Effect of dipyridamole on glomerular mesangial cell ecto-5′-nucleotidase expression

Although dipyridamole has been extensively studied as an anti-aggregating agent, its mechanism of action has not been elucidated. Cultured mesangial cells were treated with dipyridamole 1–100 M from 6–72 h. Ecto-5-nucleotidase activity approximately doubled (from 115±11 to 226±14 nmol/min/mg) after treatment with 100 M dipyridamole for 72 h. This effect was concentration- and time-dependent. Cycloheximide, an inhibitor of protein synthesis, did not alter basal 5-nucleotidase activity. However, it prevented stimulation by 5 M dipyridamole. Adenosine availability at the receptor sites was increased by dipyridamole and S-(p-nitrobenzyl)-6-thioinosine (NBTI), which inhibit adenosine uptake into the cell. Addition of dipyridamole or NBTI to the adenosine-treated mesagial cells produced an additive increase in ecto-5-nucleotidase activity. Dipyridamole, through its effect on extracellular adenosine and ecto-5-nucleotidase, may have an influence upon regulation of the glomerular microcirculation.     

The selective esterification of the 5′-hydroxyl of thymidine

Zusammenfassung Ein einfacher Weg zur selektiven Esterbildung mit der 5-Hydroxylgruppe von Thymidin wird beschrieben. Beispiele: Thymidin-5-phosphat, Thymidin-5-sulfat und Thymidin-5-trichloracetat.     

Synthesis of dinucleoside phosphates by reaction of 5′-chloro-5′-deoxynucleosides with nucleotide anions

Zusammenfassung Uridylyl-(3–5)-adenosin und Uridylyl-(3-5)-ionosin wurden durch Umsetzung eines passenden 5-chloro-5-deoxy-2, 3-O-isopropylidene-Nukleosids mit Uridin-3-phosphat synthetisiert.     

A new synthetic method of coumestan derivatives

Zusammenfassung Die Synthese von 7-Hydroxy-5,6-dimethoxybenzofurano[3,2:3,4]coumarin aus 7,2-Dihydroxy-oder 7-Benzyloxy-2-hydroxy-4,5-dimethoxyisoflavon wird beschrieben.     

Hb F in sickle cell anemia

We have reviewed the methodology for an accurate quantitation of Hb F in the blood of patients with sickle cell anemia, values observed in hundreds of patients of different (racial or ethnic) backgrounds and with differences in severity of the disease, and the various factors that affect the level of Hb F. The latter include sex, age, genetic background or chromosomal haplotypes, variations in the sequences of the locus control region(s) 5 to the -globin gene, and the presence of an chain deficiency or -thalassemia. Finally, a few remarks about agents effective in increasing the in vivo Hb F synthesis are also included.     

Evidence for the extranuclear localization of thymosins in thymus

A new radioimmunoassay has been developed for thymosin ₄ by generating rabbit polyclonal antibodies against the synthetic N-terminal peptide fragment 1–15 coupled to KLH. The synthetic analogue [Tyr¹²]-thymosin ₄ (1–15) was used as tracer. This radioimmunoassay, with a useful range of 10–1000 pmoles, showed cross-reactivity with the second homologous -thymosin of man and rat (thymosin ₁₀) but not of calf (thymosin ₉). This radioimmunoassay, together with an improved radioimmunoassay for the N-terminus of parathymosin , was employed for the measurement of the levels of thymosin ₄ and parathymosin in nuclear and extranuclear extracts of calf thymus. The bulk of these polypeptides was found in the extranuclear material whereas only traces were observed in the nuclear environment, which indicates the extranuclear localisation of - and -thymosins.     

Detection of pyrimidine 5′-nucleotidase deficiency using1H-or31P-nuclear magnetic resonance

Summary We describe here a further Japanese family with pyrimidine 5-nucleotidase (P5N) deficiency diagnosed using a nuclear magnetic resonance (NMR) spectrum, in Kumamoto prefecture where two families having the disease have been reported before. The specific spectra in¹H-NMR of P5N deficient erythrocytes were due to three methyl protons of CDP-choline at 3.22 ppm and to H-2, H-8 and ribose-1 of pyrimidine nucleotide phosphate(s) in the lower fields (at 5.82 and 8.00 ppm). The other specificities in³¹P-NMR spectra were due to CDP-choline, CDP-ethanolamine and UDP-glucose. Those spectra were not detected in other types of hemolytic anemia.     

