Aging and endothelin-1 induced vascular contractions

Summary Contractions produced by endothelin-1 (0.3–30 nM) have been investigated in aorta, renal arteries and mesenteric arteries from 2- and 24-month-old Sprague-Dawley rats. In senescent rats the EC₅₀ values of endothelin-1 for aorta and renal artery were significantly increased (aorta: from 6.2 to 12 nM; renal artery: from 5.2 to 7.8 nM). For mesenteric artery the EC₅₀ value (4.3 nM) was unchanged by aging, whereas the maximal contractile response to endothelin-1 was enhanced (from 8.3 to 11.7 mN). In contrast, there was no significant age-related difference in the maximal endothelin-1 response of aorta and renal artery. The present data demonstrate a reduced sensitivity for aorta and renal artery and an enhanced maximal response to endothelin-1 in the mesenteric artery in senescent rats.     

Aging and endothelin-1 induced vascular contractions

Contractions produced by endothelin-1 (0.3-30 nM) have been investigated in aorta, renal arteries and mesenteric arteries from 2- and 24-month-old Sprague-Dawley rats. In senescent rats the EC50 values of endothelin-1 for aorta and renal artery were significantly increased (aorta: from 6.2 to 12 nM; renal artery: from 5.2 to 7.8 nM). For mesenteric artery the EC50 value (4.3 nM) was unchanged by aging, whereas the maximal contractile response to endothelin-1 was enhanced (from 8.3 to 11.7 mN). In contrast, there was no significant age-related difference in the maximal endothelin-1 response of aorta and renal artery. The present data demonstrate a reduced sensitivity for aorta and renal artery and an enhanced maximal response to endothelin-1 in the mesenteric artery in senescent rats.     

Development of DNA probes for cytotoxin and enterotoxin genes in enteric bacteria

Summary DNA probes to identify the genes encoding toxins in enteric bacteria have been developed. Use of these probes reduces the number of animals required for toxicity testing, as suspect bacteria can be directly tested for the presence of toxin. We have augmented the gene probes available by developing probes against theEscherichia coli enterotoxin LTII and shiga toxin fromShigella dysenteriae 1.The LTII gene fromE. coli 357900 was identified and characterised and a suitable internal probe was obtained. The LTII gene was found not to be common among enterobacteriae from various geographical locations. Isolates predominately of animal origin from Nigeria and Thailand hybridized with the probe.The shiga toxin gene was isolated fromS. dysenteriae 1 by a combination of in vivo and in vitro methods. An internal probe was identified and used against different serogroups ofShigella andE. coli isolated. The probe was found to hybridize withS. dysenteriae 1 isolates and also someS. flexneri andS. sonnei strains. Representatives were tested for toxin production and found to produce toxin at low levels.     

The sperm attractant ofHormosira banksii (Phaeophyceae,Fucales), a seaweed common to Australia and New Zealand

Summary Hormosira banksii is a taxonomically isolated brown seaweed endemic to Australia and New Zealand. The sperm attractant of this species has been isolated and identified as trans-1-vinyl-2-(1E, 3Z-hexadienyl)-cyclopropane (I) (hormosirene).Hormosira is the first organism in which a cyclopropane derivative has been found to act as a hormone in sexual reproduction. The implication of this finding in relation to phylogeny and phytogeography is discussed.Acknowledgment. A substantial part of this work was supported by Deutsche Forschungsgemeinschaft (DFG).     

Length-tension dependence in vascular smooth muscle: Possible participation of endothelin

The results of experiments with isolated strips of rat portal vein, and of ‘sandwich’ experiments involving strips of the portal vein or aorta, revealed the involvement of endothelin in the development of myogenic reactions of the vascular smooth muscle, observed on distension of the vascular wall. Released by the endothelial cells, endothelin is capable of stimulating the contraction of these muscles.     

