PHYSIOLOGICAL STRESS AND COLOR POLYMORPHISM IN THE INTERTIDAL SNAIL NUCELLA LAPILLUS

The intertidal snail Nucella lapillus exhibits considerable variation in shell color both within and between populations differentially exposed to wave action. Populations from high-wave-energy shores tended to be highly polymorphic and were dominated by pigmented morphs (especially brown), while those at more sheltered locations exhibited less polymorphism and were predominantly white. Field and laboratory experiments were conducted to determine the role of physiological stress and selective predation in maintaining the observed distribution of color morphs. The results demonstrated that 1) physiological stress from high temperature and desiccation during periods of tidal emersion was greater on protected shores, 2) under similar natural conditions, brown morphs heated up faster, attained higher temperatures, desiccated more rapidly, and suffered greater mortality than did white morphs, and 3) when pairs of brown and white morphs were tethered intertidally there was virtually no mortality of either morph on the exposed shore or in shaded microhabitats on the protected shore, but brown morphs suffered much greater mortality in sunny microhabitats on the protected shore. These findings demonstrate that the interpopulation variation in shell color of N. lapillus is in part a response to a selective gradient in physiological stress. Selection for crypsis by visually hunting predators did not appear to play a prominent role; however, only adults were considered, and the predation experiments were conducted in the fall before shorebirds that prey on whelks had arrived from their summer feeding grounds. Further experimentation to quantify the effects of visual predators such as birds and fish, particularly on juvenile snails, is necessary to assess adequately the importance of predation.     

The hemoglobins of Artemia salina. IV. A model for genetic control of hemoglobin 1, hemoglobin 2, and hemoglobin X

Two loci account for all genetic variation resulting in difference in electrophoretic mobility in three hemoglobins (Hb1, Hb2, and HbX) in the hemolymph of the brine shrimp. Four alleles and nine alleles have been studied. In shrimps of all genotypes and in electrophoresis in media with varying degrees of molecular sieving, Hb2 is approximately equidistant from Hb1 and HbX. A shrimp heterozygous at both loci has a three-banded Hb1, a four-banded Hb2, and a three-banded HbX. We conclude that Hb2 contains n -polypeptides and n -polypeptides. Hb1 contains 2n -polypeptides. HbX contains 2n -polypeptides. During electrophoresis, the three native hemoglobins undergo reversible dissociation to n subunits. Subunits with the same charge reassemble to migrate as molecules of the same size as the native molecules. Although there is no evidence for an additional polypeptide in the three hemoglobins, we cannot exclude such a possibility. If it exists, it is under three constraints: (1) it must be present in equal amounts in each of the three hemoglobins; (2) it must have the same molecular weight as the - and -polypeptides; and (3) it must be free of genetic variation (detectable by electrophoresis).Supported by National Institutes of Health Grant HE-11445.     

Differential expression of enzyme activities initiating anoxic metabolism of various aromatic compounds via benzoyl-CoA

The regulation of the expression of enzyme activities catalyzing initial reactions in the anoxic metabolism of various aromatic compounds was studied at the whole cell level in the denitrifying Pseudomonas strain K 172. The specific enzyme activities were determined after growth on six different aromatic substrates (phenol, 4-hydroxybenzoate, benzoate, p-cresol, phenylacetate, 4-hydroxyphenylacetate) all being proposed to be metabolized anaerobically via benzoyl-CoA. As a control cells were grown on acetate, or aerobically on benzoate. The expression of the following enzyme activities was determined.Phenol carboxylase, as studied by the isotope exchange between ¹⁴CO₂ and the carboxyl group of 4-hydroxybenzoate; 4-hydroxybenzoyl-CoA reductase (dehydroxylating); p-cresol methylhydroxylase; 4-hydroxybenzyl alcohol dehydrogenase; 4-hydroxybenzaldehyde dehydrogenase; coenzymeA ligases for the aromatic acids benzoate, 4-hydroxybenzoate, phenylacetate, and 4-hydroxyphenylacetate; phenylglyoxylate: acceptor oxidoreductase and 4-hydroxyphenylglyoxylate: acceptor oxidoreductase; aromatic alcohol and aldehyde dehydrogenases.The formation of most active enzymes is strictly regulated; they were only induced when required, the basic activities being almost zero. The observed whole cell regulation pattern supports the postulate that the enzyme activities play a role in anoxic aromatic metabolism and that the compounds are degraded via the following intermediates: Phenol 4-hydroxybenzoate 4-hydroxybenzoyl-CoA benzoyl-CoA; 4-hydroxybenzoate 4-hydroxybenzoyl-CoA benzoyl-CoA; benzoate benzoyl-CoA; p-cresol 4-hydroxybenzaldehyde 4-hydroxybenzoate 4-hydroxybenzoyl-CoA benzoyl-CoA; phenylacetate phenylacetyl-CoA phenylglyoxylate benzoyl-CoA plus CO₂; 4-hydroxyphenylacetate 4-hydroxyphenylacetyl-CoA 4-hydroxyphenylglyoxylate 4-hydroxybenzoyl-CoA plus CO₂ benzoyl-CoA.     

