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Biomedical subjects

R J Weber

Publications and source records attributed to R J Weber.

At least 55 records · Page 3Linked to original sources
PubMed1989

Implantation of dispersed cells into primate brain.

Although several experimental therapies such as dopaminergic cell implantation in parkinsonian models and intratumoral placement of lymphokine-activated killer cells require intracerebral deposition of dispersed cell suspensions, a successful technique of needle implantation of cells into primate brain has not been demonstrated. The authors have sought to establish a stereotaxic technique to predictably deposit dispersed cells in primate brain. Human lymphocytes were cultured in recombinant interleukin-2, labeled with sodium 51 chromate (51Cr), and stereotaxically injected into the frontal white matter of six anesthetized rhesus monkeys. A 10-microliters aliquot of cell suspension (2 X 10(7) cells/ml) was deposited 16 mm deep to the dura at 5 microliters/min via Hamilton No. 22s or 26s needles. Five control aliquots were counted for each injection. Reflux out of the needle track was absorbed on gauze, and the recovered cells were counted. The animals were sacrificed 1 hour after implantation and the brain was removed and sectioned such that the cortex and white matter along the needle track were separate. The tissue sections were then counted. Recovery was expressed as the percentage of total injected radioactivity (cpm) that was present in each brain section. Two additional injected hemispheres were processed for autoradiography and histological study. Cell recovery in the brain (mean +/- standard deviation) was 87.2% +/- 13.9% (3.3% +/- 4.9% in cortex and 83.9% +/- 15.9% in white matter). The autoradiograms and histological examination showed a dense accumulation of radioactivity (cells) at the target site and minimal radioactivity (cells) in the needle track. Accurate intracerebral deposition of dispersed cells in primates was achieved with the technique described. This knowledge permits reliable stereotaxic implantation of cells into the brains of nonhuman primates and humans for investigation and therapy.

Animals↗
PubMed1988

Morphology of interleukin-2-stimulated human peripheral blood mononuclear effector cells killing glioma-derived tumor cells in vitro.

This is the first morphological study of interleukin-2-stimulated human peripheral blood mononuclear (PBM) cells resulting in lymphokine-activated killer (LAK) cell activity against human glioma-derived tumor cells in vitro, in which high-resolution differential interference video light microscopy, scanning electron microscopy, and transmission electron microscopy were used. A subset of cells within the LAK cell population are the effector cells and have an asymmetric cellular architecture characteristic of cytotoxic T lymphocytes (CTLs) and natural killer (NK) cells. Upon binding to target cells, the LAK effector cell nucleus is positioned away from the target cell, whereas the granules, Golgi apparatus, and microtubules orient toward the target cell. These LAK-glioma cell conjugates form very tight plasma membrane bonds with numerous interdigitations, and vesicles were found in the small extracellular spaces between the cells. This morphology was not observed in unstimulated PBM-glioma cell co-cultures. Glioma-derived cells react to LAK effector cells by blebbing, becoming round, and rapidly detaching from the substrate. The injured glioma-derived cells had a highly condensed cytoplasm and chromatin, lobular nucleus, and severe plasma membrane blebs, which are consistent with an apoptotic rather than an osmotic lysis mechanism of cell death. This study provides morphological evidence that supports a common cytotoxic mechanism for CTLs, NK cells, and LAK effector cells. The cytotoxic mechanism is based on the local exocytosis of vesicles by the effector cell into the small extracellular space between the effector-target cell conjugate. Granules found in CTLs, NK cells, and LAK cells contain a pore-forming protein that inserts holes in the target cell's plasma membrane through which a lethal substance(s) not yet identified is thought to enter the cell.

Cell Line↗
PubMed1988

Regional distribution and density of Thy 1.1 in rat brain and its relation to subpopulations of neurons.

We have used a radioimmunohistochemical technique employing OX-7, a monoclonal antibody to rat Thy 1.1, to determine the regional distribution and density of Thy 1 in rat brain. Thy 1.1 was found to be unevenly distributed in rat brain with distinct regional differences related to the density of neuronal perikarya. This is consistent with the previously reported findings that Thy 1.1 is found primarily on neurons. However, the relative absence of Thy 1 in some cell body-dense areas of the brain suggests that Thy 1 is expressed differentially on specific subsets of neurons which are abundant and widespread throughout the brain.

Animals↗
PubMed1988

Stereotaxic implantation of dispersed cell suspensions into brain. A systematic appraisal of cell placement and survival.

