Toxigenic fungi and the deterioration of Nigerian poultry feeds

Growth of toxigenic strains of Aspergillus clavatus Des. and Aspergillus flavus Link at 30°C on milled poultry feeds led to a considerable decrease in the protein, oil and crude fibre contents of the feed substrate. A corresponding increase in the free fatty acid fractions of the feeds due to the activities of these microbes was also recorded. Rapid degradation of the feedstuff by both species was recorded at a temperature of 25°C and 30°C and a pH range of 4.8–6.4. When grown on feed infusion broth at 30°C, the highest amounts of mycelial production with sporulation of both fungal species occurred within the 8-day incubation period. A determination of their extra-cellular enzyme profile showed the production of amylases, pectate lyase, cellulases, proteases, lipases, xyalanases, DNase and RNase.All the carbon and nitrogen sources used (except L-sorbose and DL-tryptophan), supported good mycelial growth with sporulation. An optimal CN ratio of 5.04.5 and 7.53.0 was recorded for growth and sporulation of A. clavatus. For A. flavus, a CN ratio of 7.54.5 was found best for growth and 5.03.0 for sporulation.     

Comparison of fatty acids of marine fungi using multivariate statistical analysis

Summary Ten obligate marine fungi have as their principal fatty acids 160, 180, 181n9 and 182n6. The fatty acids ranged from 14 to 22 carbons, completely dominated by those with even numbers of carbons. The amount of unsaturated fatty acids varied between 35% and 80%. Each isolate contained small amounts of the acids 183n3 and 204n6. Branched, hydroxy- or cyclic fatty acids were not detected. Multivariate statistical, i.e. principal component analysis, showed that all ten strains could be distinguished on the basis of their fatty acid composition. These results indicate that the marine fungi do not have an unusual fatty acid composition and suggest that chemometric, multivariate analysis might be employed to confirm taxonomic relationships among these organisms.     

Bioassembly of acyl lipids in microspore-derived embryos of Brassica campestris L.

The native lipid composition and the capacity of cell-free extracts to biosynthesize acyl lipids in vitro were determined for the first time using the recently reported microspore-derived (MD) embryo system from the Brassica campestris low erucic acid line BC-2 (Baillie et al. 1992). The total lipid fraction isolated from midcotyledonary stage MD embryos (21 days in culture) was composed primarily of triacylglycerol (76%) with an acyl composition quite similar to that of mature BC-2 seed. When incubated in the presence of glycerol-3-phosphate, ¹⁴C 181-CoA, and reducing equivalents, homogenates prepared from 21-day cultured MD embryos were able to biosynthesize glycerolipids via the Kennedy pathway. The maximum in vitro rate of triacylglycerol biosynthesis could more than account for the known rate of lipid accumulation in vivo. The homogenate catalyzed the desaturation of 181 to 182 and to a lesser extent, 183. The newly-synthesized polyunsaturated fatty acids initially accumulated in the polar lipid fraction (primarily phosphatidic acid and phosphatidylcholine) but began to appear in the triacylglycerol fraction after longer incubation periods. As expected for a low erucic acid cultivar, homogenates of MD embryos from the BC-2 line were incapable of biosynthesizing very long chain monounsaturated fatty acyl moieties (201 and 221) from 181-CoA in vitro. Nonetheless, embryo extracts were still capable of incorporating these fatty acyl moieties into triacylglycerols when supplied with ¹⁴C 201-CoA or ¹⁴C 221-CoA. Collectively, the data suggest that developing BC-2 MD embryos constitute an excellent experimental system for studying pathways for glycerolipid bioassembly and the manipulation of this process in B. campestris.Abbreviations CPT sn-1,2-diacylglycerol cholinephosphotransferase - DAG diacylglycerol - DGAT diacylglycerol acyltransferase - DGDG digalactosyldiacylglycerol - G-3-P glycerol-3-phosphate - G-3-PAT glycerol-3-phosphate acyltransferase - LPA lyso-phosphatidic acid - LPAT lyso-phosphatidic acid acyltransferase - LPC lyso-phosphatidylcholine - LPCAT acyl-CoA: lyso-phosphatidylcholine acyltransferase - LPE lyso-phosphatidylethanolamine - MGDG monogalactosyldiacylglycerol - PA phosphatidic acid - PA Phosphatase, phosphatidic acid phosphatase - PC phosphatidylcholine - PE phosphatidylethanolamine - PG phosphatidylglycerol - TAG triacylglycerol - 181-CoA oleoyl-Coenzyme A - 181 oleic acid, cis-9-octadecenoic acid - 182 linoleic acid, cis-9,12-octadecadienoic acid - 183 -linolenic acid, cis-9,12,15-octadecatrienoic acid - 201 cis-11-eicosenoic acid - 221 erucic acid, cis-13-docosenoic acid; all other fatty acids are designated by number of carbon atoms: number of double bonds National Research Council of Canada Publication No. 35896     

