Comparison of the effects of illumination on the melanophores of intact and eyestalkless fiddler crabs,Uca pugilator,and inhibition of the primary response by cytochalasin B

Summary Approximately 100 times more illumination is required to produce pigment dispersion in the melanophores of eyestalkless fiddler crabs (Uca pugilator) than in the melanophores of intact crabs. The pigment in melanophores of isolated legs will normally disperse in response to irradiation, but this response is inhibited by cytochalasin B.This investigation was supported by a Faculty Research Grant from Western Kentucky University toT. P. Coohill and by Grant No. GB-27497 toM. Fingerman from the National Science Foundation.     

Structure and function of coagulogen,a clottable protein in horseshoe crabs

Mammalian blood coagulation is based on the proteolytically induced polymerization of fibrinogens. Initially, fibrin monomers noncovalently interact with each other. The resulting homopolymers are further stabilized when the plasma transglutaminase (TGase) intermolecularly cross-links -(-glutamyl)lysine bonds. In crustaceans, hemolymph coagulation depends on the TGase-mediated cross-linking of specific plasma-clotting proteins, but without the proteolytic cascade. In horseshoe crabs, the proteolytic coagulation cascade triggered by lipopolysaccharides and b-1,3-glucans leads to the conversion of coagulogen into coagulin, resulting in noncovalent coagulin homopolymers through head-to-tail interaction. Horseshoe crab TGase, however, does not cross-link coagulins intermolecularly. Recently, we found that coagulins are cross-linked on hemocyte cell surface proteins called proxins. This indicates that a cross-linking reaction at the final stage of hemolymph coagulation is an important innate immune system of horseshoe crabs.Received 27 October 2003; received after revision 25 November 2003; accepted 1 December 2003     

Trends of karyological evolution in Pelobatoid frogs

Summary The Pelobatids have a basal karyotype of 26 chromosomes, with 6 pairs of large and 7 of small chromosomes (6+7). From this karyotype, shown byLeptobrachium, Pelobates andScaphiopus, those ofMegophrys andSpea (5+8) may be derived, as well as that ofL. pelodytoides, with only 24 chromosomes.Pelodytes has 24 chromosomes and a karyotype 7+5 differing from that of typical Pelobatids under several respects.Research supported by a contribution from the Italian C. N. R.     

Properties of a calcium-dependent apyrase in the saliva of the blood-feeding bug,Rhodnius prolixus

Summary The saliva of the blood-feeding bugRhodnius prolixus contains at least 11 proteins, and has Ca²⁺-dependent apyrase activity. The activity has a broad pH optimum between pH 7 and 9, and is inhibited by Mg²⁺. Polyacrylamide gel electrophoresis and gel filtration suggest the possibility of at least 2 enzymes responsible for the activity.This work was supported by the National Science and Engineering Research Council of Canada. We are grateful to D. F. Mettrick for use of equipment, and to R. Hewson for invaluable assistance.     

Etude de l'influence d'une microsporidiose sur les électrophorégrammes (protéinogramme et zymogramme) des protéines de l'hémolymphe deCarcinus mediterraneus (Crustacé, Brachyoure)

Summary The use of the method of concentration gradient of polyacrylamide gel, electrophoresis has permitted to reveal a difference in zymograms when studying the hemolymph of healthy crabs and that of crabs parasitized by the muscular microsporidaThelohania, maenadis., The zymograms of the parasitized crabs present an increase in chymotrypsin activity and variations in the zones of esterase activity.

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Contribution n^o 1 du Centre de Parasitologie et de Pathologie comparées de l'Université des Sciences et Techniques du Languedoc, F-34060 Montpellier Cédex.     

Erythropoietic effects of PGE2 and 2 endoperoxide analogs

Summary The erythropoietic effects in exhypoxic polycythemic mice of two endoperoxide analogs were assessed and compared with PGE₂. The 9a, 11a epoxymethano analog (U-44069) was found to be a much more potent erythropoietic stimulus than the 11a,9a analog (U-46619) or PGE₂.This work was supported by a Senior Research Grant-in-Aid from the American Heart Association-Louisiana (DMG) and USPHS Grant No. AM 13211 (JWF).     

