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Biomedical subjects

J R Smith

Publications and source records attributed to J R Smith.

At least 19 recordsLinked to original sources
PubMed1992

Contribution of the p51 subunit of HIV-1 reverse transcriptase to enzyme processivity.

Human immunodeficiency virus Type I reverse transcriptase is active as either the homodimer (p66/p66) or the heterodimer (p66/p51). Purified recombinant p66 and p51 expressed in yeast were reconstituted in the presence of 60 mM sodium pyrophosphate to enhance dimer formation. Comparison of the processivity of these two active reconstituted forms shows that the heterodimer is more processive than the homodimer with a cycle almost twice as long as judged by assays utilizing poly (U,G) as a challenger to primer-template. Binding assays demonstrated that the heterodimer has a higher affinity for primer-template than the homodimer and that the p51 subunit has an affinity equal to that of the heterodimer. These results suggest that the p51 subunit functions to increase processivity in the heterodimer.

Cloning, Molecular↗
PubMed1992

Common senescent cell-specific antibody epitopes on fibronectin in species and cells of varied origin.

The phenomenon of in vitro cellular senescence has been demonstrated in cultured cells derived from humans and various other species. We have previously shown that monoclonal antibodies SEN-1, SEN-2, and SEN-3 react to epitopes on fibronectin that are exposed when human diploid fibroblasts become senescent. We here present results demonstrating that exposure of these epitopes is specific to senescence for a variety of human cells: epidermal keratinocytes, mammary epithelial cells, as well as fibroblasts. Fibronectin from 11 additional species was also analyzed by Western immunoblot for ability to bind the SEN antibodies. SEN-1 bound only human and gorilla fibronectin, whereas SEN-2 and SEN-3 bound fibronectin from those two species as well as the horse, cow, sheep, goat, dog, and chick. None of the antibodies reacted with fibronectin from the rabbit, rat, or mouse. These data indicated a correlation between the ability of the SEN antibodies to bind fibronectin from a particular species and the ability of cells from that species to exhibit a stable senescent phenotype in vitro. Therefore, exposure of this region of fibronectin may be important in the establishment and maintenance of cellular senescence. In addition, the ability of the SEN antibodies to react with fibronectin from a variety of senescent cells emphasizes their usefulness as markers for cellular senescence.

Animals↗
PubMed1992

A model for focal magnetic brain stimulation.

In this study, we describe a magnetic coil designed for focal brain stimulation. We determined the distribution of magnetically induced currents using a multi-concentric spherical cranial computer model. The induced currents were primarily linearly oriented and concentrated below the coil's center. In comparison with large coils, small coils were less efficient but produced a more concentrated current distribution. Variations in conductivity among brain, scalp and skull produced secondary currents that reduced the magnetically induced current. This reduction in magnetically induced current was greater for larger coils.

Algorithms↗
PubMed1992

Epinephrine absorption from commercial gingival retraction cords in clinical patients.

Absorption of epinephrine from commercial gingival retraction cords was measured in vivo under clinical conditions. The amount of epinephrine absorbed was estimated by measurement with fluorospectrophotometry of the amount of dl-epinephrine remaining in the cords after they were removed and subtracting this value from the dl-epinephrine concentration in matching control cords. All cords lost substantial amounts of epinephrine while in the sulcus, with individual losses ranging from 64% to 94%. An average 71 micrograms of l-epinephrine was absorbed per inch of two-ply cord, and extreme variability in the dl-epinephrine concentration for individual segments of commercial cord was noted. In addition, the cords did not contain the concentration of epinephrine (500 micrograms/inch) listed on the package label but instead had a mean dl-epinephrine concentration of 225.5 micrograms/inch.

Absorption↗
PubMed1992

Expectant management after sonographic diagnosis of placental abruption.

Forty cases of placental abruption were diagnosed by sonography after 20 weeks' gestation and managed expectantly. Tocolytics were used in 18 patients and were successful in delaying delivery for 34 +/- 24 days in 10. One-third of the patients delivered at term, after delays of 12.3 +/- 5.7 weeks. The majority of the patients who delivered before term (63%) had at least one other risk factor for preterm delivery, such as twins, ruptured membranes, or cervical dilation at presentation. Preterm delivery was not correlated with any of several clinical indicators of the severity of the abruption. Although the perinatal mortality rate was 22%, all perinatal deaths except one were attributable to extreme prematurity. It concluded that mild placental abruption is often a self-limited event and can safely be managed expectantly. Most of the perinatal morbidity and mortality is associated with premature delivery and, thus, in the absence of fetal distress or maternal compromise, delivery may often be delayed until fetal maturity is attained.