Temperature adaptation of biological membranes. Compensation of the molar activity of cytochrome c oxidase in the mitochondrial energy-transducing membrane during thermal acclimation of the carp (cyprinus carpio L.)

The phospholipid composition, fatty acid pattern and cholesterol content are studied in mitochondria of red lateral muscle of carp acclimated to high and low environmental temperatures.The results of the experiments are: mitochondria from cold-acclimated carp contain higher proportions of ethanolamine phosphatides than mitochondria from warm-acclimated fish, the opposite is true for the choline phosphatides. Thus, at constant pH, the membrane phospholipids are slightly more negatively charged at low acclimation temperature. The total plasmalogen content is reduced in the cold; this reduction is caused by a decrease in the proportion of the choline plasmalogens. The ethanolamine phosphoglycerides contain approx. 20% of the alk-1-enyl acyl type, irrespective of the acclimation temperature. There is no temperature-dependent difference in the low proportion of cholesterol.The fatty acids of total mitochondrial phospholipids are characterized by large amounts of the n-3 and n-6 families. The ratio of unsaturated to saturated fatty acids and the unsaturation index are remarkably higher than those reported for comparable mammalian phospholipids. Cold acclimation of carp does not significantly increase the unsaturation of total phospholipids. A fatty acid analysis of the main isolated phospholipids, however, shows that cold acclimation considerably increases unsaturation of the neutral phosphatidylcholine, whereas it dramatically decreases unsaturation of the negatively charged cardiolipin. It is suggested that the observed fatty acid substitution in phosphatidylcholine indicates a temperature-induced fluidity adaptation within the mitochondrial lipid bilayer, whereas the inverse acclimation pattern of cardiolipin provides a suitable lipid to accommodate the temperature-dependent modifications in the dynamic surface shape of integral membrane proteins.     

Structure and properties of aCephalosporium acremonium α-galactosidase

The amino acid and sugar composition of the enzyme protein, the effect of urea, sodium dodecyl sulphate and Concanavalin A on the purified -galactosidase (EC 3.2.1.22) from the moldCephalosporium acremonium has been studied. The results obtained by gas liquid chromatography indicated the presence ofN-acetylglucosamine, mannose, galactose andN-acetylneuramic acid in the molar proportions 27311. The presence of two types of Asn-linked oligosaccharide structures in the enzyme molecule is assumed. The -galactosidase liberates (1–3), (1–4) and (1–6)-linkedd-galactose units from various synthetic and natural substrates which have been tested. The effects of pH, substrate concentration and temperature on the catalytic activity of the enzyme are described. The purified -galactosidase also exhibited a lectin activity with an affinity towards glucose, and to some extent mannose.Abbreviations p-NPG p-nitrophenyl--d-galactopyranoside - 4-MUG 4-methylumbelliferyl--d-galactopyranoside - HU hemagglutinin unit - PBS phosphate buffered saline - SDS sodium dodecyl sulphate - ConA Concanavalin A - WGA wheat germ agglutinin - LCA Lens culinaris agglutinin - PHA phytohemagglutinin fromPhaseolus vulgaris     

A novel fungal ω3-desaturase with wide substrate specificity from arachidonic acid-producing Mortierella alpina 1S-4

