Biomedical subjects

L H Young

Publications and source records attributed to L H Young.

At least 19 recordsLinked to original sources

PubMed1992

Characterization of a novel mRNA expressed by neurons in mature brain.

In previous studies, differential hybridization screening of an activated murine T-lymphocyte cDNA library identified an interleukin 2-responsive mRNA, designated F5, expressed in lymphoid tissues and brain only. We now report characterization of a full-length clone isolated from an adult mouse brain cDNA library. Neither the nucleic acid nor amino acid sequences demonstrated similarity to reported sequences. On Southern blotting, the protein coding sequence hybridized to genomic DNA from a variety of species. On Northern blotting, F5 mRNA was expressed in adult mouse brain, spinal cord, eye, and dorsal root ganglia but not in peripheral nerve. In situ hybridization studies demonstrated prominent expression by neurons in brain. F5 mRNA expression was undetectable in embryonic rat cerebral hemisphere and low until postnatal day 21. F5 is a novel mRNA selectively expressed by proliferating lymphocytes and mature neurons.

Amino Acid Sequence↗

PubMed1992

Experimental allergic encephalomyelitis and multiple sclerosis: lesion characterization with magnetization transfer imaging.

Magnetization transfer imaging (MTI) was initially performed in normal guinea pigs and human volunteers. A magnetization transfer ratio (MTR) was calculated in the normal white matter and was found to be 42%-44%, with less than 2.5% variation, which indicates the high reproducibility of the measurement. MTI was then applied to an animal model of white matter disease, acute experimental allergic encephalomyelitis (EAE). In this model of EAE, pathologically proved lesions were edematous with essentially no demyelination. MTRs decreased slightly but significantly (5%-8%) compared with the MTRs of the same tissue region measured before the onset of the lesion [corrected]. Fifteen patients with multiple sclerosis (MS) also underwent MTI. In the 15 patients with MS, all lesions (209 plaques) had a significantly decreased MTR (average, 26%). The authors believe that demyelination produced the lower MTR, and, thus, lesions varied in transfer ratio on the basis of the extent of myelin loss. In patients with MS, particularly those with chronic and/or progressive MS, the MTR of the normal-appearing white matter was significantly decreased. The data suggest that calculated MTR obtained with in vivo MTI may enable differentiation of edema from demyelination, and that MTI can demonstrate white matter abnormalities that cannot be seen with standard spin-echo or gradient-echo magnetic resonance imaging.

Adult↗

PubMed1992

Physiological hyperinsulinemia inhibits myocardial protein degradation in vivo in the canine heart.

Myocardial protein turnover in vivo was examined in anesthetized dogs following a 16- or 36-hour fast and again during a hyperinsulinemic (2 mU/kg per minute) euglycemic clamp with or without amino acid replacement or during saline infusion. We measured myocardial phenylalanine balance and rates of protein synthesis and degradation, using the extraction of intravenously infused L-[ring-2,6-3H]phenylalanine and the dilution of its specific activity across the heart at isotopic steady state. After both a 16-hour (n = 19) and 36-hour fast (n = 10), there was net myocardial release of phenylalanine indicated by the negative balances for phenylalanine of -52 +/- 9 (p less than 0.001) and -38 +/- 9 (p less than 0.005) nmol/min, respectively. Overall, the basal rate of myocardial protein degradation was lower in the 36-hour-fasted animals (81 +/- 13 versus 121 +/- 12 nmol/min, p less than 0.05). Myocardial phenylalanine balance and rates of protein synthesis and degradation did not change during insulin and glucose infusion in the 36-hour-fasted animals (n = 10). In these animals, there was a 30-40% decline in plasma amino acid concentrations, including branched chain (p less than 0.001) and essential amino acids (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

PubMed1992

Equine arch vessel anomaly associated with coarctation of the aorta.

Angiography in a 30-year-old man revealed the unique combination of aortic coarctation and an unusual arch anomaly. Proximal to the coarctation, a single arch vessel trifurcated into the brachiocephalic, left common carotid and left subclavian arteries. This anomalous arch vessel is a normal equine variant.

