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Biomedical subjects

R Chen

Publications and source records attributed to R Chen.

At least 433 records · Page 24Linked to original sources
PubMed1995

Microsatellite instability and K-ras mutations associated with pancreatic adenocarcinoma and pancreatitis.

ras oncogene mutations and microsatellite instability (MIN) have been described in pancreatic cancer studies from paraffin blocks and fresh frozen tissue. We sought to determine whether they could be detected in endoscopic retrograde cholangiopancreatography-derived pancreatic juice. ras mutations were detected in the pancreatic juice of 40% (2 of 5) of patients with pancreatic cancer and 2 of 5 patients with pancreatitis. MIN was detected at a single locus in the pancreatic juice of 40% of pancreatic cancer patients and at > or = 2 loci of 100% of pancreatitis patients. The finding of MIN in pancreatitis specimens was verified in studies performed on paraffin blocks. MIN was not detected in normal pancreas controls. All of the cancer patients who had ras mutations in their pancreatic juice also had evidence of MIN at one or more loci (P < or = 0.05), suggesting that MIN is associated with the development of a ras mutation. More importantly, the finding of MIN in pancreatitis specimens suggests that MIN can occur in nonneoplastic conditions of the pancreas and may represent the saturation of an intact mismatch repair system.

Adenocarcinoma↗
PubMed1995

Binding of d-threo-[11C]methylphenidate to the dopamine transporter in vivo: insensitivity to synaptic dopamine.

The regional distribution of [11C]d-threo-methylphenidate in mouse brain was very similar to that of [3H]WIN 35,428 ((-)-2 beta-carbomethoxy-3 beta-(4-fluorophenyl)tropane), and the two radioligands were displaced from striatum similarly after administration of the potent cocaine analog RTI-55 ((-)-2 beta-carbomethoxy-3 beta-(4-iodophenyl)tropane). However, while striatal [3H]WIN 35,428 increased between 5 and 30 min, striatal [11C]d-threo-methylphenidate halved. Thus [11C]d-threo-methylphenidate binds similarly to but more reversibly than [3H]WIN 35,428. The methyl ester of L-DOPA (L-3,4-dihydroxyphenylalanine; 200 mg/kg) plus benserazide plus clorgyline, which markedly elevates rat striatal extracellular dopamine (Wachtel and Abercrombie, 1994, J. Neurochem. 63, 108), decreased the mouse striatum-to-cerebellum ratio for [11C]d-threo-methylphenidate at 30 min by 13% (P < 0.05). In positron emission tomographic (PET) baboon studies [11C]d-threo-methylphenidate binding was insensitive to drugs expected to lower endogenous dopamine. These experiments suggest that normal synaptic dopamine does not compete for binding with [11C]d-threo-methylphenidate, and will not affect PET measures of dopamine transporter availability.

Animals↗
PubMed1995

Fractal dimension of exon and intron sequences.

In this paper, the concept of fractal is applied to describe the features of nucleotide sequences. We introduce the mapping from nucleotide sequences to two-dimensional metric space. Then we use this mapping to study quantitatively the self-similarity of exon and intron sequences in different scales. We find that self-similarity exists in the geometrical range and main range of a nucleotide sequence and define the fractal dimension in these ranges. The results show that the fractal properties of exon sequences are quite different from those of introns, reflecting their difference in structure and function. The fractal dimension of the geometrical range may be used to predict the exon regions of a raw nucleotide sequence.

Animals↗
PubMed1995

Glucose responsiveness of a reporter gene transduced into hepatocytic cells using a retroviral vector.

An MMLV-based retroviral vector containing the chloramphenicol acetyl transferase reporter gene under the control of a glucose-dependent internal promoter derived from the L-type pyruvate kinase gene was constructed. After transfection into psi-CRIP packaging cells, clones producing recombinant retrovirus were selected. These retroviruses were used to infect cultured established hepatocytic cells whose endogenous L-type pyruvate kinase gene is transcriptionally regulated by glucose. In the infected cells, the reporter gene was as responsive to glucose as the endogenous L-type pyruvate kinase gene, and the glucose gene activation was time- and concentration-dependent. The possibility to confer a glucose responsiveness on a transgene carried by a retroviral vector provides a powerful tool in the prospect of gene therapy for diabetes mellitus.

Animals↗
PubMed1995

Identification of a second corticotropin-releasing factor receptor gene and characterization of a cDNA expressed in heart.