Abruptio Placentae↗
PubMed1992

Why are transformed cells immortal? Is the process reversible?

Normal cells have finite proliferative potential in culture. In contrast, cells derived from tumors immortalized by chemical carcinogens or viruses are able to divide indefinitely. A question of major importance is the mechanism that limits the proliferative potential of normal cells, and conversely, the process by which immortal cells have escaped irreversible growth cessation. To address this question we fused a number of different normal human fibroblast cell lines with various immortal human cell lines and determined the proliferative behavior of the resulting hybrids. In all cases the hybrids had a limited ability to proliferate in culture. These results suggested that the finite proliferative capacity of normal human cells was dominant and that immortal cells had acquired recessive changes in their genetic program, which allowed them to escape senescence. We were also able to assign approximately 30 immortal human cell lines to four complementation groups for indefinite division.

Animals↗
PubMed1992

Immunophenotypic subclassification of chronic lymphocytic leukaemia (CLL).

To determine the significance of the immunophenotypic heterogeneity of B-cell chronic lymphocytic leukaemia (CLL), surface immunoglobulins (SIgs), mouse rosette assays (MR), and a panel of monoclonal antibodies for B cells, T cells and myeloid cells were performed on peripheral blood samples from 61 newly diagnosed cases. Four groups were observed: group I (SIg+, MR+, CD19/20+, CD5+, T antigen (Ag)-; 27 cases); group II (SIg+, MR+, CD19/20+, CD5+, T Ag+; 17 cases); group III (SIg+, MR+ CD19/20+, CD5-, T AG-; 12 cases); and group IV (SIg-, MR+, CD19/20+, Cd5+, T Ag-; 5 cases). Groups were compared according to French-American-British Cooperative Group subtypes, clinical and laboratory features, Rai staging, and survival. Typical CLL morphology (greater than 90% small lymphocytes) was present in 20/20 (100%) of group I cases and 23/27 (85%) group II, III and IV cases (P = 0.09). Expression of a myeloid antigen was seen in 5/27 group I cases (18%) and 1/16 group II cases (6%), but was not predictive of survival (P = 0.36). The CD5- group III had a lower haemoglobin level (P less than 0.0001), higher Rai stage (P less than 0.002), and poorer survival at 5 years (P less than 0.02) than the other groups. We conclude that at least four distinct immunophenotypic subgroups of B-cell CLL can be determined. Expression of myeloid or T-cell antigens does not appear to predict for patient survival; however, lack of CD5 antigen may be associated with more advanced stage of disease and poor patient survival.

Adult↗
PubMed1992

Magnetic resonance imaging and anatomic atlas mapping for thalamotomy.

A computerized anatomic atlas mapping technique has been developed and used to determine target sites for stereotactic thalamotomies. The digitized stereotactic atlases of Schaltenbrand and Bailey, Van Buren and Borke, and Schaltenbrand and Wahren are used in conjunction with an associated electrophysiological data base to determine the intended target site on MR images of the diencephalon. This mapping technique uses the 'Brierley and Beck' proportional hypothesis for thalamic nuclear determination. Target site determination by this method was found to be very accurate, and frequently the mere placement of the probe in the target site resulted in tremor cessation.

Brain Mapping↗
PubMed1992

Comparison of CT- versus MRI-guided, computer-assisted depth electrode implantation.

Fourteen candidates for ablative seizure surgery underwent CT-guided, computer-assisted stereotactic depth electrode implantation and 21 underwent MRI-guided, computer-assisted implantation. A hand-held computer with no graphic capability was used for CT-guided procedures. A computer work station which included a high-resolution color graphics terminal with touchscreen interfacing and software capable of simulating targets and trajectories in single or multiple views was used for MRI-guided procedures. Previous phantom studies done with a 1.5-tesla MR scanner suggested acceptable localization error. Localizing information was obtained in 10 (71.4%) of 14 of the CT-guided implants and in 16 (76.2%) of 21 of MRI-guided cases. In the CT group, 7 (70%) were seizure-free and 8 (80%) were greater than 90% improved at 1 year follow-up. In the MRI group, 8 (80%) were seizure-free and 9 (90%) were greater than 90% improved at latest follow-up.

Computer Graphics↗
PubMed1992

Differential effects of left versus right seizure focus on human hippocampal evoked responses.

