Interaction of [125I]β nerve growth factor with acidic proteins

We have demonstrated that the nerve growth factor will interact with various acidic proteins apparently nonspecifically. When¹²⁵I-labeled nerve growth factor at a concentration of 3.8×10^–10 M is incubated with an acidic protein at 2 mg/ml (4.5×10^–6–4.4×10^–5 M), a complex is formed. This complex changes the isoelectric point of the¹²⁵I-labeled nerve growth factor sufficiently so that the¹²⁵I-labeled nerve growth factor migrates anomalously in polyacrylamide gel electrophoresis. The interaction between nerve growth factor and bovine serum albumin, which appears to be complex, may be the cause of the previously reported activation of the nerve growth factor when bovine serum albumin is present in a typical bioassay.A preliminary report of this work was presented at the American Society of Biological Chemists, 71st Annual Meeting, in June 1980.     

Effects of L-Glutamate Transport Inhibition by a Conformationally Restricted Glutamate Analogue (2S,1'S,2'R)-2-(Carboxycyclopropyl)Glycine (L-CCG III) on Metabolism in Brain Tissue In Vitro Analysed by NMR Spectroscopy

(2S,1'S,2'R)-2-(Carboxycyclopropyl)glycine (L-CCG III) was a substrate of Na⁺-dependent glutamate transporters (GluT) in Xenopus laevis oocytes (IC₅₀ 13 and 2 M for, respectively, EAAT 1 and EAAT 2) and caused an apparent inhibition of [³H]L-glutamate uptake in mini-slices of guinea pig cerebral cortex (IC₅₀ 12 M). In slices (350 M) of guinea pig cerebral cortex, 5 M L-CCG III increased both the flux of label through pyruvate carboxylase and the fractional enrichment of glutamate, GABA, glutamine and lactate, but had no effect on total metabolite pool sizes. At 50 M L-CCG III decreased incorporation of ¹³C from [3-¹³C]-pyruvate into glutamate C4, glutamine C4, lactate C3 and alanine C3. The total metabolite pool sizes were also decreased with no change in the fractional enrichment. Furthermore, L-CCG III was accumulated in the tissue, probably via GluT. At lower concentration, L-CCG III would compete with L-glutamate for GluT and the changes probably reflect a compensation for the missing L-glutamate. At 50 M, intracellular L-CCG III could reach > 10 mM and metabolism might be affected directly.     

Synthesis of Gal-β-(1→4)-GlcNAc-β-(1→6)-[Gal-β- (1→3)]-GalNAc-α-OBn oligosaccharides bearing O-methyl or O-sulfo groups at C-3 of the Gal residue: specific acceptors for Gal: 3-O-sulfotransferases

Our recent studies have revealed the existence of two distinct Gal: 3-O-sulfotransferases capable of acting on the C-3 position of galactose in a Core 2 branched structure, e.g., Gal14GlcNAc16(Gal13)GalNac1OBenzyl as acceptor to give 3-O-sulfoGal14GlcNAc13(Gal13)GalNAc1OB 20 and Gal14GlcNAc16(3-O-sulfoGal13)GalNAc1OB 23. We herein report the synthesis of these two compounds and also that of other modified analogs that are highly specific acceptors for the two sulfotransferases. Appropriately protected 1-thio-glycosides 7, 8, and 10 were employed as glycosyl donors for the synthesis of our target compounds.     

Comparative residual toxicities of pesticides to the predator Agistemus industani (Acari: Stigmaeidae) on citrus in Florida