Corticotropin-releasing factor (CRF; corticoliberin) regulates the secretion of corticotropin (ACTH) and beta-endorphin and has a broad range of effects on the nervous, endocrine, reproductive, cardiovascular, gastrointestinal, and immune systems. Recently, human, rat, and mouse CRF receptors (CRF-R) have been cloned and functionally and anatomically characterized. We report here the cloning of a second CRF-R cDNA (CRF-RB), which encodes a protein of 431 amino acids, which is 16 amino acids longer and 68% similar to the previously cloned CRF-R, CRF-RA. When transiently expressed in COS-M6 cells, CRF-RB binds CRF with high affinity [Kd = 1.2 (0.57-2.5)nM] and transduces the CRF-stimulated signal of the accumulation of intracellular cAMP, which is inhibited by a CRF antagonist. Comparison of the amino acid sequences of CRF-RB and the previously cloned receptor reveals major differences in the N-terminal domain and in the extracellular loops, whereas the sequences of the intracellular loops are nearly identical. CRF-RB and related transcripts are expressed in the heart, as well as in other tissues, including the gastrointestinal tract, epididymis, and brain.

Amino Acid Sequence↗
PubMed1995

5'-[4-(Pivaloyloxy)-1,3,2-dioxaphosphorinan-2-yl]-2'-deoxy-5-fluorouridine: a membrane-permeating prodrug of 5-fluoro-2'-deoxyuridylic acid (FdUMP).

5'-[4-(Pivaloyloxy)-1,3,2-dioxaphosphorinan-2-yl]-2'-deoxy-5 -fluorouridine (1c) was designed as a potential membrane-permeable prodrug of 2'-deoxy-5-fluorouridine 5'-monophosphate (FdUMP), a putative active metabolite of the antitumor drug 5-fluorouracil (FU). It was anticipated that 1c would be hydrolyzed in vivo by carboxylate esterase (E.C. 3.1.1.1) to the labile 4-hydroxy analogue 2a, which should penetrate cells by passive diffusion and ring open to the aldehyde 3a. Spontaneous elimination of acrolein from 3a would then generate the free nucleotide, FdUMP. 1c might also penetrate cells directly and undergo the same degradation sequence after hydrolysis by cellular esterases. 1c was prepared by condensing 2-hydroxy-2-oxo-4-(pivaloyloxy)-1,3,2-dioxaphosphorinane with 2'-deoxy-5-fluorouridine (FUdR) in the presence of triphenylphosphine and diethyl azodicarboxylate. 1c was moderately stable in aqueous buffers over the pH range 1-7.4 (T1/2 > 30 h). In the presence of carboxylate esterase, however, it was degraded, in a concentration-dependent manner, to FdUMP. No intermediates were detected in the incubation mixture. In mouse plasma, 1c was degraded first to FdUMP and then to FUdR. The latter is presumably formed by dephosphorylation of FdUMP by plasma 5'-nucleotidases or phosphatases. 1c and FU inhibited the growth of Chinese hamster ovary (CHO) cells in culture at a concentration of 5 x 10(-6) M. 1c was equally potent against a CHO variant that was 20-fold resistant to FU. Administered intraperitoneally for 5 consecutive days, 1c was as effective as FU at prolonging the life span of mice bearing P-388 leukemia. In the presence of 2-mercaptoehtanesulfonic acid, an acrolein scavenger, 1c was equally effective against a P-388 mutant cell line that was resistant to FU. Collectively, these data suggest that 1c acts as a membrane-permeable prodrug of FdUMP. This prodrug strategy may be generally useful for introducing dianionic phosphates and phosphonates into cells.

Animals↗
PubMed1995

Phrenic nerve conduction study in normal subjects.

Phrenic nerve conduction studies were performed in 50 phrenic nerves from 25 normal subjects using a technique modified from previously described methods. The normal ranges for latency, amplitude, negative peak area, and duration were established. The latency correlates with age and the amplitude increases with chest circumference. With our method, the amplitude increases and the duration decreases with lung volume. We found good right-left agreement and reproducibility. Therefore, the unaffected side can be used as a reference in unilateral phrenic nerve lesions and previous studies can be used for comparison in serial studies. We recommend that phrenic nerve conduction studies be used routinely to diagnose and monitor patients with respiratory involvement from neuromuscular diseases.

Action Potentials↗
PubMed1995

Bio-affinity characterization mass spectrometry.