Abnormalities, Multiple↗

PubMed1992

Expression of cytolytic mediators by synovial fluid lymphocytes in rheumatoid arthritis.

To understand the role of cytolytic lymphocytes in the pathogenesis of rheumatoid arthritis, we investigated the expression of lymphocyte cytotoxicity mediators, perforin, and serine esterases, in lymphocytes derived from the synovial fluid of 15 patients with rheumatoid arthritis. Previous work has shown that CD8+ lymphocytes that possess markers of activation appear to be present in rheumatoid arthritis (RA). By means of in situ hybridization techniques and immunohistochemical analysis, the authors show that perforin and two serine esterases (serine esterase 1/Hanukah factor/granzyme A, and serine esterase 2/granzyme B) are expressed by subpopulations of CD8+ and CD56+ lymphocytes obtained from synovial fluid. The presence of these cytotoxic mediators suggests a possible mechanism for tissue damage, and provides evidence implicating cytolytic lymphocytes in the pathogenesis of RA.

Arthritis, Rheumatoid↗

PubMed1991

Myocardial protein turnover in patients with coronary artery disease. Effect of branched chain amino acid infusion.

The regulation of protein metabolism in the human heart has not previously been studied. In 10 postabsorptive patients with coronary artery disease, heart protein synthesis and degradation were estimated simultaneously from the extraction of intravenously infused L-[ring-2,6-3H]phenylalanine (PHE) and the dilution of its specific activity across the heart at isotopic steady state. We subsequently examined the effect of branched chain amino acid (BCAA) infusion on heart protein turnover and on the myocardial balance of amino acids and branched chain ketoacids (BCKA) in these patients. In the postabsorptive state, there was a net release of phenylalanine (arterial-cardiac venous [PHE] = -1.71 +/- 0.32 nmol/ml, P less than 0.001; balance = -116 +/- 21 nmol PHE/min, P less than 0.001), reflecting protein degradation (142 +/- 40 nmol PHE/min) in excess of synthesis (24 +/- 42 nmol PHE/min) and net myocardial protein catabolism. During BCAA infusion, protein synthesis increased to equal the degradation rate (106 +/- 24 and 106 +/- 28 nmol PHE/min, respectively) and the phenylalanine balance shifted (P = 0.01) from negative to neutral (arterial-cardiac venous [PHE] = 0.07 +/- 0.36 nmol/ml; balance = 2 +/- 25 nmol PHE/min). BCAA infusion stimulated the myocardial uptake of both BCAA (P less than 0.005) and their ketoacid conjugates (P less than 0.001) in proportion to their circulating concentrations. Net uptake of the BCAA greatly exceeded that of other essential amino acids suggesting a role for BCAA and BCKA as metabolic fuels. Plasma insulin levels, cardiac double product, coronary blood flow, and myocardial oxygen consumption were unchanged. These results demonstrate that the myocardium of postabsorptive humans is in negative protein balance and indicate a primary anabolic effect of BCAA on the human heart.

Aged↗

PubMed1991

Application of modern NMR techniques in the structure determination of microbial transformation products from sterols.

Microbial transformations of cycloartenol, 24-methylenecycloartanol and lanosterol with Mycobacterium sp. (NRRL B-3805) resulted in the isolation and characterization of five C19 steroids. Details of the tremendous structural modifications of these metabolites from their parent compounds were clarified by 2D nmr techniques. The stereochemistry of these novel metabolites was determined by nOe studies.

Lanosterol↗

PubMed1990

Perforin-mediated myocardial damage in acute myocarditis.

Endomyocardial specimens were obtained from 7 patients with acute myocarditis. Immunohistochemical examination of the mononuclear infiltrate showed mainly cytotoxic T lymphocytes and natural killer cells. Perforin (a pore-forming protein found in cytotoxic lymphocytes) was identified in this myocardial lymphocytic infiltrate and electron microscopy showed myocardial cell damage that may have been associated with these perforin containing lymphocytes. The results indicate that in acute idiopathic and viral myocarditis, myocardial damage may be due to the action of perforin-secreting lymphocytes.