A filamentous fungus, Mortierella alpina 1S-4, is capable of producing not only arachidonic acid (AA; 20:4n-6) but also eicosapentaenoic acid (EPA; 20:5n-3) below a cultural temperature of 20°C. Here, we describe the isolation and characterization of a gene (maw3) that encodes a novel 3-desaturase from M. alpina 1S-4. Based on the conserved sequence information for M. alpina 1S-4 12-desaturase and Saccharomyces kluyveri 3-desaturase, the 3-desaturase gene from M. alpina 1S-4 was cloned. Homology analysis of protein databases revealed that the amino acid sequence showed 51% identity, at the highest, with M. alpina 1S-4 12-desaturase, whereas it exhibited 36% identity with Sac. kluyveri 3-desaturase. The cloned cDNA was confirmed to encode the 3-desaturase by its expression in the yeast Sac. cerevisiae. Analysis of the fatty acid composition of the yeast transformant demonstrated that 18-carbon and 20-carbon n-3 polyunsaturated fatty acids (PUFAs) were accumulated through conversion of exogenous 18-carbon and 20-carbon n-6 PUFAs. The substrate specificity of the M. alpina 1S-4 3-desaturase differs from those of the known fungal 3-desaturases from Sac. kluyveri and Saprolegnia diclina. Plant, cyanobacterial and Sac. kluyveri 3-desaturases desaturate 18-carbon n-6 PUFAs, Spr. diclina 3-desaturase desaturates 20-carbon n-6 PUFAs and Caenorhabditis elegans 3-desaturase prefers 18-carbon n-6 PUFAs as substrates rather than 20-carbon n-6 PUFAs. The substrate specificity of M. alpina 1S-4 3-desaturase is rather similar to that of C. elegans 3-desaturase, but the M. alpina 3-desaturase can more effectively convert AA into EPA when expressed in yeast. The M. alpina 1S-4 3-desaturase is the first known fungal desaturase that uses both 18-carbon and 20-carbon n-6 PUFAs as substrates.     

Adaptation of biological membranes to temperature. The effect of temperature acclimation of goldfish upon the viscosity of synaptosomal membranes

The fluidity of synaptosomal membrane preparations isolated from goldfish acclimated to 5, 15 and 25°C and from rat has been estimated using the fluorescence polarisation technique with 1,6-diphenyl-1,3,5-hexatriene as probe. Membranes of cold-acclimated goldfish were more fluid than those of warm-acclimated goldfish when measured at an intermediate temperature, indicating a temperature-dependent regulation of this parameter. Similarly, membranes of warm-acclimated goldfish were more fluid than those prepared from rat brain. Liposomes prepared from the purified phospholipids of goldfish and rat synaptosomal preparations showed differences similar to those of the native membranes. Increased membrane fluidity of cold-acclimated goldfish was correlated with a decrease in the proportion of saturated fatty acids of the major phospholipid classes and an increased unsaturation index in choline phosphoglycerides. Rat membranes showed a substantial reduction in unsaturation index and an increase in the proportion of saturated fatty acids compared to the membranes of 25°C-acclimated goldfish. The cholesterol content of synaptosomal membranes of goldfish was unaffected by acclimation treatment.The role of homeoviscous adaptation in the compensation of the rates of membrane processes during thermal acclimation, and upon the resistance adaptation of poikilotherms to extreme temperatures is discussed.     

Developmental changes in the fatty acids of synaptic membrane phospholipids: Effect of protein malnutrition

The acyl-linked fatty acid composition of the major phospholipid species in rat cortical synaptic membranes was determined at various stages of development. For most species there was a decrease during development in the short chain saturated fatty acids, 140 and 160, an increase in 180 and 226 (n-3) and an increase in the ratio of 226 (n-3)/225 (n-6). Pups were protein deprived by feeding the dams a 12% casein diet as compared to the 24% casein control diet. Protein malnutrition markedly affected the composition of acyl-linked fatty acids in the synaptic membranes. The increases in the ratio of 226 (n-3)/225 (n-6) fatty acids were especially compromised.Abbreviations used (PC) Phosphatidylcholine - (PE) phosphatidylethanolamine - (EP) ethanolamine plasmalogen - (PS) phosphatidylserine - (PI) phosphatidylinositol - (SP) sphingomyelin - (t.l.c.) thin layer chromatography - (BHT) butylated-hydroxytoluene     

