Topology of PhoE porin: the 'eyelet' region

A model for the topology of the PhoE porin has been proposed according to which the polypeptide traverses the outer membrane sixteen times mostly as amphipathic β-sheets, thereby exposing eight loops at the cell surface. Until now, no evidence has been obtained for the surface exposure of the third loop. Recently, the structure of porin of Rhodobacter capsulatus has been determined. The proposed model of PhoE is very similar to the structure of the R capsulatus porin, which has an ‘eyelet’ region, extending into the interior of the pore. The proposed third external loop of PhoE might form a similar ‘eyelet’ region. To determine the location of the predicted third external loop of PhoE, multiple copies of an oligonucleotide linker encoding an antigenic determinant of VP1 protein of foot-and-mouth disease virus (FMDV) were inserted. All hybrid proteins were properly inserted in the outer membrane. The monoclonal antibody MA11, directed against the linear FMDV epitope, was able to bind only to intact cells expressing a hybrid PhoE protein with at least three copies of the FMDV epitope present. Antibiotic sensitivity tests and single-channel conductance measurements revealed that the insertions influenced the channel size. These results are consistent with a location of the third loop of PhoE within the pore channel.     

The trace fossils of the Baggy Beds (Upper Devonian) of North Devon,England

The trace fossils of the Baggy Beds are described. They includeTeichichnus cf.rectus Seilacher,Monocraterion cf.tentaculatum Torell,Diplocraterion yoyo sp. nov.,Arenicolites curvatus sp. nov., and other forms insufficiently well preserved for generic assignment. The protrusive and retrusive behaviour pattern ofD. yoyo is interpretated as a response to repeated phases of sedimentation and erosion and as a measure of these.A. curvatus is preserved in two modes: in the position of formation and as eroded tubes in conglomerates. The second reveals that the sediment around the tube was cemented to a thickness equal to the diameter of the tube.     

Colony Member Discrimination by Juvenile Columbian Ground Squirrels (Spermophilus columbianus)

Whether an individual strives to breed or restrains from reproducing directly but increases the fitness of another individual through its help, may be viewed as a result of a trade-off between fitness costs and benefits arising from this decision. A population of the large carpenter bee Xylocopa sulcatipes Maa was studied in southeast Israel over a two year period. Female reproductive success, in terms of genetic representation in the gene pool (genetic gain), was calculated using coefficients of relatedness and the number of offspring produced by solitary, reproductively dominant and helper bees of social nests. In one year helper females, that shared a nest with a related female accrued a higher genetic gain than did solitary females. In the subsequent year solitary females did better than the helper bees. In nests founded by unrelated females a helper bee accrued only little genetic gain. However, evidence is presented to show that subordinate behaviour can have an adaptive value and can ultimately be beneficial through nest inheritance by the subordinate bee.     

CHLOROPLAST DNA VARIATION AND THE PHYLOGENY OF HORDEUM (POACEAE)

Restriction site variation in the chloroplast genome (cpDNA) was surveyed among 37 taxa or cytotypes (40 accessions) of the genus Hordeum. Seventeen restriction enzymes were employed, and a total of 491 restriction sites were assayed. Of these, 120 were variable among the taxa, including 70 synapomorphies. The level of sequence divergence (p) among species of Hordeum varied from 0.0 to 0.017, indicating that Hordeum possesses an about-average level of cpDNA diversity as compared to most other genera of flowering plants for which data are available. Wagner and polymorphism parsimony phytogenies were constructed from the restriction site data. These analyses divided the genus into several distinct groups; 1) American taxa; 2) diploid H. marinum; 3) Asian taxa; 4) H. vulgare-H. bulbosum; and 5) the H. murinum complex. Bootstrap-based confidence limits provided statistical support for the monophylesis of the latter three groups. The cpDNA data showed remarkably good congruence with previously published isoenzymatic, molecular, cytological, and crossing data.     

Intergeneric hybridization between Hordeum and Hordelymus (Poaceae)

Intergeneric hybridization between the monotypic Eurasian genus Hordelymus and species of Hordeum was performed. Hybrids were obtained in nine combinations and the meiotic pairing was analyzed in four combinations (6 families). Hordelymus europaeus is probably an alloploid with two different genomes. The very low pairing in the hybrids with H. jubatum, H. brachyantherum , and H. pusillum indicates that these species do not have any genomes in common with Hordelymus. The hybrids with H. depressum have an intermediate pairing which suggests that there is a certain homoeology in one genome.     

Detection of renal cell carcinoma-associated markers via proteome- and other 'ome'-based analyses.

Proteome analysis has rapidly developed in the post-genome era and is now widely accepted as the complementary technology for genetic profiling. It has been shown to be a powerful tool for studying human diseases and for identifying novel prognostic, diagnostic and therapeutic markers. This review focuses on the identification of new biomarkers and therapeutic targets for renal cell carcinoma using different 'ome'-based technologies.     

