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Biomedical subjects

N Shen

Publications and source records attributed to N Shen.

70 records · Page 4Linked to original sources
PubMed1993

DNA adducts in model systems and humans.

The etiology of chemically induced cancer is thought to involve the covalent binding of carcinogens to DNA (adducts) leading to mutations in oncogenes or tumor suppressor genes, and ultimately to tumors. Thus, the DNA-carcinogen adduct has been used as a measurable biochemical endpoint in laboratory studies designed to assess carcinogen exposure, carcinogen metabolism, mutagenesis, and tumorigenesis. Unfortunately, the significance of adducts in the etiology of human cancer is still unclear. This is partially due to the difficulty detecting adducts at carcinogen exposures relevant to humans, which are often orders of magnitude lower than animal model exposures. The relationship between adducts and higher biological effects is also not known at low doses. We have been assessing the DNA damage caused by exposure to heterocyclic amine carcinogens in the diet. Using the technique of 32P-postlabeling in combination with accelerator mass spectrometry, we have determined that DNA adduction in rodents decreases linearly with decreasing dose from the high doses used in typical cancer bioassays to the low doses relevant to human exposures. For a given tissue, adduct levels are correlated with dose, but the level of DNA modification by carcinogens is tissue-specific and does not completely correlate with tumor site. This lack of correlation may be due to differences in adduct formation and repair rates among tissues. Comparison of carcinogen metabolism routes between rodents and humans also indicates that species differences could influence the amount and type of damage resulting from exposure to these carcinogens. The use of model systems to study dosimetry, species differences in adduction, and role of adducts in mutation will ultimately lead to a better understanding of the significance of adducts in human disease. This should eventually allow the use of adducts as biomarkers for estimating carcinogen exposure and individual susceptibility.

Amines↗
PubMed1993

Delivery of recombinant gene products with microencapsulated cells in vivo.

If established cultured cell lines genetically modified to secrete desired gene products could be implanted in different allogeneic recipients without immune rejection, novel gene products would be delivered more cost effectively. We tested this strategy by encapsulating mouse Ltk- cells transfected with the human growth hormone (hGH) gene in immunoprotective perm-selective alginate microcapsules. Allogeneic mice implanted with these microcapsules demonstrated hGH in their circulation (0.1-1.5 ng/ml serum) within the first 2 weeks. Control mice implanted with only the transfected cells without microcapsules did not demonstrate significant levels of circulating hGH. By about 3 weeks, antibodies against hGH developed in the microcapsule-implanted mice. The immune response was detected only against the hGH and no other secretory products from the transfected cells. The antibody titer continued to escalate for more than three months, thus demonstrating indirectly the continued delivery of the growth hormone. The persistent expression of the transgene and survival of the transfected cells were verified when the microcapsules were retrieved periodically to demonstrate that the encapsulated cells remained viable, proliferative, and productive of hGH even by 78-111 days. In conclusion, delivering gene products with genetically modified allogeneic cells in vivo has been shown feasible for prolonged periods. This technology should have potential applications in somatic gene therapy and in treatment of other somatic diseases.

Animals↗
PubMed1992

Fate and distribution of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine in mice at a human dietary equivalent dose.

2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a heterocyclic amine rodent carcinogen that is found at the ppb level in cooked meat. Most laboratory studies are at 10(4)-10(7)-fold greater concentrations than actual ingested human doses. We report the first study of the bioavailability and fate of this heterocyclic amine at a human dietary equivalent dose using the high sensitivity offered by accelerator mass spectrometry. [2-14C]PhIP was administered to C57BL/6 male mice (41 ng/kg) by gavage. Tissues and excreta were collected over the subsequent 96 h. One hundred % of the administered dose was excreted in urine (90%) and feces (10%) over the length of the study. Absorption of the radiocarbon-tagged PhIP from the gastrointestinal tract was rapid, with radiocarbon levels peaking in the whole blood and urine within 1 h of exposure. Fecal 14C levels peaked at 12 h. Tissue levels peaked by 3 h with the highest concentrations of radiolabel in the intestine, stomach, and liver, followed by the kidney, pancreas, lung, and spleen. Low levels of 14C from PhIP (0.01-0.04% of the administered dose) could be detected in the tissues 48-96 h after exposure, possibly due to covalent binding to protein or DNA. The calculated half-life of PhIP at this dose was 1.14 h. This study is the first example of how accelerator mass spectrometry can be used to gather biological information about carcinogenic compounds at environmental levels of exposure.

Animals↗
PubMed1992

Apoptosis and DNA fragmentation precede TNF-induced cytolysis in U937 cells.

