NMR spin-rotation relaxation and diffusion of methane.

The translational diffusion-coefficient and the spin-rotation contribution to the 1H NMR relaxation rate for methane (CH4) are investigated using MD (molecular dynamics) simulations, over a wide range of densities and temperatures, spanning the liquid, supercritical, and gas phases. The simulated diffusion-coefficients agree well with measurements, without any adjustable parameters in the interpretation of the simulations. A minimization technique is developed to compute the angular velocity for non-rigid spherical molecules, which is used to simulate the autocorrelation function for spin-rotation interactions. With increasing diffusivity, the autocorrelation function shows increasing deviations from the single-exponential decay predicted by the Langevin theory for rigid spheres, and the deviations are quantified using inverse Laplace transforms. The 1H spin-rotation relaxation rate derived from the autocorrelation function using the "kinetic model" agrees well with measurements in the supercritical/gas phase, while the relaxation rate derived using the "diffusion model" agrees well with measurements in the liquid phase. 1H spin-rotation relaxation is shown to dominate over the MD-simulated 1H-1H dipole-dipole relaxation at high diffusivity, while the opposite is found at low diffusivity. At high diffusivity, the simulated spin-rotation correlation time agrees with the kinetic collision time for gases, which is used to derive a new expression for 1H spin-rotation relaxation, without any adjustable parameters.

Role of internal motions and molecular geometry on the NMR relaxation of hydrocarbons.

The role of internal motions and molecular geometry on 1H NMR relaxation rates in liquid-state hydrocarbons is investigated using MD (molecular dynamics) simulations of the autocorrelation functions for intramolecular and intermolecular 1H-1H dipole-dipole interactions. The effects of molecular geometry and internal motions on the functional form of the autocorrelation functions are studied by comparing symmetric molecules such as neopentane and benzene to corresponding straight-chain alkanes n-pentane and n-hexane, respectively. Comparison of rigid versus flexible molecules shows that internal motions cause the intramolecular and intermolecular correlation-times to get significantly shorter, and the corresponding relaxation rates to get significantly smaller, especially for longer-chain n-alkanes. Site-by-site simulations of 1H's across the chains indicate significant variations in correlation times and relaxation rates across the molecule, and comparison with measurements reveals insights into cross-relaxation effects. Furthermore, the simulations reveal new insights into the relative strength of intramolecular versus intermolecular relaxation as a function of internal motions, as a function of molecular geometry, and on a site-by-site basis across the chain.

Second-order resummed thermodynamic perturbation theory for central-force associating potential: multi-patch colloidal models.

We propose a second-order version of the resummed thermodynamic perturbation theory for patchy colloidal models with arbitrary number of multiply bondable patches. The model is represented by the hard-sphere fluid system with several attractive patches on the surface and resummation is carried out to account for blocking effects, i.e., when the bonding of a particle restricts (blocks) its ability to bond with other particles. The theory represents an extension of the earlier proposed first order resummed thermodynamic perturbation theory for central force associating potential and takes into account formation of the rings of the particles. In the limiting case of singly bondable patches (total blockage), the theory reduces to Wertheim thermodynamic perturbation theory for associating fluids. Closed-form expressions for the Helmholtz free energy, pressure, internal energy, and chemical potential of the model with an arbitrary number of equivalent doubly bondable patches are derived. Predictions of the theory for the model with two patches appears to be in a very good agreement with predictions of new NVT and NPT Monte Carlo simulations, including the region of strong association.

Polymorphic electrophile response elements in the mouse glutathione S-transferase GSTa1 gene that confer increased induction.

Induced transcription of a battery of stress response genes in mammals, including several phase I and phase II drug-metabolizing enzymes, is regulated by the electrophile responsive element (EpRE). Because previous directed mutagenesis of nucleotide motifs within the large, composite EpRE were shown to affect transcription factor binding and associated induced expression of dependent genes, we hypothesized that naturally-occurring variation or polymorphism in the EpRE sequence, if found, could affect the induced expression of important protective genes like glutathione S-transferases, and that this could be an important determinant of cancer risk in humans and other mammals. To determine whether this occurred in nature, 32 strains and species of inbred mice were screened to examine the EpRE sequence present in the mGSTa1 promoter. Two species, Mus caroli and Mus spretus, showed TGAC-->TGGC mutations in the tandem TGAC motif. Inducibility (15-fold) of the variant Mus spretus EpRE sequence in a reporter gene construct in HepG2 cells was significantly increased versus the wild-type EpRE sequence (8-fold). A comparison of mGSTa1-induced expression in the livers of Mus spretus, Mus caroli, and BALB/cJ mice showed the highest level of mGSTa1 mRNA in livers from the Mus spretus and Mus carolimice. This naturally-occurring polymorphism within the EpRE domain is the first mutation with an associated phenotype to be reported within a promoter regulatory element of a drug metabolizing gene.

