PubMed Health⌕ Search

Biomedical subjects

W Resch

Publications and source records attributed to W Resch.

11 recordsLinked to original sources

Improved success of phenotype prediction of the human immunodeficiency virus type 1 from envelope variable loop 3 sequence using neural networks.

We have assembled two sets of HIV-1 V3 sequences with defined epidemiologic relationships associated with experimentally determined coreceptor usage or MT-2 cell tropism. These data sets were used for three purposes. First, they were employed to test existing methods for predicting coreceptor usage and MT-2 cell tropism. Of these methods, the presence of one basic amino acid at position 11 or 25 proved to be most reliable for both phenotypic classifications, although its predictive power for the X4 phenotype was less than 50%. Second, we used the sequence sets to train neural networks to infer coreceptor usage from V3 genotype with better success than the best available motif-based method, and with a predictive power equal to that of the best motif-based method for MT-2 cell tropism. Third, we used the sequence sets to reexamine patterns of variability associated with the different phenotypes, and we showed that the phenotype-associated sequence patterns could be reproduced from large sets of V3 sequences using phenotypes predicted by the trained neural network.

Algorithms↗

A multiple-site-specific heteroduplex tracking assay as a tool for the study of viral population dynamics.

Rapidly evolving entities, such as viruses, can undergo complex genetic changes in the face of strong selective pressure. We have developed a modified heteroduplex tracking assay (HTA) capable of detecting the presence of single, specific mutations or sets of linked mutations. The initial application of this approach, termed multiple-site-specific (MSS) HTA, was directed toward the detection of mutations in the HIV-1 pro gene at positions 46, 48, 54, 82, 84, and 90, which are associated with resistance to multiple protease inhibitors. We demonstrate that MSS HTA is sensitive and largely specific to all targeted mutations. The assay allows the accurate and reproducible quantitation of viral subpopulations comprising 3% or more of the total population. Furthermore, we used MSS HTA in longitudinal studies of pro gene evolution in vitro and in vivo. In the examples shown here, populations turned over rapidly and more than one population was present frequently. To demonstrate the versatility of MSS HTA, we also constructed a probe sensitive to changes at positions 181 and 184 of the RT coding domain. Changes at these positions are involved in resistance to nevirapine and 2',3'-dideoxy-3'-thiacytidine (3TC), respectively. This assay easily detected the evolution of resistance to 3TC. MSS HTA provides a rapid and sensitive approach for detecting the presence of and quantifying complex mixtures of distinct genotypes, including genetically linked mutations, and, as one example, represents a useful tool for following the evolution of drug resistance during failure of HIV-1 antiviral therapy.

Antiviral Agents↗

Using HIV-1 sequence variability to explore virus biology.

Human immunodeficiency virus type 1 (HIV-1) only recently established an epidemic world-wide infection in the human population. The virus persists in the human host through active replication and is able to avoid clearance by the immune system. Active replication is an important component of the rapid evolutionary potential of HIV-1, a potential which manifests itself in the evolution of immune escape variants, drug resistant variants, and variants with the ability to use different cell surface coreceptors in conjunction with CD4. Multiple zoonotic introductions, compartmentalization of virus replication in the body, and genetic bottlenecks associated with sampling during transmission, antiretroviral therapy, and geographic and/or host population isolation further contribute to the range of sequences present in extant viruses. The sum of the history of all of these phenomena is reflected in HIV-1 sequence variability, and most of these phenomena are ongoing today. Here we review the use of HIV-1 sequence variability to explore its underlying biology.

Animals↗

Imperatoxin A induces subconductance states in Ca2+ release channels (ryanodine receptors) of cardiac and skeletal muscle.