Residual toxicities of registered and selected experimental pesticides used on citrus against Agistemus industani Gonzalez (Acari: Stigmaeidae) were compared. Pesticides considered highly toxic to A. industani were: abamectin 0.15 EC at 731ml/ha+FC 435-66 petroleum oil at 46.8l/ha, pyridaben 75WP at 469g/ha, ethion 4EC at 7.01l/ha+FC 435-66 petroleum oil at 46.8l/ha, propargite 6.55 EC at 3.51l/ha, chlorfenapyr 2SC at 1.46l/ha applied alone or in combination with FC 435-66 petroleum oil at 46.8l/ha, sulphur 80DF at 16.81kg/ha, dicofol 4EC at 7.01l/ha, fenbutatin oxide 50WP at 2.24kg/ha, benomyl 50WP at 2.24kg/ha, benomyl 50WP at 1.68kg/ha+ferbam 76 GF at 5.60kg/ha, ferbam 76GF at 11.21kg/ha, neem oil 90EC at 46.8l/ha, and copper hydroxide DF (40% metallic copper) at 4.48kg metallic copper/ha+FC 435-66 petroleum oil at 46.8l/ha. Pesticides that were moderately to slightly toxic included: copper sulphate 98% at 4.48kg metallic copper/ha+FC 435-66 petroleum oil at 46.8l/ha, fenbuconazole 2F at 280ml/ha+FC 435-66 petroleum oil at 46.8l/ha, FC 435-66 petroleum oil applied alone at 46.8l/ha or 23.4l/ha, and diflubenzuron 25WP at 1.40kg/ha. Pesticides that were non-toxic included: fenbuconazole 2F at 585ml/ha, malathion 57EC at 5.85l/ha, FC 435-66 petroleum oil at 46.8l/ha, carbaryl 80S at 3.36kg/ha, chlorpyrifos 4EC at 4.68l/ha, and formetanate 92SP at 1.12kg/ha. Understanding the toxic effects of field weathered pesticides against key predacious mite species is important for effective IPM. The results of this study provide a comparison of direct and indirect toxic effects of various pesticides to A. industani under field conditions.     

Transient gene expression in electroporated Picea glauca protoplasts

The reporter gene for chloramphenicol acetyltransferase (CAT) was introduced into white spruce (Picea glauca (Moench) Voss.) protoplasts by electroporation. CAT transient gene expression was increased by increasing the concentration of pCaMVCN plasmid and was affected by the level of the applied voltage. Highest CAT activities were obtained after electroporation with a pulse of 350V.cm^–1 having an exponential decay constant of approximately 105ms. Linearized plasmid constructs gave much higher levels of CAT activity than circular plasmid. Attempts to use the Escherichia coli -glucuronidase gene (-GUS) as a marker gene revealed very high levels of -GUS-like activity in electroporated protoplasts. This activity was mainly due to a small molecule and may mask successful transformation since -GUS-like activity increased when plasmid DNA was present during electroporation.Abbreviations CAT chloramphenicol acetyltransferase - -GUS -glucuronidase - MUG 4-methyl umbelliferyl glucuronide - F microfarads NRCC No. 29150     

Finger pulse amplitude and frontalis EMG biofeedback effects of single- and two-system training

The relationship between finger pulse amplitude (PA) and frontalis EMG was studied first by looking at general physiological changes accompanying successful bidirectional PA control. Seven successful subjects were then asked to produce two patterns of PA and EMG (PAincEMGdec and PAdecEMGdec) while receiving both PA and frontalis EMG biofeedback. Results indicate subjects can easily produce the differentiation pattern of PAdecEMGdec but cannot produce the integration pattern of PAincEMGdec. These rather paradoxical results may indicate subjects were using an attentional rather than arousal strategy for controlling PA and have implications for the use of peripheral vasomotor training as a general relaxation technique.     

Conformational energy analysis of the leucine repeat regions of C/EBP, GCN4, and the proteins of the myc, jun, and fos oncogenes

It has been recently proposed that certain DNA binding proteins (including C/EBP, GCN4 and themyc, jun, andfos oncogene proteins) share a common structural motif based on helix-promoting regions containing heptad repeat sequences of leucines. It has been suggested that this structure is critical to the biological activity of these proteins, since it facilitates the formation of functional dimers held together by interdigitating leucine side-chains along the hydrophobic interfaces between long -helical regions of the polypeptide chains in a configuration termed the leucine zipper. In this paper, conformational energy analysis is used to determine the preferred three-dimensional structures of the leucine repeat regions of these proteins. The results indicate that, in all cases, the global minimum energy conformation for these regions is an amphipathic -helix with the leucine side-chains arrayed on one side in such a way to favor leucine zipper dimerization. Furthermore, amino acid substitutions in these regions (such as Pro for Leu), that are known to inhibit dimer formation and prevent DNA binding, are found to produce significant conformational changes that disrupt the amphipathic helical structure. Thus, these results provide support for the proposed leucine zipper configuration as a critical structural feature of this class of DNA binding proteins.     

