Opposing temperature responses to intrahypothalamic injections of 5-hydroxytryptamine in the pigeon exposed to cold

Summary 5-Hydroxytryptamine injected into posterior and anterior parts of the pigeon hypothalamus evoked a short lasting hyperthermia or hypothermia, respectively. Variable responses obtained within the same brain region suggest the existence of different 5-HT systems, even in rather limited hypothalamic areas.     

Thrombospondins: from structure to therapeutics

Cartilage oligomeric matrix protein, also known as thrombospondin-5 (TSP-5), is an extracellular matrix protein found primarily in cartilage and musculoskeletal tissues. TSP-5 is of interest because mutations in the gene cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (MED/EDM1). Both PSACH and EDM1 have a characteristic chondrocyte phenotype distinguished by giant rough endoplasmic reticulum (rER) cisternae containing TSP-5 and other extracellular matrix proteins such as type IX collagen and matrilin-3. The accumulation of proteinaceous material in the rER compromises cellular function and leads to premature chondrocyte death. Both in vitro and in vivo models have been generated with varying degrees of success to study the cellular mechanisms of the disease process. Here we review and discuss in vitro and in vivo PSACH and MED model systems and describe two transgenic mouse lines expressing human mutant TSP-5 protein. These model systems have revealed several important features of the PSACH cellular pathology: unfolded protein response activation, upregulation of apoptosis and inappropriate assembly of matrix network in the rER. Some of these models are valuable reagents that may be of use in testing therapeutic interventions. (Part of a Multiauthor Review).     

Studies on a compound antagonistic to 5-hydroxytryptamine

Riassunto La 1-metil-8-carbobenzilossi-aminometil-10-ergoline (MCE) impedisce la morte dei feti in ratte gravide trattate con enteramina (5-HT). L'effetto massimo è ottenuto quando l'MCE è somministrato 3 h prima della 5-HT, ma si manifesta anche 14 giorni dopo una iniezione del composto.     

High affinity binding of 5-hydroxytryptamine to some membrane preparations from cattle brain. Relationship to lysergic acid diethylamide (LSD)]

5 hydroxytryptamine binds to crude brain membrane preparations with two different affinities (KD = 1 to 2 X 10(-9) M for the highest, 1 to 2 X 10(-8) M for the lowest). LSD also binds with two affinities (KD = 3 to 4 X 10(-9) M and KD = 2 to 3 X 10(-8) M). Subcellular distribution of these sites shows that binding involves the two binding affinities in microsomal membranes but solely the high affinity binding sites are present in purified synaptosomal membranes. High affinity sites for 5 HT and for LSD are different as no direct competitive inhibition is observed in that case. On microsomal membranes, direct relationship occurs between low affinity binding for 5 HT and high affinity binding for LSD.     

Participation of DNA repair in the response to 5-fluorouracil

The anti-metabolite 5-fluorouracil (5-FU) is employed clinically to manage solid tumors including colorectal and breast cancer. Intracellular metabolites of 5-FU can exert cytotoxic effects via inhibition of thymidylate synthetase, or through incorporation into RNA and DNA, events that ultimately activate apoptosis. In this review, we cover the current data implicating DNA repair processes in cellular responsiveness to 5-FU treatment. Evidence points to roles for base excision repair (BER) and mismatch repair (MMR). However, mechanistic details remain unexplained, and other pathways have not been exhaustively interrogated. Homologous recombination is of particular interest, because it resolves unrepaired DNA intermediates not properly dealt with by BER or MMR. Furthermore, crosstalk among DNA repair pathways and S-phase checkpoint signaling has not been examined. Ongoing efforts aim to design approaches and reagents that (i) approximate repair capacity and (ii) mediate strategic regulation of DNA repair in order to improve the efficacy of current anticancer treatments. Received 08 September 2008; received after revision 25 September 2008; accepted 03 October 2008     

Monoamine pathways to the cerebellum and cerebral cortex

Zusammenfassung In kombinierter histochemischer und biochemischer Untersuchung werden Dopamin-, Noradrenalin-und 5-Hydroxytryptamin-Neurone nach Entfernung von Cortex cerebri und Cerebellum studiert. Es ergibt sich, dass die Noradrenalin-Nerventerminale, welche Cortex cerebri und Cerebellum innervieren, vermutlich von feinen Axonen her stammen, deren Zellkörper mindestens zum Teil in der Formatio reticularis der Medulla oblongata gelegen sind. Die Noradrenalin-und 5-Hydroxytryptamin-Axone, die nach dem Cortex cerebri ziehen, passieren vermutlich zur Hauptsache das Cingulum.     

A response to monoamines inPeripatopsis moseleyi (Onychophora)

Summary Preparations of longitudinal muscle respond to catecholamines if the ventral nerve cords are present, and also to 5-hydroxytryptamine. The response to 5HT is complex if the central nervous system is present, but muscle alone is probably inhibited.Acknowledgments. It is a pleasure to thank Professor A.C. Brown for providing facilities in the Zoology Department, University of Cape Town, and Mr L.R. Alexander and his colleagues for help in obtaining materials.     

