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Biomedical subjects

R Li

Publications and source records attributed to R Li.

At least 397 records · Page 22Linked to original sources
PubMed1996

Host range restrictions of oncogenes: myc genes transform avian but not mammalian cells and mht/raf genes transform mammalian but not avian cells.

The host range of retroviral oncogenes is naturally limited by the host range of the retroviral vector. The question of whether the transforming host range of retroviral oncogenes is also restricted by the host species has not been directly addressed. Here we have tested in avian and murine host species the transforming host range of two retroviral onc genes, myc of avian carcinoma viruses MH2 and MC29 and mht/raf of avian carcinoma virus MH2 and murine sarcoma virus MSV 3611. Virus vector-mediated host restriction was bypassed by recombining viral oncogenes with retroviral vectors that can readily infect the host to be tested. It was found that, despite high expression, transforming function of retroviral myc genes is restricted to avian cells, and that of retroviral mht/raf genes is restricted to murine cells. Since retroviral oncogenes encode the same proteins as certain cellular genes, termed protooncogenes, our data must also be relevant to the oncogene hypothesis of cancer. According to this hypothesis, cancer is caused by mutation of protooncogenes. Because protooncogenes are conserved in evolution and are presumed to have conserved functions, the oncogene hypothesis assumes no host range restriction of transforming function. For example, mutated human proto-myc is postulated to cause Burkitt lymphoma, because avian retroviruses with myc genes cause cancer in birds. But there is no evidence that known mutated protooncogenes can transform human cells. The findings reported here indicate that host range restriction appears to be one of the reasons (in addition to insufficient transcriptional activation) why known, mutated protooncogenes lack transforming function in human cells.

Animals↗
PubMed1996

The nu gene acts cell-autonomously and is required for differentiation of thymic epithelial progenitors.

The nude mutation (nu) causes athymia and hairlessness, but the molecular mechanisms by which it acts have not been determined. To address the role of nu in thymogenesis, we investigated whether all or part of the nude thymic epithelium could be rescued by the presence of wild-type cells in nude <--> wild-type chimeric mice. Detailed immunohistochemical analyses revealed that nude-derived cells could persist in the chimeric thymus but could not contribute to cortical or medullary epithelial networks. Nude-derived cells, present in few clusters in the medulla, expressed markers of a rare subpopulation of adult medullary epithelium. The thymic epithelial rudiment of nude mice strongly expressed these same markers, which may therefore define committed immature thymic epithelial precursor cells. To our knowledge, these data provide the first evidence that the nu gene product acts cell-autonomously and is necessary for the development of all major subpopulations of mature thymic epithelium. We propose that nu acts to regulate growth and/or differentiation, but not determination, of thymic epithelial progenitors.

Animals↗
PubMed1996

Increased plasma rifabutin levels with concomitant fluconazole therapy in HIV-infected patients.

OBJECTIVE: To determine the effect of fluconazole on rifabutin pharmacokinetics. DESIGN: An open-label, crossover, phase 1 trial. SETTING: Outpatient clinical research center at a university medical center in Washington, D.C. PATIENTS: 12 persons with human immunodeficiency virus (HIV) infection whose CD4 lymphocyte counts were between 200 and 500 cells/mm3 and who were receiving maintenance therapy with zidovudine. INTERVENTION: Fluconazole, 200 mg/d for 2 weeks; then a combination of fluconazole, 200 mg/d, and rifabutin, 300 mg/d, for 2 weeks; and then rifabutin, 300 mg/d, for the final 2 weeks of the study. MEASUREMENTS: Blood and urine samples were obtained at regular intervals for 24 hours at the end of each 2-week dosing period to ascertain concentrations of fluconazole and rifabutin and the 25-desacetyl metabolite of rifabutin, LM565. RESULTS: Fluconazole significantly increased the plasma concentrations of both rifabutin and LM565. Mean increases in the area under the plasma concentration curve compared with the time curve over a 24-hour dosing interval were 82% (5442 +/- 2404 ng.h/mL compared with 3025 +/- 1117 ng.h/mL; P less than or equal to 0.05) for rifabutin and 216% (959 +/- 529 ng.h/mL compared with 244 +/- 141 ng.h/mL; P less than or equal to 0.05) for LM565. CONCLUSIONS: Fluconazole significantly increases the systemic exposure of both rifabutin and LM565. This pharmacokinetic interaction offers a mechanism that may explain the changes reported in both the efficacy and toxicity of rifabutin with concomitant fluconazole therapy.