Utilization and degradation of lindane by soil microorganisms

Of 147 microorganisms isolated from a loamy sand, 71 showed good growth with lindane (-1,2,3,4,5,6-hexachlorocyclohexane) and produced chloride in an aqueous medium. Thirteen soil microorganisms were selected to study the utilization of lindane. Lindane was metabolized by the microbes to -2,3,4,5,6-pentachloro-1-cyclohexene (-PCCH), -3,4,5,6-tetrachloro-1-cyclohexene (-TCCH), -3,4,5,6-tetrachloro-1-cyclohexene (-TCCH), -3,4,5,6-tetrachloro-1-cyclohexene (-TCCH), and pentachlorobenzene (PCB). Cells of Pseudomonas sp. No. 62 grown on lindane simultaneously adapted to -PCCH, -TCCH, -TCCH, -TCCH, PCB, 1,2,3,4-tetrachlorobenzene (1,2,3,4-TCB) and 1,2,4,5-tetrachlorobenzene (1,2,4,5-TCB). The bacteria degraded each of these chemicals at least partially as indicated by an increased rate of oxygen consumption.Abbreviations Lindane -1,2,3,4,5,6-hexachlorocyclohexane - -PCCH -2,3,4,5,6-pentachloro-1-cyclohexene - -TCCH -3,4,5,6-tetrachloro-1-cyclohexene - -TCCH -3,4,5,6-tetrachloro-1-cyclohexene - -TCCH -3,4,5,6-tetrachloro-1-cyclohexene - PCB pentachlorobenzene - 1,2,3,4-TCB 1,2,3,4-tetrachlorobenzene - 1,2,3,5-TCB 1,2,3,5-tetrachlorobenzene - 1,2,4,5-TCB 1,2,4,5-tetrachlorobenzene - 1,2,3-tCB 1,2,3-trichlorobenzene - 1,2,4-tCB 1,2,4-trichlorobenzene - 1,3,5-tCB 1,3,5-trichlorobenzene - 1,2-DCB 1,2-dichlorobenzene - 1,3-DCB 1,3-dichlorobenzene - 1,4-DCB 1,4-dichlorobenzene - MCB monochlorobenzene Contribution No. 631, Research Institute, Agriculture Canada, University Sub Post Office, London, Ontario N6A 5B7     

β-Ether cleavage of the lignin model compound 4-ethoxy-3-methoxyphenylglycerol-β-guaiacyl ether and derivatives by Phanerochaete chrysosporium

The white rot basidiomycete Phanerochaete chrysosporium metabolized 4-ethoxy-3-methoxyphenyl-glycerol--guaiacyl ether (V) in low nitrogen, stationary cultures under which conditions the ligninolytic enzyme system is expressed. 4-Ethoxy-3-methoxyphenylglycerol XIII, guaicol and 4-ethoxy-3-methoxybenzyl alcohol (II) were isolated as metabolic products. Exogenously added XIII was rapidly converted to 4-ethoxy-3-methoxybenzyl alcohol indicating that it is an intermediate in the metabolism of V. P. chrysosporium also metabolized 1-(4-ethoxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)-3-hydroxypropane VI. The degradation pathway for this dimer also included initial -ether cleavage and -hydroxylation of the diol product 1-(4-ethoxy-3-methoxyphenyl) 2,3 dihydroxypropane (XI) to yield the triol XIII which was cleaved at the , bond to yield 4-ethoxy-3-methoxybenzyl alcohol. Finally P. chrysosporium also cleaved the dimer 1-(4-ethoxy-3-methoxyphenyl)-2-(2-methoxyphenoxy)-1-hydroxypropane (VIII) at the -ether linkage yielding 1-(4-ethoxy-3-methoxyphenyl) 1,2 dihydroxypropane (IX) which was subsequently cleaved at the , bond to yield II. All of the results indicate that oxidative -ether cleavage is an important initial reaction in the metabolism of -aryl ether lignin substructure dimeric compounds. Metabolities were identified after comparison with chemically synthesized standards by gas liquid chromatography-mass spectrometry.Abbreviations GLC Gas liquid chromatography - TMSi trimethylsilyl - TLC thin layer chromatography     