Effects of photoperiod,temperature, food and relative humidity on the induction of diapause in the predatory miteAmblyseius potentillae

Aspects of the induction of diapause were studied in a Dutch strain of the phytoseiid miteAmblyseius potentillae. The photoperiodic response curve was of the long-day type, with a sharply defined critical daylength of 14.5 h. Critical daylength varied only little at temperatures between 15.0 and 22.5°C.All post-embryonic and possibly even late-embryonic stages of development were found to be sensitive to photoperiod; sensitivity appeared to be maximal during the protonymphal stage.It is shown that -carotene is necessary for some early step in the physiological mechanism of photoperiodic induction, and not (or not exclusively) for the expression of the diapause response.Two points of sensitivity to light could be demonstrated in the nights ofld 1311 andld 1212 long-night regimes, using 1-h night interruptions. These results are similar to those obtained in lightbreak experiments with spider mites and insects. However, no effect was found with light interruptions applied during the dark phase of anld 1014 long-night regime.In resonance experiments with a constant photophase (12 h) and a variable scotophase, a weak rhythmic response was found at 22.5°C; at 19.0°C this effect was completely absent.The relative humidity experienced by the mites during diapause induction as well as during diapause development influenced the rate of diapause completion under long days (ld 168). Diapause duration appeared to be shortest when the mites experienced low relative humidity (35±5%) during diapause induction and high relative humidity (75±5%) during diapause termination, and longest under the reverse conditions.     

The role of α-amylase of symbiotic microflora in digestion by lower cestodes and their fish hosts

The symbiotic microflora associated with the digestive-transport surfaces of the pike intestine and the parasitic cestodes Triaenophorus nodulosus proved capable of the initial stages of carbohydrate hydrolysis mediated by -amylase. The products of hydrolysis by -amylase can be used by both the host and the parasite, which decreases energy expenditures of the macroorganisms. The levels of the bacterial -amylase activity are comparable to those of the analogous enzyme absorbed on the mucosa of the intestine and on the cestode tegument, which indicates a considerable contribution of enzymes of the symbiotic microflora to digestion by the host and the parasite. Apparently, this contribution depends on the fish diet.Translated from Izvestiya Akademii Nauk, Seriya Biologicheskaya, No. 2, 2005, pp. 208–213.Original Russian Text Copyright © 2005 by Izvekova, Komova.     

Temperature acclimation in brook trout muscle: Adenine nucleotide concentrations, phosphorylation state and adenylate energy charge

Summary Cold acclimation in fish is associated with an elevation in metabolic rate. The present study investigates the role of adenine nucleotides and related compounds in metabolic regulation following temperature acclimation. Brook trout (Salvelinus fontinalis) were acclimated for 10 weeks to either +4°C or +24°C. Both groups of fish were exercised at 2.5 body lengths s^–1 for 2 weeks prior to sacrifice in order to control for differences in spontaneous activity.Concentrations of ATP, ADP, AMP, P _i and PC were approximately 2-fold higher in white than red muscles. Temperature acclimation had little effect on total adenine nucleotide concentration in either muscle type. In white fibres acclimation to 4°C results in a 39% increase in [ADP] and [AMP], a 35% decrease in [PC] (phosphorylcreatine), and no significant change in [P _i ]. In contrast temperature has little effect on concentrations of these compounds in red muscle.Parameters of metabolic control — adenylate energy charge ([ATP]+0.5 [ADP]/[ATP]+[ADP]+[AMP]), phosphorylation state ([ATP]/[ADP]·[P _i ]), and the ratios [ATP][ADP] and [ATP][AMP] — were significantly lower in cold- than warm-acclimated white muscle. The observed changes in phosphorylation state and [ATP][AMP] are consistent with an increase in mitochondrial respiration and glycolysis, respectively.In conclusion, changes in metabolites may be an important factor in producing an enhanced metabolic rate in cold-acclimated fish.     