AIDS-Related Opportunistic Infections↗
PubMed1996

Identification of Gas6 as a growth factor for human Schwann cells.

Schwann cells are one of the principal components of the peripheral nervous system. They play a crucial role in nerve regeneration and can be used clinically in the repair of injured nerves. We have established serum-free, defined culture conditions that rapidly expand adult human Schwann cells without fibroblast growth. We find that Gas6, a ligand for the Axl and Rse/Tyro3 receptor protein tyrosine kinase family, stimulates human Schwann cell growth, increasing both cell number and thymidine incorporation. Gas6 has synergistic effects with the other known human Schwann cell mitogens, heregulin/glial growth factor and forskolin. Addition of Gas6 causes phosphorylation of Axl and Rse/Tyro3 simultaneously and results in ERK-2 activation. A combination of Gas6 with heregulin and forskolin, on a defined background, supports maximal Schwann cell proliferation, while preserving the typical Schwann cell morphology and expression of the Schwann cell markers S-100, glial fibrillary acidic protein, and low-affinity nerve growth factor receptor. Gas6 mRNA is present in both spinal motor neurons and large neurons of the dorsal root ganglia, and neural injury has been reported to upregulate Rse/Axl in the schwann cell. This is the first demonstration of a potentially important biological role for the human Gas6/Rse-Axl system.

Animals↗
PubMed1996

Deficiencies in progenitor cells of multiple hematopoietic lineages and defective megakaryocytopoiesis in mice lacking the thrombopoietic receptor c-Mpl.

Mice with a null mutation in the thrombopoietin (TPO) receptor c-Mpl were generated by gene targeting. c-mpl-deficient mice developed normally but were deficient in megakaryocytes and severely thrombocytopenic. The hematocrit and numbers of mature circulating leukocytes were normal in mpl-/- mice, as was the distribution of morphologically identifiable precursors in hematopoietic tissues. Bone marrow and spleen cells of adult mpl-/- mice lacked specific binding sites for TPO, were unresponsive to TPO in culture, and displayed a marked deficiency in progenitor cells with megakaryocytic potential. Significantly, total hematopoietic progenitor cell numbers were also reduced in mpl-/- mice including multipotential, blast cell, and committed progenitors of multiple lineages. The megakaryocyte deficiency was evident as early as 14 days of gestation in mpl-deficient mice, although reductions in progenitor cell numbers arose only later in development. The data suggest that the critical function of c-Mpl signalling in megakaryocytopoiesis is in maintenance of mature megakaryocyte numbers through control of progenitor cell proliferation and maturation. Moreover, our results also imply an important role for TPO and c-Mpl in the production of primitive pluripotent progenitor cells as well as progenitor cells committed to nonmegakaryocytic lineages.

Amino Acid Sequence↗
PubMed1996

Subcellular distribution of p21 and PCNA in normal and repair-deficient cells following DNA damage.

BACKGROUND: The p21 protein binds to both cyclin-dependent kinases (Cdks) and the proliferating cell nuclear antigen (PCNA). In mammalian cells, DNA damage results in an increase in the level of p53 protein, which stimulates expression of the gene encoding p21, which in turn leads to an inhibition of Cdk activity. Biochemical studies have shown that the direct interaction between p21 and PCNA blocks the latter's function in DNA replication but not in DNA repair. In addition to the p53-dependent damage response, the stimulation of quiescent cells with serum can also cause a p53-independent elevation in p21 gene expression. It is not clear, however, whether the induction of p21 protein under these two circumstances serves the same purpose. In this study, we have investigated the kinetics of p21 induction by DNA damage and serum stimulation and the consequent effects on cell-cycle progression. Using both normal and repair-deficient human cells, we have also analyzed the nuclear distribution of p21 in relation to that of PCNA. RESULTS: In vivo immunofluorescence staining experiments indicate that, following UV damage, DNA repair is not inhibited by the presence of a large amount of p21 protein in the nucleus; in contrast, cells undergoing DNA replication during S phase contain very low amounts of p21. The addition of serum induced a transitory elevation of p21 levels, whereas UV damage to cells resulted in a sustained, high level of p21 that was more tightly associated with the nuclear structure. Interestingly, cells deficient in global nucleotide excision-repair displayed a distinct pattern of detergent-insoluble p21 that co-localized with PCNA. CONCLUSIONS: The in vivo studies presented here, which are consistent with our previous findings in vitro, indicate that p21 has a differential effect on DNA replication and DNA repair, and that the induction of p21 by serum and DNA damage may have different consequences. Furthermore, the co-localization of p21 and PCNA in the nucleus of normal and repair-deficient human cells indicates that p21 and PCNA interact during post-damage events.

