Solid-state 15N NMR of oriented lipid bilayer bound gramicidin A'

Highly oriented samples of lipid and gramicidin A' (8:1 molar ratio) have been prepared with the samples extensively hydrated (approximately 70% water v/w). These preparations have been shown to be completely in a bilayer phase with a transition temperature of 28 degrees C, and evidence is presented indicating that the gramicidin is in the channel conformation. An estimate of the disorder in the alignment of the bilayers parallel with the glass plates used to align the bilayers can be made from the asymmetry of the nuclear magnetic resonances (NMR). Such an analysis indicates a maximal range of disorder of +/- 3 degrees. Uniformly 15N-labeled gramicidin has been biosynthesized by Bacillus brevis grown in a media containing 15N-labeled Escherichia coli cells as the only nitrogen source. When prepared with labeled gramicidin, the oriented samples result in high-resolution 15N NMR spectra showing 12 resonances for the 20 nitrogen sites of the polypeptide. The frequency of the three major multiple resonance peaks has been interpreted to yield the approximate orientation of the N-H bonds in the peptide linkages with respect to the magnetic field. These bond orientations are only partially consistent with the extant structural models of gramicidin.     

Reversible hyperphosphorylation and reorganization of vimentin intermediate filaments by okadaic acid in 9L rat brain tumor cells.

Okadaic acid (OA), a protein phosphatase inhibitor, was found to induce hyperphosphorylation and reorganization of vimentin intermediate filaments in 9L rat brain tumor cells. The process was dose dependent. Vimentin phosphorylation was initially enhanced by 400 nM OA in 30 min and reached maximal level (about 26-fold) when cells were treated with 400 nM OA for 90 min. Upon removal of OA, dephosphorylation of the hyperphosphorylated vimentin was observed and the levels of phosphorylation returned to that of the controls after the cells recovered under normal growing conditions for 11 h. The phosphorylation and dephosphorylation of vimentin induced by OA concomitantly resulted in reversible reorganization of vimentin filaments and alteration of cell morphology. Cells rounded up as they were entering mitosis in the presence of OA and returned to normal appearance after 11 h of recovery. Immuno-staining with anti-vimentin antibody revealed that vimentin filaments were disassembled and clustered around the nucleus when the cells were treated with OA but subsequently returned to the filamentous states when OA was removed. Two-dimensional electrophoresis analysis further revealed that hyperphosphorylation of vimentin generated at least seven isoforms having different isoelectric points. Furthermore, the enhanced vimentin phosphorylation was accompanied by changes in the detergent-solubility of the protein. In untreated cells, the detergent-soluble and -insoluble vimentins were of equal amounts but the solubility could be increased when vimentins were hyperphosphorylated in the presence of OA. Taken together, the results indicated that OA could be involved in reversible hyperphosphorylation and reorganization of vimentin intermediate filaments, which may play an important role in the structure-function regulation of cytoskeleton in the cell.     

Seed Germination and Seedling Growth in Sopubia delphinifolia - a Hemi-root Parasite: Germination Requirements

Requirements for seed germination (emergence of radicle) andseedling formation (emergence of both radicle and cotyledons)of a hemi-root parasite Sopubia delphinifolia were studied inthe absence of any host stimulus, in Petri dish and asepticcultures. Both water washing in trickling tap water as wellas cold treatment were effective in inducing/stimulating germinationand seedling formation in the light. Although ethrel, an ethylenereleasing compound, stimulated radicle emergence it inhibitedthe emergence of the cotyledons. Light was found to be essentialfor germination; none of the growth substances could replacethe light requirement. Light responses seem to be mediated throughthe phytochrome system. The results indicate that the emergenceof the radicle, its further growth into the root and the emergenceof cotyledons are controlled by different factors. Sopubia delphinifolia, hemi-root parasite, seed germination requirements, pretreatment, seedling formation     

Enzyme histochemical studies of the general body surface apocrine glands in the domestic cat]

With enzyme histochemical methods, the distribution and the activities of various oxidative and hydrolytic enzymes in the apocrine glands of the general body surface of the domestic cat were investigated. The results obtained support the view that these glands are clearly active in function. On the whole, however, the relatively weak enzyme activities in the secretory portion of the glands point to only unimportant secretion production rates.     

Fusion of lipid bilayers: a model involving mechanistic connection to HII phase forming lipids.

A model for the molecular mechanism of the fusion of lipid bilayers is described. A crucial feature of this model and related to the lamellar-->hexagonal phase HII transition is a novel, hypothetical lipid conformation, tentatively referred to here as extended. During fusion this conformation could manifest itself in the contact site between two vesicles in close proximity and involves the extension of the acyl chains of a phospholipid molecule in opposite directions, i.e. embedded into the two opposing bilayers while maintaining the headgroup in the interface. Although evidence for the occurrence of the extended conformation for phospholipids is sparse this conformation appears to be compatible with currently available experimental data. Of importance also is that the extended conformation allows for the fusion of two bilayer membranes to proceed with minimal exposure of the lipid hydrocarbon chains to water. It can also account for other features of membrane fusion such as lipid mixing in the intermediate state without mixing of the vesicle contents as well as for the molecular basis of the action of fusogenic lipids.     

Sexual behavior and serum hormone levels in aging rhesus males: effects of environmental change

Old (N = 9) and middle-aged (N = 9) rhesus males (Macaca mulatta) that were housed in the same type of home cage and the same room for several years were moved to a new location in an adjacent building. Their sexual behavior and hormone levels in the weeks preceding the move were compared to those after the move. The hypothesis that the nonspecific sensory stimulation provided by the move would increase levels of sexual behavior was not supported by the data. To the contrary, the percentages of tests with contacts, mounts, and penile erections, and the rate of contacting the female, decreased significantly for both old and middle-aged males. The intromission rate of middle-aged, but not of old, males also decreased. In consonance with previous findings, middle-aged males displayed significantly higher levels of sexual activity than did old males. The differences were observed both before and after the move. Serum testosterone levels did not differ between middle-aged and old males, and the move did not produce significant changes in testosterone levels. Serum cortisol levels were significantly higher immediately after the move (1 hr) but returned to premove levels the following day.     

Aggression persists after ovariectomy in female rats

Aggression occurs not only in males but also in females, however, under different sex-specific stimulus and endocrine conditions. After being housed with males, female rats exhibit frequent and intense aggressive behavior toward unfamiliar rats. However, the female residents primarily attack female intruder rats, while the male residents attack males and not females. Altering the hormonal condition of the intruders can modify the behavior that they provoke from the residents. Castration of the male intruders reduces aggression from male residents, but ovariectomy of the female intruders does not alter the behavior of the female residents. Treatment of the gonadectomized intruders with gonadal steroids significantly alters the response of the male residents. Resident-intruder aggressive behavior depends on the presence of the testes in the male residents but not on the ovaries or on lactation in the female residents. Even 7 weeks after ovariectomy the female residents continue to show aggressive behavior toward female intruders. In the same time period the castrated male residents show a marked decrease in aggressive and sexual behavior.