Influence of Androgens on Circulating Adiponectin in Male and Female Rodents

Several endocrine factors, including sex-steroid hormones are known to influence adiponectin secretion. Our purpose was to evaluate the influence of testosterone and of the synthetic non-aromatizable/non-5α reducible androgen 17β-hydroxyestra-4,9,11-trien-3-one (trenbolone) on circulating adiponectin and adiponectin protein expression within visceral fat. Young male and female F344 rats underwent sham surgery (SHAM), gonadectomy (GX), or GX plus supraphysiologic testosterone-enanthate (TE) administration. Total circulating adiponectin was 39% higher in intact SHAM females than SHAM males (p<0.05). GX increased total adiponectin by 29–34% in both sexes (p<0.05), while TE reduced adiponectin to concentrations that were 46–53% below respective SHAMs (p≤0.001) and ablated the difference in adiponectin between sexes. No differences in high molecular weight (HMW) adiponectin were observed between sexes or treatments. Adiponectin concentrations were highly and negatively associated with serum testosterone (males: r = −0.746 and females: r = −0.742, p≤0.001); however, no association was present between adiponectin and estradiol. In separate experiments, trenbolone-enanthate (TREN) prevented the GX-induced increase in serum adiponectin (p≤0.001) in young animals, with Low-dose TREN restoring adiponectin to the level of SHAMs and higher doses of TREN reducing adiponectin to below SHAM concentrations (p≤0.001). Similarly, TREN reduced adiponectin protein expression within visceral fat (p<0.05). In adult GX males, Low-dose TREN also reduced total adiponectin and visceral fat mass to a similar magnitude as TE, while increasing serum HMW adiponectin above SHAM and GX animals (p<0.05). Serum adiponectin was positively associated with visceral fat mass in young (r = 0.596, p≤0.001) and adult animals (r = 0.657, p≤0.001). Our results indicate that androgens reduce circulating total adiponectin concentrations in a dose-dependent manner, while maintaining HMW adiponectin. This change is directionally similar to the androgen-induced lipolytic effects on visceral adiposity and equal in magnitude between TE and TREN, suggesting that neither the aromatization nor the 5α reduction of androgens is required for this effect.     

An AFM study of solid-phase bilayers of unsaturated PC lipids and the lateral distribution of the transmembrane model peptide WALP23 in these bilayers

An altered lipid packing can have a large influence on the properties of the membrane and the lateral distribution of proteins and/or peptides that are associated with the bilayer. Here, it is shown by contact-mode atomic force microscopy that the surface topography of solid-phase bilayers of PC lipids with an unsaturated cis bond in their acyl chains shows surfaces with a large number of line-type packing defects, in contrast to the much smoother surfaces observed for saturated PC lipids. Di-n:1-PC (n = 20, 22, 24) and (16:0,18:1)-PC (POPC) were used. Next, the influence of an altered lipid environment on the lateral distribution of the single α-helical model peptide WALP23 was studied by incorporating the peptide in the bilayers of di-n:1-PC (n = 20, 22, 24) and (16:0,18:1)-PC unsaturated lipids. The presence of WALP23 leads to an increase in the number of packing defects but does not lead to the formation of the striated domains that were previously observed in bilayers of saturated PC lipids and WALP. This is ascribed to the less efficient lateral lipid packing of the unsaturated lipids, while the increase in packing defects is probably an indirect effect of the peptide. Finally, the fact that an altered lipid packing affects the distribution of WALP23 is also confirmed in an additional experiment where the solvent TFE (2,2,2-trifluorethanol) is added to bilayers of di-16:0-PC/WALP23. At 3.5 vol% TFE, the previous striated ordering of the peptide is abolished and replaced by loose lines.     

Crystal structures of substrate-free and nitrosyl cytochrome p450cin: implications for o(2) activation

The crystal structure of the P450cin substrate-bound nitric oxide complex and the substrate-free form have been determined revealing a substrate-free structure that adopts an open conformation relative to the substrate-bound structure. The region of the I helix that forms part of the O(2) binding pocket shifts from an α helix in the substrate-free form to a π helix in the substrate-bound form. Unique to P450cin is an active site residue, Asn242, in the I helix that H-bonds with the substrate. In most other P450s this residue is a Thr and plays an important role in O(2) activation by participating in an H-bonding network required for O(2) activation. The π/α I helix transition results in the carbonyl O atom of Gly238 moving in to form an H-bond with the water/hydroxide ligand in the substrate-free form. The corresponding residue, Gly248, in the substrate-free P450cam structure experiences a similar motion. Most significantly, in the oxy-P450cam complex Gly248 adopts a position midway between the substrate-free and -bound states. A comparison between these P450cam and the new P450cin structures provides insights into differences in how the two P450s activate O(2). The structure of P450cin complexed with nitric oxide, a close mimic of the O(2) complex, shows that Gly238 is likely to form tighter interactions with ligands than the corresponding Gly248 in P450cam. Having a close interaction between an H-bond acceptor, the Gly238 carbonyl O atom, and the distal oxygen atom of O(2) will promote protonation and hence further reduction of the oxy complex to the hydroperoxy intermediate resulting in heterolytic cleavage of the peroxide O-O bond and formation of the active ferryl intermediate required for substrate hydroxylation.     

