Purification of platelet-derived endothelial cell growth inhibitor and its characterization as transforming growth factor-β type 1

In 1986, Brown and Clemmons (Proc. natl Acad. Sci. USA83 (1986) 3321) showed that platelets contain a substance, platelet-derived growth inhibitor (PDGI), that inhibits in vitro endothelial cell replication. Although platelets are rich in transforming grwoth factor (TGF-), PDGI was considered not to be related to TGF-, on the basis of its reported properties (extraction from platelets at neutral pH, binding to heparin-Sepharose). However, we purified PDGI to near homogeneity and showed that on the basis of HPLC retention behavior, in vitro growth inhibitory activities with several cell types, receptor binding, and immunoneutralization of growth inhibitory activity with specific anti-TGF- type 1 antibodies, PDGI is most probably identical with TGF- type 1.     

Recent evolutionary origin within the primate lineage of two pseudogenes with similarity to members of the transforming growth factor-β superfamily

Using a search engine called Motifer, we searched the public database of the human genome for genes matching a consensus pattern of cysteine residues derived from members of the transforming growth factor-beta (TGF-) superfamily. We identified two genes (named MDF451 and MDF628) that display sequence similarity to members of the TGF- superfamily in the arrangement of six conserved cysteine residues. Phylogenetic analyses revealed that MDF451 and MDF628 constitute a distinct subgroup within the TGF- superfamily, distantly related to the GDNF subfamily of ligands. Both genes could be identified in several primate species in addition to human, including chimpanzee, gorilla, guereza, and green and gray monkey, but not in rodents or other non-primate mammals, and appear not to be present in the genomes of mouse, rat or zebrafish. RNAs for MDF451 and MDF628 were expressed at low levels within distinct regions of the human central nervous system, including adult cerebellum, adult spinal cord and fetal brain. Despite expression at the RNA level, both genes presented a transcribed upstream stop codon that would prevent translation of the TGF--like reading frame. The coding potential of alternative reading frames was not immediately apparent. The two genes may represent TGF--like pseudogenes that have recently appeared in evolution in a common ancestor of the primate lineage by duplication from a GDNF/TGF--like ancestral gene.Received 9 October 2003; received after revision 13 November 2003; accepted 2 December 2003     

Butyrate-induced reactivation of the fetal globin genes: A molecular treatment for theβ-hemoglobinophaties

The inherited -hemoglobinopathies (sickle cell disease and thalassemia) are the result of a mutation in the adult () globin gene. The fetal globin chain, encoded by the globin genes, can substitute for the mutated or defective globin chain, but expression of the globin gene is developmentally inactivated prior to birth. Reinducing expression of the normal fetal globin genes is a preferred method of ameliorating sickle cell disease and the thalassemias. Stimulation of as little as 4–8% fetal globin synthesis in the bone marrow can produce >20% fetal hemoglobin in the peripheral circulation, due to enhanced survival of red blood cells containing both sickle and fetal hemoglobin, compared to those containing sickle hemoglobin alone. Butyric acid and butyrate derivatives are generally safe compounds which induce fetal hemoglobin production by stimulating the promoter of the fetal globin genes. An initial trial with the parent compound, delivered as Arginine Butyrate, has demonstrated rapid stimulation of fetal globin expression to levels that have been shown to ameliorate these conditions. Phase 1 trials of an oral butyrate derivative with a long plasma half-life have just begun. These agents now provide a specific new apporach for ameliorating these classic molecular disorders and merit further investigation in larger patient populations.     

The molecular structure of the benzilic acid rearrangement product of 3α, 17β-diacetoxy-11-hydroxy-12-oxo-5β-androst-9(11)-ene

Zusammenfassung Die Struktur desp-Brombenzoates des Benzilsäureumlagerungsproduktes von 3, 17-Diacetoxy-11-hydroxy-12-oxo-5-9(11)-androsten wurde durch dreidimensionale Röntgenstrukturanalyse eines Einkristalls als 3-p-Brombenzoat des 11-Carboxy-3, 11, 17-trihydroxy-13-C-nor-5-androstan 11a, 17-Laktons erkannt.     

Selective protection of cells against X-irradiation. Isoproterenol protects only those cells that possesβ-adrenoreceptors

Summary In mixed culture of Chinese hamster fibroblatst, clone 431, and transformed murine L fibroblasts, clone B-82, isoproterenol was found to protect only 431 cells against ionizing radiation. It was shown that 431 cells, in contrast to B-82 cells, possess-adrenoreceptors, and the radioprotective effect of isoproterenol can be realized only if this agent interacts with-adrenoreceptors coupled with the cAMP system. Since malignization often causes the disappearance of-adrenergic and other hormone receptors, the combined culturing and irradiation of the cells studied can be regarded as a model of the growth of malignant cells (B-82) among normal tissue cells (431 cells) under conditions of radiation therapy. A possibility of selective protection against radiation damage of normal tissue cells, with retention of the former radiosensitivity of tumor cells, is discussed.     

