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A Bax

Publications and source records attributed to A Bax.

At least 19 recordsLinked to original sources
PubMed2004

Incidence of cervical carcinoma in a high-risk, non-screened area results of a retrospective analysis on the Dutch Caribbean Antilles from 1983 to 1998.

Cancer of the uterine cervix is one of the most common malignancies worldwide. The average annual incidence of cervical cancer varies widely per geographical area. Some of the highest rates have been reported in Latin American and Caribbean countries. Previous research into the frequency of cervical carcinoma in Curaçao, the main island of the Dutch Caribbean, has reported predominantly far-advanced stages of the disease. The objectives of this study were to determine: whether there are indications of changes in the incidence of cervical carcinoma, whether these changes are a result of increased clinical attention paid by general practioners (GPs) and gynaecologists, and whether less advanced stages of cervical cancer were found, indicating earlier diagnosis. Data on patients with cervical carcinoma, obtained from records kept in the Department of Pathological Anatomy at the St Elisabeth Hospital in Curaçao were retrospectively analyzed for the period 1983 to 1998. To study the incidence rate over time, cumulative and incidence rates, standardized to the World Standard Population, were calculated and compared to the rates in other Latin American and Caribbean countries. The age-standardized incidence rate for Curaçao was 14.3 per 100 000 women. The cumulative rate up to 74-years-of-age was 1.6%. Sub-division into the three groups showed a trend towards a decrease in the incidence and cumulative rates. A comparison of clinical staging International Federation of Obstetrics and Gynaecology (FIGO) showed a trend towards an increase in time of almost 48% to 59% in stage IA (ie less severe cases). A significant increase (p < 0.0001) was found in the numbers of smears performed in the complete period of 1983-1998, compared to a previously examined period of 1972-1982. The incidence of cervical cancer in Curaçao appears to be decreasing, and is lower than in other areas in the region. The number of smears GPs performed over time increased However the incidence is still relatively high compared to western countries and cases are still presenting in relatively advanced stages of the disease. Although the introduction of a screening programme might not influence the actual incidence of cervical cancer dramatically, nonetheless it may contribute to earlier diagnosis of cervical carcinoma.

Carcinoma, Squamous Cell↗
PubMed2001

Morphology of three lyotropic liquid crystalline biological NMR media studied by translational diffusion anisotropy.

The morphologies of three dilute liquid crystalline phases, which are widely used for biological NMR spectroscopy, are investigated by the study of tracer self-diffusion. The aqueous liquid crystalline media investigated include the common phospholipid bicelle medium, a phase consisting of a mixture of pentaethyleneglycol mono dodecyl ether and hexanol, and a medium containing cetylpyridinium bromide and hexanol. Threonine and water were used as tracer molecules for probing the aqueous environment, and tetramethylsilane (TMS) was for probing the lipophilic environment. Pulsed field gradient NMR was used to measure tracer self-diffusion rates in three orthogonal directions. Although results for the water-soluble tracers in bicelle media do not contradict the widely accepted disk-shaped bicelle model, the high TMS diffusion rate observed in the bilayer plane requires extensive transient edge-to-edge contacts of such disks. This morphology is essentially that of a heavily perforated lamellar bilayer phase and explains why this medium remains liquid crystalline well below the Onsager limit for disk-shaped nematogens. Below 25 degrees C, a bicelle mixture consisting of dimyristoyl phosphatidyl choline and dihexanoyl phosphatidyl choline remains isotropic, but tracer diffusion obstruction indicates that the particles are significantly oblate. The diffusion anisotropy in the penta(ethyleneglycol) mono dodecyl ether liquid crystals confirms the previously proposed alpha-lamellar phase. However, weak inhibition of aqueous-phase self-diffusion in the z direction points to the presence of bridge- or caplike obstructions, and the bilayers appear slightly permeable to water. If the previously proposed concentric cylinder superstructure of bilayers applies, the diffusion data indicate that the most outer cylinder must have a diameter greater than 50 microm. The tracer self-diffusion data for the cetylpyridinium bromide/hexanol medium is only compatible with a planar alpha-lamellar phase, with its local director orthogonal to the magnetic field, and a very large domain size over which the director remains parallel.