Oxaliplatin-induced damage of cellular DNA.

Damage to cellular DNA is believed to determine the antiproliferative properties of platinum (Pt) drugs. This study characterized DNA damage by oxaliplatin, a diaminocyclohexane Pt drug with clinical antitumor activity. Compared with cisplatin, oxaliplatin formed significantly fewer Pt-DNA adducts (e.g., 0.86+/-0.04 versus 1.36+/- 0.01 adducts/10(6) base pairs/10 microM drug/1 h, respectively, in CEM cells, P<.01). Oxaliplatin was found to induce potentially lethal bifunctional lesions, such as interstrand DNA cross-links (ISC) and DNA-protein cross-links (DPC) in CEM cells. As with total adducts, however, oxaliplatin produced fewer (P<.05) bifunctional lesions than did cisplatin: 0.7+/-0.2 and 1.8+/-0.3 ISC and 0.8+/-0.1 and 1.5+/-0.3 DPC/10(6) base pairs/10 microM drug, respectively, after a 4-h treatment. Extended postincubation (up to 12 h) did not compensate the lower DPC and ISC levels by oxaliplatin. ISC and DPC determinations in isolated CEM nuclei unequivocally verified that oxaliplatin is inherently less able than cisplatin to form these lesions. Reactivation of drug-treated plasmids, observed in four cell lines, suggests that oxaliplatin adducts are repaired with similar kinetics as cisplatin adducts. Oxaliplatin, however, was more efficient than cisplatin per equal number of DNA adducts in inhibiting DNA chain elongation ( approximately 7-fold in CEM cells). Despite lower DNA reactivity, oxaliplatin exhibited similar or greater cytotoxicity in several other human tumor cell lines (50% growth inhibition in CEM cells at 1.1/1.2 microM, respectively). The results demonstrate that oxaliplatin-induced DNA lesions, including ISC and DPC, are likely to contribute to the drug's biological properties. However, oxaliplatin requires fewer DNA lesions than does cisplatin to achieve cell growth inhibition.

Neurotensin receptor-mediated inhibition of pancreatic cancer cell growth by the neurotensin antagonist SR 48692.

Second messenger calcium responses to the neuropeptide neurotensin and its non-peptide antagonist SR 48692 were studied in relation to the proliferation of pancreatic cancer cells. Neurotensin caused a transient increase in intracellular calcium in two pancreatic lines, MIA PaCa-2 and PANC-1, with EC50 values of 4.6 and 11.4 nM and peak calcium concentrations of 190% and 470% of basal levels, respectively. SR 48692 inhibited these calcium changes with an IC50 (at 25 nM neurotensin) of 4.9 and 4.1 nM in MIA PaCa-2 and PANC-1 cells, respectively. In MIA PaCa-2 cells, SR 48692 may act as an inverse agonist as it depressed basal calcium. SR 48692 inhibited growth of both MIA PaCa-2 and PANC-1 cells. Only in MIA PaCa-2 cells did neurotensin overcome this inhibition or stimulate proliferation. The results imply that, in MIA PaCa-2 cells, the neurotensin antagonist SR 48692 inhibits growth in a neurotensin receptor-mediated fashion.

Induction of AT-specific DNA-interstrand crosslinks by bizelesin in genomic and simian virus 40 DNA.