Cell Cycle↗
PubMed1996

Lymphoblastoid transmembrane potential in bipolar patients, their siblings, and unrelated healthy comparison subjects.

Bipolar illness appears to be characterized by alterations in ionic homeostasis that are related to mood state. This is reflected by reports of altered intracellular sodium and calcium concentrations, altered sodium pump (Na+,K+-adenosine triphosphatase), and calcium pump (Ca2+-adenosine triphosphatase) activity. Recent ex vivo studies with fresh lymphocytes and in vitro studies with cultured lymphoblasts suggest that there may be an enduring trait-related abnormality in Na+,K+-adenosine triphosphatase as well. We have previously found that the lymphocyte transmembrane potential (TMP) varies with mood state. To examine the question of trait-related changes in TMP, we studied TMP of immortalized lymphoblasts from 14 patients with bipolar illness, nine unaffected siblings, and eight normal subjects. TMP was the same in all three groups. These preliminary data suggest that TMP is a state-- rather than a trait-related marker.

Adult↗
PubMed1996

Thyromimetic action of the peroxisome proliferators clofibrate, perfluorooctanoic acid, and acetylsalicylic acid includes changes in mRNA levels for certain genes involved in mitochondrial biogenesis.

The effects of three peroxisome proliferators on the mRNA levels for some mitochondrial inner-membrane proteins in rat liver were investigated. Clofibrate, perfluorooctanoic acid, and acetylsalicylic acid all increased the mRNA levels for the mitochondrial-encoded respiratory-chain components cytochrome c oxidase subunit I and NADH dehydrogenase subunit I. Mitochondrial 16S rRNA was also induced by clofibrate. The mRNA levels for the nuclear-encoded mitochondrial inner-membrane proteins adenine nucleotide translocator and cytochrome c1 were selectively induced by the different peroxisome proliferators. Malic enzyme, which is induced by thyroid hormone, was also induced by the three peroxisome proliferators tested. These effects are in some ways similar to those obtained with thyroid hormone.

Animals↗
PubMed1996

Hlx homeo box gene is essential for an inductive tissue interaction that drives expansion of embryonic liver and gut.

The divergent murine homeo box gene Hlx is expressed in restricted hematopoietic cell types and, during embryogenesis, prominently in visceral mesenchyme of the developing liver, gall bladder, and gut. Targeted disruption of the gene has now established that it plays a key role in visceral organogenesis. Embryos homozygous for the mutation died around embryonic day 15 with anemia and severe hypoplasia of the liver and gut. Liver ontogeny commenced normally with formation of the liver diverticulum and differentiation of hepatocytes, but the organ failed to expand and reached only 3% of normal size. The apparent liver hypoplasia was not associated with a notable increase in apoptotic cells. Gut development also began normally, but the intestines failed to undergo extensive elongation and looping and reached only a quarter of normal length. The anemia resulted from a deficiency in the fetal form of hematopoiesis, which occurs in the liver, but no intrinsic defect in Hlx-/- hematopoietic cells was observed in vitro, and liver-derived Hlx-/- hematopoietic stem cells that were transplanted to irradiated normal mice could fully reconstitute hematopoiesis. The impaired fetal hematopoiesis therefore reflects insufficient support function provided by the minute liver. Hlx is normally expressed in visceral mesenchyme lying adjacent to the developing liver and gut epithelia affected by the mutation, but not in the epithelia themselves. Hence, Hlx regulates a mesenchymal-epithelial interaction that drives a vital growth phase in visceral organogenesis. Moreover, because mutation of Hlx blocked liver growth but not its specification, early morphogenesis, or differentiation, development of this organ appears to occur by step-wise inductive interactions under separate genetic control.

Animals↗
PubMed1996

Structural characterization and in vivo immunosuppressive activity of neuroblastoma GD2.

