Heritable variation in the phaseolin protein of nondomesticated common bean,Phaseolus vulgaris L.

Summary Crude protein extracts from single seeds of nondomesticated Mexican bean accessions were analysed by SDS polyacrylamide gel electrophoresis for variability in phaseolin protein. Six new phaseolin types; M1, M2, M3, M4, M5, M6, which contained polypeptides within the same range of molecular weights (51,000 to 45,000 daltons) as occur in the S, T and C phaseolin types of cultivated beans were identified. No T and C types were found among the non-domesticated Mexican accessions, and the S type occurred in less than 7% of the seeds screened. Genetic analyses of F₂ progenies from crosses between Sanilac (S), and five of the M types showed that each M phaseolin phenotype was allelic to the S type and expressed codominantly.     

Idioms of distress: Kinship and sickness among the people of the Kingdom of Tonga

Idioms of distress refers to the popular expression of emotional tension that arises in the relationship between sickness and kinship. By reference to case studies and discussions among the Polynesian people of Tonga, the author shows where such tension arises and how it influences the sickness process. Sickness is necessarily a collective phenomenon which can best be understood not simply as a clinical event, but as an experience that is part of the experience of family. Various ways of expressing distress as a reflexive encounter between personal and cultural meaning systems are reviewed, as are several new concepts such as doing sickness as kinship, and turning in the process of decision making in the kinship management of sickness. The explanatory models of sickness in Tonga are shown to encompass culturally sanctioned expressions of distress as part of the adaptive coping mechanisms in that society. Distress frequently emerges in somatic form, as a number of studies have shown. However, the author emphasizes the kinship meaning of sickness, kinship management and sickness therapy, the adaptive process of idiomatic expressions of distress, which are expanded here and offered as potential avenues for elaboration in other cultural milieu. Two aspects of the notion idioms of distress are noted, and the phenomenon is understood as a process which acts as a prime mover in social change.     

The glycolipid specificity ofErythrina cristagalli agglutinin

The interaction of¹²⁵I-labeledErythrina cristagalli agglutinin (ECA) with neutral glycosphingolipids on thin layer chromatograms was examined by the overlay technique followed by radioautography. The lectin bound topara-globoside with a sensitivity about 10 times higher than to lactosylceramide or globoside, in agreement with the specificity of the lectin forN-acetyllactosamine. The lower limit of detection ofpara-globoside was about 0.66 nmol. The specific binding of ECA to this glycolipid was confirmed by a highly sensitive enzyme-linked lectin assay (ELLA), utilizing the horseradish peroxidase-avidin-biotin system for detection of bound lectin. Overlays of neutral glycosphingolipid extracts from human erythrocyte membranes and from human granulocytes with ECA demonstrated that the lectin can be employed for the detection of small amounts ofpara-globoside in biological materials also in the presence of excess globoside. No staining was obtained when thin layer chromatograms of neutral glycosphingolipid extracts from rabbit erythrocyte membranes were overlayed with¹²⁵I-ECA. Afterin situ treatment of the chromatograms with -galactosidase, the lectin bound to several components, one of which had a mobility corresponding to that of the pentahexosylceramide Gal3Gal4GlcNAc3Gal4Glc1Cer, the major neutral glycosphingolipid of rabbit erythrocytes, thus providing further evidence for the specificity of ECA forpara-globoside.Abbreviations GSL glycosphingolipid(s) - CDH lactosylceramide, Gal4Glc1Cer - CTH trihexosylceramide, Gal4Gal4Glc1Cer - GLOB globoside, GalNac3Gal4Gal4Glc1Cer - PG para-globoside, Gal4GlcNAc3Gal4Glc1Cer - AsG_M1 asialo-G_M1, Gal3GalNAc4Gal4Glc1Cer - FORS Forsmann antigen, GalNAc3GalNAc3Gal4Gal4Glc1Cer - CPH pentahexosylceramide, Gal3Gal4GlcNAc3Gal4Glc1Cer - ECA Erythrina cristagalli agglutinin - SBA soybean agglutinin - PBS phosphate-buffered saline - PVP-40 polyvinylpyrrolidone M.W. 40000 - BSA bovine serum albumin - HRP-avidin horseradish peroxidase conjugated to avidin - ELLA enzyme-linked lectin assay - ELISA enzyme-linked immunosorbent assay - PMNL polymorphonuclear leukocytes - HPTLC high performance thin layer chromatography     