Temperature adaptation of biological membranes. The effects of acclimation temperature on the unsaturation of the main neutral and charged phospholipids in mitochondrial membranes of the carp (cyprinus carpio L.)

The phospholipid composition, fatty acid pattern and cholesterol content are studied in mitochondria of red lateral muscle of carp acclimated to high and low environmental temperatures.The results of the experiments are: mitochondria from cold-acclimated carp contain higher proportions of ethanolamine phosphatides than mitochondria from warm-acclimated fish, the opposite is true for the choline phosphatides. Thus, at constant pH, the membrane phospholipids are slightly more negatively charged at low acclimation temperature. The total plasmalogen content is reduced in the cold; this reduction is caused by a decrease in the proportion of the choline plasmalogens. The ethanolamine phosphoglycerides contain approx. 20% of the alk-1-enyl acyl type, irrespective of the acclimation temperature. There is no temperature-dependent difference in the low proportion of cholesterol.The fatty acids of total mitochondrial phospholipids are characterized by large amounts of the $\text{n-3}\text{and}\text{n-6}$ families. The ratio of unsaturated to saturated fatty acids and the unsaturation index are remarkably higher than those reported for comparable mammalian phospholipids. Cold acclimation of carp does not significantly increase the unsaturation of total phospholipids. A fatty acid analysis of the main isolated phospholipids, however, shows that cold acclimation considerably increases unsaturation of the neutral phosphatidylcholine, whereas it dramatically decreases unsaturation of the negatively charged cardiolipin. It is suggested that the observed fatty acid substitution in phosphatidylcholine indicates a temperature-induced fluidity adaptation within the mitochondrial lipid bilayer, whereas the inverse acclimation pattern of cardiolipin provides a suitable lipid to accommodate the temperature-dependent modifications in the dynamic surface shape of integral membrane proteins.     

Calcium regulatory proteins and temperature acclimation of actomyosin ATPase from a eurythermal teleost (Carassius auratus L.)

Summary Goldfish (Carassius auratus) were acclimated for 5 months at temperatures of either 2°C or 31°C. Natural actomyosin was prepared from white myotomal muscle and its Mg²⁺Ca²⁺ ATPase activity determined. Temperature acclimation results in adaptations in substrate turnover number and thermodynamic activation parameters of the ATPase. When assayed at 31°C the Mg²⁺Ca²⁺ ATPase of natural actomyosin was 4 times higher in 31°C than 2°C acclimated fish. Arrhenius plots of natural actomyosin ATPase from cold acclimated fish show a break in slope at 15–18°C. In contrast, the temperature dependence of warm acclimated actomyosin was linear. Activation enthalpy (H ) of the ATPase, calculated over the range 0–16°C, was approximately 8,000 cal/mole lower in 2°C than 32°C acclimated fish.In contrast, desensitised actomyosins from which the calcium regulatory proteins have been removed show a linear temperature dependence in the range 0–32°C and have similar properties in 2°C and 31°C acclimated fish. Cross-hybridisation of regulatory proteins (tropomyosin-troponins complex) from cold-acclimated fish to desensitised actomyosin from warm-acclimated fish alters the ATPase towards that of cold-acclimated natural actomyosin and vice versa. The results suggest that the regulatory proteins can influence the kinetics of the ATPase and, furthermore, that they are involved in the acclimation of the actomyosin to different cell temperatures.     

Testing for non-randomness in sizes and habitats of West Indian lizards: choice of species pool affects conclusions from null models