Acute Disease↗

PubMed1990

Granulated metrial gland cells of pregnant mouse uterus are natural killer-like cells that contain perforin and serine esterases.

The mouse uterus during pregnancy contains a large population of lymphoid cells termed granulated metrial gland (GMG) cells. Our observations suggest that these cells are highly activated cytolytic lymphocytes related to NK or lymphokine-activated killer cells. Immunostaining demonstrated asialo GM1 and Thy-1 on GMG cells, both of which are expressed by NK cells. Decidua basalis tissue and isolated GMG cells contained three proteins that are characteristic of activated cytolytic lymphocyte granules: perforin, serine esterase 1, and serine esterase 2. These mediators were demonstrated in GMG cells by Western blot analysis using polyclonal antisera and by Northern blot analysis using specific cDNA probes for their mRNA. The proteins were not detected in normal spleen or liver or in asialo GM1+ cells isolated from those organs, consistent with the absence of these mediators from resting cytolytic cells. The amount of perforin in GMG cells was similar to that present in cloned, IL-2-stimulated, CTL shown previously to contain a large amount of this protein. A large population of NK cells bearing the surface marker LGL-1 was demonstrated at the implantation site by labeling with monoclonal antibody 4D11, but T cells were not detected. Many LGL-1+ cells at the implantation site expressed the GMG cell markers asialo GM1, Thy-1, and perforin. Staining intensities were inversely correlated, with LGL-1-bright cells showing little or no staining of GMG cell markers and LGL-1-faint cells showing more obvious staining of GMG cell markers. This suggests that LGL-1+ NK cells may differentiate in situ to GMG cells, losing LGL-1 and gaining a high concentration of GMG cell markers in the process. Activated cytolytic cells related to NK or lymphokine-activated killer cells may function in the pregnant rodent uterus to intercept and kill aberrant placental or embryonic cells that might otherwise enter the female and proliferate.

Animals↗

PubMed1990

Metabolic and functional effects of perfluorocarbon distal perfusion during coronary angioplasty.

Myocardial lactate metabolism and left ventricular function were studied in 12 patients during angioplasty of the left anterior descending artery performed with distal coronary perfusion (oxygenated and nonoxygenated Fluosol) and by conventional technique without distal perfusion. Before balloon inflation there was net lactate extraction by the heart (31 +/- 6%). During balloon inflations performed with distal perfusion there was net lactate release into the great cardiac vein while the balloon was inflated; the great cardiac vein lactate concentration was approximately 25% lower during perfusion with oxygenated versus nonoxygenated Fluosol (p less than 0.02) indicating less myocardial lactate release. After balloon deflation washout of lactate into the great cardiac vein (net myocardial release) was observed in all 3 protocols. Left ventricular ejection fraction measured by echocardiography decreased markedly during nonperfused (53 +/- 3 to 36 +/- 3%, p less than 0.001) and nonoxygenated Fluosol (52 +/- 2 to 30 +/- 3%, p less than 0.001) inflations. This dysfunction was largely prevented by oxygenated Fluosol where only a minimal decrease in ejection fraction (51 +/- 2 vs 48 +/- 2%, p less than 0.02) occurred. Analysis of regional contractile function yielded similar results. Although oxygenated perfluorocarbons decrease cardiac lactate release during angioplasty, this study provides evidence for the onset of lactate production even when ventricular function is preserved.

Aged↗

PubMed1990

Immunocytochemical localization of gentamicin in the rabbit retina following intravitreal injection.

The localization of gentamicin in the retina after a single intravitreal injection in the rabbit eye was examined by indirect immunofluorescent staining with goat antigentamicin antiserum. Eight hours after the injection of 400 micrograms of gentamicin, staining was observed in the ganglion cell layer, the inner plexiform layer, the inner nuclear layer, and the photoreceptors. By 12 hours, the staining was also observed in the retinal pigment epithelium. Howeever, by 24 hours the staining was predominantly found in the retinal pigment epithelium and choriocapillaris, and only occasional staining was seen scattered in the neurosensory retina. At 36 to 48 hours, the labeling was confined to the retinal pigment epithelium and choriocapillaris. Electron microscopy confirmed the cytoplasmic localization of gentamicin in the retina.