Shedding of neuroblastoma gangliosides is positively correlated with tumour progression in patients with neuroblastoma. In assessing the biological activity of these ganglioside molecules, we recently found that total human neuroblastoma gangliosides inhibit cellular immune responses. Here, we have studied the major neuroblastoma ganglioside, GD2. GD2 was purified by high performance liquid chromatography and structurally characterized by mass spectrometry. Immunoregulatory effects of GD2 in vivo were then determined in an established murine model. GD2 significantly downregulated the local cellular immune response to an allogeneic cell challenge; the usual increase in mass of the lymph node draining the injection site was reduced by 88%, from 1.52 to 0.19 mg (control versus GD2-treated mice; p < 0.01). In parallel, lymphocyte recovery from each node was also reduced from 2.4 to 1.2 x 10(6) cells, and lymphocyte DNA synthesis was reduced to half of the control level. These results show that certain shed tumour gangliosides, such as GD2, function as intercellular signalling molecules, downregulate the cellular immune response, and may thereby enhance tumour formation and progression.

Animals↗
PubMed1996

Effect of domestically-made levonorgestrel-releasing intrauterine device on the endocrine system and menstruation in monkeys.

Effect of domestically-made levonorgestrel-releasing intrauterine device (LNG, release rate, 6 micrograms/day) on the endocrine system and menstruation in monkeys was investigated. The results showed that the Estradiol (E2) and progesterone (P) levels were significantly decreased 2 ovulatory cycles after insertion of the devices as compared with those before insertion in 3 monkeys (P > 0.001), suggesting an evident ovulation-suppressing effect. In 1 monkey, the E2 and P levels were the same before and after insertion, and expulsion of device was found in the monkey later. The plasma LNG concentration in 3 animals was 493.69-454.60 pg/ml and plasma LNG was not detected in 1 monkey. The hormone level returned to normal level after removal of the devices.

Animals↗
PubMed1996

Effects of chronic episodic hypoxia on monoamine metabolism and motor activity.

Chronic episodic hypoxia produces a wide array of cardiovascular dysfunctions in rats, including increases in blood pressure, heart rate, and sympathetic nerve activity. The action of episodic hypoxia might be related to low oxygen itself (hypoxemia) and/or combined with low CO2 (hypocapnia) resulting from hyperventilation. It is unknown whether or not the cardiovascular abnormalities are related to alterations in the central nervous system (CNS) that may be manifested as neurotransmitter and/or behavioral changes. In this study, we investigated effects of episodic eucapnic and hypocapnic hypoxia on monoamine metabolism in both CNS and adrenal glands, and on motor behavioral activity. Thirty-five male rats were divided into 3 groups. Experimental rats were exposed 8 h daily to varying fractional concentrations of inspired oxygen (FiO2) and carbon dioxide (FiCO2) for 35 days. These consisted of brief exposures (3-6s) of episodic (twice every min) eucapnic (3.5% FiO2 and 10% FiCO2, n = 6) or hypocapnic (3.5% FiO2 and 0% FiCO2, n = 14) hypoxia, or room air (21% FiO2 and 0.03% FiCO2, n = 15). Norepinephrine, dopamine, serotonin, and their metabolites in the hypothalamus, hippocampus, and adrenal glands were measured by high-performance liquid chromatography (HPLC). Spontaneous behavioral activity was assessed for 30 min by automated activity monitors. Episodic hypocapnic hypoxia produced a decrease in dopamine turnover and eucapnic hypoxia increased norepinephrine levels in the hypothalamus. Animals exposed to hypocapnic hypoxia also exhibited a consistent increase in horizontal (walking) and vertical (rearing) activity, as well as in total activity time. From these results, it is concluded that episodic eucapnic and hypocapnic hypoxia may affect metabolism of different neurotransmitters in the CNS.

Amines↗
PubMed1996

Reproducibility of extracranial carotid atherosclerotic lesions assessed by B-mode ultrasound: the Atherosclerosis Risk in Communities Study.

The reproducibility in the identification of carotid artery lesions using B-mode ultrasound was studied in a large random sample selected from the Atherosclerosis Risk in Communities (ARIC) Study. Carotid lesions were defined as plaque with or without acoustic shadowing (indicative of mineralization). A weighted kappa (kappa w) statistic was used as a chance-adjusted measure of repeatability. In the ARIC baseline survey, the kappa w values for the assessment of lesions on repeat reading were 0.47, 0.60 and 0.69 in the left common carotid artery, the carotid bifurcation and the internal carotid artery, respectively. In a repeat scanning, the kappa w values ranged from 0.59 to 0.79 in the left carotid segments. The results were similar in the left and right carotid arteries. Covariates (age, race, gender, body mass index, study center) did not influence the reproducibility. Similar results were also found in both the baseline survey and the first follow-up examination. In conclusion, reproducibility in the assessment of carotid lesions by B-mode ultrasound can be achieved in multicenter studies at fair to good levels of agreement.