The trefoil protein TFF1 is bound to MUC5AC in humangastric mucosa

The trefoil protein TFF1 is expressed principally in the superficial cells of the gastric mucosa. It is a small protein and forms homo- and hetero-dimers via a disulphide bond through Cys58 which is located three amino acids from the C terminus. TFF1 is co-expressed with the secreted mucin MUC5AC in superficial cells of the gastric mucosa suggesting that it could be involved in the packaging or function of gastric mucus. We have previously shown that TFF1 co-sediments with mucin glycoproteins on caesium chloride gradients. To extend this observation we have now used gel filtration under physiological conditions, immunoprecipitation and Western transfer analysis to characterise the interaction of TFF1 with gastric mucin glycoproteins. We show that TFF1 co-elutes with MUC5AC but not MUC6 on gel filtration and that immunoprecipitation and Western transfer analysis confirms that TFF1 interacts with MUC5AC. We also demonstrate that the TFF1 dimer is the predominant molecular form bound to MUC5AC. Salt and chelators of divalent cations such as EDTA and EGTA disrupted the TFF1- MUC5AC interaction and increased the degradation of MUC5AC, whereas calcium increased the amount of TFF1 bound to MUC5AC. These data support the contention that TFF1 is pivotal in the packaging and function of human gastric mucusa.Received 24 March 2004; received after revision 14 May 2004; accepted 7 June 2004     

The impact of cyclin-dependent kinase 5 depletion on poly(ADP-ribose) polymerase activity and responses to radiation

Cyclin-dependent kinase 5 (Cdk5) has been identified as a determinant of sensitivity to poly(ADP-ribose) polymerase (PARP) inhibitors. Here, the consequences of its depletion on cell survival, PARP activity, the recruitment of base excision repair (BER) proteins to DNA damage sites, and overall DNA single-strand break (SSB) repair were investigated using isogenic HeLa stably depleted (KD) and Control cell lines. Synthetic lethality achieved by disrupting PARP activity in Cdk5-deficient cells was confirmed, and the Cdk5^KD cells were also found to be sensitive to the killing effects of ionizing radiation (IR) but not methyl methanesulfonate or neocarzinostatin. The recruitment profiles of GFP-PARP-1 and XRCC1-YFP to sites of micro-irradiated Cdk5^KD cells were slower and reached lower maximum values, while the profile of GFP-PCNA recruitment was faster and attained higher maximum values compared to Control cells. Higher basal, IR, and hydrogen peroxide-induced polymer levels were observed in Cdk5^KD compared to Control cells. Recruitment of GFP-PARP-1 in which serines 782, 785, and 786, potential Cdk5 phosphorylation targets, were mutated to alanines in micro-irradiated Control cells was also reduced. We hypothesize that Cdk5-dependent PARP-1 phosphorylation on one or more of these serines results in an attenuation of its ribosylating activity facilitating persistence at DNA damage sites. Despite these deficiencies, Cdk5^KD cells are able to effectively repair SSBs probably via the long patch BER pathway, suggesting that the enhanced radiation sensitivity of Cdk5^KD cells is due to a role of Cdk5 in other pathways or the altered polymer levels.     

Azelastine inhibits bronchial hyperreactivity to acetylcholine in guinea pigs

Contractile responses to acetylcholine were measured using isolated tracheae obtained from actively sensitized guinea pigs 0.5, 1, 5, 20, 24, 48 and 72 h after antigen challenge. Tracheal contractions to acetylcholine and to histamine were significantly increased 20 h but not 0.5, 1, 5, 24, 48 and 72 h after antigen challenge indicating bronchial hyperreactivity. When animals were pretreated with azelastine and then exposed to antigen challenge, concentration-response curve to acetylcholine did not differ from that obtained in control (non-challenged) tracheae. It is likely that azelastine is able to inhibit bronchial hyperresponsiveness to chemical mediators of bronchial asthma.     

Difference in the sensitivity to Ca ion among glycerinated skeletal, cardiac and smooth muscles

Glycerinated smooth muscle was contracted almost maximally with 15 mM Mg and 5 mM ATP, while extracted skeletal and cardiac muscles needed Ca ion with 15 mM Mg and 5 mM ATP for producing contraction.     

The timing of cyclophosphamide therapy in tumor-bearing rats affects the resistance to tumor challenge in survivors

Summary Cyclophosphamide given to rats 2 or 5 days after an injection of Yoshida ascites sarcoma cured approximately the same proportion of animals, but the resistance to a subsequent tumor challenge was found only in rats treated with the drug 5 days after tumor injection.     