Anisotropy↗
PubMed2001

A model for the abrogation of the SOS response by an SOS protein: a negatively charged helix in DinI mimics DNA in its interaction with RecA.

DinI is a recently described negative regulator of the SOS response in Escherichia coli. Here we show that it physically interacts with RecA and prevents the binding of single-stranded DNA to RecA, which is required for the activation of the latter. DinI also displaces ssDNA from a stable RecA-DNA cofilament, thus eliminating the SOS signal. In addition, DinI inhibits RecA-mediated homologous DNA pairing, but has no effect on actively proceeding strand exchange. Biochemical data, together with the molecular structure, define the C-terminal alpha-helix in DinI as the active site of the protein. In an unusual example of molecular mimicry, a negatively charged surface on this alpha-helix, by imitating single-stranded DNA, interacts with the loop L2 homologous pairing region of RecA and interferes with the activation of RecA.

Amino Acid Sequence↗
PubMed2001

Measurement of homonuclear proton couplings from regular 2D COSY spectra.

An interactive computer procedure is described which determines (1)H--(1)H couplings from fitting the cross-peak multiplets in regular phase-sensitive COSY spectra. The robustness and simplicity of the method rely on the fact that a given cross-peak intensity is not an independent variable in the fitting procedure, making it possible to measure couplings accurately even from individual cross peaks with unresolved multiplet structure.

Computer Graphics↗
PubMed2001

Measurement of homonuclear proton couplings based on cross-peak nulling in CT-COSY.

A method in which 1H-1H scalar and dipolar couplings are obtained from the cross-peak nulling condition in a series of constant-time (CT) COSY spectra, as a function of the duration of the CT period, is described. The method is best suited for measurement of 1H-1H couplings in the range 5-20 Hz. It is shown, however, that results can be sensitive to cross-correlated relaxation effects. Also, artifactual resonances, resulting from strong coupling, can be quite pronounced in CT-COSY spectra, even when /J(AB)/(deltaA-deltaB)/<0.1. The experiments are demonstrated for the DNA dodecamer d(CGCGAATTCGCG)2, both in isotropic solution and in a liquid crystalline phase.

Computer Graphics↗
PubMed2001

H...N hydrogen bond lengths in double stranded DNA from internucleotide dipolar couplings.

The ratio of the internucleotide dipolar coupling and the corresponding one-bond imino 15N-1H dipolar coupling provides a measure for the N...H/H-N distance ratio. Measurements were carried out for a dodecamer, d(CGCGAATTCGCG)2, in which a C-G and an A-T basepair were uniformly enriched in 15N. When assuming H-bonds to be perfectly linear, dipolar data indicate time-averaged hydrogen bond lengths of 1.80 +/- 0.03 A for A-T and 1.86 +/- 0.02 A for C-G. When using H-bond orientations from high resolution X-ray data, H-bond lengths are about 0.1 A shorter.

Crystallography, X-Ray↗
PubMed2001

Characterization of molecular alignment in aqueous suspensions of Pf1 bacteriophage.

The phase diagram of Pf1 solutions has been studied indirectly by observation of 2H quadrupole splittings of the solvent signal and measurement of dipolar couplings in solute macromolecules. At low volume fractions of Pf1 and at high ionic strength, alignment of both the phage and the solute depends strongly on the strength of the magnetic field. Both the theoretical and experimentally determined phase diagram of Pf1 show that at low concentrations and high ionic strengths the solution becomes isotropic. However, just below the nematic phase boundary the behavior of the system is paranematic, with cooperative alignment which depends on the strength of the applied magnetic field. Above 16 mg/ml Pf1 is fully nematic up to 600 mM NaCl. Alignment of proteins with a significant electric dipole moment, which tends to be strong in Pf1, can be reduced by either high ionic strength or low phage concentration. Because ionic strength modulates both the orientation and magnitude of the alignment tensor in Pf1 medium, measurement at two ionic strengths can yield linearly independent alignment tensors.