Molecular cloning of fragments of bacteriophage T4 DNA

Summary Non-glucosylated T4 DNA was digested with R. EcoRI and the resulting fragments covalently joined to vectors. The genetic content of each -T4 hybrid was determined by marker-rescue tests. The isolation of many recombinants containing partialdigestion products of T4 DNA provided the overlapping sequences necessary to order fragments within the T4 genome. The present analyses include parts of the early region between genes 42 and 46, and much of the late region between genes 50 and 29. T4 cytosine-DNA digested to completion by R.EcoRI was used to identify the fragments of DNA within the -T4 recombinants. The T4 cytosine-DNA was also sensitive to R.HindIII and R.Xho but not to R.BamH1.     

Partial purification and characterization of a folatebinding protein from human choroid plexus

A folate-binding protein (binder) from human choroid plexus was solubilized with Triton X-100 and partially purified in three steps: (1) affinity chromatography, (2) Sephadex G-200 column chromatography, and (3) polyacrylamide gel electrophoresis. When the partially purified binder was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the binding activity was located in the region of the gel with a molecular weight between 45,000 and 60,000. The specific activity of the binder after the three purification steps was 1.2 g folic acid/mg protein, a 316-fold purification. Binding activity of the partially purified binder decreased below pH 6.0 and above pH 8.0 was unaffected by treatment with ribonuclease or deoxyribonuclease, but was abolished with trypsin, chymotrypsin, or protease (Streptomyces griesus). The binding of folic acid to the human binder was inhibited by folate > H₄-folate > methyl-H₄-folate dihydrofolate pteroic acid methotrexate aminopterin.     

Experimental effects of elevated salinity on three benthic invertebrates in Pyramid Lake,Nevada

Salinity of Pyramid Lake increased from 3.7 to 5.5 between 1933 and 1980. Concern over future reductions in overall species richness prompted experiments to assess responses of dominant lake organisms to elevated salinity. Salinity tolerances of three important benthic invertebrates, Hyalella aztecta, Chironomus utahensis, and Heterocypris sp., were tested in controlled laboratory bioassays and also in a semi-natural environment consisting of large (47 m³) mesocosms.Densities of H. azteca in mesocosms were significantly lower at salinities of 8.0 and 11.0 compared with 5.6 controls in year one, but not in 8.5 salinity mesocosms in year two. The 96-h LC₅₀ for H. azteca was high at 19.5. Short-term mortalities of C. utahensis were 100% at salinities of 13.3 and greater. Fifty-seven percent fewer larvae matured from third to fourth instar at 8.9 than at 5.5 salinity in 17 day subacute bioassays. Furthermore, larval chironomid densities and emergence of adults from mesocosms were significantly reduced at salinities of 8.0 and higher compared with controls. Mortality of Heterocypris sp. was 50% at a salinity of 18.6 in laboratory bioassays and populations in mesocosms ranged between 40 and 100% lower at salinities of 8.0 and 11.0 than in controls.Multiple generation mesocosm experiments indicated all three invertebrates were more sensitive to elevated salinity than results of short-term bioassays. Our studies suggest populations of these invertebrates may be reduced from present levels if Pyramid Lake's salinity were to double, although none are expected to be extirpated. Food habit shifts and reduced production of lake fishes are likely consequences of salinity-induced disruption in the benthic invertebrate forage base.     

