Chromatophore vesicles of Rhodobacter capsulatus contain on average one F(O)F(1)-ATP synthase each

ATP synthase is a unique rotary machine that uses the transmembrane electrochemical potential difference of proton (Delta(H(+))) to synthesize ATP from ADP and inorganic phosphate. Charge translocation by the enzyme can be most conveniently followed in chromatophores of phototrophic bacteria (vesicles derived from invaginations of the cytoplasmic membrane). Excitation of chromatophores by a short flash of light generates a step of the proton-motive force, and the charge transfer, which is coupled to ATP synthesis, can be spectrophotometrically monitored by electrochromic absorption transients of intrinsic carotenoids in the coupling membrane. We assessed the average number of functional enzyme molecules per chromatophore vesicle. Kinetic analysis of the electrochromic transients plus/minus specific ATP synthase inhibitors (efrapeptin and venturicidin) showed that the extent of the enzyme-related proton transfer dropped as a function of the inhibitor concentration, whereas the time constant of the proton transfer changed only marginally. Statistical analysis of the kinetic data revealed that the average number of proton-conducting F(O)F(1)-molecules per chromatophore was approximately one. Thereby chromatophores of Rhodobacter capsulatus provide a system where the coupling of proton transfer to ATP synthesis can be studied in a single enzyme/single vesicle mode.     

High resolution NMR analysis of the seven transmembrane domains of a heptahelical receptor in organic-aqueous medium

The NMR properties of seven peptides representing the transmembrane domains of the alpha-factor receptor from Saccharomyces cerevisiae were examined in trifluoroethanol/water (4:1) at 10 to 55 degrees C. The parameters extracted indicated all peptides were helical in this membrane mimetic solvent. Using chemical shift indices as the criterion, helicity varied from 64 to 83%. The helical residues in the peptides corresponded to the region predicted to cross the hydrocarbon interior of the bilayer. A study of a truncated 25-residue peptide corresponding to domain 2 gave evidence that the helix extended all the way to the N-terminus of this peptide, indicating that sequence and not chain end effects are very important in helix termination for our model peptides. Both nuclear Overhauser effect spectroscopy (NOESY) connectivities and chemical shift indices revealed significant perturbations around prolyl residues in the helices formed by transmembrane domains 6 and 7. Molecular models of the transmembrane domains indicate that helices for domains 6 and 7 are severely kinked at these prolyl residues. The helix perturbation around proline 258 in transmembrane domain 6 correlates with mutations that cause phenotypic changes in this receptor.     

Host discrimination by two desert fleas using an odour cue

We compared the responses of two fleas, Xenopsylla dipodilli and Parapulex chephrenis simultaneously exposed to the odours of their rodent hosts, Gerbillus dasyurus (specific host ofX. dipodilli ) and Acomys cahirinus (specific host of P. chephrenis). We hypothesized that fleas are able to discriminate between host species by using an odour cue and predicted that X. dipodilli andP. chephrenis would select an odour of an appropriate host species. Xenopsylla dipodilli choseG. dasyurus significantly more often than A. cahirinus, whereas P. chephrenis choseA. cahirinus significantly more often than G. dasyurus. The ability to select an appropriate host species did not differ significantly either between flea species or between individuals of different sex or age classes within flea species. No X. dipodilli, but 67 of 150 P. chephrenis, refused to choose a host. The latency to move in an experimental maze was significantly shorter for X. dipodilli than P. chephrenis. The flea species also differed in the time taken from the beginning of the movement to the choice of a host, withX. dipodilli being faster than P. chephrenis. Neither flea sex nor age affected this parameter in either species. Females of both flea species produced significantly more eggs when they fed on their specific host than when they fed on the other host species. Copyright 2002 The Association for the Study of Animal Behaviour. Published by Elsevier Science Ltd. All rights reserved.     

Myofibroblasts and mechano-regulation of connective tissue remodelling

During the past 20 years, it has become generally accepted that the modulation of fibroblastic cells towards the myofibroblastic phenotype, with acquisition of specialized contractile features, is essential for connective-tissue remodelling during normal and pathological wound healing. Yet the myofibroblast still remains one of the most enigmatic of cells, not least owing to its transient appearance in association with connective-tissue injury and to the difficulties in establishing its role in the production of tissue contracture. It is clear that our understanding of the myofibroblast its origins, functions and molecular regulation will have a profound influence on the future effectiveness not only of tissue engineering but also of regenerative medicine generally.     

Affinity modification of EcoRII DNA methyltransferase by the dialdehyde-substituted DNA duplexes: mapping the enzyme region that interacts with DNA

Affinity modification of EcoRII DNA methyltransferase (M x EcoRII) by DNA duplexes containing oxidized 2'-O-beta-D-ribofuranosylcytidine (Crib*) or 1-(beta-D-galactopyranosyl)thymine (Tgal*) residues was performed. Cross-linking yields do not change irrespective of whether active Crib* replaces an outer or an inner (target) deoxycytidine within the EcoRII recognition site. Chemical hydrolysis of M x EcoRII in the covalent cross-linked complex with the Tgal*-substituted DNA indicates the region Gly268-Met391 of the methylase that is likely to interact with the DNA sugar-phosphate backbone. Both specific and non-specific DNA interact with the same M x EcoRII region. Our results support the theoretically predicted DNA binding region of M x EcoRII.     