Antigens, Bacterial↗
PubMed2001

Multiplet component separation for measurement of methyl 13C-1H dipolar couplings in weakly aligned proteins.

A simple spectral editing procedure is described that generates separate subspectra for the methyl 13C-[1H3] multiplet components of 1H-13C HSQC spectra. The editing procedure relies on co-addition of in-phase and antiphase spectra and yields 1H-coupled constant-time HSQC subspectra for the methyl region that have the simplicity of the regular decoupled CT-HSQC spectrum. Resulting spectra permit rapid and reliable measurement of 1H-13C J and dipolar couplings. The editing procedure is illustrated for a Ca2+-calmodulin sample in isotropic and liquid crystalline phases.

Algorithms↗
PubMed2001

Measurement of 1H3'-31P dipolar couplings in a DNA oligonucleotide by constant-time NOESY difference spectroscopy.

The ratios of cross peak intensities in a selective constant-time NOESY experiment, recorded with and without 31P decoupling, yield values for the sum of the H3'-P scalar and dipolar couplings. The selective refocusing of H3' resonances in this experiment results in excellent resolution and sensitivity, even in the liquid crystalline phase where the 1H spectrum is broadened by unresolved homonuclear dipolar couplings. The vicinal H3'-P scalar and dipolar couplings in the DNA oligomer d(CGCGAATTCGCG)2 were measured in both isotropic solution, and in a liquid crystalline phase. Isotropic values are in good agreement with values reported previously. Dipolar couplings are in excellent agreement with the NMR structure for this dodecamer, and to a somewhat lesser extent with the X-ray structures.

Crystallography, X-Ray↗
PubMed2001

A simple apparatus for generating stretched polyacrylamide gels, yielding uniform alignment of proteins and detergent micelles.

Compressed and stretched polyacrylamide hydrogels previously have been shown to offer a robust method for aligning proteins. A simple, funnel-like apparatus is described for generating uniformly stretched hydrogels. For prolate-shaped proteins, gels stretched in the direction of the magnetic field yield two-fold larger alignment than gels compressed to the same aspect ratio in this direction. Empirically, protein alignment is found to be proportional to (c-2.3)2 [(d(o/dN)3-1], where do and dN are the diameters of the cylindrical gels before and after stretching, respectively, and c is the polyacrylamide weight fraction in percent. Low gel densities, in the 4-7% range, are found to have minimal effects on macromolecular rotational correlation times, tauc, and no effect of the compression ratio on tauc could be discerned over the range studied (do/dN < or = 1.4). Application is demonstrated for a sample containing the first Ig-binding domain of protein G, and for a detergent-solubilized peptide.

Acrylic Resins↗
PubMed2001

Solution structure of Ca(2+)-calmodulin reveals flexible hand-like properties of its domains.

The solution structure of Ca(2+)-ligated calmodulin is determined from residual dipolar couplings measured in a liquid crystalline medium and from a large number of heteronuclear J couplings for defining side chains. Although the C-terminal domain solution structure is similar to the X-ray crystal structure, the EF hands of the N-terminal domain are considerably less open. The substantial differences in interhelical angles correspond to negligible changes in short interproton distances and, therefore, cannot be identified by comparison of NOEs and X-ray data. NOE analysis, however, excludes a two-state equilibrium in which the closed apo conformation is partially populated in the Ca(2+)-ligated state. The difference between the crystal and solution structures of Ca(2+)-calmodulin indicates considerable backbone plasticity within the domains of calmodulin, which is key to their ability to bind a wide range of targets. In contrast, the vast majority of side chains making up the target binding surface are locked into the same chi(1) rotameric states as in complexes with target peptide.