The application of several recent advances in cell biology, brain implantation, and cell-mediated tumor immunotherapy requires successful and reproducible placement of viable cell suspensions into brain. Stereotaxic implantation is being used to inject cytotoxic lymphocytes into gliomas and to replace dopaminergic cells in parkinsonian models. Systematic assessment of the factors that influence success in implantation of cell suspensions into solid tissues is needed. A model was developed for investigation of stereotaxic implantation using radiolabeled rat lymphokine-activated killer (LAK) cells. Anesthetized rats received microliter injections of cell suspension into the right caudate nucleus. The injection volume, cell concentration, infusion rate, and needle size were varied systematically. The animals were sacrificed 1 hour after injection; the brain was removed and sectioned, and the radioactivity was counted. Three aliquots of the suspension were injected into counting tubes for control analysis. Recovery of radioactivity was expressed as the percent of mean counts per minute (cpm) in the right frontal lobe/mean cpm in the three control tubes. To assess the viability of implanted cells, the right frontal region was mechanically dissociated in media and centrifuged, and the pellet and supernatant were counted. By using small needles and slow infusion of volumes of 10 microliters or less, 85% to 90% of the radioactivity was recovered in the caudate nucleus. At least half of the implanted cells were viable. Consistent, accurate implantation of dispersed cells into brain over a range of volumes, cell concentrations, infusion rates, and needle sizes was achieved.

Animals↗
PubMed1987

Increased expression of the B lymphocyte receptor for transferrin is stimulated by in vivo crosslinking of cell surface IgD.

Expression of a receptor for the serum protein transferrin has been shown to be a characteristic of several cell lineages and increased transferrin receptor (TFR) expression to reflect cellular activation. In vitro studies of human B lymphocytes stimulated with antibodies to IgM have demonstrated that these cells have the ability to express TFR and that increased B-cell TFR expression is seen first sometime after these cells enter the G1 phase of the cell cycle. It also has been shown that TFR expression is necessary for proliferation to occur and may be regulated by a factor produced by mitogen-activated T lymphocytes. To examine expression of TFR by activated B lymphocytes in vivo, and to determine the kinetics of induction of TFR expression, we have studied the effects of injecting mice with an affinity-purified goat antibody to mouse IgD (GaM delta) on TFR expression. This antibody previously has been shown to activate polyclonally mouse splenic B cells in vivo in a T-independent fashion. Results show that there is a small but definite quantity of TFR on resting splenocytes, at 24 hr after injection nearly all B cells but not T cells express increased amounts of TFR, TFR is increased to nearly the same extent in congenitally athymic BALB/c nu/nu mice as in their normal nu/+ littermates and therefore GaM delta-induced increased B lymphocyte TFR expression is relatively T independent, TFR expression increases as early as 3 hr after injection of 800 micrograms of GaM delta and increases steadily until it peaks 24-48 hr later, and TFR expression in GaM delta-injected mice increases concomitantly with surface Ia antigen and cell size.

Animals↗
PubMed1986

Influence of cardiopulmonary bypass on nitroglycerin clearance.

The effect of cardiopulmonary bypass on the clearance of nitroglycerin (NTG) was studied in seven patients scheduled for coronary artery bypass graft surgery. Intravenous NTG was administered through nonadsorbing tubing at a starting dosage of 5-10 micrograms/min and was adjusted as needed. Blood samples were obtained from the radial artery and antecubital vein before bypass and from the arterial outlet of the oxygenator during bypass at least 30 minutes apart during a constant dosage or at least 30 minutes after a dosage change. Serum concentrations were analyzed for NTG by gas chromatography. Venous NTG concentrations were always lower than concurrent arterial concentrations, with an average arteriovenous extraction of 67.2%. Serum concentrations of NTG were generally within the range associated with a therapeutic response in congestive heart failure patients. Consistent with other reports, NTG concentrations varied widely among patients and considerable intrasubject fluctuations in drug concentrations were seen. The mean +/- SD apparent clearance of NTG before bypass of 0.044 +/- 0.02 L/kg/min increased 20% to 0.052 +/- 0.02 L/kg/min during bypass (P = .05). These results suggest that cardiopulmonary bypass increases the clearance of NTG; however, the magnitude appears to be small and only partially explains the reported increased dosage needed during cardiopulmonary bypass.

Adult↗
PubMed1985

Direct evidence that endogenous GM1 ganglioside can mediate thymocyte proliferation.

The B subunit of cholera toxin, which is multivalent and binds exclusively to a specific ganglioside, GM1, was mitogenic for rat thymocytes. When exposed to the B subunit, the cells proliferated, as measured by 3H-labeled thymidine incorporation. Mitogenesis depended on the direct interaction of the B subunit with GM1 on the surface of the cells. This demonstrates that endogenous plasma membrane gangliosides can mediate proliferation in lymphocytes.

Adenylyl Cyclases↗
PubMed1985

Benzodiazepine receptor-mediated chemotaxis of human monocytes.

Benzodiazepines, which are widely prescribed for their antianxiety effects, are shown to be potent stimulators of human monocyte chemotaxis. The chemotactic effects of benzodiazepine receptor agonists were blocked by the peripheral benzodiazepine receptor antagonist PK-11195, suggesting that these effects are mediated by the peripheral-type benzodiazepine receptor. Diazepam was also active in inducing chemotaxis. Binding studies on purified monocytes revealed high-affinity peripheral benzodiazepine receptors, and the displacement potencies of various benzodiazepines correlated with their relative potencies in mediating chemotaxis. The demonstration of functional benzodiazepine receptors on human monocytes, together with recent evidence of receptor-mediated monocyte chemotaxis by other psychoactive peptides (such as opiate peptides), suggests a biochemical substrate for psychosomatic communication.

