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Biomedical subjects

N Chen

Publications and source records attributed to N Chen.

At least 181 records · Page 10Linked to original sources

Peroxisomal membrane protein PMP68 of mouse liver: cloning of a cDNA encompassing the nucleotide binding fold and epitope mapping of monoclonal antibodies to the expressed protein.

We have isolated and sequenced a cDNA which encodes 376 amino acids toward the carboxy-terminus, and encompassing the putative nucleotide binding fold, of PMP68 (mouse liver peroxisomal integral membrane protein of 68 kDa) the major integral membrane protein of mouse liver peroxisomes. The protein sequence predicted from this cDNA shows 97.9% amino acid identity to this same region of rat liver PMP70, a member of the ATP-binding cassette protein superfamily (K. Kamijo, S. Taketani, S. Yokota, T. Osumi, and T. Hashimoto, 1990, J. Biol. Chem. 265, 4534-4540). The section of the cDNA encoding the hydrophilic and putative cytoplasmic domain of PMP68 was expressed as a recombinant fusion protein in bacteria. Two monoclonal antibodies raised against this protein have been epitope-mapped to peptides generated by cyanogen bromide cleavage of the fusion protein. Antibody 1A4 recognizes a peptide whose sequence contains the first motif of the putative nucleotide binding fold of PMP68, and antibody 8F11 recognizes a carboxy-terminal peptide which includes the second motif of this nucleotide binding fold. These antibodies are expected to be useful in the elucidation of the biological function of this putative membrane transporter.

ATP-Binding Cassette Transporters↗

Enhanced type 2 and diminished type 1 cytokines in neonatal tolerance.

We examined the cytokine profiles associated with tolerance and rejection using the mouse model of neonatal tolerance. BALB/c mice primed with CAF1 splenocytes during the neonatal stage showed increased A/J skin graft survival of > 60 days and failed to develop anti-A/J cytotoxic responses, but rejected third-party C57BL/6 grafts. Lymph node cells that drained A/J grafts on neonatal-primed mice produced allospecific immune cytokine responses characterized by high IL-4 and low IFN-gamma levels. In contrast, lymph node cells that drained either rejected third-party grafts or rejected A/J grafts placed on adult controls produced less IL-4 and more IFN-gamma. Tolerogen-specific immune responses from neonatal-primed mice made up to 100 times higher IL-4 to IFN-gamma ratios than did controls. Alloantigen priming during the immediate neonatal stage induced constitutive expression of IL-4 mRNA in the spleen without IFN-gamma mRNA, whereas alloantigen stimulation during adulthood induced the opposite pattern. IL-4 production from neonatal primed mice was confined to the CD4 population. The altered cytokine profile of enhanced IL-4/IFN-gamma in neonatal primed mice persisted for up to 12 weeks after priming in in vitro secondary MLR assays, which suggests that the initial timing of antigen stimulation critically influenced CD4 maturation. The results support a model of immunoredirection as a mechanism of tolerance and provide rationale for examining the therapeutic use of cytokines in transplantation.

Aging↗

Effect of organic modifier concentrations on electrokinetic migrations in micellar electrokinetic chromatography.

The effect of low concentrations of organic modifiers on the electroosmotic mobility, mueo, and electrophoretic mobility of the micelle, muep, mc, in micellar electrokinetic chromatography (MEKC) with sodium dodecyl sulfate (SDS) solutions was investigated. The results showed that mueo was mainly influenced by the viscosity of the running solution. A linear relationship between mueo and the concentration of the organic modifier was found in MEKC. A newly defined parameter, the mobility ratio, Rm, mc, which is the ratio of the electrophoretic mobility of the micelle to the electroosmotic mobility, and which characterizes the surface charge density of the micelle in MEKC system with organic modifiers, was introduced. A linear relationship between Rm, mc and the organic modifier concentration was observed. The parameter Rm, mc was found to be useful to investigate the changes in the micellar phase with the addition of organic modifier. The measured muep, mc values showed different and interesting characteristics among different organic modifiers.

1-Propanol↗

A quantitative study on the effect of organic modifiers in micellar electrokinetic chromatography.