Animals↗
PubMed2000

Evaluation of cross-correlation effects and measurement of one-bond couplings in proteins with short transverse relaxation times.

Various strategies are described and compared for measurement of one-bond J(NH) and J(NC') splittings in larger proteins. In order to evaluate the inherent resolution obtainable in the various experiments, relaxation rates of (15)N-(1)H(N) coupled and heteronuclear decoupled resonances were measured at 600- and 800-MHz field strengths for both perdeuterated and protonated proteins. A comparison of decay rates for the two (15)N-¿H(N)¿ doublet components shows average ratios of 4.8 and 3.5 at 800- and 600-MHz (1)H frequency, respectively, in the perdeuterated proteins. For the protonated proteins these ratios are 3.2 (800 MHz) and 2.4 (600 MHz). Relative to the regular HSQC experiment, the enhancement in TROSY (15)N resolution is 2.6 (perdeuterated; 800 MHz), 2.0 (perdeuterated; 600 MHz), 2.1 (protonated; 800 MHz), and 1.7 (protonated; 600 MHz). For the (1)H dimension, the upfield (1)H(N)-¿(15)N¿ component on average relaxes slower than the downfield (1)H(N)-¿(15)N¿ component by a factor of 1.8 (perdeuterated; 800 MHz) and 1.6 (perdeuterated; 600 MHz). The poor resolution for the upfield (15)N-¿(1)H¿ doublet component in slowly tumbling proteins makes it advantageous to derive the J(NH) splitting from the difference in frequency between the narrow downfield (15)N doublet component and either the (1)H-decoupled (15)N resonance or the peak position in an experiment which J-scales the frequency of the upfield doublet component but maintains some of the advantages of the TROSY experiment.

Magnetic Resonance Spectroscopy↗
PubMed2000

Measurement of one-bond 15N-13C' dipolar couplings in medium sized proteins.

A simple and accurate method is described for measurement of 1J(C'N) splittings in isotopically enriched proteins. The method is of the quantitative J correlation type, and the 1J(C'N) splitting is derived from the relative intensity in two 3D TROSY-HNCO spectra with 1J(C'N) dephasing intervals of approximately 1/(2 1J(C'N)) (reference intensity) and approximately 1/1J(C'N) (residual intensity). If the two spectra are recorded under identical conditions and with the same number of scans, the random error in the 1J(C'N) value extracted in this manner is inversely related to the signal-to-noise (S/N) in the reference spectrum. A S/N of 30:1 in the reference spectrum yields random errors of less than 0.2 Hz in the extracted 1J(C'N) value. Dipolar couplings obtained from the difference in 1J(C'N) splitting in the isotropic and liquid crystalline phase for the C-terminal domain of calmodulin are in excellent agreement with its 1.68-A crystal structure, but agree considerably less with the 2.2-A structure.

Animals↗
PubMed2000

Measurement of dipolar couplings in a transducin peptide fragment weakly bound to oriented photo-activated rhodopsin.

Rhodopsin-containing disks, isolated from rod outer segments of bovine retina, align at high magnetic fields with their membrane normal parallel to the magnetic field. After light-activation of rhodopsin, transient binding of the C-terminal transducin undecapeptide, selectively labeled with 15N at Leu5 and Gly9, results in residual dipolar contributions to the 1J(NH) splittings for these two residues. Both residues show 1J(NH) splittings which are smaller than in the dark-adapted or rhodopsin-free sample, and return to their isotropic values at a rate determined by the decay of the meta II state of rhodopsin. The dipolar couplings indicate that in the bound state, N-H vectors of Leu5 and Gly9 make angles of 48+/-4 degrees and 40+/-8 degrees, respectively, with the disk normal. These 'transferred' dipolar couplings potentially offer a useful method for studying the conformation and orientation of flexible, low affinity ligands when bound to oriented integral membrane receptors.

Amino Acid Sequence↗