The inner membrane protein YhiM is necessary for Escherichia coli growth at high temperatures and low osmolarity.

To survive, Escherichia coli must be able to survive in rapidly changing environmental conditions including changes in temperature and osmolarity. We have studied the role of the inner membrane protein YhiM in changing environmental conditions. Our data indicate that YhiM is required for normal growth at 37 and 41 °C but not 21 °C. YhiM-deficient cells grown at high temperatures spend more time in lag phase and stop growing at lower cell densities in comparison with their wild-type counterparts. They also have growth defects in low NaCl medium at 37 °C and do not grow at all at 41 °C. The effects of low NaCl can be rescued by addition of KCl or sucrose to the low salt medium. Finally, YhiM-deficient cells fail to grow in dilute medium at 41 °C. These data suggest that YhiM may be important in protecting the cells from changes in temperature and osmolarity.

Formation of amino acids and nucleotide bases in a Titan atmosphere simulation experiment.

The discovery of large (>100 u) molecules in Titan's upper atmosphere has heightened astrobiological interest in this unique satellite. In particular, complex organic aerosols produced in atmospheres containing C, N, O, and H, like that of Titan, could be a source of prebiotic molecules. In this work, aerosols produced in a Titan atmosphere simulation experiment with enhanced CO (N(2)/CH(4)/CO gas mixtures of 96.2%/2.0%/1.8% and 93.2%/5.0%/1.8%) were found to contain 18 molecules with molecular formulae that correspond to biological amino acids and nucleotide bases. Very high-resolution mass spectrometry of isotopically labeled samples confirmed that C(4)H(5)N(3)O, C(4)H(4)N(2)O(2), C(5)H(6)N(2)O(2), C(5)H(5)N(5), and C(6)H(9)N(3)O(2) are produced by chemistry in the simulation chamber. Gas chromatography-mass spectrometry (GC-MS) analyses of the non-isotopic samples confirmed the presence of cytosine (C(4)H(5)N(3)O), uracil (C(5)H(4)N(2)O(2)), thymine (C(5)H(6)N(2)O(2)), guanine (C(5)H(5)N(5)O), glycine (C(2)H(5)NO(2)), and alanine (C(3)H(7)NO(2)). Adenine (C(5)H(5)N(5)) was detected by GC-MS in isotopically labeled samples. The remaining prebiotic molecules were detected in unlabeled samples only and may have been affected by contamination in the chamber. These results demonstrate that prebiotic molecules can be formed by the high-energy chemistry similar to that which occurs in planetary upper atmospheres and therefore identifies a new source of prebiotic material, potentially increasing the range of planets where life could begin.

Chemical characterization of Titan's tholins: solubility, morphology and molecular structure revisited.

In this work Titan's atmospheric chemistry is simulated using a capacitively coupled plasma radio frequency discharge in a N(2)-CH(4) stationnary flux. Samples of Titan's tholins are produced in gaseous mixtures containing either 2 or 10% methane before the plasma discharge, covering the methane concentration range measured in Titan's atmosphere. We study their solubility and associated morphology, their infrared spectroscopy signature and the mass distribution of the soluble fraction by mass spectrometry. An important result is to highlight that the previous Titan's tholin solubility studies are inappropriate to fully characterize such a heterogeneous organic matter and we develop a new protocol to evaluate quantitatively tholins solubility. We find that tholins contain up to 35% in mass of molecules soluble in methanol, attached to a hardly insoluble fraction. Methanol is then chosen as a discriminating solvent to characterize the differences between soluble and insoluble species constituting the bulk tholins. No significant morphological change of shape or surface feature is derived from scanning electron microscopy after the extraction of the soluble fraction. This observation suggests a solid structure despite an important porosity of the grains. Infrared spectroscopy is recorded for both fractions. The IR spectra of the bulk, soluble, and insoluble tholins fractions are found to be very similar and reveal identical chemical signatures of nitrogen bearing functions and aliphatic groups. This result confirms that the chemical information collected when analyzing only the soluble fraction provides a valuable insight representative of the bulk material. The soluble fraction is ionized with an atmospheric pressure photoionization source and analyzed by a hybrid mass spectrometer. The congested mass spectra with one peak at every mass unit between 50 and 800 u confirm that the soluble fraction contains a complex mixture of organic molecules. The broad distribution, however, exhibits a regular pattern of mass clusters. Tandem collision induced dissociation analysis is performed in the negative ion mode to retrieve structural information. It reveals that (i) the molecules are ended by methyl, amine and cyanide groups, (ii) a 27 u neutral moiety (most probably HCN) is often released in the fragmentation of tholin anions, and (iii) an ubiquitous ionic fragment at m/z 66 is found in all tandem spectra. A tentative structure is proposed for this negative ion.

Photoionization cross section of Xe+ ion in the pure 5p5 2P3/2 ground level.

Coupling an ion trap with synchrotron radiation is shown here to be a powerful approach to measure photoionization cross sections on ionic species relaxed in their ground state. The photoionization efficiency curve of Xe+ ions stored in a Fourier transform ion cyclotron resonance ion trap was recorded at ELETTRA in the 20-23 eV photon energy range. Absolute cross sections were derived by comparison of the photoionization yield of Xe+ with measurements from the ASTRID merged-beam experiment. Multiconfiguration Dirac-Fock calculations were performed for the interpretation of these new data.

Gas-phase properties and fragmentation behavior of cationic, dinuclear iron chloride clusters Fe(2)Cl(n)()(+) (n = 1-6).