Myeloid or lymphoid promiscuity as a critical step in hematopoietic lineage commitment

We demonstrate here that "promiscuous" expression of myeloid or lymphoid genes precedes lineage commitment in hematopoiesis. Prospectively purified single common myeloid progenitors (CMPs) coexpress myelo-erythroid but not lymphoid genes, whereas single common lymphoid progenitors (CLPs) coexpress T and B lymphoid but not myeloid genes. Genes unrelated to the adopted lineage are downregulated in bipotent and monopotent descendants of CMPs and CLPs. Promiscuous gene expression does not alter the biological potential of multipotent progenitors: CMPs with an activated endogenous M lysozyme locus yield normal proportions of myelo-erythroid colonies, and CLPs expressing the pre-T cell receptor alpha gene differentiate into normal numbers of B cells. Thus, the accessibility for multiple myeloid or lymphoid programs promiscuously may allow flexibility in fate commitments at these multipotent stages.     

Effect of natural b-carotene supplementation in children exposed to radiation from the Chernobyl accident

Attempts were made to evaluate 709 children (324 boys and 385 girls) who had been exposed long-term to different doses of radiation during and after the Chernobyl accident and had moved to Israel between 1990 and 1994. Upon arrival, all of them underwent a check-up for most common clinical disorders and were then divided into three groups according to their residences (distance from the reactor) and the level of irradiation exposure: no radiation, <5 Ci/m², and >5 Ci/m², respectively. Blood serum analyses for total carotenoids, retinol, α-tocopherol and oxidized conjugated dienes in 262 of the children showed increased HPLC levels of conjugated dienes, indicating increased levels of oxidation of in vivo blood lipids in children from the contaminated areas. The levels were higher in girls than in boys. Some 57 boys and 42 girls were given a basal diet with a diurnal supplementation of 40 mg natural 9-cis and all-trans equal isomer mixture β-carotene in a capsulated powder form of the alga Dunaliella bardawil, for a period of 3 months. Blood serum analyses were regularly conducted before supplementation to determine the baseline effect of radiation exposure to the children, after 1 and 3 months of natural β-carotene supplementation. After supplementation, the levels of the oxidized conjugated dienes decreased in the children's sera without any significant changes in the level of total carotenoids, retinol or α-tocopherol. Other common blood biochemicals were within the normal range for all tests and no statistical differences before or after supplementation of β-carotene were noted. High pressure liquid chromatography (HPLC) analyses for carotenoids in the blood detected mainly oxycarotenoids, and to a lesser extent, all-trans β-carotene, α-carotene, but not 9-cis β-carotene. The results suggest that irradiation increases the susceptibility of lipids to oxidation in the Chernobyl children and that natural β-carotene may act as an in vivo lipophilic antioxidant or radioprotector. Received: 20 March 1998 / Accepted in revised form: 1 August 1998     

Phytoplankton development and turbulent mixing in Lake Kinneret (1992-1996)

We have utilized data from a recently developed three-dimensionalvelocity fluctuation meter to compute the dissipation of turbulentkinetic energies (TKE) and the intensity of turbulent mixingin horizontal and vertical planes in the pelagic, epilimnicwater of Lake Kinneret, Israel. These characteristics of wind-inducedturbulent movement have been monitored from January 1992 throughDecember 1996. The turbulence parameters were strongly correlatedto wind energy inputs, calculated daily as 5 day cumulativeinputs. There have been dramatic changes in the annual and seasonaldevelopment of phytoplankton, together with unusually high levelsof primary production in this lake since 1994. We observed differentpatterns of vertical and horizontal turbulent movement and ofTKE dissipation rates during the years when ‘unusual’phytoplankton development occurred (1994–1996) comparedto ‘normal’ years (1992, 1993). The first appearanceof the filamentous cyanobacterium Aphanizomenon in this lakein August–September 1994 coincided with a period of markedlylower rates of TKE dispersion and a shift from vertical to horizontaldominance of the turbulent eddy spins. The absence of a regularwinter-spring bloom of the dinoflagellate, Peridinium, in 1996occurred when dissipation rates of TKE were extremely high,while record high amounts of dinoflagellates (1994, 1995) appearedwhen dissipation rates were very low. Correlations were shownbetween phytoplankton parameters (chlorophyll, primary productionand the ratio of primary production to chlorophyll) and boththe dissipation rate of TKE and the intensity of water turbulentmixing in the vertical plane. We suggest that the changes inthe ‘turbulence climate’ of Lake Kinneret were animportant factor in determining shifts in phytoplankton successionand the population composition of the algal assemblage.