Relationship between action potential, contraction-relaxation pattern, and intracellular Ca2+ transient in cardiomyocytes of dogs with chronic heart failure

Abnormalities of contractile function have been identified in cardiomyocytes isolated from failed human hearts and from hearts of animals with experimentally induced heart failure (HF). The mechanism(s) responsible for these functional abnormalities are not fully understood. In the present study, we examined the relationship between action potential duration, pattern of contraction and relaxation, and associated intracellular Ca²⁺ transients in single cardiomyocytes isolated from the left ventricle (LV) of dogs (n = 7) with HF produced by multiple sequential intracoronary microembolizations. Comparisons were made with LV cardiomyocytes isolated from normal dogs. Action potentials were measured in isolated LV cardiomyocytes by perforated patch clamp, Ca²⁺ transients by fluo 3 probe fluorescence, and cardiomyocyte contraction and relaxation by edge movement detector. HF cardiomyocytes exhibited an abnormal pattern of contraction and relaxation characterized by an attenuated initial twitch (spike) followed by a sustained contracture ('dome') of 1 to 8 s in duration and subsequent delayed relaxation. This pattern was more prominent at low stimulation rates (58% at 0.2 Hz, n = 211, 21% at 0.5 Hz, n = 185). Measurements of Ca²⁺ transients in HF cardiomyocytes at 0.2 Hz manifested a similar spike and dome configuration. The dome phase of both the contraction/relaxation pattern and Ca²⁺ transients seen in HF cardiomyocytes coincided with a sustained plateau of the action potential. Shortening of the action potential duration by administration of saxitoxin (100 nM) or lidocaine (30 μM) reduced the duration of the dome phase of both the contraction/relaxation profile as well as that of the Ca²⁺ transient profile. An increase of stimulation rate up to 1 Hz caused shortening of the action potential and disappearance of the spike-dome profile in the majority of HF cardiomyocytes. In HF cardiomyocytes, the action potential and Ca²⁺ transient duration were not significantly different from those measured in normal cells. However, the contraction-relaxation cycle was significantly longer in HF cells (314 ± 67 ms, n = 21, vs. 221 ± 38 ms, n = 46, mean ± SD), indicating impaired excitation-contraction uncou pling in HF cardiomyocytes. The results show that, in cardiomyocytes isolated from dogs with HF, contractile abnormalities and abnormalities of intracellular Ca²⁺ transients at low stimulation rates are characterized by a spike-dome configuration. This abnormal pattern appears to result from prolongation of the action potential. Received 22 January 1998; received after revision 16 March 1998; accepted 27 March 1998     

Sérotonine et scglycémie chez les crustacés

Summary Injection of 0.2 cm³ of 5-HT 0.1% inCarcinus maenas has a powerful hyperglycaemic effect and determines at the same time a dispersion of the melanin granules in chromatophores of eyestalkless crabs. In both instances, 5-HT mimics the effects of the injection of an extract from the eyestalks.     

Rotationssinn nach dem Bogengangsprinzip bei Crustaceen

Summary Blinded crabs (Carcinus maenas andMaja verrucosa) show compensatory eye stalk movements on rotation about a vertical axis, in water and in air. On a sudden arrest of the turntable typical after-effects are observed, which show a striking resemblance to those known from vertebrates. All these reactions are abolished after bilateral elimination of the so-called thread hairs within the statocysts (experiments onMaja). In fresh preparations these hairs can be seen to follow the slightest movements of the surrounding fluid, swaying around their point of attachment.Elimination of the thread hairs does not diminish, but does markedly delay the compensatory eye stalk movements of a blinded crab on rotation about horizontal axes. The latter reactions are abolished if in addition the hook hairs (i.e. the sense hairs of the statolith-bearing type) are destroyed.It is concluded that the statocyst of crabs is a staticdynamic sense organ: the hook hairs are position receptors, the thread hairs react to angular displacements about all three main body axes; the group hairs may have no sensory function at all.Rotation sense may be expected to occur in other Crustacea possessing thread hairs or similar receptors in their statocysts as well.     