Analysis of the effect of rye chromosomes 4R, 6R and telomeric heterochromatin on 7R on homologous chromosome pairing in pentaploid hybrids (AABBR,AABBD)

Summary Meiotic pairing in Triticum turgidum cv. Ma (4x) with a mean chiasmata frequency of 27.16 per cell was compared with chiasmata frequencies in its hybrids with several triticale strains, Chinese Spring wheat and its addition lines for Imperial rye chromosomes 4R and 6R. In hybrids between Ma and x Triticosecale cv. Rosner the chiasmata frequency was marginally reduced by an average of 1.25%, by 8.8% in hybrids with x Triticosecale cv. DRIRA HH and by 6.7% with DRIRA EE (lacking 90% telomeric heterochromatin from chromosome arm 7RL). In pentaploid hybrids between Ma and T. aestivum cv. Chinese Spring the reduction was an average of 10.30%, while addition lines with rye chromosome 6R reduced chiasmata frequencies by an average of 7.4% and rye addition line for 4R showed the greatest depression in chiasmata frequency in hybrids by a 25.04% reduction. An interchange difference involving long chromosome segments was observed between Ma and Rosner.Contribution No. 819 Ottawa Research Station     

11 S seed storage proteins fromHelianthus species (Compositae): Biochemical,size and charge heterogeneity

The seeds of 19 sunflower species were compared on the basis of their protein contents and the relative proportions of their protein fractions. The globulin content varied from 50% to about 70% and the albumin content from 18% to 35% according to the species. The level of intermediateMr polypeptides showed a great variability (9.6 to 24.3%). Comparative studies onMr polymorphism were carried out by means of sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of non reduced and/or reduced samples using both mono- and bidimensional procedures. Polypeptide constituents of helianthinin were compared including both number and molecular size (cultivatedH. annuus was used as a standard). Studies focused on differences observed between the major two (Mr 38 000), (Mr 32 000) and (Mr 25 500), (21 000) polypeptides families constituting the main A, B, and C subunits. and polypeptides analyses permit to discriminate easilyH. petiolaris from the other species. Charge polymorphism was studied using isoelectric focusing (IEF) and IEF-PAGE in mono and bidimensional procedures in the presence or absence of 2-mercaptoethanol (2-ME). Only a specific ₄ polypeptide enables an easy discrimination betweenH. petiolaris and all the other species. Detailed nomenclature of the , and , polypeptides constituting the different helianthinin globulin subunits is given via the results of pI andMr analyses. Monodimensional IEF patterns of the more basic albumins (pI > 8.0) appear to provide a more valuable approach to identifying specific protein markers.     

A general strategy for the isolation of carbohydrate chains fromN-,O-glycoproteins and its application to human chorionic gonadotrophin