Benzodiazepinones↗
PubMed1985

Transferrin receptors in rat brain: neuropeptide-like pattern and relationship to iron distribution.

We have characterized and visualized the binding of 125I-labeled transferrin to sections of rat brain. This saturable, reversible, high-affinity (Kd = 1 X 10(-9) M) binding site appears indistinguishable from transferrin receptors previously characterized in other tissues. Moreover, a monoclonal antibody raised to rat lymphocyte transferrin receptors could immunoprecipitate recovered intact transferrin solubilized from labeled brain slices, indicating that labeling was to the same molecular entity previously characterized as the transferrin receptor. The pattern of transferrin receptor distribution visualized in brain with both 125I-labeled transferrin and an anti-transferrin receptor monoclonal antibody are almost indistinguishable but differ from the pattern of iron distribution. Iron-rich brain areas generally receive neuronal projections from areas with abundant transferrin receptors, suggesting that iron may be transported neuronally. However, many brain areas with a high density of transferrin receptors appear unrelated to iron uptake and neuronal transport and form a receptor distribution pattern similar to that of other known neuropeptides. This "neuropeptide-like" distribution pattern suggests that transferrin may have neuromodulatory, perhaps behavioral, function in brain.

Animals↗
PubMed1985

The intensive care unit syndrome: causes, treatment, and prevention.

The psychological assessment and management of the critically ill patient is often overlooked as a part of the patient care plan. The intensive care unit (ICU) syndrome is a type of organic brain syndrome manifested by a variety of psychological reactions, including fear, anxiety, depression, hallucinations, and delirium. Causes, treatment modalities, and a multidisciplinary approach to preventing the ICU syndrome are presented. Causative factors that should be assessed in the psychological evaluation of ICU patients include: (1) preadmission history; (2) past ability to adapt to stress; (3) past and current medications; (4) current clinical status; and (5) environmental factors. The treatment of the ICU syndrome includes: (1) the correction or elimination of causative factors; (2) the appropriate choice, dose, and route of administration of anxiolytic and antipsychotic agents; (3) reduction or elimination of sources of environmental stress; and (4) frequent patient and family communication. Finally, the prevention of the ICU syndrome through the involvement of physicians, nurses, and pharmacists is stressed.

Aging↗
PubMed1985

Neuropeptides and their receptors: a psychosomatic network.

A major conceptual shift in neuroscience has been wrought by the realization that brain function is modulated by numerous chemicals in addition to classical neurotransmitters. Many of these informational substances are neuropeptides, originally studied in other contexts as hormones, "gut peptides," or growth factors. Their number presently exceeds 50 and most, if not all, alter behavior and mood states, although only endogenous analogs of psychoactive drugs like morphine, Valium, and phencyclidine have been well appreciated in this context. We now realize that their signal specificity resides in receptors (distinct classes of recognition molecules), rather than the close juxtaposition occurring at classical synapses. Rather precise brain distribution patterns for many neuropeptide receptors have been determined. A number of brain loci, many within emotion-mediating brain areas, are enriched with many types of neuropeptide receptors suggesting a convergence of information at these "nodes." Additionally, neuropeptide receptors occur on mobile cells of the immune system; monocytes can chemotax to numerous neuropeptides via processes shown by structure-activity analysis to be mediated by distinct receptors indistinguishable from those found in brain. Neuropeptides and their receptors thus join the brain, glands, and immune system in a network of communication between brain and body, probably representing the biochemical substrate of emotion.

Animals↗
PubMed1984

The molecular mechanism of cryoimmunoglobulin precipitation--II. Thermodynamic basis for self-association as determined by fluorescence polarization.

Protein MAT is a homogeneous human IgM (lambda) cryoprecipitating cold agglutinin wherein the Fabmu and Fcmu5 fragment interaction is facilitated at low temps. Thermal dependence of the association of 2,5 DNS labeled Fab MAT with unlabeled Fcmu5 MAT was examined by fluorescence depolarization. The association data, treated by the method of van't Hoff, showed a nonlinear increase in binding with decreased temp, suggestive of certain dynamic changes in the system. Temperature effects on the rotational diffusion of five different 1,5 DNS labeled Fabmu fragments (including two derived from cryoglobulins) were also examined by fluorescence polarization. The linear nature of the Perrin plots derived from the data failed to reveal temp-induced hydrodynamic changes in any of the Fabmu fragments studied. Involvement of carbohydrates in the low-temp self-association of protein MAT was established by the finding that glycopeptides isolated from another IgM molecule (BAZ) could inhibit (as judged by depolarization of fluorescence) the interaction of 2,5 DNS labeled Fabmu MAT and unlabeled Fcmu5 BAZ fragments. These findings indicate that, although cryoprecipitation of protein MAT seemingly involves an antigen-antibody-like reaction between a site on the Fab region and carbohydrate moieties on the Fcmu5 region, no direct evidence for a low-temp-induced conformational change in the Fab region was obtained.

Agglutinins↗