Animals↗

PubMed1990

How lymphocytes kill.

Cytotoxic T lymphocytes and natural killer cells are potent killers of target cells. These lymphocytes have large cytoplasmic granules containing cytotoxic peptides and other factors. Several of these molecules have been isolated and their functions elucidated. These molecules may be directly involved in the killing of virus-infected and transformed cells as well as in the development of cell-mediated autoimmune disorders.

Animals↗

PubMed1990

In vivo measurement of myocardial protein turnover using an indicator dilution technique.

We applied a nondestructive tracer technique, previously developed for measuring skeletal muscle protein turnover, to the measurement of myocardial protein turnover in vivo. During a continuous infusion of L-[ring-2,6-3H]phenylalanine to anesthetized, overnight-fasted dogs, we measured the uptake of radiolabeled phenylalanine from plasma and the release of unlabeled phenylalanine from myocardial proteolysis using arterial and coronary sinus catheterization and analytic methods previously applied to skeletal muscle. Using these measurements, together with a model of myocardial protein synthesis that assumes rapid equilibration of tracer specific activity between myocardial phenylalanyl-tRNA and circulating phenylalanine, we estimated the rates of heart protein synthesis and degradation. The rate of heart protein synthesis was also estimated directly from the incorporation of labeled phenylalanine into tissue protein. The use of [3H]phenylalanine was compared with L-[1-14C]leucine in the measurement of heart protein turnover in dogs given simultaneous infusion of both tracers. Leucine uptake and release by the myocardium exceeded that of phenylalanine by 3.1 +/- 0.4- and 1.7 +/- 0.3-fold, respectively, consistent with leucine's 2.4-fold greater abundance in heart protein and its metabolism via other pathways. Phenylalanine is the preferred tracer for use with this method because of its limited metabolic fate in muscle. One theoretical limitation to the method, slow equilibration of circulating labeled phenylalanine with myocardial phenylalanyl-tRNA, was resolved by comparison of these specific activities after a 30-minute infusion of labeled phenylalanine in the rat. A second, empirical limitation involves precision in the measurement of the small decrements in phenylalanine specific activity that occur with each pass of blood through the coronary circulation. This was addressed by improving the precision of both the measurements of phenylalanine concentration and phenylalanine specific activity using high-performance liquid chromatography. We conclude that the in vivo measurement of phenylalanine tracer exchange across the myocardium permits the nondestructive estimation of heart protein turnover in the intact animal.

Animals↗

PubMed1990

A monoclonal antibody specific to Müller cells and selective synaptic sites in the retina.

We have produced a monoclonal antibody which stains the Müller cells in the region of the photoreceptors, nerve terminals surrounding the horizontal cells, and nerve terminals in the inner plexiform layer in carp, goldfish, and white perch retinas. Electron microscopy showed that the staining in the outer and inner retina was confined to Müller cells and presynaptic terminals, respectively. In the teleost brain, the antibody stained only the optic tectum and efferent fibers from the stratum album centrale. Biochemical characterization by immunoblotting showed that this monoclonal antibody recognizes an approximately 70-kD molecule in both whole retina and brain homogenates, suggesting that the antibody recognizes an identical molecule. No staining was noted in the spleen or the liver. This monoclonal antibody appears to be specific to a molecule common to the Müller cells and presynaptic terminals in the teleost retina, and although it is present in other parts of the central nervous system, it is confined to the visual pathway.

Animals↗

PubMed1990

In vivo expression of perforin by natural killer cells during a viral infection. Studies on uveitis produced by herpes simplex virus type I.