Concluding remarks

On the basis of symposium contributions onChlorella, Hibbertia, Eucalyptus, Ambrosia and on numerical approaches some fundamental problems of (bio)systematics, evolution, and taxonomic categories are discussed: Methods available for analysing affinities; conflicting evidence from phenetic, biochemical, cytogenetic and other analyses; further classification problems in cases of intermediacy, etc. While sibs of various levels and their natural hierarchy often can be objectively defined, this appears impossible for particular taxonomic levels itself (e. g. species). A single objective taxonomic system of organisms is unrealistic. Certain guiding lines for relative and practicable concepts of species and genus are proposed.Presented at the symposium Speciation and the Species Concept during the XIIth International Botanical Congress, Leningrad, July 8, 1975.     

Calcium restriction induces cardenolide accumulation in cell suspension cultures of Digitalis thapsi L.

Summary The removal of calcium ions from Murashige and Skoog culture medium induced a marked increase in the accumulation of cardenolides in cell suspension cultures of Digitalis thapsi. Cell viability was not affected although growth was slightly reduced. Strontium ions could substitute for calcium in inhibiting cardenolide production, this effect of calcium being reversed by the addition of LaCl₃ or ethyleneglycol-bis-(-aminoethyl ether)-N,N-tetraacetic acid. The results suggest that calcium, apart from its general effects on growth, may play a role in the regulation of cardenolide metabolism in a concentration dependent manner.Abbreviations BA 6-benzylaminopurine - 2 4-D 2,4-dichlorophenoxyacetic acid - EGTA ethyleneglycol-bis-(-aminoethyl ether)-N,N-tetraacetic acid - FW fresh weight - MS Murashige and Skoog (1962)     

Evaluation of an exotic maize population adapted to a locality

Summary An exotic Zea mays L. population (Tuxpeno) was adapted to North Carolina conditions by first introducing genes for adaptability from two North Carolina varieties ([(Jarvis X Indian Chief)Tuxpeno]Tuxpeno) including four generations of intermating, and then selecting for adaptability using maturity as the primary measure. The study evaluated selection for adaptability and the diversity available between adapted Tuxpeno and the local varieties, Jarvis and Indian Chief. Analytical procedures were developed to quantify the diversity between populations and the complementation of local varieties by introduced germ plasms. The analyses utilized the specific effects available from the diallel mating design.Three replicate selections responded similarly under simple recurrent mass selection (1/10) for the earliest disease-free plants initially and additionally for plant types (primarily height) in the final generation. The 1/4 local germ plasm permitted rapid adaptation of Tuxpeno gene pool to local conditions. The adapted Tuxpeno populations yielded similarly to the local populations with an average heterosis for grain yield of 28% when crossed to the local populations used as source of genes for adaptability. The diversity found between adapted Tuxpeno lines and these local varieties based on genes affecting grain yield was 1.5 to 2.5 times that measured between the local varieties (Jarvis and Indian Chief). Diversity lost through intergradation with local material was a reasonable investment. Yield genes introduced from Tuxpeno complemented local gene pools through nonadditive, primarily dominance-associated, gene effects. Reassortment of major gene blocks apparently occurred leading to significant divergence among replicate selections involving both additive-associated and dominance-associated gene effects.Paper No. 6355 of the North Carolina Agri. Res. Ser., Raleigh, NC. Research supported in collaboration with the Rockefeller Foundation and CIMMYT, D.F. (Mexico)     