Cross-resistance of trichomonads to 5-nitroimidazole-derivatives

Summary The strainT. foetus KV 1/M 100, a sub-strain ofT. foetus Kv 1, was made highly resistant to metronidazole by several passages in mice under increasing drug pressure. This drug resistance was accompained by a remarkable cross-resistance to all 5-nitroimidazole-derivatives tested in vitro and in vivo.     

Reduced deformability of erythrocytes exposed to hypercapnia.

The effect of hypercapnia on the deformability of erythrocytes was studied by means of a nuclepore membrane filter method. A decrement of the deformability by 20--40% was observed when PCO2 was increased from 50 mm Hg to 200 mm Hg, accompanied with an increment of 5% in hematocrit value.     

Thiopental binding to serum albumin

Zusammenfassung Die Bindungskapazität und die Anlagerungskonstante von Thiopental an Rinderalbumin wurde bei pH 7,42 in Trispuffer bei einer Temperatur von 7°C nach der Methode der Gleichgewichtsdialyse gemessen. Die Bindungskapazität betrug 5 und die Vergleichskonstante war 12000. Durch Zugabe von EDTA wurde die Bindungskapazität auf 4 reduziert; gleichzeitig verringerte sich die Anlagerungskonstante auf 10000.     

Galectin-3 stabilizes heterogeneous nuclear ribonucleoprotein Q to maintain proliferation of human colon cancer cells

Comparative analysis of proteomes using 5-fluorouracil (5-FU)-resistant human colon cancer cell line revealed that decreased galectin-3 expression was significantly associated with retarded proliferation. However, in the presence of 5-FU proliferation rate of cells with suppressed galectin-3 expression did not differ from that of cells with normal galectin-3 expression, even galectin-3 suppression augmented apoptosis. Mechanism by which galectin-3 regulates cancer cell proliferation has been identified in immunoprecipitates of the anti-galectin-3 antibody. Heterogeneous nuclear ribonucleoprotein Q (hnRNP Q) was identified as a protein interacting with galectin-3. Interestingly, while galectin-3 protein was not affected by the hnRNP Q level, its suppression was accompanied by a decrease in hnRNP Q expression. The present study demonstrates that galectin-3 stabilizes hnRNP Q via complex formation, and reduction in the hnRNP Q level leads to slow proliferation and less susceptibility to 5-FU. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. B.C.Yoo; S-H.Hong; These two authors contributed equally to this work. Received 10 September 2008; received after revision 19 October 2008; accepted 07 November 2008     

Action of puromycin and 5-fluorouracil on the adrenal responsiveness to corticotrophin

Zusammenfassung Puromycin und 5-Fluorouracil beeinträchtigen in Nebennierenschnitten der Ratte die durch ACTH und cyclisches 3, 5-Adenosinphosphat stimulierte Corticosteronausscheidung. Die durch TPN plus Glukose-6-phosphat stimulierte Corticosteronausscheidung bleibt unbeeinflusst. Dagegen bewirkt Puromycin bei Zusatz von TPN allein eine Hemmung. Die Wirkung von Puromycin und 5-Fluorouracil dürfte daher eher auf die Beeinflussung des Energiestoffwechsels als auf eine Hemmung der Neubildung induzierter Enzyme zurückzuführen sein.     

Resistance of cold-exposed rats to aflatoxin

Summary Male rats kept at a temperature of 4°–5°C were refractory to a lethal dose of aflatoxin compared to animals at 20°–21°C which exhibited a high mortality and marked liver damage. It is suggested that this decreased susceptibility is mediated through a stimulated microsomal drug-metabolizing system in cold environment.     

Axillary 5 alpha-androst-16-en-3-one in men and women: relationships with olfactory acuity to odorous 16-androstenes

Axillary 5 alpha-androst-16-en-3-one (5 alpha-androstenone) levels were found to be significantly higher in men than in women but do not vary between left and right axillae, are not related to age, handedness or degree of hirsutism (in women) nor to anosmia to this steroid. In men (but not in women), levels are related linearly to axillary cholesterol concentrations but not to squalene. Olfactory thresholds for 5 alpha-androstenone varied widely, the lowest recorded being 0.2 ppb, but there was no difference in thresholds between men and women. Women (70%) found the smell 'repellant' but anosmia did not differ greatly between men and women (9-20%). Anosmia to the smell of 5 alpha-androst-16-en-3 alpha-ol was most marked in women (90%) rather than in men (45%). Axillary 5 alpha-androstenone values were generally consistent with the 'musky' or 'strong' smells of male axillary extracts, compared with the 'sweet' smell of those from female subjects.     

Difference in the sensitivity to Ca ion among glycerinated skeletal,cardiac and smooth muscles

Summary Glycerinated smooth muscle was contracted almost maximally with 15 mM Mg and 5 mM ATP, while extracted skeletal and cardiac muscles needed Ca ion with 15 mM Mg and 5 mM ATP for producing contraction.Acknowledgments. I thank Prof. T. Suzuki and Dr H. Nakanishi for helpful suggestions, and Mr G. Ito for technical assistance.