A quantitative study was made on the effect of organic modifiers in micellar electrokinetic chromatography (MEKC). A linear relationship between 1/k' and organic modifier concentration ([L]) at lower concentration range was derived and experimentally verified. This linearity was also examined at higher concentrations of organic modifier. The results showed that the linear relationship between 1/k' and [L] can only be observed under the situation that no significant change on micellar structure occurs upon the addition of organic modifiers. The mobility ratio of the solute which characterizes the properties of the solute was found to be mainly independent of [L]. A resolution equation for the neutral solutes was also derived in term of mobility ratio.

1-Propanol↗

Effects of long-term acclimatization in lowlanders migrating to high altitude: comparison with high altitude residents.

The physiological response to submaximal and maximal exercise was assessed in lowlanders and Tibetans at low (500 m above sea level) and high altitude (HA, 3,680 m). The times spent at HA by the lowland migrators was 8 days (n = 60), 7 months (n = 60, same group), 15 months (n = 29) and 27 months (n = 29). After the 15-month stay at HA, the maximal oxygen uptake (VO2max) and maximal heart rate of the lowland migrators almost reached those of the HA native residents (Tibetans, n = 57), but their total work capacity and the gross efficiency (eta) of mechanical work remained lower than those of the Tibetans. The rate of VO2max achieved at 90 W by the Tibetans was lower than that of the lowland migrators. It was concluded that, at HA, the lowlanders regained much of the aerobic capacity which they had lost initially. However, they did not attain the same gross mechanical efficiency as the Tibetans, who seemed to be at an advantage in respect of work at HA.

Acclimatization↗

Experimental topical tetracycline-induced neuritis in the rat.

PURPOSE: Recent studies have reported that tetracycline may induce chronic inflammatory responses in the adjacent soft tissues. The purpose of the present study was to determine if dry, powdered tetracycline evoked an inflammatory response in nerve tissue. MATERIALS AND METHODS: Twenty male Sprague-Dawley rats were anesthetized and bilateral mental nerves were exposed. Half of the 40 nerves were intentionally injured by removing the epineurium, and the remaining nerves were uninjured. In a prospective, randomized, double-blind manner, equal volumes of color-matched, precoded tetracycline powder or Gelfoam (Upjohn, Kalamazoo, MI) powder control was placed in intimate contact with the nerves and sealed in polyethylene tubes. Forty-five days later the animals were killed and 5-micron sections of the nerves were prepared for histologic examination. The presence of epineurium, fascicular number, and fascicular area were measured. An inflammatory response in nerve tissue and the intrafascicular and epifascicular inflammatory index was measured by counting darkly stained nuclei with a Jandel Video Analysis System (Jandel Scientific, Corte Madera, CA). RESULTS: Experimental injury (ie, epineurectomy) of nerves resulted in a statistically significant increase in fascicular number (P = .034), but not in fascicular area. For the intrafascicular inflammatory index, there was a significant main effect for drug (P = .002) and injury (P = .002). Experimental injury in both Gelfoam control and tetracycline grouped nerves resulted in an increase in intrafascicular inflammation. There was no significant increase in intrafascicular inflammation in either Gelfoam control or tetracycline grouped nerves when the epineurium was intact. The combination of both tetracycline and epineurectomy resulted in the largest increase in intrafascicular inflammation found among the groups studied. For epifascicular inflammatory index, there was a significant main effect for drug (P = .003) and injury (P = .001) that mirrored the intrafascicular inflammatory pattern. CONCLUSION: The results of the present study suggest that dry, powdered tetracycline accentuates the inflammatory response in intrafascicular and epifascicular nerve tissue following epineurectomy. The maintenance of epineurium inhibited the inflammatory response in intrafascicular and epifascicular nerve tissues. Gel-foam was an inert material when placed against exposed nerve tissue. These findings suggest that tetracycline should not be placed in extraction sockets when injury of the inferior alveolar and/or lingual nerves are present.

Administration, Topical↗

Echographic evaluation of primary cysts of the iris pigment epithelium.