Single-channel and [3H]ryanodine binding experiments were carried out to examine the effects of imperatoxin activator (IpTxa), a 33 amino acid peptide isolated from the venom of the African scorpion Pandinus imperator, on rabbit skeletal and canine cardiac muscle Ca2+ release channels (CRCs). Single channel currents from purified CRCs incorporated into planar lipid bilayers were recorded in 250 mM KCl media. Addition of IpTxa in nanomolar concentration to the cytosolic (cis) side, but not to the lumenal (trans) side, induced substates in both ryanodine receptor isoforms. The substates displayed a slightly rectifying current-voltage relationship. The chord conductance at -40 mV was approximately 43% of the full conductance, whereas it was approximately 28% at a holding potential of +40 mV. The substate formation by IpTxa was voltage and concentration dependent. Analysis of voltage and concentration dependence and kinetics of substate formation suggested that IpTxa reversibly binds to the CRC at a single site in the voltage drop across the channel. The rate constant for IpTxa binding to the skeletal muscle CRC increased e-fold per +53 mV and the rate constant of dissociation decreased e-fold per +25 mV applied holding potential. The effective valence of the reaction leading to the substate was approximately 1.5. The IpTxa binding site was calculated to be located at approximately 23% of the voltage drop from the cytosolic side. IpTxa induced substates in the ryanodine-modified skeletal CRC and increased or reduced [3H]ryanodine binding to sarcoplasmic reticulum vesicles depending on the level of channel activation. These results suggest that IpTxa induces subconductance states in skeletal and cardiac muscle Ca2+ release channels by binding to a single, cytosolically accessible site different from the ryanodine binding site.

Animals↗

[Stress to the lateral ankle joint ligament complex in confined imaging with an a.-p. radiation path].

Physical definition and laws of rotating systems will be stated. The application for supination in ankle joint shows, that lever action increases the force applied up to four times in case of stress X-rays in the a.-p. view. As quoted in literature, the test power of up to 25 kp means a loading of the lateral collaterals up to 100 kp. An additional iatrogenic rupture of already traumatically partially sprained structures seems possible. The critical judgement of five different methods producing stress X-rays of the ankle joint shows the weak points of this test system. Beck's method seems sufficient for precise statements and judgement of the stress X-ray tests without endangering ligaments so far unsprained.

Ankle Injuries↗

[Carbon monoxide in mother and newborn infant from smoking during pregnancy].

COHb-values of 24 newborn infants investigated (cord blood) were higher altogether than the corresponding maternal concentrations (venous) at birth. COHb in percent of saturable Hb ranged from 0.3 to 0.7 in 10 infants of nonsmokers and from 0.8-11.9 in 14 infants of smokers. Correlations between maternal and infant COHb were statistically significant. At birth the mean infant/maternal COHb ratio was 2.5. The significant correlations between maternal COHb during pregnancy (32.-38. week) and infant COHb at birth (cord blood) is an indication for the predictive value of such tests.

Carbon Monoxide↗

Nucleic acid and protein content of rat cecum mucosa in dietary adaptation-growth by cellular hyperplasia.

The deoxyribonucleic acid (DNA), ribonucleic acid (RNA) and protein content of the isolated cecum mucosa was determined in control rats and rats with adaptive cecum growth induced by dietary polyethylene glycol. Feeding the polymer resulted in an increase in the total amount of mucosal nucleic acids and protein which was statistically significant after 2-4 days and reached twice the control value after 2 months. During growth, the protein/DNA, protein/RNA and RNA/DNA ratios remained unaltered, indicating hyperplasia and not hypertrophy of the epithelial cells. The water content of the mucosa increased from 75 to 81% during adaptation, possibly related to the stimulation in active sodium transport reported earlier. Recalculation of parallel Na-K-ATPase data on the basis of DNA demonstrated a doubling of Na-K-ATPase activity per mg DNA in the fully adapted cell.

Adaptation, Physiological↗

[A rapid carboxyhemoglobin determination by means of non-dispersive ultra-red gas analysis (author's transl)].

A rapid and reliable method of carboxyhemoglobin determination is described. Hemoglobin-bound carbon monoxide is released chemically into a continuous nitrogen stream and transported to an infrared gas analyser. The total amount of released CO is determined electronically. 100% CO Hb values are obtained in the same way from diluted blood samples after flushing with pure CO in special saturation vessels. The method described yields results with a standard deviation of +/- 2.5%.

Carbon Monoxide↗