Bizelesin is a bifunctional AT-specific DNA alkylating drug. Our study characterized the ability of bizelesin to induce interstrand crosslinks, a potential lethal lesion. In genomic DNA of BSC-1 cells, bizelesin formed from approx. 0.3 to 6.03+/-0.85 interstrand crosslinks per 106 base pairs, at 5-100 nM drug concentration, respectively, comparable to the number of total adducts previously determined in the same system (J.M. Woynarowski, M.M. McHugh, L.S. Gawron, T.A. Beerman, Biochemistry 34 (1995) 13042-13050). Bizelesin did not induce DNA-protein crosslinks or strand breaks. A model defined target, intracellular simian virus 40 (SV40) DNA, was employed to map at the nucleotide level sites of bizelesin adducts, including potential interstrand crosslinks. Preferential adduct formation was observed at AT tracts which are abundant in the SV40 matrix associated region and the origin of replication. Many sites, including each occurrence of 5'-T(A/T)4A-3', co-mapped on both DNA strands suggesting interstrand crosslinks, although monoadducts were also formed. Bizelesin adducts in naked SV40 DNA were found at similar sites. The localization of bizelesin-induced crosslinks in AT-rich tracts of replication-related regions is consistent with the potent anti-replicative properties of bizelesin. Given the apparent lack of other types of lesions in genomic DNA, interstrand crosslinks localized in AT-rich tracts, and to some extent perhaps also monoadducts, are likely to be lethal effects of bizelesin.

Sequence- and region-specificity of oxaliplatin adducts in naked and cellular DNA.

Oxaliplatin is a clinical anticancer drug with a pharmacological profile distinct from that of cisplatin. Our studies compared site- and region-specificity of lesions induced by oxaliplatin and cisplatin in naked and intracellular DNA, respectively. Oxaliplatin adducts in naked Simian virus 40 (SV40 DNA) were mapped by repetitive primer extension. The sites of oxaliplatin adducts were nearly identical to the sites of cisplatin adducts and were focused in G clusters and GNG motifs probably reflecting intrastrand cross-links. Although alkaline agarose electrophoresis of specific SV40 fragments showed that oxaliplatin formed interstrand cross-links, the levels of this lesion type were low. Drug-induced lesions in discrete loci of cellular DNA were assessed by the polymerase chain reaction stop assay in human tumor A2780 cells. Oxaliplatin at 200 microM induced approximately 1300, approximately 1500, approximately 800, and approximately 300 lesions/10(6) bp in the human beta-globin, c-myc, and HPRT genes and in mitochondrial DNA, respectively. Cisplatin formed two to six times more lesions in the same regions. For both drugs, lesion frequencies seem to parallel the density of drug-binding motifs in the nuclear regions, whereas mitochondrial DNA was disproportionately less affected. Despite less potent induction of DNA lesions, oxaliplatin was more cytotoxic than cisplatin against A2780 cells. Because our findings clearly demonstrate that oxaliplatin forms covalent adducts with a similar sequence- and region-specificity to that of cisplatin, other properties of oxaliplatin adducts, factors other than DNA binding, or both determine the unique features of the mechanism of action of oxaliplatin.

Regression analysis of changes in blood pressure with oral contraceptive use.

Measurements of blood pressure were obtained on 2,673 women from East Boston, Massachusetts, an urban, working class neighborhood, in surveys conducted in both 1973 and 1976-1977. Of these, 927 women participated in a third survey in 1978. The women were 16 to 49 years of age in 1973, premenopausal throughout the study, and not on blood pressure medication. Regression analyses were performed of blood pressure change between the first and second as well as between the second and third surveys on initial blood pressure, age, weight, and patterns of oral contraceptive use. For systolic pressure the effect of starting oral contraceptive use was an increase of 4.1 mmHg (p less than 0.0001), while the effect of discontinuing use relative to continued use was a 4.4 mmHg decrease (p less than 0.0001). These changes were not affected by duration of use or time since last use among past users. For diastolic pressure the average effect of starting use between surveys was an insignificant 1.0 mmHg increase, but diastolic pressure level seemed to increase with duration of use (0.5 mmHg/year, p = 0.0009). The effect of discontinuing use relative to continued use was a drop of 2.7 mmHg in diastolic pressure (p = 0.0004), which was uninfluenced by time since last use.

Stimulation by heterotypic antigens of foot-and-mouth disease virus antibodies in vaccinated cattle.