Summary Several null models are proposed for testing whether size or habitat differences in West IndianAnolis lizards are greater than expected by chance. The models differ primarily in choice of the pool from which species are sampled to form random communities. Regardless of choice of pool, size differences in the Lesser Antilles are greater than null models predict; the pool using species on the known source (Puerto Rico) gives a greater variance in ratios but about the same mean ratio (for males), or a greater mean ratio (for females), compared with the pool composed of species on the islands being tested (the Stronget al., 1979, Galápagos procedure). On satellite islands of the Greater Antilles, sizes do not differ more than expected from null models. Pools composed of mainland-source species give null communities with more small or more large ratios than those composed of island species, depending upon whether four-species islands are included or excluded, respectively. Colwell and Winkler's unmodified Narcissus hypothesis is contradicted by these results in procedures where species not likely to be able to occur on small islands are included in the species pool. Using the most biologically reasonable, but not other, choices of source pool, species on satellite islands of the Greater Antilles differ more in structural habitat then expected by chance. In contrast to some of the results on size, here mainland-source pools are more likely to produce a statistically significant difference between real and random communities, as predicted by the Narcissus hypothesis. However, exclusion of structural habitat categories not found on satellite islands is necessary to achieve this significance.     

Glycerolipid-fatty-acid desaturase deficiencies in chloroplasts from fruits of Capsicum annuum L.

Chloroplasts from fruits and leaves of Capsicum annuum cv. Bell Tower were purified on sucrose gradients, and the lipids were separated by column and thin-layer chromatography. The glycerolipids mono- and digalactosyldiacylglycerol (MGDG, DGDG), sulfoquinovosyldiacylglycerol (SQDG), and phosphatidylglycerol (PG) were quantified, and the fatty-acid composition at the 1 and 2 positions of the glycerol moiety (sn-1 and sn-2) was determined after hydrolysis with position-specific lipases. In fruit chloroplasts, ³-trans hexadecenoate (trans-3-161) was absent and replaced by palmitate (160) at sn-2 of PG, and ^7,10,13-hexadecatrienoate (163) at sn-2 of MGDG was greatly reduced and largely replaced by linoleate (182). The ratio of 182 to linolenate (183) was consistently greater in glycerolipids from fruit compared with leaf chloroplasts. The lower percentage of C-16 fatty acids at sn-2 indicated that prokaryotic molecular species were reduced by 15% in DGDG, 40% in SQDG, and 90% in MGDG, in fruit compared with leaf chloroplasts. The MGDGDGDG ratios in fruit and leaf chloroplasts were 1.21 and 2.21, respectively. Taken together, the data indicate that chloroplasts in Capsicum fruit are deficient in three desaturases: those that convert 1) 160 to ³-trans-161 at sn-2 of PG, 2) 160 to ⁷–cis-161 at sn-2 of MGDG, and 3) 182 to 183 at both sn-1 and sn-2 of various chloroplast glycerolipids.Abbreviations Chl chlorophyll - DGDG digalactosyldiacylglycerol - FS free sterol - GL galactolipid - MGDG monogalactosyldiacylglycerol - PE phosphatidyl ethanolamine - PG phosphatidylglycerol - PL phospholipid - SQDG sulfoquinovosyldiacylglycerol We are grateful to Dr. Roger Calza for providing us with the tobacco gt11 cDNA expression library and to Dr. Eric Huttner for his advice throughout the screening procedure. We also wish to thank M. Gosse for his assistance in growing and maintaining our plants. T.W.B. was supported by a BAP research grant from the Commission of the European Communities.     

Cellular fatty acid composition of six fastidious,gramnegative, xylem-limited bacteria from plants

Composition of cellular fatty acids was determined for strains of fastidious, Gramnegative, xylem-limited bacteria causing or associated with Pierce's disease of grapevine, phony disease of peach, plum leaf scorch, stunt of ragweed, elm leaf scorch, and periwinkle wilt. The most abundant fatty acids were straight-chain 150, 160, 170, and 180, unsaturated 161, 181, and unsaturated 17-and 19=carbon homologs. Minor fatty acids included straightchain 120, 140, 190, and 200; an unsaturated 15-carbon homolog; hydroxy-substituted 2-OH 120, 3-OH 120, and 3-OH 140; and branched chain iso-140 and iso-200. Cyclopropane acids were not detected. Physiological age had no effect on fatty acid composition. Class analysis of data indicated relative uniformity within the group. Saturated even-carbon chains comprised 31%–42%, unsaturated acids 41%–52%, saturated odd-carbon chains 10%–18%, hydroxysubstituted chains 2%–7%, and branched-chains 1%–4% of total fatty acids. The ratio of saturated-unsaturated acids ranged from 0.8 to 1.2.     