A potent cytolytic pore-forming protein (PFP, perforin, or cytolysin) is associated with the cytoplasmic granules of cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. The role of PFP/perforin in cytolytic reactions carried out in vivo is still unclear. Here, the authors performed immunohistochemical analysis using antibodies monospecific for perforin and made use of a murine uveitis model produced by intracameral inoculation of herpes simplex virus I (HSV-I). The main cell infiltrate found in the anterior segment of virus-inoculated eyes consisted of Thy-1+/asialo GM1+/CD8-/CD4- cells, presumably representing NK cells. Perforin staining was detected mainly in cells bearing this phenotype. Perforin was only detected in cells displaying the large granular lymphocyte morphology. A small number of perforin-positive cells (less than 5%) colabeled as CD8+, indicating that these cells could have belonged to the CTL lineage. These observations show for the first time the presence of perforin-containing NK cells in tissues of animals undergoing acute viral infections.

Animals↗

PubMed1989

In vivo expression of perforin by CD8+ lymphocytes in autoimmune disease. Studies on spontaneous and adoptively transferred diabetes in nonobese diabetic mice.

A potent cytolytic pore-forming protein (perforin or cytolysin) has previously been found to be associated with the cytoplasmic granules of CTL and NK cells. Inasmuch as all previous studies on perforin have been conducted with cultured CTL and NK cell lines, it is not clear whether perforin may play a role in the cytotoxicity mediated by CTL that have been primed in vivo. In this study, we investigated the presence of perforin in pancreata from nonobese diabetic (NOD) mice, which have been studied as a model of autoimmune, insulin-dependent (type I) diabetes mellitus. Whereas adult NOD mice spontaneously develop diabetes, it is possible to induce diabetes in young, irradiated NOD mice by adoptive transfer of splenocytes obtained from diabetic donors. By means of immunohistochemical analysis, we were able to detect perforin Ag in a small subpopulation of CD8+/Thy-1+/asialo GM1-/CD4- lymphocytes in the pancreatic islets of animals undergoing both spontaneous and adoptive transfer-mediated insulitis. Perforin+/CD8+ lymphocytes were found in small clusters and were observed to display the morphology of large granular lymphocytes. These observations show for the first time the presence of perforin-containing CD8+ lymphocytes in tissues of animals undergoing autoimmune disease.

Animals↗

PubMed1989

Localization of cyclic adenosine monophosphate in the teleost retina: effects of dopamine and prolonged darkness.

Localization of cyclic adenosine monophosphate (cAMP) in the white perch retina was carried out with immunohistochemical and autoradiographic methods. Following exposure to dopamine or prolonged darkness, cAMP staining was observed by immunohistochemistry in the distal part of the inner nuclear layer, i.e. in the horizontal cells. After exposure to dopamine, increased levels of cAMP were also observed by autoradiography in many horizontal cells. Finally, increased levels of cAMP staining were observed immunohistochemically following incubation with dopamine in all types of cone-related horizontal cells that had been isolated and maintained in culture.

Animals↗

PubMed1989

In vivo expression of perforin by CD8+ lymphocytes during an acute viral infection.

CTL and NK cells cultured in vitro have been shown to contain a cytolytic pore-forming protein (PFP/perforin/cytolysin). To date, it has not been determined whether perforin is expressed by CTL that have been primed in vivo. Here, we have infected mice with two strains of lymphocytic choriomeningitis virus (LCMV), one of which mainly produces choriomeningitis and, the other, hepatitis. Brain and liver cryostat sections obtained from LCMV-infected mice were stained for various lymphocyte markers, including perforin. We were able to detect a large accumulation of perforin antigen in CD8+/Thy-1+/asialo GM1+/CD4- lymphocytes, which in fact represent the main infiltrating cell type found in brain and liver sections obtained during the late acute stage of LCMV infection. Perforin was also detected in a smaller population of CD8-/asialo GM1+/NK 1.1+/F480- cells, presumably corresponding to NK cells. Perforin-positive cells were found to have the morphology of blasts or large granular lymphocytes (LGL). These observations, together with in vitro studies performed in the past, indicate that perforin may be associated exclusively with LGL-like CTL blasts and NK cells. Our results demonstrate for the first time the presence of perforin in CTL that have been primed in vivo and suggest that perforin-positive CTL may be directly involved in producing the immunopathology associated with the LCMV infection.

Acute Disease↗