Immunisation of cattle with foot-and-mouth disease virus failed to raise a level of antibody that provides protection against heterotypic challenge. Further the 12S substructure, produced from the 146S particle, was ineffective in providing protection against challenge by homotypic virus. These findings suggest considerable antigenic differences in the virus serotypes and between the virus and its substructure. Inoculation of homologous 12S and heterologous 1246S and 12S antigens into vaccinated cattle, however, revealed antigenic relationship between different serotypes, and between serotypes and their substructures.

Serological and immunological relations between the 146S and 12S particles of foot-and-mouth disease virus.

Intact 146S particles of the seven serotypes of foot-and-mouth disease virus (FMDV) produce type-specific precipitating, complement-fixing and neutralizing antibodies in cattle and guinea-pigs. However, the 12S structural subunit, produced from the virus particle by mild acid treatment (pH 6) or by heating at 56 degrees C, although stimulating the production of precipitating and complement-fixing antibodies, produces only low levels of neutralizing antibody. Nevertheless, 12S antibody in guinea-pigs primed with a vaccine prepared from 146S particles. Moreover, heterotypic 12S and 146S particles also boosted the neutralizing antibody response to the first virus. These results point to an antigenic similarity between the 146S particles of each type and to a close antigenic relationship between the 146S and 12S particles.

A blastogenic test for foot-and-mouth disease.

A blastogenic test to detect peripheral blood leukocytes specifically sensitized to foot-and-mouth disease virus antigen is described. The test is carried out in microtitre plates and optimum conditions were found by titration. These employed 7.5 x 10(5) cells/well and 20 complement fixing units of antigen. Peak [3H]thymidine incorporation was found to take place at 2-3 days.

Bacteriuria in a population-based cohort of women.

A survey for bacteriuria was conducted in a community-wide, unselected population of women 16--69 years old. The overall prevalence of bacteriuria was 3.5%. The prevalence of bacteriuria increased with age with a linear trend, but with a significant nonlinear component as well. Bacteriuria was associated with parity after correction for the effects of age. Current symptoms of dysuria and a history of previous urinary tract infection were slightly but significantly more common in women with bacteriuria. The population described should serve as an adequate base for continuing studies of the possible consequences of bacteriuria.

Oral-contraceptive use and bacteriuria in a community-based study.

PIP: The relationship between oral-contraceptive (OC) use and bacteriuria in a population-based cohort of 2390 women below age 50 was investigated. 5.6% of the 482 OC users and 4% of the 1908 nonusers were bacteriuric. The summary age-adjusted relative risk in users was 1.8 as compared with nonusers. After adjustment for possible confounding effects of age, parity, and weight, the users had a relative risk of 1.5.^ieng

Swine vesicular disease: attempts to transmit infection to cattle and sheep.

Cattle and sheep were housed with infected pigs for 11 days. Small amounts of virus were recovered intermittently from the pharynx, milk and rectal swabs of the cattle, but no evidence of subclinical infection was found. Some indication of virus growth in the sheep was obtained in that large amounts of virus were recovered from the pharyngeal region 4 to 7 days after exposure and six of the eight sheep developed significant titres of neutralizing antibody which were maintained in four animals for at least 6 weeks.

Current concepts in the management of Parkinson's disease

L-Dopa has now undoubtly proved itself the drug of choice in the vast majority of cases of Parkinson's Disease. When used in combination with a decarboxylase inhibitor it would appear that significant therapeutic advantages are gained (Lancet 5th May, 1973, Marsden et al 1973). The metabolism, use and side effects of L-Dopa are outlined and the role of other forms of treatment discussed (AU)

Inactivated rabies vaccine produced from the Flury LEP strain of virus grown in BHK-21 suspension cells.

Suspension cultures of BHK-21 cells maintained at 32 to 33 C were infected with the Flury LEP strain of rabies virus. By using a cell concentration of 2.0 x 10(6) to 2.5 x 10(6) cells per ml infected at a multiplicity of 0.05, high titers of extracellular virus were reached in 96 to 120 h, and potent inactivated vaccines were prepared from culture fluids harvested between 96 to 168 h. The addition of 1% bovine serum to the maintenance medium resulted in an increase in virus yields and vaccine potency. Estimation of the number of infected cells by immunofluorescent procedures proved a rapid and reliable guide to the virus content of suspension cultures.