A new approach, bio-affinity characterization mass spectrometry (BACMS), aimed at providing a more rapid, sensitive and potentially more flexible alternative to techniques presently employed for the characterization of noncovalent interactions in mixtures, such as would be encountered in combinatorial chemistry, in presented. BACMS avoids some of the difficulties and potential artifacts associated with affinity chromatography since the noncovalent associations occur in solution; thus, BACMS avoids the requirement of solid support media and the development of non-interfering linker species. This paper describes the conceptual basis for the methodology and its potential use in applications which include the screening of high affinity ligands in support of new drug development. BACMS exploits new Fourier-transform ion cyclotron resonance (FTICR) mass spectrometry technologies which, when coupled to electrospray ionization (ESI), allow the investigation of specific noncovalent complexes formed in solution. BACMS utilizes the well-known attributes of FTICR, such as the high resolution mass analysis and (MS)n (n > or = 2) capabilities; however, it is even more directly a result of recently developed techniques involving quadrupolar excitation, such as selected-ion accumulation. These tools are demonstrated and the results illustrate the extraordinary sensitivity achievable (solution concentration of 1 x 10-9 M without the use of separations prior to ESI). Thus, the new capabilities demonstrated here, in conjunction with ESI, will be useful for the investigation of very low relative concentration noncovalent association directly from solution, and promote a faster alternative for combinatorial mixture screening and analysis.

Animals↗
PubMed1995

Long-lasting inhibition of in vivo cocaine binding to dopamine transporters by 3 beta-(4-iodophenyl)tropane-2-carboxylic acid methyl ester: RTI-55 or beta CIT.

Cocaine analogs such as 3 beta-(4-iodophenyl)tropane-2 beta-carboxylic acid methyl ester (RTI-55 or beta CIT) with a higher affinity for the dopamine transporter (DAT) may be potentially useful in interfering with cocaine's actions in brain. This study evaluates the time course of the effects of RTI-55 on cocaine binding in baboon brain using PET and [11C]cocaine. [11C]Cocaine binding was measured prior to, and 90 minutes, 24 hours, 4-5 days and 11-13 days after RTI-55 (0.3 mg/kg i.v.). Parallel studies with [3H]cocaine and RTI-55 (0.5 mg/kg i.v. or 2 mg/kg i.p.) were performed in the mouse. RTI-55 significantly inhibited [11C]cocaine binding at 90 minutes and 24 hours after administration. The half-life for the clearance of RTI-55 from the DAT was estimated to be 2 to 3 days in the baboon brain. In the mouse brain, RTI-55 significantly inhibited [3H]cocaine binding at 60 and 180 minutes after administration and recovery was observed at 12 hours. These results document long-lasting inhibition of cocaine binding by RTI-55 and corroborate that binding kinetics of RTI-55 in striatum observed in imaging studies with [123I]RTI-55 represents binding to DATs.

Animals↗
PubMed1995

Studies of diagnosis and pathogenesis of Wilson's disease.

We have studied the copper and metallothionein (MT) in culture skin fibroblasts of patients and heterozygotes with Wilson's disease (WD) and controls (5 cases each) after incubation in mediums containing various concentrations of copper (C1: 15.74 mumol/L, C2: 78.70 mumol/L, C3: 157.38 mumol/L, C4: 314.76 mumol/L). The results are as follows: 1. In each of the three groups, the copper/protein ratio (Cu/P) in cytosols of 1-5 passages is significantly higher than that of 6-10, 11-15, 16-20 passages. There are no significant differences among 6-10, 11-15 and 16-20 passages. 2. In standard medium, Cu/P in cytosols of three groups are not significantly different, but Cu/P of patients is significantly higher than that of the other two groups after incubation in C4 medium for 12 or 24 hours. 3. After incubation in C1, C2, C3 and C4 mediums respectively, the Cu/P in cytosols of the three groups only increased in C4 medium with time (within 72 hours). It is higher in the patient group than the other two groups. 4. In the three groups, cytosol copper distributed similarly in two peaks after Sephadex G-75 chromatography, which are on high molecular weight (HMW) proteins and MT fractions respectively. It remained the same after incubation in various concentrations of copper. The copper content found in MT fractions in WD patients is much higher than that of controls and heterozygotes. It is even higher after incubation in various concentrations of copper but no changes was found in controls and heterozygotes.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1995

Effect of stopping fluoride administration on the distribution profiles of fluoride in three different kinds of rat bones.

The aim of this work was to explore the reduction of fluoride concentrations in the skeleton after stopping experimental fluoride administration. Fluoride was administered to the rats at varying doses (0, 50, 100 ppm in drinking water) and for different lengths of time (4, 13, 25 weeks). A series of fluoride concentrations across the full thickness of humerus, parietal bone, and vertebra arch in rats were measured by means of an abrasive micro-sampling technique. The distribution profiles of fluoride from periosteal to endosteal surfaces, which were apparently related to the histological structure of these bones, were U shaped in the humerus, V shaped in the parietal bone, and W shaped in the vertebra arch. The average fluoride concentrations in the bones increased significantly with each increasing dose and length of fluoride administration. The relative increments were similar between the different regions or the different bones. After stopping fluoride administration, on the other hand, the relative reduction of the average fluoride concentrations in the bones were 30-100%. They were greatly related to the length after stopping fluoride administration and the dose and length of fluoride administration, but also dependent upon the type of bone and the region examined.