APP dimer formation is initiated in the endoplasmic reticulum and differs between APP isoforms

The amyloid precursor protein (APP) is part of a larger gene family, which has been found to form homo- or heterotypic complexes with its homologues, whereby the exact molecular mechanism and origin of dimer formation remains elusive. In order to assess the cellular location of dimerization, we have generated a cell culture model system in CHO-K1 cells, stably expressing human APP, harboring dilysine-based organelle sorting motifs [KKAA-endoplasmic reticulum (ER); KKFF-Golgi], accomplishing retention within early secretory compartments. We show that APP exists as disulfide-bonded dimers upon ER retention after it was isolated from cells, and analyzed by SDS-polyacrylamide gel electrophoresis under non-reducing conditions. In contrast, strong denaturing and reducing conditions, or deletion of the E1 domain, resulted in the disappearance of those dimers. Thus we provide first evidence that a fraction of APP can associate via intermolecular disulfide bonds, likely generated between cysteines located in the extracellular E1 domain. We particularly visualize APP dimerization itself and identified the ER as subcellular compartment of its origin using biochemical or split GFP approaches. Interestingly, we also found that minor amounts of SDS-resistant APP dimers were located to the cell surface, revealing that once generated in the oxidative environment of the ER, dimers remained stably associated during transport. In addition, we show that APP isoforms encompassing the Kunitz-type protease inhibitor (KPI) domain exhibit a strongly reduced ability to form cis-directed dimers in the ER, whereas trans-mediated cell aggregation of Drosophila Schneider S2-cells was isoform independent. Thus, suggesting that steric properties of KPI-APP might be the cause for weaker cis-interaction in the ER, compared to APP695. Finally, we provide evidence that APP/APLP1 heterointeractions are likewise initiated in the ER.     

A new alarm pheromone (2-decen-1-yl-acetate) isolated from the stings ofApis dorsata andApis florea (Hymenoptera: Apidae)

Summary From sting extracts ofApis dorsata andA. florea, a substance was isolated which is active in alarm behaviour of both of these species but not ofA. mellifera andA. cerana. The active substance was identified as 2-decen-1-yl-acetate.     

Sociétés animales et effet de groupe

Summary In the present note, the author is reviewing recent researches dealing with the action carried by the group on the individual taken isolated. He distinguishes two kinds of effects: 1.Mass effect, owing above all to a modification of the medium by the population itself; 2.group effect, induced chiefly by sensorial stimuli which occurs without requiring the gathering of a large number of individuals.Some examples of group effects are given. Individuals living in society are physiologically different from isolated. Correlation, coordination and social facilitation are to be included among group effects. To-day, we are allowed to speak of social physiology.     

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     

Myotoxic phospholipases A from snake venom,Pseudechis colletti,producing myoglobinuria in mice

Summary 2 proteins producing myoglobinuria in mice were isolated from the venom of the Australian elapid snakePseudechis colletti and identified as phospholipases A showing close similarities in amino acid composition to a similarly acting enzyme from a sea snake venom (Enhydrina schistosa).     

Anthraquinone pigments from a conidiating mutant ofTrichoderma viride

Summary Two pigments responsible for the yellow-orange color of a brown conidiating mutant of the deuteromyceteTrichoderma viride were isolated and spectroscopically identified as 1,3,6,8-tetrahydroxyanthraquinone (I) and 1-acetyl-2,4,5,7-tetrahydroxy-9,10-anthracenedione (II). Both compounds are known substances but were not yet reported as metabolites of this fungal species. Their relationship to other anthraquinones produced byT. viride is discussed.Dedicated to Dr Z. Vanek, on the occasion of his 60th birthday.Upper case letters denote multiplicity arising from the directly bonded protons, lower case letters that from the geminal and vicinal couplings.     

Chronic sensory denervation reduces thrombin-stimulated endothelin release from aortic endothelial cells

The long-term (trophic) influence of perivascular nerves on the endothelium was investigated by measuring changes in thrombin-stimulated release of the potent vasoconstrictor, endothelin, after selective chronic denervation. Rat pups were treated with either guanethidine or capsaicin to destroy sympathetic or sensory nerves, respectively. The abdominal aortas from the rats at three months of age (5 pooled per experiment) were incubated with 4U thrombin/ml in medium for 24 h at 37°C, and the amount of endothelin released from the preparation determined by immunoassay. After neonatal sensory denervation there was a significant reduction in the thrombinstimulated release of endothelin compared to the controls (0.012±0.012 (4) compared to 0.063±0.012 (6), pmol/cm²/24 h, p<0.02). There was no change in endothelin release after sympathetic denervation. In summary, sensory nerves play a trophic role in the expression of endothelin in endothelial cells of the intima.     