The hypothesis that activation of apoptosis and DNA fragmentation is involved in TNF-mediated cytolysis of U937 tumor cells was investigated. Morphological, biochemical, and kinetic criteria established that TNF activates apoptosis as opposed to necrosis. Within 2-3 h of exposure to TNF, U937 underwent the morphological alterations characteristic of apoptosis. This was accompanied by cleavage of DNA into multiples of nucleosome size fragments. Both of these events occurred 1-2 h prior to cell death as defined by trypan blue exclusion or 51Cr release. DNA fragmentation was not a non-specific result of cell death since U937 cells lysed under hypotonic conditions did not release DNA fragments. The percentage of cells undergoing apoptosis depended on the concentration of TNF and was augmented by the addition of cycloheximide. A TNF-resistant variant derived from U937 did not undergo apoptosis in response to TNF, even in the presence of cycloheximide. Furthermore, TNF could still activate NFkB in this variant, suggesting that this pathway is not involved in TNF-mediated cytotoxicity. Two agents known to inhibit TNF-mediated cytotoxicity, ZnSO4 and 3-aminobenzamide, were shown to inhibit TNF-induced apoptosis. Taken altogether, these data support the hypothesis that activation of apoptosis is at least one essential step in the TNF lytic pathway in the U937 model system.

Benzamides↗
PubMed1992

Effects of calcium on recombinant bovine chromogranin A.

Bovine chromogranin A, the acidic calcium-binding protein characteristic of endocrine secretory vesicles, has been expressed in Escherichia coli using the pET3a vector system under T7 polymerase control. The expressed protein is located in the bacterial cytosol and can be purified from bacterial proteins by a heat treatment step, followed by gel filtration, anion-exchange, and reversed-phase chromatography. The purified recombinant chromogranin A has an apparent M(r) of ca. 72,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, in spite of its 432-amino acid polypeptide chain, consistent with observations on natural chromogranin A. The primary structure has been confirmed by mass spectral analysis of tryptic peptides, by Edman degradation of the intact protein, and by immunoreactivity with sequence-specific antibodies. Analysis by circular dichroism spectroscopy shows pH- and concentration-dependent spectra. The spectra are Ca2(+)-dependent from 5 to 40 microM.

Amino Acid Sequence↗
PubMed1992

[Relationship between height, collagen metabolism, hair zinc and excessive fluoride intake].

After eliminating confounding factors, the study was made on the relationship between height, collagen metabolism, hair zinc and excessive fluoride intake. 140 schoolchildren aged 12-13 years born and reared in endemic fluorosis areas were surveyed. The results were as follows: 1. The average height of children with dental fluorosis III degree (DF III degree) was appreciably smaller than that of children without dental fluorosis. Among children with excessive fluoride intake, a negative correlation between the height and fluoride level in staple foods was seen. 2. The more the fluoride ingested, the higher the urinary THP excreted, showing that fluoride intoxication interfered with the collagen metabolism. 3. Among children with excessive fluoride intake, the height showed negative correlation with urinary THP/Cr, suggesting that the effect of fluoride on collagen metabolism indicated the mechanism of height retardation. 4. As compared to control group with the excessive fluoride intake but without dental fluorosis group, there was a significant reduction in hair zinc in group with DF III degree, suggesting that the zinc in the body decreased because of zinc metabolism disturbance by excessive fluoride intake. But among cases with excessive fluoride intake, no appreciable correlation between hair zinc and height was found. Therefore, it could not be confirmed that the effect of fluoride on zinc metabolism affected the height development.

Adolescent↗
PubMed1990

[Protective effects of gypenosides on experimental myocardial infarction].

In the model of myocardial infarction produced by occlusion of left anterior descending coronary artery (LAD) in rabbit, gypenosides (GP 100 but not 50 mg/kg, ip) reduced myocardial infarct size and decreased serum free fatty acid (FFA). In rat model of myocardial infarction, GP and the fractions of GP of non ginsenosides (FGNG) both in dose of 100 mg/kg, ip, protected significantly myocardial superoxide dismutase (SOD) activity and decreased the myocardial malondialdehyde (MDA). The results indicate that the protective effect of GP on myocardial infarction may be correlated with its prevention of myocardial lipid peroxidation, and attributed to the amelioration of FFA metabolic deterioration.

Animals↗
PubMed1989

T cell determinants of myelin basic protein include a unique encephalitogenic I-E-restricted epitope for Lewis rats.

The major encephalitogenic epitope for Lewis rats is the 72-89 sequence of guinea pig basic protein (GP-BP) or rat basic protein (Rt-BP). T cells responsive to this epitope are I-A restricted and preferentially express the V alpha 2:V beta 8 gene combination in their TCR. In this work, we describe for the first time the delayed appearance of T cells specific for additional discrete determinant of BP, the nonencephalitogenic 55-68 sequence of GP-BP restricted by I-A, and the encephalitogenic 87-99 sequence of Rt-BP restricted by I-E. The TCR V alpha 2:V beta 8 gene combination was expressed by both encephalitogenic GP-BP S72-89 and Rt-BP S87-99 T cell specificities but not by GP-BP 44-68-specific T cells. This is the first demonstration of I-E-restricted encephalitogenic T cells in Lewis rats and supports the conclusion that the I-E class II locus is involved in autoimmune diseases.