Ligand-specific dose-response of heterologously expressed olfactory receptors.

Primary olfactory neuronal cultures exposed to odorant stimulation have previously exhibited concentration-related effects in terms of intracellular cAMP levels and adenylate cyclase activity [Ronnett, G.V., Parfitt, D.J., Hester, L.D. & Snyder, S.H. (1991) PNAS88, 2366-2369]. Maximal stimulation occurred for intermediate concentrations, whereas AC activity declined for both low and high odorant concentrations. We suspected that this behavior might be ascribed to the intrinsic response of the first molecular species concerned by odorant detection, i.e. the olfactory receptor itself. In order to check this hypothesis, we developed an heterologous expression system in mammalian cells to characterize the functional response of receptors to odorants. Two mammalian olfactory receptors were used to initiate the study, the rat I7 olfactory receptor and the human OR17-40 olfactory receptor. The cellular response of transfected cells to an odorant stimulation was tested by a spectrofluorimetric intracellular calcium assay, and proved in all cases to be dose-dependent for the known ligands of these receptors, with an optimal response for intermediate concentrations. Further experiments were carried out with the rat I7 olfactory receptor, for which the sensitivity to an odorant, indicated by the concentration yielding the optimal calcium response, depended on the carbon chain length of the aldehydic odorant. The response is thus both ligand-specific and dose-dependent. We thus demonstrate that a differential dose-response originates from the olfactory receptor itself, which is thus capable of efficient discrimination between closely related agonists.     

Lactate and epinephrine during exercise in altitude natives

Kayser, Bengt, Roland Favier, Guido Ferretti, DominiqueDesplanches, Hilde Spielvogel, Harry Koubi, Brigitte Sempore, and HansHoppeler. Lactate and epinephrine during exercise in altitudenatives. J. Appl. Physiol. 81(6):2488-2494, 1996.We tested the hypothesis that the reported lowblood lactate accumulation ([La]) during exercise inaltitude-native humans is refractory to hypoxia-normoxia transitions byinvestigating whether acute changes in inspiredO₂ fraction(FI_O2) affect the[La] vs. power output ()relationship or, alternatively, as reported for lowlanders, whetherchanges in [La] vs. on changes inFI_O2 are related tochanges in blood epinephrine concentration ([Epi]). Altitude natives [n = 8, age 24 ± 1 (SE) yr, body mass 62 ± 3 kg, height 167 ± 2 cm]in La Paz, Bolivia (3,600 m) performed incremental exercise with twolegs and one leg in chronic hypoxia and acute normoxia (AN). Submaximalone- and two-leg O₂ uptake (O₂) vs. relationships were not altered byFI_O2. AN increased two-legpeak O₂ by 10% and peak by 7%. AN paradoxically decreasedone-leg peak O₂ by 7%,whereas peak remained the same. The[La] vs. relationships were similar tothose reported in unacclimatized lowlanders. There was a shift to theright on AN, and maximum [La] was reduced by 7 and 8% forone- and two-leg exercises, respectively. [Epi] and[La] were tightly related (mean r = 0.81) independently ofFI_O2. Thus normoxiaattenuated the increment in both [La] and [Epi]as a function of , whereas the correlation between[La] and [Epi] was unaffected. These data suggest loose linkage of glycolysis to oxidative phosphorylation under influence from [Epi]. In conclusion, high-altitudenatives appear to be not fundamentally different from lowlanders with regard to the effect of acute changes inFI_O2 on [La] during exercise.

     

Influence of Androgenicity on Adipocytes and Precursor Cells in Female Rats

We examined the effects of overexposure of testosterone (T) on fat cell morphology and adipocyte precursor pools in inguinal and retroperitoneal fat depots of ovariectomized rats. In both tissues peripubertal T decreased weights without affecting adipocyte mean cell size or the size distribution profiles, but adipocyte number was decreased by 65% in the inguinal and by 38% in the retroperitoneal depots. Immunofluorescent flow cytometry utilizing a specific antibody to rat adipose tissue lipoprotein lipase was used to quantify regional precursor cell populations. T sharply reduced the percentages of differentiated and undifferentiated preadipocytes in the inguinal depot, from 43.2 ± 5.3 to 23.5 ± 2.1% and from 57.7 ± 4.0 to 43.6 ± 5.3%, respectively, with a concomitant increase in fibroblasts from 1.6 to 32.9%. On the other hand, T had no effect on retroperitoneal preadipocyte pools. Perinatal andro-genization exacerbated the decline in the inguinal weight (1.4 ± 0.1 vs. 2.2 ± 0.1g) but otherwise did not influence the actions of peripubertal T. Androgens may thus act in a tissue-specific manner to regulate fat cell growth potential in the femoral region in the female.