Life Cycle of the Tick Haemaphysalis Leporis-palustris (Acari: Ixodidae) Under Laboratory Conditions

The life cycles of two separate populations (colonies A and B) of the rabbit tick, Haemaphysalis leporis-palustris, were studied under laboratory conditions. Domestic New Zealand rabbits, Oryctolagus cuniculus, and wild rabbits, Sylvilagus brasiliensis, were used as hosts for ticks from colony B and only O. cuniculus rabbits were used as hosts for ticks from colony A. Developmental periods were observed in an incubator at 27±1°C and RH 90±5%. Larvae from colonies A and B fed for 8.0±3.7 days and 8.5±1.3 days, respectively, on O. cuniculus. On S. brasiliensis larvae from colony B fed for 7.2±1.3 days. Nymphs from colony A fed for 8.1±1.4 days on O. cuniculus and nymphs from colony B fed for 8.1±1.0 days on S. brasiliensis. Only one engorged nymph from colony B was recovered from O. cuniculus. Females from colony A fed for 20.9±5.9 days on O. cuniculus and females from colony B fed for 18.6±2.4 days on O. cuniculus and 18.7±3.7 days on S. brasiliensis. Engorged larvae from colony A required 13.7±3.7 days to molt while engorged larvae from colony B required 11.8±3.0 and 11.5±1.8 days to molt, after having fed on O. cuniculus and S. brasiliensis, respectively. Engorged nymphs from colonies A and B required 16.3±1.9 days and 14.7±1.4 days to molt, respectively. Engorged females from colonies A and B required 4–7 and 3–5 days, respectively, to start oviposition. Mean egg incubation periods lasted for 33–34 days. For ticks from colony B, host species accounted for significant differences (p<0.05) in larval and nymphal feeding periods, oviposition weights and CEIs. Significant differences (p<0.05) between the two colonies when ticks fed on O. cuniculus were observed for larval and nymphal feeding and premolt periods, engorged female and oviposition weights and conversion efficiency indexes (CEI). S. brasiliensis were always a more suitable host for H. leporis-palustris than O. cuniculus. Significantly more larvae and nymphs engorged and molted when fed on S. brasiliensis (p<0.001). Females fed S. brasiliensis were more successful to lay fertile eggs and showed the highest engorged and egg mass weights, and the highest CEIs. Data of H. leporis-palustris fed on wild rabbits (one of its natural host species) are reported for the first time.     

Steroid-induced regulation of 3α,20β- and 3β,17β-hydroxysteroid dehydrogenase activity in wild type and mutants of Streptomyces hydrogenans

The effect of estradiol, hydrocortisone and progesterone on 3,20-and 3,17-hydroxysteroid dehydrogenase (HSD) in mutants of Streptomyces hydrogenans was compared to the steroid response of the wild type. Mutants were defective in arginine biosynthesis and/or aerial mycelial formation and lacked both enzymes or only 17-HSD. Some 17-HSD mutants had lost the ability to be induced by estradiol, by progesterone or by both. Some 20-HSD mutants had lost the ability to be induced by hydrocortisone, by progesterone or by both. Non-inducibility of 17-and 20-HSD by progesterone was not co-ordinate. An additional study of the growth phase-dependent enzyme activity of the wild type after induction with estradiol, hydrocortisone and progesterone was performed.Non-standard abbreviations 17-HSD 3,17-Hydroxysteroid dehydrogenase (EC 1.1.1.51) - 20-HSD 3,20-hydroxysteroid dehydrogenase (EC 1.1.1.53) - AO acridine orange - EBr ethidium bromide - EMS ethyl methanesulfonate - MNNG N-methyl-N-nitro-N-nitrosoguanidine     

Microaerophily

Objections to previous definitions of anaerobiosis and micro-aerophily are discussed, and a new definition of micro-aerophily is proposed. A survey of physiological characteristics of the micro-aerophilic bacterium Campylobacter sputorum spp. bubulus is given, and the concept of micro-aerophily is illustrated by the results of a study on the oxygen metabolism and oxygen sensitivity of this bacterium.     