Antiproliferative effects of 4′, 5′-unsaturated adenine nucleosides on leukemia L 1210 cells in vitro

Summary Several 4, 5-unsaturated adenine nucleosides were shown to have antiproliferative activity against L 1210 leukemia cells in vitro. The active nucleosides were cytotoxic to the L 1210 cells as demonstrated by Trypan Blue uptake. The cytotoxicity was not induced by alterations in the ribonucleoside and deoxyribonucleoside triphosphate levels of the L 1210 cells.     

The synthesis of a natural dinucleoside phosphate derivative with the aid of a purine cyclonucleoside

Zusammenfassung Die Behandlung von 8-5-Anhydro-8-mercapto-2-o-mesylguanosin mit (1,2 Äquivalent) Tributylammonium-uridylat in siedendem DMF lieferte das Dinucleosidphosphat Uridyl-(3-5)-8-mercapto-2-o-mesylguanosin. Weitere Behandlung mit Raney Ni führte zum Uridyl-(3-5)-2-o-mesylguanosin.     

Microbiological transformation of biflavone

Summary Incubation of 5,5-7,7-tetramethoxy-8,8-biflavone withAspergillus niger results in the formation of 4,4-dihydroxy-5,5-7,7-tetramethoxy-8,8-biflavan.Part of this work was done at Allahabad University, Chemical Laboratories.     

Über das Vorkommen verschieden substituierter Chalkone während der Mikrosporogenese beiTulipa

Summary During the development of anthers inTulipa cv. Apeldoorn the following chalcones were isolated: 2, 3, 4, 4, 6-pentahydroxychalcone as the main component and 2, 4, 4, 6-tetrahydroxy-3-methoxychalcone as well as 2, 4, 4, 6-tetrahydroxychalcone in a very small amount.

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Die Untersuchungen wurden mit Mitteln der Deutschen Forschungsgemeinschaft unterstützt.     

The tonically acting pulmonary receptors innervated by C fibres

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

Summary Axillary 5-androst-16-en-3-one (5-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-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-androst-16-en-3-ol was most marked in women (90%) rather than in men (45%). Axillary 5-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.Supported by the Herbert Dunhill Trust.     

Effect of urochordamines on larval metamorphosis of ascidians

Two pteridine-containing bromophysostigmine alkaloids, urochordamine A and B, which were isolated from ascidians as larval metamorphosis promoters, were converted to more polar compounds, urochordamine A and B, respectively, when left standing in protic solvents. These four compounds promoted larval metamorphosis of the ascidianHalocynthia roretzi in the order A>A>B>B, and induced metamorphosis of the pediveliger larvae of the musselMytilus edulis galloprovincialis.     

The syntheses of axillarin and its related compounds

Zusammenfassung Axillarin (5, 7, 3, 4-Tetrahydroxy-3,6-dimethoxyflavon). 5,3,4-Trihydroxy-3,6,7-trimethoxyflavon und 5,7,3,4-Tetrahydroxy-3,8-dimethoxyflavon wurden synthetisiert.     

Peroxides of diisopropylether

Zusammenfassung Stellt man 2,2-Dihydroperoxy-2, 2-Diisopropyläther nachIvanov et al. aus Isopropyläther her, so entsteht nebenbei auch noch 2, 2-Dihydroperoxy-2, 2-Diisopropyl-Peroxid. Beide Peroxide kann man papier- und dünnschichtchromatographisch nachweisen und mit präparativer Dünnschichtchromatographie rein isolieren.     