For the structural analysis of the carbohydrate chains ofN-,O-glycoproteins a straightforward strategy was developed based on the cleavage of theN-linked chains with immobilized peptide-N ⁴-(N-acetyl--glucosaminyl) asparagine amidase-F (PN-Gase-F) fromFlavobacterium meningosepticum, followed by alkaline borohydride treatment of the remainingO-glycoprotein material. This methodology was applied to the isolation of the Asn- and Ser-linked carbohydrate chains of human chorionic gonadotrophin. The structures of the isolated oligosaccharides were verified by 500-MHz¹H-NMR spectroscopy. The Asn-linked sugar chains were shown to be: NeuAc2-3Gal1-4GlcNAc1-2Man1-6[NeuAc2-3Gal1-4GlcNAc1-2Man1-3]Man 1-4GlcNAc1-4[Fuc1-6]_0-1GlcNAc and Man1-6[NeuAc2-3Gal1-4GlcNAc1-2Man 1-3]Man1-4GlcNAc1-4GlcNAc. Also some minor constituents occurred. The structures of the Ser-linked oligosaccharides were established in the form of their oligosaccharide-alditols as: NeuAc2-3Gal1-3[NeuAc2-6]GalNAc, NeuAc2-3Gal 1-3GalNAc and NeuAc2-3Gal1-3[NeuAc2-3Gal1-4GlcNAc1-6]GalNAc.Abbreviations hCG human chorionic gonadotrophin - hCG- -subunit - hCG- -subunit - ElA enzyme immunoassay - PNGase-F peptide-N ⁴-(N-acetyl--glucosaminyl)asparagine amidase-F (EC 3.5.1.52) - SDS sodium dodecyl sulphate - GalNAc N-acetylgalactosamine - GlcNAc N-acetylglucosamine - NeuAc N-acetylneuraminic acid - Man mannose - Gal galactose - Fuc fucose     

Adjacent 2 meiotic disjunction. Report of a case resulting from a familial 13q;15q balanced reciprocal translocation and review of the literature

Summary An abnormal short-lived female infant with almost complete trisomy 13 (pterq32 or 33) and partial monosomy 15 (pterq14 or 15) resulting from an adjacent 2 meiotic disjunction of a paternal reciprocal translocation is described. Cases with monosomy of chromosome 15 material are reviewed. It appears likely that monosomy of an interstitial long arm segment, approximating to 15q2124, imparts the lethality associated with the full monosomic condition. Adjacent 2 disjunction in man has been further characterised by reviewing the literature.     

Microcephalic cerebrum with hypomyelination in the congenital goiter mouse (cog)

2,3-Cyclic nucleotide 3-phosphohydrolase activity in the cerebrum of the congenital goiter mouse (cog/cog) is reduced in comparison with the normal heterozygote (cog/+). The weight of thecog/cog cerebrum and cerebellum were significantly less than those of the normal controls, 89.0% less for the cerebrum, and 81.1% less for the cerebellum. However, no differences were observed with regard to DNA and RNA content and the RNA/DNA ratio. The results of this study indicate that hypomyelination in the congenital goiter mouse is restricted to the cerebrum, and is not related to arrested glial proliferation.     

Genetic analysis of resistance to whitebacked planthopper in twenty-one varieties of rice,Oryza sativa L.

Summary The inheritance of resistance to whitebacked planthopper Sogatella furcifera (Horvath) was studied in 21 rice varieties. Reactions of F₁; F₂ and F₃ progenies of the crosses of 21 resistant varieties with the susceptible variety TN 1 revealed that a single dominant gene governs resistance in Mushkan 41, Santhi, Siahnakidar 195, SM2-34, Tirisurkh 251, Zirijowaian 245, 18, 24A, 39, 76 S, 78, 180, 213 B, 267, 293, CI 6037-4, NP97, S39 JKW and Bansphul. In varieties 65 and 274 A, resistance is governed by one dominant and one recessive gene which segregate independently of each other. Tests for allelism with the Wbph 1 gene originally identified in N 22 revealed that the dominant gene present in all the test varieties is the same as Wbph 1. Further studies are required to determine the allelic relationships of the recessive gene found in varieties 65 and 274 A.     