The frequency of 47,+21, 47,+18, and 47,+13 at the uppermost extremes of maternal ages: results on 56,094 fetuses studied prenatally and comparisons with data on livebirths

Summary We examined the proportions (or so called rates) of fetuses with 47,+21, 47,+18, or 47,+13 diagnosed prenatally in women at the upper extremes of age. Our analysis was prompted by results from a large scale European study of amniocentesis which indicated that after increasing exponentially from age 35 years, the proportions of the autosomal trisomies reached a peak at a specific age and then leveled off or declined at the upper end of the age range. We analyzed North American data on 56,075 fetuses studied because of no known cytogenetic risk factor (aside from maternal age). This is the largest series to data. For 47,+21, the data from amniocentesis studies provide no evidence for any drop in the rate of change of proportion with maternal age up to 49 years. There is, if anything, a trend in our data to a steepening in the exponential rate of change at the upper extreme of age (above 46 years). Data from livebirths on the Down syndrome phenotype are at least consistent with an exponential rate of increase in proportion affected up to age 49 years. For 47,+18 our data from prenatal diagnoses are more consistent with an exponential increase up to age 43 years and a level proportion (or rate) after that. For 47,+13 no cases were observed above age 42 years, consistent with the drop in proportion affected above this age observed in the European series. We emphasize the possible effect of sampling fluctuation and reporting error upon these apparent trends.     

Root exudation and physiological status of a root-colonizing fluorescent pseudomonad in mycorrhizal and non-mycorrhizal pepper (Capsicum annuum L.)

Ultramafic (serpentine) soils from and adjacent to the Dun Mountain Ophiolitic Belt, South Island, New Zealand were analysed for 11 elements in order to establish to what degree edaphic factors influenced the character of the overlying vegetation. Using principal components analysis with a mutual plot of the first two principal components, involving the total elemental concentrations in the soils, it was possible to divide the soils into 6 virtually non-overlapping fields, each of which represented a specific vegetation community. Component 1 was essentially an ultramafic plot with heavy loadings from the elements chromium, cobalt, iron, magnesium, manganese, and nickel. Component 2 was a non ultramafic plot with heavy loadings from aluminium, copper and zinc. For elements extracted from the soils at pH 5.9, discrimination was somewhat poorer but confirmed the great importance of magnesium and nickel as controlling elements for the serpentine vegetation. It was concluded that the results indicated the overriding importance of edaphic factors in controlling the serpentine vegetation.     

The cytology of phytophthora infestans

Summary Phytophthora infestans has three kinds of somatic nuclei: an oval shaped nucleus (approx. 3.1×2.7 ) which stains diffusely except for a crescent shaped Feulgen positive cap which stains intensely; a granular nucleus whose contents are organized into a fairly constant number of stained bodies, and, a deeply staining condensed nucleus. The capped nucleus is thought to be metabolic or resting and the granular nucleus is thought to be dividing as it is most commonly found in hyphal tips. Attenuated forms of all three kinds of nuclei are found.Nuclear division is mitotic and intranuclear. Eight—ten chromosomes are seen at metaphase.Sporangia have a mean of 6.3 nuclei which is constant for age and strain of culture. Sporangia become multinucleate as a result of nuclear migration and not by division in the developing sporangium. Zoospores are usually uninucleate.The nuclear cap is persistent throughout nuclear division when it also divides. It is associated with flagella production and nuclear migration and has some of the properties of a blepharoplast.     

Preparation of 5′-GMP-rich yeast extracts from spent brewer's yeast

Spent brewer's yeast was autolysed and used as a raw material for the preparation of 5-GMP-rich yeast extracts. Malt rootlets were used as a source of 5-phosphodiesterase. The crude enzyme was extracted from malt rootlets and pretreated to inactivate 5-nucleotidase. The optimum pretreatment conditions were heating at 65 °C for 30 min or 70 °C for 7 min. The effects of autolysis time, phosphodiesterase concentration and incubation period on 5-GMP content were examined. The suitable autolysis time was 8 h. The preferable enzyme treatment period was in the range of 8–14 h. Longer autolysis and enzyme incubation periods caused a decrease in the 5-GMP content from 0.7–0.9% (w/w) to 0.2–0.4% (w/w). The 5-GMP content in extracts from debittered and non-debittered yeast was similar. The highest 5-GMP content in yeast extract was 0.93% (w/w), obtained with a phosphodiesterase concentration of 1.6unit/ml of yeast extract (5% solids content).