Hippocampal evoked potentials were recorded using the P3 tonal oddball paradigm in 30 patients with unilateral temporal lobe seizure focus. Spectral power of the evoked potentials was decreased on the side of seizure focus, but this reduction was much greater when the focus was on the left. The effect of left/right focus does not appear to be due to group differences in age, sex, seizure duration, or operative pathology. Remote or finer left/right structural differences or differential left/right hippocampal processing for the sequential tonal task are possible explanations.

Adolescent↗
PubMed1992

An E.S.R. investigation of the reactive intermediate generated in the reaction between FeII and H2O2 in aqueous solution. Direct evidence for the formation of the hydroxyl radical.

The technique of E.S.R. spectroscopy, when employed in conjunction with a continuous flow system, provides direct evidence for the nature of free radicals formed from organic substrates in the presence of FeII and H2O2 in aqueous solution. It is shown, both via the identification of hydroxyl-radical adducts to alkenes and via the observed site-selectivity of radical attack, that the hydroxyl radical is formed as the reactive intermediate in the presence of various chelators (e.g. EDTA, DTPA). This approach also allows the rate constants for the FeII-H2O2 reaction in the presence of the different chelates to be determined; values obtained are in reasonable agreement with most of those measured by other methods. Examples of radical oxidation (by FeIII) and reduction (by FeII) are revealed.

Electron Spin Resonance Spectroscopy↗
PubMed1992

Cytokine-induced differentiation of IgA B cells: studies using an IgA expressing B-cell lymphoma.

Cytokines such as interleukin-5 (IL-5) and transforming growth factor beta 1 (TGF beta 1) increase IgA production by heterogeneous populations of lipopolysaccharide (LPS)-activated murine B cells. We have used IgA expressing murine B-lymphoma cells CH12.LX.C4.4F10 (4F10) to define the activity of these and other cytokines on IgA secretion at the single-cell level, membrane IgA expression, IgA polymerization and cell growth. IL-5 as well as LPS significantly increases IgA secretion of 4F10 cells, whereas TGF beta 1, a cytokine known to stimulate isotype switching to IgA among surface IgM-bearing B cells, inhibits IgA secretion. When tested alone, IL-1 beta, IL-2, IL-4, IL-6 and interferon-gamma (IFN-gamma) do not significantly alter IgA secretion. However, there is a synergistic increase in IgA secretion when 4F10 cells are co-stimulated with IL-5 and IL-4, while IFN-gamma inhibits IL-5-stimulated up-regulation of IgA secretion. In parallel with increased IgA secretion after cytokine stimulation, 4F10 cells display less membrane IgA. Increased J-chain steady-state mRNA levels after IL-5 or LPS stimulation are paralleled by increased mRNA levels for secreted IgA, but are not accompanied by alterations in the ratio of monomeric to polymeric IgA. IL-5 and LPS initially stimulated but later inhibited 4F10 cell proliferation suggesting an inverse relationship between proliferation and differentiation in this cell line. 4F10 cells are a useful model for the characterization of discrete aspects of IgA B-cell differentiation, since the secretory and membrane Ig and proliferative responses of this IgA B-cell line to cytokines and LPS appear to parallel those of freshly isolated murine B cells.

Animals↗
PubMed1991

Senescent cells fail to express cdc2, cycA, and cycB in response to mitogen stimulation.

Senescent human diploid fibroblasts (HDF) contain no detectable cdc2 mRNA or p34cdc2 protein. Similarly, young quiescent HDF have only low levels of cdc2 mRNA and protein. After serum stimulation, quiescent HDF accumulate increasing amounts of cdc2 mRNA and protein and go through DNA synthesis and mitosis. In contrast, serum-stimulated senescent HDF fail to accumulate detectable amounts of cdc2 mRNA and protein and fail to enter S phase. Mitosis is likewise deficient in senescent cells even when they have been induced to synthesize DNA by simian virus 40 large tumor antigen. Since p34cdc2 or its homologues appear to be required for DNA synthesis and mitosis in eukaryotes, a lack of these molecules in serum-stimulated senescent HDF could be an important reason for their inability to enter S phase or mitosis. Nuclear microinjection of cdc2 DNA into senescent HDF causes rounding up of the cells but no induction of DNA synthesis. Since cyclins A and B are important cofactors of the protein kinase activity of p34cdc2 or its homologues, we analyzed expression of these genes in serum-stimulated senescent HDF and determined that they contain little or no cycA or cycB mRNA. These deficiencies may be relevant to the lack of DNA synthesis and mitosis in senescent HDF.

Amino Acid Sequence↗