Animals↗
PubMed1995

Chronic pulmonary infection caused by Mycoplasma pneumoniae leading to pulmonary arteriole remodeling and pulmonary hypertension in rats.

The pulmonary arteriole remodeling in Wistar rats with respiratory infection induced by mycoplasma pneumoniae was observed using light microscopy and morphometry. The pulmonary artery pressure (PAP) and index of right ventricular hypertrophy (RVHI) were measured. The intimal and medial hypertrophy can be seen in the pulmonary arterioles, leading to vessel wall thickening and narrowing of the lumina. The total number of the pulmonary arterioles decreased (P < 0.01), and both pulmonary hypertension (Ppa 4.11 +/- 0.19 kPa) and right ventricular hypertrophy (RVHI = 34.96 +/- 3.91%) occurred. In addition, an interstitial pulmonary fibrosis (IPF) was found, in which the content of collagen in the lung tissue changed, i. e., type I collagen increased whereas type III one decreased, and the ratio of type I collagen to type III one increased. It suggested that respiratory infection induced by repeated MP may result in remodeling of pulmonary arterioles and are closely related to pulmonary hypertension.

Animals↗
PubMed1995

Tb3+ binding to human erythrocyte spectrin resulting in conformation change and aggregation.

The Tb3+ binding to spectrin tetramer (SPT) was studied by Tb3+ fluorescence titration and CD spectra. The results indicated that the total high-affinity Tb3+ binding sites are n1 = 330, with average Kd = 3.6 x 10(-6) M. Among them, ca. 90 sites are of higher affinity and are probably more specific to Tb3+ than the remaining sites. There are 520 low affinity Tb3+ binding sites with average Kd = 1.5 x 10(-5) M. Fluorescence and CD spectra revealed that the alpha-helix content of SPT decreased with Tb3+ binding to specific sites and further binding did not result in conformation change. Tb3+ binding to SPT and the subsequent reactions were studied by employing stopped-flow fluorescence and light scattering methods. The studies demonstrate that this is a multistep reaction assembly: high-affinity terbium binding-conformation change-aggregation-low-affinity terbium binding--the second conformation change. The critical Tb3+ concentration-induced spectrin dimer (SPD) aggregation was determined with a light scattering method.

Binding Sites↗
PubMed1995

Isolation, purification, and characterization of immunosuppressive compounds from tripterygium: triptolide and tripdiolide.

Crude extracts derived from the root of Tripterygium wilfordii Hook f (TWHf) have previously been demonstrated to have immunosuppressive properties and have been used as anti-rheumatic therapy in Chinese traditional medicine. Although these extracts contain a large number of chemical components, the precise nature of the compound(s) responsible for this therapeutic effect has not been established with certainty. An aqueous extract of TWHf was resolved into chemical components by medium-pressure and high-performance liquid chromatography. Immunosuppressive fractions were identified with a mixed lymphocyte reaction (MLR) and chemically characterized by nuclear magnetic resonance spectroscopy and mass spectrometry. Two major peaks of immunosuppressive activity were identified. These were the closely related diterpenoid triepoxides, triptolide and tripdiolide. No other immunosuppressive compounds were identified using MLR as the biologic screening assay. Triptolide and tripdiolide may be responsible for the anti-rheumatic properties of crude aqueous extracts of TWHf and represent a novel class of immunosuppressive drugs with potential clinical utility.

Animals↗
PubMed1995

Changes in the characteristics of clients admitted to an Australian therapeutic community, "The Buttery", 1980-92.

Data are presented on the characteristics of clients admitted to an Australian therapeutic community, The Buttery, between 1980 and 1992. The typical client was a 28-year-old male with a primary opioid drug problem complicated by polydrug use, particularly of alcohol and stimulants. Prior treatment experience was common, with one in three having been enrolled in methadone maintenance treatment. The average age of clients increased by about 7 months per year, and there was an increase in the prevalence of alcohol, stimulant and polydrug problems over the period of study. There was also an increase in the exposure of clients to methadone maintenance treatment prior to admission to the Buttery. Overall, the characteristics of patients in this drug-free treatment programme were strikingly similar to those observed among patients in methadone maintenance treatment over the same period.

Journal Article↗