Na+-dependent accumulation of sulfate and thiosulfate in marine sulfate-reducing bacteria

Uptake of ³⁵S-labelled sulfate and thiosulfate was studied in twenty sulfate-reducing bacteria. Micromolar additions of these substrates were highly accumulated by washed cells of freshwater and marine strains. In marine strains accumulation required Na⁺. Generally, the uptake capacity was increased after sulfate limitation during growth. With two marine species, Desulfovibrio salexigens and Desulfobacterium autotrophicum, the effects of various ionophores and inhibitors affecting the transmembrane pH or Na⁺ gradient or the membrane potential were studied. In both strains transport was reversible. There was no discrimination between sulfate and thiosulfate. With increasing additions the amount taken up increased, while the accumulation factor (C_in/C_out) decreased. Uptake was not directly correlated with the ATP level inside the cells. From these results and the action patterns of the inhibitors tested it is concluded that marine sulfate-reducing bacteria accumulate sulfate and thiosulfate electrogenically in symport with Na⁺ ions, while in freshwater strains protons are symported. The high-accumulating systems are induced only at low sulfate concentration, while low-accumulating systems are active at sulfate-sufficient conditions.Abbreviations CCCP carbonyl cyanide m-chlorophenylhydrazone - DCCD dicyclohexylcarbodiimide - ETH 157 N, N-dibenzyl-N,N-diphenyl-1,2-diphenylendioxydiacetamide - TCS 3,3,4,5-tetrachlorosalicylanilide     

Further genetic variation at the esterase loci of Drosophila virilis

Reexamination of the electrophoretic mobilities of esterases encoded by the Est- and the Est- alleles of Drosophila virilis was carried out in detail using both thin-layer agar gel and polyacrylamide slab gel electrophoresis. Many allelic products with fine differences in their electrophoretic mobilities were found and designated by a new system. Some esterases separable by the agar gel method were indistinguishable using the polyacrylamide gel method. But the polyacrylamide gel method uncovered two multiband homozygotes, ^(d).77 and ^(d)1.28. Some allelic frequencies on the basis of the new designation were estimated in two natural populations. As a result, it is proposed that the total scope of allelic variation at the two esterase loci of Drosophila virilis is composed of discrete distribution patterns of gene frequencies, each histogram of which shows a bell-shaped pattern.     

Characterization of the specificity of binding ofMoluccella laevis lectin to glycosphingolipids

The specificity ofMoluccella laevis lectin was investigated by analysing its binding to glycosphingolipids separated on thin-layer chromatograms or adsorbed on microtitre wells. The binding activity of the lectin was highest for glycosphingolipids with terminal -linkedN-acetylgalactosamine, both in linear structures, as the Forssman glycosphingolipid, GalNAc3GalNAc3Gal4Gal4Glc1Cer, and in branched structures, as glycosphingolipids with the blood group A determinant, GalNAc3(Fuc2)Gal. In addition, the lectin bound, though considerably more weakly, to linear glycosphingolipids with terminal -linked galactose. When considering the use of theM. laevis lectin for biochemical and medical purposes this cross-reactivity may be of importance. Nomenclature: The glycosphingolipid nomenclature follows the recommendations by the IUPAC-IUB Commission on Biochemical Nomenclature (CBN for Lipids:Eur J Biochem (1977)79:11–21,J Biol Chem (1982)257:3347–51, andJ Biol Chem (1987)262:13–18). It is assumed that Gal, Glc, GlcNAc, GalNAc, and NeuAc are of thed-configuration, Fuc of thel-configuration, and all sugars present in the pyranose form.     

Production of a novel syrup containing neofructo-oligosaccharides by the cells of Penicillium citrinum

A novel syrup containing neofructo-oligosaccharides was produced from sucrose (Brix 70) by whole cells of Penicillium citrinum. The efficiency of fructo-oligosaccharides production was more than 55% and those of the main carbohydrate components, 1-kestose (Fruf 21Fruf 21 Glc), nystose (Fruf 21Fruf 21 Fruf 21 Glc) and neokestose (Fruf 26 Glc12 Fruf), were 22, 14 and 11%, respectively.     