Activation of immunoglobulin control elements in trasgenic mice

To assess the role interleukins and mitogens play in regulating immunoglobulin (Ig) gene expression via the Ig enhancer and promoter, transgenic mice carrying two different Ig gene regulatory regions were generated. One, EkCAT, contains the Ig heavy chain enhancer (E) and the light chain promoter driving the chloramphenicol acetyltransferase (CAT) gene. In the other, EkCAT, CAT is under the control of the promoter alone. E and relative activity were assessed by CAT assay. In EkCAT mice, low CAT expression was consistently found in spleen, bone marrow, mesenteric lymph node, and thymus but not in brain, lung, or kidney. In EkCAT mice, CAT expression was detectable just above background in lymphoid tissues, suggesting a basic level of tissue specificity in the absence of the enhancer. Whole spleen cell cultures prepared from the mice were treated with lymphokines and mitogens. Lipopolysaccharide (LPS), concanavilin A (Con A), interleukin 6 (IL-6), and interferon- (IFN-) increased CAT expression to varying extents in cells derived from EkCAT mice but not in spleen cells prepared from EkCAT mice. Thus, the presence of E, in addition to the promoter, is essential for the stimulation of CAT expression mediated by these factors. B cells from EkCAT mice were separated by density into populations of small and large cells. In untreated small B cells, no CAT expression was detected and only addition of LPS resulted in an increase in CAT expression. In large B cells, CAT was expressed at a low level without addition of exogenous factors. Incubation with LPS, IL-6, Con A and IFN- caused CAT expression to increase several-fold. This transgenic system provides a means to identify exogenous factors that activate Ig enhancers and promoters.This work has been submitted in partial fulfillment of the requirements for the doctoral degree from the George Washington University.     

Inhibition of axonal transport in vitro in frog sciatic nerves by chlorpromazine and lidocaine

Summary The effects of chlorpromazine hydrochloride (CPZ HCl) and prochlorperazin-metansulfonate (PCPZ) on the fast axonal transport of labelled proteins were examined in vitro in a peripheral frog nerve.A 0.1 mM concentration of CPZ HCl and PCPZ reduced the amount of transported proteins by more than 50 per cent. An almost complete block was obtained with a 0.5 mM concentration of these two drugs. The lower concentration hardly affected the protein synthesis. The transport inhibiting effect of 0.1 mM of the drugs was reversible but not that of the higher concentration.The number of microtubuli was strongly decreased and the number of filaments increased at the transport inhibiting concentrations. The ultrastructural changes induced by 0.1 mM of the phenothiazine tranquilizer were largely reversible. The local anesthetics lidocaine (18.3 mM) and tetracaine (3.3 mM) both caused similar changes, i.e. a reduction in the number of microtubuli. No ultrastructural effects were observed after treatment with 1 mM ouabain. These three drugs are known to block the axonal flow in the present system at the above mentioned concentrations.The biochemical and ultrastructural results are discussed in relation to those induced by other drugs affecting axonal transport.The present work was supported by grants from Statens Naturvetenskapliga Forskningsråd (No. 2535-8), C.-B. Nathorsts Vetenskapliga och Allmännyttiga Stiftelser, the Swedish Medical Research Council (B73-12X-2543-05B), H. Hierta's Stiftelse and W. och M. Lundgrens Stiftelse. Thanks are due to Mrs B. Egnér, Mrs E. Fjällstedt, Mrs. E. Norström and Mrs U. Svedin for expert technical assistance.     

Interrelationship between chromocentres and chiasmata in radish (Raphanus sativus L.)