Uptake of D-glucose anomers by rat retina

Summary Uptake ofd-glucose anomers by isolated rat retina was studied. After 3 min incubation at 37°C in the presence of or anomer (750 g/ml), a significantly greater uptake (1.32 mg/g wet tissue) of -anomer was observed compared with that of -d-glucose (1.11 mg/g wet tissue). This result and other data suggest that the carrier ford-glucose transport in the retina prefers the -anomer stereospecifically.Acknowledgment. The authors are grateful to Mr S. Suzuki for his technical assistance.     

Proteolytic generation and aggregation of peptides from transmembrane regions: lung surfactant protein C and amyloid β-peptide

The formation of amyloid fibrils is associated with several devastating diseases in humans and animals, including e.g. Alzheimers disease (AD) and the spongiform encephalopathies. Here, we review and discuss the current knowledge on two amyloid peptides: lung surfactant protein C (SP-C) and the amyloid -peptide (A), implicated in human lung disease and in AD, respectively. Both these hydrophobic peptides are derived from the transmembrane region of their precursor protein, and can transit from a monomeric -helical state to a -sheet fibril. The helices of SP-C and A are composed of amino acid residues with inherently higher propensities for strand than helix conformation. Their helical states are stabilized by a membrane environment, and loss of membrane association thus promotes structural conversion and fibril formation. We speculate that the loss of structural context for sequences with a high propensity for formation of sheets may be a common feature of amyloid formation in general.Received 9 July 2003; received after revision 15 August 2003; accepted 3 September 2003     

Zur Konstitution des Resibufogenins und Artebufogenins

Summary The structure of Resibufogenin is unambiguously shown to be 3-hydroxy-14, 15-oxido-bufadien-(20, 22)-olide on the basis of new degradation work. Artebufogenin A and Artebufogenin B are 3-hydroxy-14-15-keto-bufadien-(20, 22)-olide and 3-Hydroxy-14-14-keto-bufadien-(20, 22)-olide, respectively.     

Hemmung der Phosphodiesterase aus Rattenherzen durch Intensain

Summary Intensain causes in vitro a significant and apparently competitive inhibition of phosphodiesterase, which transforms the 3,5-AMP to 5-AMP. These results explain some intensain effects, especially the period of latency to reach the maximum effect, the inotropic action and its reaction under the influence of-blocking and-stimulating agents^10–13.

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3-(-Diäthylamino-äthyl)-4-methyl-7-carbäthoxy-methoxy-2-oxo-(1,2-chromen), Carbochromen.     

Thrombospondin co-localises with TGFβ and IGF-I in the extracellular matrix of human osteoblast-like cells and is modulated by 17β estradiol

Thrombospondin (TSP) is a multifunctional glycoprotein which is synthesised by several cell types including osteoblasts, and incorporated into the extracellular matrix (ECM) of these cells. The function and regulation of TSP in bone is not clear. In this study, using a long term culture model of human osteoblast-like cells, we examined the distribution of TSP in the ECM and its modulation by added estradiol. In this model the osteoblast-like cells form a regular multilayer which continues to increase in depth up to 50 days post confluence. In the ECM of these cultures and in 19-week fetal bone, the bone markers osteocalcin and alkaline phosphatase were diffusely distributed in the matrix. In contrast, labelling for TSP was concentrated, confined to the banded collagen and its immediately adjacent ECM. This pattern of labelling resembled that of the growth factors transforming growth factor-I (TGF), and insulin-like growth factor-I (IGF-I), with which TSP label co-localised. Labelling intensities were comparable between fetal bone and the in vitro material for TSP, TGF and IGF-I. TSP label was present by 10 days post confluence, reached a maximum by 20 days, and declined slowly thereafter, a time course which was similar to that of IGF-I. Incubation of osteoblast-like cell cultures with 17 estradiol resulted in an increase in multilayer depth and a maximal 3-fold increase in TSP labeling at 30 days as well as approximately 2-fold increases for TGF and IGF-I. The dose-response relationship for these responses to estradiol treatment was biphasic with maximal increases at 10^–10 M–10^–11 M of added estradiol. Treatment with 17 estradiol produced labelling intensities that were not significantly different from controls. Studies with other cell types have suggested that TSP may be involved in modulation of growth factor activity. The similarities between TSP, TGF and IGF-I, in terms of their distribution and regulation by 17 estradiol treatment, may indicate a role for TSP in modulating bone cell proliferation and function through interaction with local growth factors.     