EPR and Mössbauer spectroscopic studies on the tetrameric,NAD-linked hydrogenase of Nocardia opaca 1b and its two dimers: 1. The βδ-dimer—a prototype of a simple hydrogenase

The cytoplasmic, tetrameric NAD-linked hydrogenase from Nocardia opaca 1b can be separated in two dimeric substructures, an -dimer with NADH:electron acceptor oxidoreductase (diaphorase) activity and a -dimer which displays hydrogenase activity with artificial electron carriers. These two dimers were preparatively isolated by a FPLC Mono Q procedure in the absence of nickel and at alkaline pH values. The hydrogenase-active -dimer contained, as analyzed by inductively coupled plasma mass spectrometry (ICP-MS), 3.5–3.9 iron atoms and 1.3–1.7 nickel atoms per dimer molecule. EPR and Mössbauer spectra indicated the presence of a [4Fe-4S] cluster. This center turned out to be extremely labile towards oxidants. Oxidation led to irreversible convertion into a [3Fe-4S] form, thus representing an artifact and not a regulatory state of the cluster. The midpoint redox potential of the [4Fe-4S] cluster was determined to be -385 mV. Very weak EPR Ni signals of the -dimer were detectable in the oxidized as well as in the reduced state. The diaphorase-active -dimer was free of nickel and the iron content corresponded to 11.2–12.8 Fe atoms per dimer molecule. From EPR and Mössbauer measurements it was concluded that this dimer contained two [4Fe-4S] clusters, one [2Fe-2S] and one [3Fe-4S] cluster. In accordance with the results obtained for the dimer proteins, for the whole enzyme an iron content of 15.8–16.2 atoms per enzyme molecule have been determined. EPR spectra and spectrum simulations of the native hydrogenase corroborate the cluster assignments of the two dimers: in total the enzyme contains one [2Fe-2S] cluster, one [3Fe-4S] cluster and three [4Fe-4S] clusters.     

Selection of peach cells for insensitivity to culture filtrates of Xanthomonas campestris pv. pruni and regeneration of resistant plants

Summary Individual callus cultures were initiated from 400 immature embryos of bacterial leaf spot-susceptible Sunhigh peach. Each was subjected to several selection cycles of a toxic culture filtrate produced by Xanthomonas campestris pv. pruni, the causal agent of leaf spot of peach. Progressively higher concentrations of the filtrate were used in each cycle. Two calli survived, and two plants were regenerated from each of the surviving calli. Each of the four clones was propagated in vitro and tested for whole plant resistance to X. c. pv. pruni. Results from bioassays on greenhouse-grown plants indicated that two out of the four selected clones were significantly more resistant to X. c. pv. pruni than the parental cv Sunhigh. In addition, one clone was significantly more resistant than the moderately resistant cv Redhaven.     

Graphical evaluation and partitioning of turgor responses to drought in leaves of durum wheat

The relationship between relative water content (R) and turgor potential (_p) may be derived from pressure-volume (PV) curves and analyzed in various ways. Fifty PV curves were measured with the pressure chamber on leaves of durum wheat (Triticum durum L.). The plots of _p versus R were highly variable and could not be adequately described by a single mathematical function. The area below the curve was therefore determined by means of an area meter. This procedure gave the integral of turgor from full saturation to the turgor-loss point. Responses to drought treatment could thus be quantified and partitioned into effects of osmotic adjustment and elastic adjustment. These two adjustment responses, which are probably of different metabolic origin, together improve turgor maintenance in durum wheat considerably.Abbreviations and symbols PV pressure-volume - R relative water content - T_i turgor integral between full saturation and turgor-loss point - _p turgor (pressure) potential     

Adding teeth to wave action: the destructive effects of wave-borne rocks on intertidal organisms

Summary Observations in rocky intertidal areas demonstrate that breaking waves throw rocks and cobbles and that these missiles can damage and kill organisms. Targets in the intertidal were dented by impacts from wave-borne rocks. New dents/day in these targets was positively correlated with the daily maximum significant wave height and with new patches/day in aggregations of the barnacle Chthamalus fissus. Impact frequency was highest in the upper intertidal and varied dramatically between microhabitats on individual boulders (edges, tops and faces). These patterns were reflected in the microhabitat abundances of old and young barnacles. Comparisons were made of the survivorship and the frequency of shell damage in two populations of the limpet Lottia gigantea living in habitats which differed primarily in the number of moveable rocks (i.e. potential projectiles). The mortality rate and frequency of shell damage were significantly higher in the projectilerich habitat. In addition only in this habitat did the frequency of shell damage covary significantly with seasonal periods of high surf. Investigation of the response of limpet shells to impacts suggests that shell strength varies between species and increases with shell size. Species-specific patterns of non-fatal shell breakage may have evolved to absorb the energy of impacts. In two of the intertidal habitats studied, wave-borne rock damage was chronic and, at least in part, may have governed the faunal makeup of the community by contributing to the physical boundaries of the environment within which the inhabitants must survive.     