Ca2+-dependence and metabolic status of an obligate thermophile,Thermoactinomyces vulgaris,under shake culture conditions

Summary Ca²⁺ stimulates germination ofT. vulgaris spores. There is a higher mycelial yield as well as higher protein, DNA, RNA and free Pi content in cultures grown in the presence of Ca²⁺ as compared to those grown in the absence of this divalent cation.Grateful thanks are due to Professor R. P. Roy for providing the necessary facilities. V. P. S. acknowledges with thanks the receipt of a Post Doctoral Research Fellowship of the Council of Scientific and Industrial Research, New Delhi.     

Increase in the total reducing substances in the hemolymph of the freshwater crab,Barytelphusa guerini,produced by a pesticide (DDT) and an indolealkylamine (serotonin)

Summary DDT and serotonin produced significant increases in the total reducing substances in the hemolymph of intact crabs,Barytelphusa guerini, apparently by triggering release of the hyperglycemic hormone.     

(±)9, 10-dihydroxy-Δ 6a(10a)-tetrahydrocanabinol and (±)8, 9-dihydroxy-Δ 6a(10a)-tetrahydrocannabinol: 2 new cannabinoids fromCannabis sativa L.

Summary The structures of 2 new polyhydroxylated cannabinoids, (±)9, 10-dihydroxy- ^6a(10a)-tetrahydorcannabinol and (±)8, 9-dihydroxy- ^6a(10a)-tetrahydrocannabinol, obtained from a hexane extract of an IndianCannabis variant were determined by spectral means and correlation with cannabinol.Acknowledgments. Supported by contract No. HSM-42-70-109 from the National Institute on Drug Abuse and the Research Institute of Pharmaceutical Sciences, School of Pharmacy, University of Mississippi. The authors are grateful to Mrs Glenda Lewis (Research Institute) for running the gas chromatographic analyses and to Mr C. Versluis (Analytical Chemistry Laboratory, University Of Utrecht, The Netherlands) for recording an element list of both compounds.Cannabis Herbarium specimens are stored in the herbarium, Department of Pharmacology, School of Pharmacy, University of Mississippi, University, MS 38677, USA.     

Pineal function in the sheep: evidence for a possible mechanism mediating seasonal reproductive activity

Summary The 24-h profiles of plasma melatonin in the intact ewe in natural light indicate that a bimodal pattern of secretion is frequently present in oestrus, whereas a single dark-phase peak is characteristic of anoestrus. Based on these findings, a mechanism for the possible pineal-mediated control of seasonal breeding is proposed.This work was supported by the Medical Research Council and the Agricultural Research Council of Great Britain. Preliminary results from this study were reported in the proceedings of the 1st meeting of the European Pineal Study Group; J. Arendt, Prog. Brain Res.52, 249 (1979).We are grateful for the excellent technical assistance of A. Marston and J. Bradtke.     

Toxicity ofPhytophthora infestans andAlternaria solani to chick embryos

Summary None of the extracts and culture filtrates from growth of 9 races ofPhytophthora infestans on living potato tissue and potato dextrose broth are toxic to chick embryos. Chloroform extracts of 4 out of 10 isolates ofAlternaria solani grown on potato dextrose broth showed some toxicity to chick embryos.Acknowledgment. This research was supported by grant FD-00683-03 from the Office of Research Grants, Food and Drug Administration.     

CSTX-9, a toxic peptide from the spider Cupiennius salei: amino acid sequence, disulphide bridge pattern and comparison with other spider toxins containing the cystine knot structure