PURPOSE: Primary cysts of the iris pigment epithelium frequently appear to be focal elevations of the peripheral iris. To differentiate them from other anterior segment tumors requires the ability to assess the distribution, tissue characteristics, and progression of such lesions, which we accomplished by using anterior segment echography. METHODS: We reviewed the medical records of 49 consecutive patients referred to the Doheny Eye Institute for echographic evaluation of focal bulging of the peripheral iris. Echography of the anterior segment was performed on each patient by using an immersion technique. Clinical examination was also performed on 20 (41%) patients at the time of initial examination. RESULTS: We identified 52 iris pigment epithelial cysts in 52 eyes of the 49 patients studied. All lesions were single, localized to the posterior iris surface, and demonstrated a well-circumscribed, echolucent structure on B-scan echography. Average maximal sagittal thickness was 1.6 +/- 0.4 mm. Cysts were most frequently located in the inferotemporal quadrant (31%). Minimal increase in size or regression of lesions was noted in the nine patients (ten eyes) for whom follow-up echographic examinations were available. CONCLUSIONS: Primary iris pigment epithelial cysts possess characteristic features with regard to location, size, and internal structure, which can be accurately documented with echography. Clinically evident lesions tend to be between 1 and 4 mm in maximum sagittal thickness, conform to the contour of posterior chamber structures without distortion, produce anterior bulging of the peripheral iris, are preferentially located in the inferotemporal quadrant, and demonstrate limited potential for growth.

Adolescent↗

A Giardia duodenalis gene encoding a protein with multiple repeats of a toxin homologue.

A gene, CRP136, from the intestinal protozoan parasite Giardia duodenalis, expressed at a high level in a metronidazole-resistant line, encodes a 136 kDa protein with 23 copies of a 40 amino acid repeat. The protein is cysteine-rich and has the typical membrane-spanning region and CXXC amino acid motifs of a family of Giardia cysteine-rich surface proteins (CRSPs). The repeat unit in CRP136, shares 57% homology with the gene encoding the precursor of the sarafotoxins, a group of snake toxins from the burrowing adder known to cause symptoms similar to those of humans acutely infected with Giardia. The sarafotoxins are low molecular weight sulphydryl cross-linked peptides which are proteolytically cleaved from a precursor polyprotein. CRP136 has homology over the entire length of the sarafotoxin precursor, and the repeats are of the same length. Thus CRP136 represents the first evidence for a potential Giardia toxin. The genomic copy number of CRP136 appears to be the same in both the parent and drug-resistant lines and expression of this gene, and at least one other, is associated with a conserved partial duplication, but not amplification, of one chromosome.

Amino Acid Sequence↗

Orthogonal array design as a chemometric method for the optimization of analytical procedures. Part 5. Three-level design and its application in microwave dissolution of biological samples.

The theory and methodology of a three-level orthogonal array design as a chemometric method for the optimization of analytical procedures were developed. In the theoretical section, firstly, the matrix of a three-level orthogonal array design is described and orthogonality is proved by a quadratic regression model. Next, the assignment of experiments in a three-level orthogonal array design and the use of the triangular table associated with the corresponding orthogonal array matrix are illustrated, followed by the data analysis strategy, in which significance of the different factor effects is quantitatively evaluated by the analysis of variance (ANOVA) technique and the percentage contribution method. Then, a quadratic regression equation representing the response surface is established to estimate each factor that has a significant influence. Finally, on the basis of the quadratic regression equation established, the derivative algorithm is used to find the optimum value for each variable considered. In the application section, microwave dissolution for the determination of selenium in biological samples by hydride generation atomic absorption spectrometry is employed, as a practical example, to demonstrate the application of the proposed three-level orthogonal array design in analytical chemistry.

Chemistry Techniques, Analytical↗

[Inclusion compounds of jiuxin oil with beta-cyclodextrin].

The inclusion compounds of Jiuxin Oil with beta-cyclodextrin were prepared. The influence of temperature, ratio of host and guest molecules, stirring time on inclusion were observed. The quantitative determination was performed by gas chromatography. Inclusion compounds of Jiuxin Oil were confirmed by differential scanning calorimetry and powder X-ray diffractometry.

Cyclodextrins↗

Effects of linear polyacrylamide concentrations and applied voltages on the separation of oligonucleotides and DNA sequencing fragments by capillary electrophoresis.