A comparative study of the number and distribution of chromocentres in interphase nuclei and mean chiasma frequency at diakinesis has been made in three varietal populations of radish (Raphanus sativus L.), Scarlet Globe, Japanese White and Chinese White. The study showed a significant difference between the varietal populations in mean chiasma frequency and number of chromocentres (P<0.001), indicating that these nuclear characters are genotypically controlled. The correlation analysis revealed a significant negative correlation between chromocentres and chiasma frequency (r= -0.87). It was concluded that an increase in the amount of constitutive heterochromatin, as inferred by chromocentre counts, adversely affects the chiasma frequency and, consequently, genetic recombination in radish.     

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.     

Sterols of male and female flowers of Cucumis sativus

Sterols of male and female flowers of Cucumis sativus L. were similar in composition. The principal compound was 24-ethyl-5-cholesta-7,22-dien-3-ol. Five other 5-⁷ were detected: 24-methyl-7-ene, 24-ethyl-7-ene, 24-ethyl-7,24(28)Z-diene, 24-ethyl-7,25-diene and 24-ethyl-7,22,25-triene. Small amounts of ⁵ (cholesterol, 24-methylcholesterol and 24-ethylcholesterol) were detected. The possible significance of these sterols is discussed.Abbreviations GLC gas-liquid chromatography - GC-MS combined gas chromatography-mass spectrometry     

The ∈ subunit of the chloroplast ATP synthase of Pisum sativum

In contrast to the well-characterized spinach ( Spinacea oleracea) chloroplast ATP synthase (CF1–CFo), the properties of the chloroplast ATP synthase from pea (Pisum sativum ) have not been as intensively studied. Preliminary data suggested that the regulatory properties of the two enzymes differ. In the absence of activating treatments the ATPase activity of pea thylakoids in the dark was higher than that in spinach thylakoids. When assayed in the presence of sulfite, the MgATPase activity of pea thylakoids was inhibited to a maximum of 67% by tentoxin, indicating that the dark ATPase activity is in part catalyzed by CF1–CFo. The ATPase activity of purified pea CF1 was also higher than that of spinach CF1 in the absence of activating treatments. These differences could result from the different regulatory properties of the pea or subunit or both. The pea subunit was less effective in binding to or inhibiting the ATPase activity of pea o r spinach CF1 deficient in (CF1-). Spinach inhibited the ATPase activity of pea CF1- at lower concentrations than pea . The gene encoding the pea subunit was cloned and over-expressed. Recombinant pea did not restore low proton permeability to spinach thylakoid membranes reconstitituted with spinach CF1-, although pea was effective when tested with pea thylakoids reconstitituted with pea CF1-. These results confirm earlier suggestions that the C-terminal region of is important in -CF1 and -CFo interactions.     

Mutants of Arabidopsis thaliana with altered phototropism

Thirty five strains of Arabidopsis thaliana (L.) Heynh. have been identified with altered phototropic responses to 450-nm light. Four of these mutants have been more thoroughly characterized. Strain JK224 shows normal gravitropism and second positive phototropism. However, while the amplitude for first positive phototropism is the same as that in the wild-type, the threshold and fluence for the maximum response in first positive phototropism are shifted to higher fluence by a factor of 20–30. This mutant may represent an alteration in the photoreceptor pigment for phototropism. Strain JK218 exhibits no curvature to light at any fluence from 1 mol·m^-2 to 2700 mol·m^-2, but shows normal gravitropism. Strain JK345 shows no first positive phototropism, and reduced gravitropism and second positive phototropism. Strain JK229 shows no measurable first positive phototropism, but normal gravitropism and second positive phototropism. Based on these data, it is suggested that: 1. gravitropism and phototropism contain at least one common element; 2. first positive and second positive phototropism contain at least one common element; and 3. first positive phototropism can be substantially altered without any apparent alteration of second positive phototropism.Abbreviation WT wild-type     

Comparison of the apoptotic processes induced by the oxysterols 7β-hydroxycholesterol and cholesterol-5β,6β-epoxide