Neurosteroid enhances glutamate release in rat prelimbic cortex via activation of α1-adrenergic and σ1 receptors

The present paper studied the effect and mechanism of neurosteroid pregnenolone sulfate (PREGS) on spontaneous glutamate release using electrophysiological and biochemical methods combined with a pharmacological approach. The results suggested that PREGS had a selective enhancing effect on spontaneous glutamate release in the prelimbic cortex and the hippocampus but not in the striatum. The effect of PREGS in the prelimbic cortex appeared to be via modulation of ₁-adrenergic and ₁ receptors, but in the hippocampus it might be dependent on ₁ receptors only. The activation of ₁-adrenergic receptors synergized ₁ receptor activation in the prelimbic cortex. Intracellular calcium released from the endoplasmic reticulum, protein kinase C, adenylyl cyclase and protein kinase A played a key role in the effect of PREGS. Intracellular calcium, protein kinase C and adenylyl cyclase might be upstream events in the activation of protein kinase A after PREGS.Received 7 January 2005; received after revision 19 February 2005; accepted 22 February 2005 Available online 29 March 2005     

Hydra vulgaris ω10-lipoxygenase is used in vivo to synthesize new α-linolenic acid metabolites

Previous studies conducted in cytosolic extracts of the freshwater hydrozoanHydra vulgaris led to the finding of an abundant 11(R)-lipoxygenase catalyzing the peroxidation of polyunsaturated fatty acid (PUFAs) on the tenth carbon atom from the aliphatic end (10 peroxidation). Here we describe experiments aimed at identifying the actual metabolites generated in vivo by such enzymic activity. Homogenates ofH. vulgaris polyps were analyzed by HPLC. This showed the presence of three major components chromatographically identical to three metabolites obtained when incubating the homogenates with exogenous -linolenic acid (-LA). The presence, in extracts of polyps prelabelled with [¹⁴C]--linolenic acid, of radioactive metabolites displaying the same chromatographic properties, substantiated the hypothesis that the natural products isolated in vivo are derived from -LA. Gas chromatographic analyses revealed that this was the most abundant PUFA in both free and phosphoglyceride-bound fatty acid pools. [¹H]-NMR analysis of the endogenous substances, carried out in comparison with products obtained from exogenously incubated -LA, indicated that their structures were those of 9-hydroxy-, 9-hydroperoxy- and 9-keto-octadeca-10E-12Z-15Z-trienoic acids (9--HOTrE,-HPOTrE and-KOTrE).Hydra homogenates transformed 9--HPOTrE partly into 9--HOTrE and partly into 9--KOTrE. Chiral phase HPLC conducted on 9--HOTrE established that this metabolite was composed mostly of theR anantiomer. These observations, and the finding that the presence of exogenous arachidonic acid in incubated homogenates significantly reduces the production of -LA metabolites, provide strong evidence that these compounds are produced by an enzymic activity identical to the previously-describedH. vulgaris (R)-10-lipoxygenase. Further experiments suggested that -LA, acting as the native substrate for this enzyme, is mainly esterified on the 2 position ofHydra phosphoglycerides, and that the production of the -LA metabolites described here for the first time from natural sources, can be potentially enhanced in vivo by stimuli activating phospholipase A₂.     

Correlation between 3′,5′, c-AMP levels and thyrotropin in separated rat pituitary thyrotropic cells

Summary The correlation between 3,5, c-AMP levels, TSH content and secretion of separated thyrotropic cells was studied. Incubation of the separated cells with 1, 10 and 100 ng of TRH does not change the 3,5, c-AMP levels, despite the significant rises of the TSH level. Dibutyryl c-AMP causes rise in TSH content, with no indication of its secretion. PGE₂ 10^–5 increased 3, 5, c-AMP levels with no change in the content or secretion of TSH in separated thyrotropic cells.     

Efficient cell proliferation and predominant accumulation of ɛ-globin mRNA in human leukemic K562 cells which produce mostly Hb Gower 1

Summary Long-term cultures of K562(S) cells in 50–75 M hemin allow the selection of hemin-resistant K562 cells together with cells which proliferate efficiently while fully induced to express the human embryonic globin genes, as the hemoglobin Gower 1 (₂₂) is the predominant hemoglobin produced. Our experiments demonstrate that these K562 cells accumulate mostly -globin mRNA (-globin mRNA/-globin mRNA=2.9) suggesting that the control of hemoglobin expression is at a pretranslational level.We thank Dr Irene Bozzoni (Centro degli Acidi Nucleici, Università di Roma) for the pXCR7 probe. Address for reprint request: R.G. Centro Studi Biochimici sul Morbo di Cooley, Via Borsari 46, I-44100 Ferrara.