Oligonucleotides and DNA sequencing fragments have been separated by capillary electrophoresis employing linear polyacrylamide (LPA) as a sieving matrix. A commercially available apparatus equipped with a laser-induced fluorescence (LIF) detection system has been utilized, but the capillary cartridge has been modified to position the capillaries without coiling. The performance of the separation, the relationship between resolution and analysis time, has been examined using poly(dT)16-500 by changing LPA concentration, capillary length, and electric field strength. It was found that, for large DNA fragments, the migration time interval between bands decreases linearly as DNA fragment size increases. This implies that there exists a maximum base number to be resolved, irrespective of the band width (we named the maximum base number Nmax). The higher value of Nmax is obtained when the applied field strength is lower, but this accompanies longer analysis time with a concomitant increase in band width. Simple experimental equations have been proposed to calculate resolution and migration times of DNA fragments separated in our system at given electrophoretic conditions. Using 9% T LPA and an electric field strength of 100 V/cm, single-base resolution of M13mp10 DNA fragments up to 520 nucleotides has been obtained.

Acrylic Resins↗

Retinoblastoma protein expression is frequently altered in chronic lymphocytic leukemia.

The detection of abnormalities at the retinoblastoma (RB) locus by cytogenetics, Southern blot, and fluorescence in situ hybridization studies suggests that the RB gene has a role in chronic lymphocytic leukemia (CLL). To further study this role, we determined the level of RB protein present in the mononuclear cell fraction derived from peripheral blood or bone marrow samples from 74 patients with CLL, by Western blotting. Compared to similarly prepared samples from the peripheral blood of normal individuals, the level of RB in CLL cells was less than normal in 42% of patients, equal to normal in 22% of patients, and in excess of normal in 36% of patients. Regardless of whether the source of the sample was blood or marrow or if the patients were untreated or previously treated, similar rates of low, normal, and elevated RB levels were observed. RB protein in the CLL patient samples was never phosphorylated. RB levels showed no correlation with the lymphocyte doubling time or with proliferating cell nuclear antigen levels. Low RB levels could arise from genetic alterations of the RB gene or altered regulation of expression. To determine which was occurring, we stimulated the cells from 27 CLL patients in culture with either phytohemagglutinin or pokeweed mitogen in an attempt to induce RB expression and phosphorylation. Among patients with low levels of RB, expression was induced in 46% (6 of 13), and phosphorylation of RB was seen in 31% (4 of 13). Increased expression of phosphorylated RB was induced in 80% (4 of 5) of patients with normal levels of RB and in 78% (7 of 9) of patients with high levels of RB. This study demonstrates that absent RB expression occurs commonly in patients with CLL. Intrinsic abnormalities of the RB gene may be present in those patients with low levels of RB that could not be stimulated by mitogens, while regulatory abnormalities located in trans to the RB gene may occur in the other half. Given the importance that RB levels play in other cancers, the prognostic implication of low RB levels should be studied in CLL.

Blotting, Western↗

Physical map of a 2 Mb chromosome of the intestinal protozoan parasite Giardia duodenalis.

The protozoan parasite, Giardia duodenalis, is regarded as the most primitive eukaryote. The two apparently identical nuclei presumably carry the same chromosomes but the number of different chromosomes in the organism is unknown. A genome map of G. duodenalis is required to resolve this issue and mapping studies were initiated using chromosome 5. This chromosome was estimated to be approximately 2 Mb when Giardia chromosomes were separated by contour-clamped homogeneous electric field gel electrophoresis. A plasmid library of chromosome 5-specific DNA sequences was constructed from gel-extracted chromosome 5 and selected probes were used as markers to identify NotI DNA segments derived from chromosome 5. Fifty-nine unique copy markers were used to identify thirteen NotI segments which ranged in size from 47 kb to 400 kb. The sum of the NotI segments was 1.78 Mb which indicated that most, if not all, of the chromosome was accounted for and that chromosome band 5 of the cloned line WB-1B, used in this study comprised only one chromosome type. The NotI segments were ordered on the map by comparison of hybridization patterns of the markers with partial NotI cleavages of whole chromosomes. Chromosome rearrangements occur readily in Giardia, and in two drug-resistant lines selected for resistance to different drugs, partial conservative duplications of chromosome 5 were observed in addition to the original, full length chromosome 5. Both duplications retained the central region of chromosome 5 but were deleted at different termini resulting in one duplication of 1.5 Mb and the other of 1.3 Mb.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of chorda-lingual nerve injury and repair on human taste.

Citric acid detection threshold and magnitude response were measured on the anterior tongue in 10 patients with unilateral chorda-lingual nerve transections before and after repair. Fungiform taste buds were analysed by videomicroscopy. Preliminary data suggests that humans can regenerate fungiform taste buds and recover some taste sensitivity after repair.