CSTX-9 (68 residues, 7530.9 Da) is one of the most abundant toxic polypeptides in the venom of the wandering spider Cupiennius salei. The amino acid sequence was determined by Edman degradation using reduced and alkylated CSTX-9 and peptides generated by cleavages with endoproteinase Asp-N and trypsin, respectively. Sequence comparison with CSTX-1, the most abundant and the most toxic polypeptide in the crude spider venom, revealed a high degree of similarity (53% identity). By means of limited proteolysis with immobilised trypsin and RP-HPLC, the cystine-containing peptides of CSTX-9 were isolated and the disulphide bridges were assigned by amino acid analysis, Edman degradation and nanospray tandem mass spectrometry. The four disulphide bonds present in CSTX-9 are arranged in the following pattern: 1-4, 2-5, 3-8 and 6-7 (Cys₆-Cys₂₁, Cys₁₃-Cys₃₀, Cys₂₀-Cys₄₈, Cys₃₂-Cys₄₆). Sequence comparison of CSTX-1 with CSTX-9 clearly indicates the same disulphide bridge pattern, which is also found in other spider polypeptide toxins, e.g. agatoxins (ω-AGA-IVA, ω-AGA-IVB, μ-AGA-I and μ-AGA-VI) from Agelenopsis aperta, SNX-325 from Segestria florentina and curtatoxins (CT-I, CT-II and CT-III) from Hololena curta. CSTX-1/CSTX-9 belong to the family of ion channel toxins containing the inhibitor cystine knot structural motif. CSTX-9, lacking the lysine-rich C-terminal tail of CSTX-1, exhibits a ninefold lower toxicity to Drosophila melanogaster than CSTX-1. This is in accordance with previous observations of CSTX-2a and CSTX-2b, two truncated forms of CSTX-1 which, like CSTX-9, also lack the C-terminal lysine-rich tail. Received 23 July 2001; accepted 31 July 2001     

Elimination and metabolism of dimethylnitrosamine (DMN) byXenopus laevis and other amphibians

Summary The elimination of (¹⁴C)-DMN after i.p. injection intoXenopus was measured, as was the metabolism in vitro of (¹⁴C)-DMN by liver fromXenopus and 9 other amphibian species. In view of its rapid elimination from the body and low rate of metabolism byXenopus liver in vitro, DMN is unlikely to be toxic or carcinogenic inXenopus.This work was supported by a grant from the Cancer Research Campaign. We are also grateful to Dr P. F. Swann, both for the supply of DMN and DEN and for many useful discussions.     

Methionine enkephalin inhibits the bursting activity of the Br-type neuron inHelix pomatia L.

Summary The present study demonstrates that methionine enkephalin can inhibit the normal bursting activity pattern of the RPal or Br-type neuron and this inhibition can be blocked by prior treatment with naloxone, the selective opiate antagonist. The study demonstrates indirectly the presence of opiate-like receptors inHelix pomatia.Acknowledgments. This work was funded by grant RR 08171 Division of Research Resources and NIMH, and a National Academy of Sciences Travel grant and a Hungarian Academy of Sciences research support award to G.B.S. We also gratefully acknowledge thoughtful comments from Dr J. Salanki.     

High K+-induced release of somatostatin from the cortical preparation of rat brain

Summary Release of endogenous somatostatin (SRIF) from the rat cerebral cortical slices incubated in Krebs-bicarbonate buffer was increased from the basal rate of 3.4±0.6% of the total SRIF content in 15 min at [K⁺]_o=5.6 mM, to 13.1±1.6% upon raising the [K⁺]_o to 56.6 mM. The high-K⁺ evoked SRIF release was absent when Ca⁺⁺ in the medium was replaced by Mn⁺⁺. The isolated synaptosomes from rat cerebral cortex contain 13.2±3.1 ng SRIF/mg protein compared to 0.33±0.01 ng/mg protein in the cortical tissue as a whole, suggesting that nerve terminals are the main source of the peptide released upon membrane depolarization.The study was supported by a grant from the Medical Research Council of Canada. Results of this work have been published in part as abstracts: Can. Physiol.9, 45 (1978), and Fedn Proc.37, 665 (1978).The authors are greatly indebted to Dr M. Gotz and the Ayerst Research Laboratories for the most generous supply of the synthetic somatostatin.     

An Argentine ant aggregation factor

Summary (Z)-9-Hexadecenal, a general aggregation factor of the Argentine ant,Iridomyrmex humilis, has been isolated and chemically characterized. It is implicated as a component of the trail-pheromone complex on evidence that it is a constitutent of the ventral gland secretion, and that it both activates and attractsI. humilis workers.Acknowledgments. We thank the Australian Research Grants Committee, and the Reserve Bank of Australia, Rural Credits Development Fund for financial support.