Amino Acid Sequence↗
PubMed1989

Both rat and mouse T cell receptors specific for the encephalitogenic determinant of myelin basic protein use similar V alpha and V beta chain genes even though the major histocompatibility complex and encephalitogenic determinants being recognized are different.

Prospects for specific immune intervention in T cell-mediated autoimmune disease via anti-idiotypic regulation depend on the degree of diversity of the responder cell antigen receptor repertoire. A highly heterogenous response against self epitopes offers little chance for such regulation. We report here that the Lewis rat autoimmune disease experimental allergic encephalomyelitis, generally considered to be a model of human multiple sclerosis, is caused by T cells that use a limited set of TCR V genes. We have cloned the rat TCR alpha and beta chain cDNAs from the Lewis rat x mouse T cell hybridoma 510, which retains the rat specificity for the encephalitogenic determinant of myelin basic protein (MBP). Using Northern blot analysis of T cell RNA with the cloned V region probes, we have found a specific, and near perfect, correlation between expression of TCR message hybridizing to the V alpha 510 and VB510 probes and specificity for the encephalitogenic determinant of MBP in both T cell hybridomas and encephalitogenic T cell clones. This restricted V gene usage provides a basis for observed idiotypic regulation of auto-reactive T cells, and possible therapy for autoimmune disease. A curious and unexplained observation is that the Lewis rat V alpha/V beta combination that dominates the encephalitogenic response to the 68-88 peptide of MBP is precisely the same V alpha/V beta combination used by the B10.PL mouse response to the encephalitogenic response to the 1-9 peptide of MBP.

Amino Acid Sequence↗
PubMed1989

Major outer membrane proteins unique to reproductive cells of Hyphomonas jannaschiana.

Separation on the basis of molecular weight resolved three proteins specific to the swarmer cell of Hyphomonas jannaschiana. In the reproductive cell, 4 major proteins were identified as cytoplasmic and 10 were identified as envelope. Of these envelope proteins, one was common to both the inner and outer membranes, four were common to the inner membrane, and five were common to the outer membrane. Four of these outer membrane proteins were specific to the reproductive cell, and two of these proteins, with apparent molecular weights of 116,000 and 29,000, constituted 19% of the total cell protein and 54% of the outer membrane protein.

Bacterial Outer Membrane Proteins↗
PubMed1987

Protective effects of ginsenosides on anoxia/reoxygenation of cultured rat myocytes and on reperfusion injuries against lipid peroxidation.

Lactic acid dehydrogenases (LDH) release from cultured rat myocytes during anoxia and reoxygenation were significantly reduced by ginsenosides (G) and its components Rb+Ro, both in 83 micrograms/ml, but not by Rg. G in higher concentration (250 micrograms/ml) enhanced the LDH release. G and Rb+Ro both 8.3 micrograms/ml in the perfusate, also significantly reduced CPK release from isolated rat heart. Rb component 50 mg/kg protected rat myocardial infarct in vivo against lipid peroxidation signified by significant reduction of malondialdehyde (MDA) and increase of superoxide dismutase (SOD) in rat myocardium. It was concluded that G in adequate dose can protect myocyte anoxia and reoxygenation as well as reperfusion damage in isolated rat heart, and the mechanism of protection may partly attribute to its action on oxygen free radical formation and lipid peroxidation. The active components of myocardial protection are Rb+Ro, and not Rg.

Animals↗
PubMedDate not supplied

Determination of two oxy-pyrimidine metabolites of diazinon in urine by gas chromatography/mass selective detection and liquid chromatography/electrospray ionization/mass spectrometry/mass spectrometry.

An analytical method was developed for the determination in urine of 2 metabolites of diazinon: 6-methyl-2-(1-methylethyl)-4(1H)-pyrimidinone (G-27550) and 2-(1-hydroxy-1-methylethyl)-6-methyl-4(1H)-pyrimidinone (GS-31144). Two of the urine sample preparation procedures presented rely on gas chromatography/mass selective detection (GC/MSD) in the selected ion monitoring mode for determination of G-27550. For fast sample preparation and a limit of quantitation (LOQ) of 1.0 ppb, urine samples were purified by using ENV+ solid-phase extraction (SPE) columns. For analyte confirmation at an LOQ of 0.50 ppb, classical liquid/liquid partitioning was used before further purification in a silica SPE column. An SPE sample preparation procedure and liquid chromatography/electrospray ionization/mass spectrometry/mass spectrometry (LC/ESI/MS/MS) were used for both G-27550 and GS-31144. The limit of detection was 0.01 ng for G-27550 with GC/MSD, and 0.016 ng when LC/ESI/MS/MS was used for both G-27550 and GS-31144. The LOQ was 0.50 ppb for G-27550 when GC/MSD and the partitioning/SPE sample preparation procedure were used, and 1.0 ppb for the SPE only sample preparation procedure. The LOQ was 1.0 ppb for both analytes when LC/ESI/MS/MS was used.

Cholinesterase Inhibitors↗