Projecting population-level response of purple sea urchins to lead contamination for an estuarine ecological risk assessment

As part of an ecological risk assessment casestudy at the Portsmouth Naval Shipyard (PNS), Kittery,Maine, USA, the population level effects of leadexposure to purple sea urchin, Arbaciapunctulata, were investigated using a stage-classifiedmatrix population model. The model divided the lifehistory of A. punctulata into five classes,incorporating both, the developmental stages of thisspecies and the endpoints from a laboratory bioassay. Finite population growth rate () was themetric relating population level impact to leadexposure. An inverse relationship was observed betweenlead tissue residues in A. punctulata and. Bioassay treatments which resulted insignificant impacts on fertilization success and zygoteviability did not translate into significant effects on, unless those treatments also negativelyimpacted adult survival. These results paralleled theelasticity (relative sensitivity) analysis of themodel, which indicated that was mostsensitive to adult and subadult survival and wasrelatively insensitive to fecundity, fertilizationsuccess, or zygote survival. Model results indicatedthat the environmental lead levels observed at PNSshould not pose significant ecological risk to seaurchin populations. Additionally, the model resultsindicated that impacts to the early life stagesroutinely used in toxicity testing do not necessarilytranslate directly into impacts at the populationlevel.     

New cytoplasmic male sterility sources in common wheat: Their genetical and breeding considerations

Summary Nuclei from Triticum aestivum L. cultivars Penjamo 62 and Siete Cerros 66 were introduced into the cytoplasms of different species of Aegilops and some subspecies (varieties) of T. dicoccoides by backcrossing. The sterile alloplasmic lines obtained were compared with the normal cultivars used as the recurrent pollen parents. According to the cytoplasmic effect, these cytoplasms were subdivided into three main groups. The first group possesses C^u type cytoplasm, the second one possesses M type and the third group includes S, C and G type. Promising male sterile cytoplasms for hybrid wheat production were found in Ae. mutica, Ae. triuncialis and T. dicoccoides var. spontaneovillosum. Based on these results and other information some conjectures were made concerning hybrid wheat breeding and phylogenetic differentiations of the cytoplasm.     

Expression of cell adhesion molecules,their ligands and tumour necrosis factor alpha in the liver of patients with metastatic gastrointestinal carcinomas

The expression of the following cell adhesion molecules, their 1 and 2 integrin ligands and the cytokine tumour necrosis factor- (TNF-) was investigated by light and electron microscope immunohistochemistry in the liver tissue in 20 patients with colorectal and gastric cancer also presenting with liver metastases: intercellular adhesion molecule-1 (ICAM-1), vascular endothelial adhesion molecule-1 (VCAM-1), E-selectin, leucocyte function-associated antigen-1 (LFA-1), macrophage antigen-1 (Mac-1), and very late antigen-4 (VLA-4). We have found a parallel enhancement of the adhesion molecules and of TNF- in liver sinusoids surrounding metastases. The expression of ICAM-1 was enhanced on sinusoidal cells in all zones of the acinus. VCAM-1 immune reactivity was diffuse but less intensive in the lobule. E-selectin expression was observed in sinusoidal cells attached to metastases. In tumour metastases the expression of ICAM-1, VCAM-1, and E-selectin was visible on the tumour vascular endothelium. Tumour infiltrating host cells sowing positive immunoreactivity for ICAM-1, VCAM-1, LFA-1, Mac-1, and VLA-4 were located mainly at the boundary between liver parenchyma and the metastasis. At the ultrastructural level, ICAM-1-positive immune deposits were observed on the cellular membrane and in some transport vesicles of gastric metastatic cells. Further, the expression of all adhesion molecules was confirmed to sinusoidal endothelial cells and tumour vessels. It is concluded that the enhanced expression of adhesion molecules in liver sinusoids could be a marker for the assessment of the ability of sinusoidal endothelial cells to control the recruitment of leukocytes and monocytes to the metastatic site. They could also direct the adhesion of new circulating tumour cells to sinusoidal endothelium.