Heterokaryon Identification Through Simultaneous Fluorescence of Tetramethylrhodamine Isothiocyanate and Fluorescein Isothiocyanate Labelled Protoplasts

Protoplasts were separately stained with the fluorescent dyes fluorescein iso-thiocyanate (FITC) and tetramethylrhodamine isothiocyanate (TRITC). Following fusion, doubly stained heterokaryons were identified under fluorescence microscopy by using the Zeiss filter set 48 77 05 (excitation filter 450-490 nm, dichroic reflector 510 nm, and barrier filter 520 nm) which allowed simultaneous fluorochrome emissions. Previously, either emisson spectrum, but not both, was possible for any single filter set.     

Brettanomyces naardenensis,a new yeast from soft drinks

A new species of yeast,Brettanomyces naardenensis, is described. The strains studied were all isolated from soft drinks with a pH of 2.6–3.2. This species differs from all the presently acceptedBrettanomyces species by assimilating soluble starch.     

IsBombus inexspectatus (Tkalcu) a workerless obligate parasite? (Hym. Apidae)

Summary Bombus inexspectatus (Tkalc) is now known from 50 males, 36 females and two individuals which have been described as workers; the species has been found in the mountains of Austria, France, Italy, Spain and Switzerland at altitudes between 1,181 m. (N.W. Spain) and 2,100 m. (N.E. Italy). More than 10,000 bumblebees have been captured in localities whereB.inexspectatus is known to occur yet no further worker individuals have been discovered. The capture of some very undersized females in spring long before workers of any other species were flying puts the status of the two so-called workers in considerable doubt and examination of these two specimens points very strongly to their being females and not workers. The possibility that the femaleB. inexspectatus behaves like aPsithyrus and lives in the nest of another bee seems a not improbable explanation of the situation though proof may come only by chance discovery. Certain peculiarities of the female, notably absence of any indication, in pinned specimens, of either wax secretion or pollen collecting and at least the suggestion that the pollen collecting equipment may be degenerate if not entirely non-functional are added pointers to the necessity of a parasitic mode of living.Bombus ruderarius Müller is suggested as a likely host.

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Zusammenfassung Bombus inexspectatus (Tkalc), ist jetzt nach 50 Männchen, 36 Weibchen und 2 als Arbeiter beschriebenen Exemplaren bekannt; die Art ist in den Gebirgen Österreichs, Frankreichs, Italiens, Spaniens und der Schweiz in Höhen zwischen 1181 m. (N.W. Spanien) und 2100 m. (N.O. Italien) gefunden worden. Mehr als 10000 Hummeln sind an den Fundorten gefangen worden, wo das Vorkommen vonB. inexspectatus bekannt ist, doch hat man keine weiteren Arbeiter entdeckt. Das Sammeln einiger sehr kleinen Weibchen am Anfang des Frühlings — lange vor der Flugzeit der Arbeiter anderer Arten — lässt den Status der zwei Arbeiter bezweifeln, und eine Untersuchung zeigt, dass sie wahrscheinlich Weibchen und keine Arbeiter sind. Es ist durchaus möglich, dass das Weibchen vonB. inexspectatus sich wie einPsithyrus verhält und im Nest einer anderen Hummel lebt, doch vielleicht könnte eine zufällige Entdeckung diese Hypothese bestätigen. Gewisse Eigentümlichkeiten des Weibchens sind weitere Anzeiger der Notwendigkeit einer parasitischen Lebensweise: besonders die Abwesenheit jeder Andeutung entweder von Waschsabscheidung oder von Pollensammeln und die Möglichkeit, dass der Pollensammelapparat wenigstens degeneriert, wenn nicht ganz funktionlos geworden, ist.Bombus ruderarius Müller wird als wahrscheinlicher Wirt vorgeschlagen.

     

Selenotila peltata comb. n.

A brief historical survey of the genusSelenotila Lagerheim is given andTorulopsis peltata Yarrow is transferred to this genus as a new combination.I would like to thank Dr. I. P. Babjeva for bringing the genusSelenotila to ray attention and Miss J. Buijze for translating the Russian text.     

Absence of mitochondrial and chloroplast DNA recombinations in Brassica napus plants regenerated from protoplasts,protoplast fusions and anther culture

Summary Over 400 Brassica napus plants regenerated from individual protoplasts, from protoplast fusions and from anther culture were analysed for chloroplast and mitochondrial genome rearrangements by restriction fragment length polymorphisms. None were detected, attesting to the fidelity of the tissue culture procedures employed. In the majority of protoplast fusion products, the cytoplasmic organelles had completely sorted out at the callus stage but three regenerated plants possessed mixed parental populations of mitochondrial genomes and one regenerant contained mixed chloroplast genomes. In all four examples, the cytoplasmic genome sorted out in planta in favor of one parental type which was faithfully maternally transmitted to progeny.