasmid replication in E. coli     

Biogeochemical interactions between iron and sulphate in freshwater wetlands and their implications for interspecific competition between aquatic macrophytes

1. Wetlands are threatened by desiccation, eutrophication and changing water quality, generally leading to greatly altered biogeochemical processes. Sulphate pollution can lead to severe eutrophication and sulphide toxicity, but may also interact with the availability of iron and other metals. 2. In the present study, we examined the biogeochemical interactions between sulphate and iron availability, and their effects on aquatic macrophytes, in a field experiment with enclosures. The natural iron supply by groundwater was mimicked by adding iron to the sediment, and the effect of increased sulphate concentrations in the surface water was also studied. The enclosure experiment was performed in a mesotrophic, anaerobic ditch in a peat meadow reserve in the Netherlands. In all enclosures, three Stratiotes aloides plants were introduced to serve as indicator species. 3. Addition of sulphate led to the mobilisation of phosphate, whereas addition of iron or both iron and sulphate did not affect P mobilisation. Growth of S. aloides was decreased by both iron addition and sulphate addition (sulphide toxicity). Addition of iron under sulphidic conditions, however, led to mutual detoxification of both toxicants (iron and sulphide) and did not decrease S. aloides growth. The uptake of metals was highest in the treatment involving sulphate addition, probably as a result of increased mineralisation of the peat soil. 4. Growth of Elodea nuttallii, which grew naturally in the enclosures, was stimulated by iron or iron plus sulphate addition. It did not, however, grow in the enclosures with sulphate addition, as a result of sulphide toxicity or sulphide‐induced iron deficiency. Under iron‐rich conditions, E. nuttallii appeared to be a better competitor than S. aloides and depressed the growth of the latter species. 5. We propose that the growth of S. aloides is directly regulated by interactions between sulphide and iron and indirectly by the effects of both compounds on the competitive strength of E. nuttallii. In general, we conclude that biogeochemical interactions between sulphate and iron can have a strong influence on plant species composition in freshwater wetlands, because of direct effects or changes in the competitive strength of plant species related to differential sensitivity to either iron or sulphide.     

Inhibition of the development of the reproductive tract in parasitized snails

Summary

Myoblasts, muscle cells with the capacity to divide, have been detected in “Anlagen” of the male copulation organ of Lymnaea stagnalis. They only occur in the apical part of the penis. Here they could be found throughout life. Mitotic activity of these cells can be demonstrated by using an antiserum to a S-phase specific cell cycle marker, PCNA [see, e.g., Baserga (1991)]. The number/percentage of PCNA positive myoblasts is a good parameter for growth of this male copulation organ and hence also for inhibition of its growth and development as occurs in parasitized snails. In transplantation experiments, “Anlagen” of the copulation organ were used from snails 7–9 weeks after being parasitized as they can be excised in this stage and transplanted into either parasitized or nonparasitized snails. These experiments have indicated that humoral, parasitic excretory/secretory factors can be responsible for the inhibition of growth and differentiation of the copulation organ in parasitized snails as reflected by a relatively low number of PCNA positive myoblasts compared to the controls. Data obtained in in vitro experiments showed a significant decrease of the number of myoblasts in “Anlagen” cultured in the presence of parasitic E/S products. The fact that no significant effect was found on the relative low number of PCNA positive myoblasts is discussed. The effect of parasitic E/S products on these myoblasts appeared to be exerted in a direct way, not mediated by CNS-derived factors or by factors from cells in the connective tissue sheath around the CNS. Although it appears possible to use transplantation and/or in vitro culturing of these “Anlagen” as a bioassay for identification of the parasitic factor(s) responsible for the inhibitory effects on myoblasts, the methods are very laborious and do not seem very appropriate for testing many fractions of E/S products.