Oxysterols have been shown to induce apoptosis in a variety of cell lines. The mechanism of oxysterol-induced apoptosis is mainly known at the post-mitochondrial level. The aim of the present study was to compare the pathway of apoptosis induced by the oxysterols 7-hydroxycholesterol (7-OH) and cholesterol-5,6-epoxide (-epoxide) in U937 cells. To this end, we employed a range of inhibitors of apoptosis; a broad-spectrum caspase inhibitor, a specific caspase-3 inhibitor and an inhibitor of cytochromec release and the antioxidants; trolox, ebselen and resveratrol. The three inhibitors of apoptosis prevented cell death induced by 7-OH; however, in -epoxide-treated cells, the inhibitor of cytochromec release did not protect against apoptosis. The cellular antioxidant glutathione was depleted in 7-OH-treated cells but not in cells incubated with -epoxide. Trolox, a water-soluble synthetic analogue of -tocopherol, prevented 7-OH-induced apoptosis but did not protect against cell death induced by -epoxide. Ebselen and resveratrol did not protect U937 cells against apoptosis induced by either 7-OH or -epoxide. Our results suggest that differences occur in the pathways of apoptosis induced by 7-OH and -epoxide in U937 cells.     

Polymorphisms and diversity of T-cell receptor-γ proteins expressed in mouse spleen

Bulk populations of T-cell receptor (Tcr) -expressing splenocytes from different inbred strains of mice were examined for the diversity of Tcr proteins. Immunoprecipitations with anti-C1/2, anti-C4, and anti-V1 sera demonstrated that splenocytes from B10.BR, C57BL/6, and C57L strains of mice expressed the same array of Tcr proteins, namely V1-C2, V1-C4, and V2-C1, although the Tcr heterodimers observed for each of these strains were biochemically distinct. Examination of bulk splenic Tcr heterodimers from several other inbred strains of mice demonstrated that each of the strains could be categorized into one of three basic phenotypes. For several reasons, the differences observed between the strains appeared to be solely dependent on polymorphisms of the Tcrg loci. First, F₁ mice co-expressed both parental Tcr phenotypes. Second, the distinguishing polymorphism between mice of phenotype 1 and phenotypes 2 or 3 was due to the presence of an N-linked glycosylation site within the Tcrg-C1 gene segment, previously described for BALB.B and C57BL/6 Tcrg-C1 genes. Finally, the V1-C4 polymorphism between mice of phenotype 3 and phenotypes 1 or 2 was due to differences in core protein size. Furthermore, the three defined Tcr chains were expressed independently of the major histocompatibility complex (MHC) haplotype. Although no striking qualitative differences in Tcr heterodimers were observed between strains (including those with autoimmune disorders), a quantitative difference in the relative amount of C4-encoded proteins was observed on Tcr splenocytes from both newborn euthymic and adult athymic mice when compared to adult Tcr splenocytes from euthymic mice. These results demonstrate that genetic polymorphisms exist among different mouse strains and suggest that selective developmental pressures may govern Tcr expression. Offprint requests to: J. A. Bluestone     

Interactions of Aβ(1–40) with Glycerophosphocholine and Intact Erythrocyte Membranes: Fluorescence and Circular Dichroism Studies

Deposition of amyloid peptide in human brain in the form of senile plaques is a neuropathological hallmark of Alzheimers disease (AD). Levels of a phospholipid breakdown product, glycerophosphocholine (GPC), also increase in AD brain. The effect of GPC on amyloid (1–40) peptide (A) aggregation in PBS buffer was investigated by circular dichroism and fluoresence spectroscopy; interactions of A and GPC with the intact erythrocyte membrane was examined by fluoresence spectroscopy. Fluorescamine labeled A studies indicate GPC enhances A aggregation. CD spectroscopy reveals that A in the presence of GPC adopts 14% more -sheet structure than does A alone. Fluorescamine anisotropy measurements show that GPC and A interact in the phospholipid head-group region of the erythrocyte membrane. In summary, both soluble A and GPC insert into the phospholipid head-group region of the membrane where they interact leading to -sheet formation in soluble A which enhances A aggregation.     