Aged↗
PubMed1996

Structure-based design of parasitic protease inhibitors.

To streamline the preclinical phase of pharmaceutical development, we have explored the utility of structural data on the molecular target and synergy between computational and medicinal chemistry. We have concentrated on parasitic infectious diseases with a particular emphasis on the development of specific noncovalent inhibitors of proteases that play a key role in the parasites' life cycles. Frequently, the structure of the enzyme target of pharmaceutical interest is not available. In this setting we have modeled the structure of the relevant enzyme by virtue of its sequence similarity with proteins of known structure. For example, we have constructed a homology-based model of falcipain, the trophozoite cysteine protease, and used the computational ligand identification algorithm DOCK to identify in compuo enzyme inhibitors including oxalic bis(2-hydroxy-1-naphthyl-methylene)hydrazide (1) [Ring, C. S.; Sun, E.; McKerow, J. H.; Lee, G.; Rosenthal, P. J., Kuntz, I. D.; Cohen, F. E., Proc. Natl Acad. Sci. U.S.A. 1993, 90, 3583]. Compound 1 inhibits falcipain (IC50 6 microM) and the organism in vitro as judged by hypoxanthine uptake (IC50 7 microM). Following this lead, to date, we have identified potent bis arylacylhydrazides (IC50 150 nM) and chalcones (IC50 200 nM) that are active against both chloroquine-sensitive and chloroquine-resistant strains of malaria. In a second example, cruzain, the crystallographically determined structure of a papain-like cysteine protease, resolved to 2.35 A, was available. Aided by DOCK, we have identified a family of bis-arylacylhydrazides that are potent inhibitors of cruzain (IC50 600 microM). These compounds represent useful leads for pharmaceutical development over strict enzyme inhibition criteria in a structure-based design program.

Aldehydes↗
PubMed1996

Polymorphism and crystallization behavior of Abbott-79175, a second-generation 5-lipoxygenase inhibitor.

The crystal polymorphism of Abbott-79175, a potent second-generation 5-lipoxygenase inhibitor, has been studied. Crystallization from an ethyl acetate/heptane system in which moisture content was not controlled and from an ethanol/water system produced a crystal form I, which by differential scanning calorimetry, thermogravimetric analysis, and Karl Fischer analysis was likely a hemihydrate. This hemihydrate form of the compound was demonstrated to reversibly dehydrate/hydrate with the use of low heat and vacuum followed by exposure to water vapor (100% relative humidity). The dehydrated version of I was designated IA. However, it was shown that a relative humidity approaching 100% was required for the water to re-enter the crystal structure. X-ray powder diffraction and solid state 13C NMR data on polycrystalline samples crystallized from seven single organic solvents suggested two nonsolvated forms of the compound, II and III. The heptane addition rate to a dry ethyl acetate solution of the drug was shown to greatly affect the crystal habit, the crystallite size, and specific surface area upon its precipitation. Solution calorimetry of forms I, IA, and II showed that I had the most endothermic heat of solution, with IA and II having lower values. Initial dissolution rates of compressed disks of these forms in water showed rates of IA > II approximately I. Considering all data, including evidence of conversion of II to I in aqueous suspension, it is suggested that the relative thermodynamic stability (at least when water was present) in decreasing order is I > II > IA, although I and II are similar energetically. The lower terminal dissolution rate for form IA was due to, in part, conversion to the less soluble Form I.

Anti-Inflammatory Agents↗
PubMed1996

A comparison of amino acid sequences of hepatitis B virus S gene in 46 children presenting various clinical features for immunoprophylaxis.

We compared amino acid sequences of hepatitis B virus (HBV) S protein deduced from analyzed DNA sequence in 46 children who received immunoprophylaxis to prevent mother-to-child transmission of HBV. They were classified into 6 groups by their clinical features. The antibody escape mutants were found in 8 cases among 46 cases. We studied the difference in clinical features in these cases and speculated that 126 Ser or 140 Ser-strain may have a different behavior in relation to antibody to hepatitis B surface antigen from 126 Asn or 145 Arg-strain.

Amino Acid Sequence↗