Sector-field mass spectrometry is used to probe the fragmentation patterns of cationic dinuclear iron chloride clusters Fe(2)Cl(n)()(+) (n = 1-6). For the chlorine-rich, high-valent Fe(2)Cl(n)()(+) ions (n = 4-6), losses of atomic and molecular chlorine prevail in the unimolecular and collision-induced dissociation patterns. Instead, the chlorine deficient, formally low-valent Fe(2)Cl(n)()(+) clusters (n = 1-3) preferentially undergo unimolecular degradation to mononuclear FeCl(m)()(+) ions. In addition, photoionization is used to determine IE(Fe(2)Cl(6)) = 10.85 +/- 0.05 eV along with appearance energy measurements for the production of Fe(2)Cl(5)(+) and Fe(2)Cl(4)(+) cations from iron(III) chloride vapor. The combination of the experimental results allows an evaluation of some of the thermochemical properties of the dinuclear Fe(2)Cl(n)()(+) cations: e.g., Delta(f)H(Fe(2)Cl(+)) = 232 +/- 15 kcal/mol, Delta(f)H(Fe(2)Cl(2)(+)) = 167 +/- 4 kcal/mol, Delta(f)H(Fe(2)Cl(3)(+)) = 139 +/- 4 kcal/mol, Delta(f)H(Fe(2)Cl(4)(+)) = 113 +/- 4 kcal/mol, Delta(f)H(Fe(2)Cl(5)(+)) = 79 +/- 5 kcal/mol, and Delta(f)H(Fe(2)Cl(6)(+)) = 93 +/- 2 kcal/mol. The analysis of the data suggests that structural effects are more important than the formal valency of iron as far as the Fe-Cl bond strengths in the Fe(2)Cl(n)()(+) ions are concerned.

Insights into the C(4)H(8)(+.) potential energy surface: fourier transform ion cyclotron resonance studies of ion-molecule reactions of D-labeled partners

Reactions of [ethylene](+.) with ethylene and of [acetylene](+.) with ethane were studied by Fourier transform ion cyclotron resonance spectrometry using labeled reactants. The results confirm and clarify the different steps of the mechanism proposed previously and elaborated with other methods. The [[acetylene](+.), ethane] system can either dissociate to give the ethyl cation product, or isomerize into [[ethylene](+.), ethylene]. The latter system can either dissociate to yield ionized ethylene or convert into ionized but-2-ene, which undergoes a complete H-exchange prior to dissociation, leading to methyl radical, hydrogen radical and ethylene losses. The transfers of labeled atoms and the existence of H-exchange prior to formation of the products were used as a probe to check the different steps of the mechanism. The influence of the initial energy of the system on the reaction pathway is discussed. Copyright 1999 John Wiley & Sons, Ltd.

Differential regulation of Dk and Kk major histocompatibility complex class I proteins on the cell surface after infection of murine cells by pseudorabies virus.

After pseudorabies virus (PRV) infection of murine L929 cells, the cell surface expression of major histocompatibility complex (MHC) class I proteins changes such that the total amount of MHC class I molecules remains relatively constant but the levels of the individual alleles Dk and Kk vary. This is an active process involving at least three PRV gene products that act in an allele-specific manner such that cell surface expression of MHC class I Dk is decreased and that of Kk is increased. Our results indicate that an early gene product mediates the overall reduction in Dk protein and a late gene product which is mutant in the attenuated PRV strain Bartha mediates the increase in Kk protein. We provide additional evidence for a third gene product involved in the regulation of the synthesis of both the Dk and Kk proteins. In addition, we show that the early decrease in the Dk protein is not due to a block in synthesis or processing of the complex through the secretory system.

Isolation of a gene encoding a Chlamydia sp. strain TWAR protein that is recognized during infection of humans.

Chlamydia sp. strain TWAR is a unique Chlamydia sp. that causes acute respiratory disease. A gene bank consisting of TWAR isolate AR-39 DNA in pUC19 was screened with anti-AR-39 rabbit immune sera. Two positive clones were isolated that contained 7.3-kilobase (pLC1) and 14.9-kilobase (pLC2) plasmids. Restriction mapping and hybridization studies showed that both pLC1 and pLC2 contained a common 4.2-kilobase PstI fragment. Plasmids were used as templates of in vitro transcription-translation. All three plasmids had a novel protein product of ca. 75 kilodaltons not found in the vector alone. Western blots showed that this protein reacted with anti-TWAR rabbit immune sera and with human immune serum from an individual who had proven TWAR infection. Whole-cell lysates of TWAR demonstrated a protein having the same molecular weight and immunoreactivity as the recombinant gene product. This protein was also recognized by rabbit immune serum against Chlamydia psittaci or Chlamydia trachomatis. Southern hybridizations with the cloned fragment as a probe of digests of other Chlamydia spp. showed weakly hybridizing fragments. These results suggest that we have isolated a gene encoding a protein recognized during human TWAR infection that contains some sequences shared among Chlamydia spp.

[The effectiveness of stimulation of skin receptors in the treatment of prolonged pain (author's transl)]. / Uber die Wirksamkeit der Stimulation von Hautrezeptoren bei der Behandlung anhaltender Schmerzzustände.

In two groups of 30 patients pain relief due to transcutaneous nerve stimulation was compared with a placebo, under similar external conditions. The two groups were approximately similar in age, sex, and the origin of pain. Only patients with continuous pain of the limbs or trunk were included in the investigation. There was no statistically significant difference in the success rate of the two groups.