Architecture of the Alzheimer’s AβP Ion Channel Pore

We have proposed that the cytotoxic action of Alzheimers amyloid beta protein might be initiated by the interaction with the neuronal cell membrane, and subsequent formation of toxic ion channels. Consequently, AP toxicity can be explained on the basis of harmful ion fluxes across AP channels. The conformation of AP in membranes is not known. However, several models suggests that a transmembrane annular polymeric structure is responsible for the ion channel properties of the membrane-bound AP. To identify that portion of the AP molecule making up the conducting pore we have hypothesized that the region of the AP sequence in the vicinity of the hypothetical pore might interact with complementary regions in the adjacent AP subunits. We have further hypothesized that an interaction by a peptide segment would block AP conductance. To test this hypothesis we synthesized peptides that encompass the histidine dyad (H-H) previously hypothesized to line the pore. We report here that peptides designed to most closely match the proposed pore are, in fact, the most effective at blocking ion currents through the membrane-incorporated AP channel. As previously shown for Zn²⁺ blockade, peptide blockade is also asymmetric. The results also provide additional evidence for the asymmetric insertion of the AP molecules into lipid membranes, and give support to the concept that rings of histidines line the entry to one side of the AP pore.     

Identification of a cDNA encoding a plant Lewis-type alpha1,4-fucosyltransferase

Recently, it has been found that plants, including tomato (Lycopersicon esculentum), express the Lewis-a epitope, Gal1,3(Fuc1,4)GlcNAc, on some N-glycans. By searching the EST database, it was possible to identify a tomato cDNA encoding a protein, designated FucTC, of 413 amino acids with homology to plant and mammalian 1,3/4-fucosyltransferases. The cDNA was expressed in Pichia pastoris and the recombinant enzyme was found to transfer fucose from GDP-Fuc (K_m 16 M) to lacto-N-tetraose (Gal1,3GlcNAc1,3Gal1,4Glc; K_m 80 M) as well as to 1,3- and 1,4-galactosylated N-glycans. It is concluded that FucTC is responsible for the biosynthesis of Lewis-a on N-glycans in tomato.     

Determination of three enzyme polymorphisms in one electrophoretic step on horizontal polyacrylamide gels

Summary A method that allows separation and determination of three enzyme polymorphisms in one polyacrylamide gel is described. Common phenotypes of adenylate kinase, adenosine deaminase and phosphoglucomutase are diagnosed without difficulties. There are small migration differences between PGM isozymes e and d.

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Zusammenfassung Es wird eine Methode beschrieben, die Trennung und Darstellung von drei Enzympolymorphismen in einem Polyacrylamid-Gel ermöglicht. Die Normtypen der Adenylatkinase, Adenosindeaminase und Phosphoglucomutase sind zweifelsfrei zu differenzieren. Geringe Wanderungsunterschiede ergeben sich zwischen den Zonen e und d der PGM Isoenzyme.

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(Direktor: Prof. Dr. W. Janssen)     

Enzymatic syntheses of GlcNAcβ1-2Man and Galβ1-4GlcNAcβ1-2Man as components of complex type sugar chains

GlcNAc1-2Man and GlcNAc1-6Man were synthesized using the reverse hydrolysis activity of -N-acetylglucosaminidase from both jack beans and Bacillus circulans. In turn, Gal1-4GlcNAc1-2Man and Gal1-4GlcNAc1-6Man were synthesized regioselectively using the transglycosylation activity of -galactosidase from Diplococcus pneumoniae and B. circulans, respectively. These di- and trisaccharides are important components of complex type sugar chains and will be used as intermediates in our synthetic studies. Abbreviations: pNp--GlcNAc, p-nitrophenyl 2-acetamido-2-deoxy--D-glucopyranoside; pNp--Gal, p-nitrophenyl -D-galacto-pyranoside