Effect of extracellular calcium on the contractility of warm-and cold-acclimated crucian carp heart

Crucian carp (Carassius carassius L.) were acclimated for at least 4 weeks to 2°C or 22°C, and the consequences of thermal acclimation on force development, time-course of contraction and action potential duration of the ventricular myocardium were studied. In cold-acclimated fish contraction was activated at much lower external [Ca] than in warm-acclimated fish: [Ca] for half-maximal force was 0.9±0.15 and 3.1±0.92 mmol·l^-1 (P<0.05) for cold- and warm-acclimated fish, respectively. Durations of contraction and relaxation were significantly longer in fish acclimated to 2°C than in fish acclimated to 22°C, especially at [Ca] below 2 mmol·l^-1. In low-Ca solution ventricular action potential was prolonged both in cold- and warm-acclimated fish. In 0.5 mmol·l^-1 Ca action potential duration at zero voltage level was longer in cold- than warm-acclimated fish. Although lengthening of action potential was evident in both acclimation groups, a marked prolongation of contraction duration by low-Ca solutions occurred only in cold-acclimated fish. This suggests that a plateau component of contraction is present in cold-acclimated fish but less well developed in warm-acclimated fish hearts. Contractions were strongly inhibited by sarcolemmal Ca-channel blocker, cadmium (100 and 300 mol·l^-1), in both warm- and cold-acclimated crucian carp hearts. However, the sarcoplasmic reticulum Ca release channel blocker, ryanodine (10 mol·l^-1), had no effect on the force of contraction in either acclimation group. These results suggest that the contraction of crucian carp heart is controlled by sarcolemmal mechanisms without contribution by sarcoplasmic reticulum Ca release. Since the Ca sensitivity of myofilaments was not altered by thermal acclimation, the results indicate that thermal acclimation alters Ca activation of contraction of the crucian carp heart at the level of sarcolemma.Abbreviations AP action potential - EGTA ethyleneglycol-bis-(-aminoethylether)-N,N,N,N-tetra-acetic acid - F _max maximum force - F _max maximum rate of contraction - F _min maximum rate of relaxation - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulphonic acid - pCa log [Ca] - Pl action potential plateau - SL sarcolemma - SR sarcoplasmic reticulum - TPF time to peak force - T1/2R time to half relaxation from the peak force     

Diverse effects of essential (n−6 andn−3) fatty acids on cultured cells

Fatty acids (FAs) have long been recognized for their nutritional value in the absence of glucose, and as necessary components of cell membranes. However, FAs have other effects on cells that may be less familiar. Polyunsaturated FAs of dietary origin (n–6 andn–3) cannot be synthesized by mammals, and are termed essential because they are required for the optimal biologic function of specialized cells and tissues. However, they do not appear to be necessary for normal growth and metabolism of a variety of cells in culture. The essential fatty acids (EFAs) have received increased attention in recent years due to their presumed involvement in cardiovascular disorders and in cancers of the breast, pancreas, colon and prostate. Manyin vitro systems have emerged which either examine the role of EFAs in human disease directly, or utilize EFAs to mimic thein vivo cellular environment. The effects of EFAs on cells are both direct and indirect. As components of membrane phospholipids, and due to their varying structural and physical properties, EFAs can alter membrane fluidity, at least in the local environment, and affect any process that is mediated via the membrane. EFAs containing 20 carbons and at least three double bonds can be enzymatically converted to eicosanoid hormones, which play important roles in a variety of physiological and pathological processes. Alternatively, EFAs released into cells from phospholipids can act as second messengers that activate protein kinase C. Furthermore, susceptibility to oxidative damage increases with the degree of unsaturation, a complication that merits consideration because lipid peroxidation can lead to a variety of substances with toxic and mutagenic properties. The effects of EFAs on cultured cells are illustrated using the responses of normal and tumor human mammary epithelial cells. A thorough evaluation of EFA effects on commercially important cells could be used to advantage in the biotechnology industry by identifying EFA supplements that lead to improved cell growth and/or productivity.Abbreviations AA arachidonic acid (20 carbons: 4 double bonds,n–6) - BHA butylated hydroxyanisole - BHT butylated hydroxytoluene - cAMP cyclic adenosine monophosphate - CHO Chinese hamster ovary - DAG diacylglycerol - DGLNA dihomo--linolenic acid (203,n–6) - DHA docosahexaenoic acid (226,n–3) - EFA essential fatty acid - EGF epidermal growth factor - EGFR epidermal growth factor receptor - EPA eicosapentaenoic acid (205,n–3) - FA fatty acid - FBS fetal bovine serum - GLNA -linolenic acid (183,n–6) - LA linoleic acid (182,n–6) - LNA -linolenic acid (183,n–3) - LT leukotriene - MDA malondialdehyde - NAD nicotinamide adenine dinucleotide - NDGA nordihydroguaiaretic acid - OA oleic acid (181,n–9) - PG prostaglandin - PKC protein kinase C - PUFA polyunsaturated fatty acid - SFM serum-free medium - TX thromboxane     