High sensitivity of porcine cerebral arteries to endothelin

Summary The threshold concentration of endothelin to induce contractions in porcine cerebral arteries (anterior cerebral, Willis ring and basilar artery) was lower than those for coronary and renal arteries. The median effective concentrations (ED₅₀) of endothelin in cerebral arteries were also significantly lower than those for coronary and renal arteries. There was no significant difference in the sensitivity to endothelin among cerebral arteries, or between coronary and renal arteries. The maximal percentage of contractions induced by endothelin, as compared to that induced by 10^–1 M potassium chloride, was not significantly different between the arteries.     

Cyanobacterial symbiont biosynthesis of chlorinated metabolites fromDysidea herbacea (Porifera)

The tropical marine spongeDysidea herbacea contains large numbers of a symbiotic filamentous cyanobacterium identified on the basis of a detailed ultrastructural study asOscillatoria spongeliae. We report the flow-cytometric separation of the symbiont from the sponge cells, and demonstrate by chemical analyses that a unique group of polychlorinated compounds isolated from the whole sponge tissue is limited to the cyanobacterial filaments, whereas the accompanying sesquiterpenoids are found only in the sponge cells. This is the first demonstration that secondary metabolites ascribed to a sponge are localized in prokaryotic symbiont cells.     

Antimalarial drugs: recent advances in molecular determinants of resistance and their clinical significance

Molecular determinants of antimalarial drug resistance are useful and informative tools that complement phenotypic assays for drug resistance. They also guide the design of strategies to circumvent such resistance once it has reached levels of clinical significance. Established resistance to arylaminoalcohols such as mefloquine and lumefantrine in SE Asia is mediated primarily by gene amplification of the P. falciparum drug transporter, pfmdr1. Single nucleotide polymorphisms in pfmdr1, whether assessed in field isolates or transfection experiments, are associated with changes in IC₅₀ values (to arylaminoalcohols and chloroquine), but not of such magnitude as to influence clinical treatment outcomes. Recently described emerging in vitro resistance to artemisinins in certain areas correlates with mutations in the SERCA-like sequence PfATP6 and supports PfATP6 as a key target for artemisinins. Received 13 February 2006; revised after revision 7 March 2006; accepted 29 March 2006     

An update on the biology and physiology of resistin

Resistin is a newly discovered adipocyte hormone. It is related to resistin-like molecules alpha, beta and gamma in structure and function. Resistin is produced by white and brown adipose tissues but has also has been identified in several other tissues, including the hypothalamus, pituitary and adrenal glands, pancreas, gastrointestinal tract, myocytes, spleen, white blood cells and plasma. The tissue level of resistin is decreased by insulin, cytokines such as tumour necrosis factor alpha, endothelin-1 and increased by growth and gonadal hormones, hyperglycaemia, male gender and some proinflammatory cytokines, such as interleukin-6 and lipopolysaccharide. Resistin antagonizes insulin action, and it is downregulated by rosiglitazone and peroxisome proliferator-activated receptor gamma agonists. Since evidence of a direct link between resistin genotype and human diabetes is still weak, more molecular, physiological and clinical studies are needed to determine the role of resistin in the aetiology of type 2 diabetes.     

Penaeidins, a family of antimicrobial peptides from penaeid shrimp (Crustacea, Decapoda)

The production of antimicrobial peptides represents a first-line host defense mechanism of innate immunity that is widespread in nature. Only recently such effectors were isolated in crustacean species, whereas numerous antimicrobial peptides have been characterized from other arthropods, both insects and chelicerates. This review presents findings on a family of antimicrobial peptides, named penaeidins, isolated from the shrimp Penaeus vannamei. Their structure and antimicrobial properties as well as their immune function will be discussed through analyses of penaeidin gene expression and peptide distribution upon microbial challenge. Received 21 January 2000; received after revision 10 March 2000; accepted 10 March 2000     

Alkaloidal oviposition stimulants for a danaid butterfly,Ideopsis similis L., from a host plant,Tylophora tanakae (Asclepiadaceae)

Chemicals releasing oviposition by an Asclepiadaceae feeder,Ideopsis similis, were identified from a host plant,Tylophora tanakae. A strong positive response was evoked by a methanolic extract of the plant, which proved to contain multiple stimulants. A mixture of two phenanthroindolizidine alkaloids, (+)-isotylocrebrine and (–)-7-demethyltylophorine, isolated from organic fractions, elicited significant ovipositional responses from females.