Cardenolide glycosides from the adults and eggs ofChrysolina fuliginosa (Coleoptera: Chrysomelidae)

Besides the known sarmentogenin 3-O--D-allopyranoside (1) and digitoxigenin-3-O-[-D-xylopyranosyl-(14)-2, 3-di-O-acetyl--D-allopyranoside] (4), four new cardiac glycosides (2, 3, 5 and6) have been isolated from the defensive glands of adults ofChrysolina fuliginosa. The structures of the new compounds were determined by¹H NMR at 600 MHz and FABMS. Compounds1–6, present in the adults ofC. fuliginosa, have also been identified in the eggs.     

Unique occurrence of fenchol in the animal kingdom

Summary (–)--Fenchol, a compound occurring in plants but not heretofore reported to occur in an animal product, was isolated from volatiles released by adult male Mediterranean fruit flies,Ceratitis capitata Wiedemann, with the male's previously identified sex pheromones. It neither attracts females nor synergizes the pheromones, and its function remains unknown.The editors do not hold themselves responsible for the opinions expressed in the authors' brief reports. — Les auteurs sont seuls responsables des opinions exprimées dans ces brèves communications. — Für die Kurzmitteilungen ist ausschliesslich der Autor verantwortlich.—Per le brevi comunicazioni è responsabile solo l'autore. — . — Solo los autores son responsables de las opiniones expresadas en estas communicationes breves.Mention of a proprietary product in this paper does not constitute a recommendation or an endorsement of the product by the US Department of Agriculture.Acknowledgments. We thank C. Harding for assistance with extractions and fractionations, R.M. Waters for spectral analyses, M.S. Fujimoto and H. Higa for collecting the volatiles and conducting laboratory and cage bioassays, and T. Urago and S. Nakagawa for conducting the field tests.     

Chromosomes and some issues of the evolution of the ground squirrel genusCitellus (Rodentia: Sciuridae)

Citellus (s. str.) relictus, C. dauricus C. pygmaeus (2=36, NF=72). . (Colobotis) erythrogenys . fulvus -. . (Urocitellus) undulatus (2n=32, NF=64) . columbianus. Citellus, , Citellus, . , .     

26-Hydroxy-β-ecdysone,a metabolite of β-ecdysone in the blowfly,Calliphora erythrocephala

Summary 26-Hydroxy--ecdysone has been identified as a metabolite of the moulting hormone, -ecdysone inCalliphora erythrocephala.     

A new chalcone glucoside and cernuoside from the flowers ofAcacia dealbata

Summary A new chalcone glucoside has been isolated from the flowers ofAcacia dealbata and shown to be 4,2,4,6-tetrahydroxy-3-methoxychalcone 2-O--D-glucoside (1) by chemical degradations and spectroscopic methods. Cernuoside (4,6,3,4-tetrahydroxyaurone 4-O--D-glucoside), (2) has also been found in this plant material.Acknowledgments. The author thanks Prof. H. Wagner, Institut für pharmazeutische Arzneimittellehre der Universität München, for a sample of homoeriodictyol, and Prof. J.B. Harborne, University of Reading, for a sample of aureusidin.     

Eubotriol and eubol,new diterpenes fromSideritis euboea

Summary 2 new diterpenes Eubotriol (ent-kaur-16-ene-7, 15, 18triol) (I), and eubol (ent-kaur-16-ene-7-acetoxy-15, 18 diol) (II) have been isolated from Sideritis euboea Helder.This work was supported by National Research Council (C.N.R.), Roma.     

Responses of resistance and capacitance vessels at various frequency electrical stimulation of sympathetic nerves

Conclusion (1) The maximal reactions of cerebral resistance vessels arise at the stimulation frequency of 30 cps, the maximal reactions of capacitance vessels at the frequency of 10 cps. (2) Reactions of resistance vessels in the pulmonary lobe increase in the range of stimulation frequencies from 11–50 cps, maximal reactions of pulmonary capacitance vessels are observed at the frequency of 30–40 cps. (3) Reactions of resistance vessels situated below the abdominal aorta bifurcation increase progressively in the range of frequencies from 6–50 cps, the magnitude of capacitance vessel reaction reaching its maximum value at 20 cps.

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30 /, — 10 /. 50 /. 30–40 /, 20 /.

     

The stimulating action of gastrin pentapeptide and histamine on adenyl cyclase activity in rat stomach

, 3',5'-, in vitro . , , 3',5'- HCl .