Modification of the properties of a translocation in the German cockroach by selection

Summary A stock of Blattella germanica bearing the interchange T(3; 12)/3;12 was subjected to close inbreeding with selection for random disjunction at metaphase I. After 3–4 generations of selection, interchange quadrivalent chiasma frequency decreased, variability in free bivalent chiasma frequency increased sharply, and individuals with either random or directed disjunction were present in the stock. Random disjunction was modified from a ratio of 2112 (adj.-1; alt.-1; adj.-2; alt.-2) to a ratio of 1111. After 7–8 generations of selection, chiasma frequency appeared to stabilize at lower than normal levels and variability decreased for both quadrivalents and free bivalents. Directed disjunction was modified from a ratio of 2114 to 1112, and no individuals with the original high level of directed disjunction were detected. Chains-of-four tended to orient randomly, especially in individuals where the ring quadrivalents showed directed disjunction. Relaxation of inbreeding, but not selection, produced an increase in chiasma frequency and variability in both free bivalents and quadrivalents, but the modified ratios for both random and directed disjunction were retained. These results are discussed with respect to inbreeding effects and genetic control of chiasma frequency and metaphase I disjunction in interchange quadrivalents.     

Changes of carp myosin subfragment-1 induced by temperature acclimation

Summary Heavy meromyosin subfragment-1 (S1) was prepared by -chymotrypsin from myosin of carp acclimated to either 10°C or 30°C for a minimum of 5 weeks. The objective of these studies was to document thermally-induced changes in the myosin molecule and to extend previous observations. Ca²⁺- and K⁺ (EDTA)-ATPase activities of cold-acclimated carp S1 were 1.1 and 0.8 mol P_i·min^-1·mg^-1, respectively, and these values did not differ significantly from those of warm-acclimated carp. The inactivation rate constant (K_D) of S1 from cold-acclimated carp was 32.1x10^-4· s^-1, compared to 13.2x10^-4·s^-1 for warm-acclimated carp. The maximum initial velocity of acto-S1 Mg²⁺-ATPase activity at pH 7.0 in 0.05 M KCl was 9.3 s^-1 with cold-acclimated carp, about 3.7 times higher than that for warm-acclimated carp. However, no significant difference was observed in the apparent affinity of S1 to actin. Peptides maps of the heavy chain of S1 were different and suggested distinct isoforms for the myosins from warm- and cold-acclimated muscle.Abbreviations ATPase adenosine 5-triphosphatase - DTNB 5,5-dithiobis (2-nitrobenzoic acid) - DTT dithiothreitol - EDTA ethylenediaminetetraacetic acid - EGTA ethyleneglycol bis (-aminoethylether)-N,N,N,N-tetraacetic acid - K _D inactivation rate constant - K _m apparent dissociation constant - P _i inorganic -phosphate - PMSF phenylmethane-sulfonyl fluoride - S ₁ heavy meromyosin subfragment-1 - SDS sodium dodecyl sulfate - SDS-PAGE SDS-polyacrylamide gel electrophoresis - TPCK N-tosyl-l-phenylalanyl chloromethyl ketone - V _max maximum initial velocity