Adult↗

Transcription of a human neurotropic virus promoter in glial cells: effect of YB-1 on expression of the JC virus late gene.

We have isolated a partial recombinant cDNA clone from a HeLa expression library which encodes a protein capable of binding to the central region of the human neurotropic JC virus (JCV) enhancer/promoter, termed the B region. Sequence analysis revealed a complete homology of the partial cDNA clone to the N-terminal region, of a previously described DNA-binding protein, termed YB-1. Band shift analyses have indicated that the bacterially produced YB-1 interacts specifically with the double-stranded B oligonucleotide as well as the corresponding single-stranded DNA fragment representing the early promoter sequence. Further analysis indicated that the YB-1 protein binds specifically to the C/T-rich sequence of the B domain, which is located in close proximity to the TATA box within the virus enhancer/promoter. Results from cotransfection experiments demonstrated that the full-length (YB-1) but not the partial cDNA enhances expression of the JCV late (JCVL) promoter in glial cells. Cointroduction into glial cells of a recombinant expressing the YB-1 and JCVL deletion mutants indicated that removal of the C/T-rich sequence of the B domain reduces the level of activation of the virus promoter by YB-1. Further cotransfection experiments revealed that the virus transactivating protein T antigen appears to diminish the ability of YB-1 to activate JCVL gene expression. RNA studies indicated that YB-1 is expressed in several cell types and tissues. Examination of YB-1 RNA from mouse brain at various stages of development revealed high levels of YB-1 RNA at early stages of development and lower levels at all subsequent developmental stages.

Antigens, Viral, Tumor↗

Prevention of paraquat toxicity in suspensions of alveolar type II cells by paraquat-specific antibodies.

1. The herbicide, paraquat, is accumulated by the energy-dependent polyamine uptake pathway of alveolar type II cells. There it undergoes redox cycling that results in an amplified production of toxic reactive oxygen species and depletion of NADPH and other reducing equivalents. These processes account for the lung being the major target organ for paraquat toxicity. 2. We postulated that paraquat-specific antibodies would inhibit the uptake of the herbicide by type II cells and prevent its toxicity. Accordingly, we examined the effects of paraquat-specific monoclonal antibodies and Fab fragments on the uptake, efflux and cytotoxicity of 50 microM paraquat in suspensions of alveolar type II cells isolated from the rat. 3. The uptake of paraquat was linear over 40 min. Over this time, the uptake rate was inhibited significantly (% inhibition, 73-89) by IgG (25 or 50 microM) or Fab fragments (50 or 100 microM). 4. The apparent efflux rate of paraquat, studied over 16 h, was increased significantly from 0.12 h-1 for the control cells in medium to 0.17 h-1 by paraquat-specific Fab fragments but was unaffected by the specific IgG. 5. Cytotoxicity was determined by measuring the release of 51Cr from the cells. The cytotoxicity of 50 microM paraquat was decreased significantly (percent decrease, 56-80%) in the presence of specific antibodies. 6. These studies in vitro suggest some potential for immunotherapy in selected cases of paraquat poisoning.

Animals↗

[Changes in serum lipoprotein cholesterol in rabbit model of calcium bilirubinate cholangiolithiasis].

The lithogenic process of cholangiolithasis is still not understood clearly. In this study, several serum lipoprotein cholesterol, the bile BA level, etc. in the rabbit model of calcium bilirubinate cholangiolithiasis was examined. The main results included: (1) The concentrations of HDL-ch and HDL2-ch in the serum decreased in all the experimental groups (P < 0.01; P < 0.05), except HDL-ch in the BO 1W group (P > 0.05) compared with the control group. The concentrations of HDL2-ch in the BOI 1W and the BOI 6W groups were lower than those in the BO 1W and the BO 6W groups respectively (P < 0.05); the both concentrations of Fch and HDL3-ch in the BOI and the BO groups were much higher than those in the control group (P < 0.01). (2) The bile T-BA level was lower in the both BOI groups than that in the control group (P < 0.05). It began to decrease in the BO group during the first week, but it was not significant in statistical difference (P > 0.05). During the 6th week the level of the bile acid (BA) was much lower than that in the control group (P < 0.05). According to the above results, it may be concluded that in the formation of calcium bilirubinate gallstones, caused by the biliary obstruction and infection, there is a disorder in the lipoprotein metabolism.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