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Biomedical subjects

J Tan

Publications and source records attributed to J Tan.

At least 235 records · Page 13Linked to original sources
PubMed1994

Functional expression and characterization of the assimilatory-type sulfite reductase from Desulfovibrio vulgaris (Hildenborough).

By use of a conjugational transfer method, the gene encoding the assimilatory-type sulfite reductase (ASiR) from Desulfovibrio vulgaris (Hildenborough) has been functionally expressed in Desulfovibrio hosts using the broad-host-range plasmid pDSK519. Production is increased greater than 50-fold relative to natural expression levels. Recombinant enzyme has been characterized by standard biochemical methods, nuclear magnetic resonance, electron paramagnetic resonance, electronic absorption, and activity measurements. It cannot be distinguished from the native enzyme on the basis of spectroscopic characteristics or enzyme activity.

Cloning, Molecular↗
PubMed1994

N-Phosphonomethyl dipeptides and their phosphonate prodrugs, a new generation of neutral endopeptidase (NEP, EC 3.4.24.11) inhibitors.

Inhibitors of the zinc protease neutral endopeptidase (NEP, EC 3.4.24.11) offer significant therapeutic interest as antihypertensives due to their ability to potentiate the biological action of the circulating natriuretic hormone ANF (atrial natriuretic factor). N-Phosphonomethyl dipeptides bearing a central (4-phenyl)phenylalanine residue have been designed to exert potent and selective NEP inhibition. In particular, (S)-3-[N-[2- [(phosphonomethyl)amino]-3-(4-biphenylyl)propionyl]amino]propionic acid (10a) (CGS 24592) displayed high inhibitory potency in vitro (IC50 = 1.9 +/- 0.1 nM) and a long plasma half-life in rats but lacked oral bioavailability. This drawback was overcome by using esterase-sensitive (acyloxy)alkyl phosphonates. More remarkable, several diaryl phosphonate derivatives of 10a also performed as effective prodrugs. Specifically, the structurally simple diphenyl phosphonate 18 (CGS 25462) induced potent inhibition of NEP ex vivo for at least 8 h after oral administration to rats (30 mg/kg). Its antihypertensive effect was demonstrated in DOCA-salt rats. At 30 mg/kg orally, 18 caused a significant reduction in mean arterial pressure measuring -35 +/- 7 mmHg at 5-h postdosing. The alpha-aminomethyl phosphonate 18 represents a new generation of selective NEP inhibitors that combine high potency, long duration of action, and oral bioavailability. Therefore, it holds promise as a novel therapeutic agent for the treatment of human hypertension and congestive heart failure.

Animals↗
PubMed1994

Activation of the leu-500 promoter by adjacent transcription.

The leu-500 mutation is an A-to-G point mutation in the -10 region of the promoter controlling the leuABCD operon of Salmonella typhimurium. Suppression of the leu-500 mutation in an S. typhimurium topA mutant has demonstrated the functional dependency of this mutated promoter on negative supercoiling. A plasmid bearing a minimal leu-500 promoter region (positions -80 to +87) failed to restore its expression in the S. typhimurium topA mutant. We showed that transcription-mediated local negative supercoiling can activate the leu-500 promoter on a plasmid. The coupled transcription and translation process is required for this activation, but peptide-mediated membrane anchorage may not be involved in this activation. Although the effect of negative supercoiling generated during transcription away from the promoter is limited to a short distance of 250 bp, it can activate the negative-supercoiling-dependent leu-500 promoter from positions either 5' or 3' of the leu-500 promoter. In the presence of a parallel-oriented lac promoter which transcribed away from the 3' end of the leu-500 promoter, transcriptional activation of the leu-500 promoter is a strong indication of cooperativity during the transcriptional initiation process.

Base Sequence↗
PubMed1994

Functional and anatomic organization of three-dimensional eye movements in rabbit cerebellar flocculus.

1. The three-dimensional, binocular eye movements evoked by electrical microstimulation of the cerebellar flocculus of alert, pigmented rabbits were recorded using the scleral search coil technique. The components of these eye movements were obtained in reference to an orthogonal coordinate system consisting of a vertical axis and two horizontal axes at 45 degrees and 135 degrees azimuth. The azimuth coordinate was taken to increase to both sides from the 0 degrees reference in the direction of the nose. 2. Eye movements were evoked most readily by stimulation (0.2-ms pulses at 200 Hz for 1 s, intensity < or = 20 microA) at loci in the deep granular layer and the white matter. They consisted of slow (5-20 deg/s) movements. The responses were either binocular, with the eye ipsilateral to the stimulated flocculus usually having the larger amplitude, or were monocular, in which case they were restricted to the ipsilateral eye. 3. The evoked responses were classified according to the combination of the largest measured component of rotation for the two eyes and its sense of rotation (clockwise, CW, or counterclockwise, CCW). Seventy-eight percent of the evoked eye movements could be placed in one of two classes. For one of these classes the largest response component was a short-latency abduction of the ipsilateral eye about its vertical axis (19%), whereas for the other class (59%), the largest response component was a short-latency CCW rotation of the ipsilateral (left) eye about its 135 degrees axis. This response was frequently (50%) accompanied by a smaller short-latency CW rotation of the contralateral (right) eye about its 45 degrees axis. 4. The two main classes of three-dimensional eye movements are associated differentially with anatomically distinguishable compartments that are revealed by acetylcholinesterase histochemistry. Of the five anatomically distinguishable compartments in the floccular white matter, three are predominant. The middle of these three compartments is associated with the vertical axis class of movements, whereas the two adjacent compartments are associated with the 135 degrees class of eye movements. The eye movement relation of the other two, smaller compartments, was not determined. 5. The spatial orientation of the rotation axes of the two main classes of evoked eye movements closely corresponds to that of the preferred axes of the visual climbing fiber input to the flocculus. This suggests that both are organized in a similar coordinate system.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1994

p53 mutations in bladder carcinoma cell lines.

Point mutations and deletions in the p53 tumor suppressor gene occur frequently in advanced stage bladder tumors. To extend these observations to an in vitro model of bladder tumorigenicity, we have evaluated the presence of p53 mutations in a panel of bladder carcinoma cell lines. p53 alleles were cloned using the reverse transcriptase-polymerase chain reaction method, and exons 2-11 were sequenced. Of 11 cell lines examined, 5 cell lines had missense point mutations, and each overexpressed p53 protein on western blot analysis. Except for the HT-1197 cell line, these point mutations occurred in evolutionarily conserved domains, which are characteristic hot spots for mutations. HT-1197 encodes an unusual C-terminal point mutation in codon 365, within the basic motif tetramerization domain, suggesting a linkage between induction of a mutant p53 conformation and alterations in protein oligomerization. Six of 11 cell lines had wild-type levels of p53 expression, with 4 producing p53 proteins having either internal deletions or truncations, and 2 producing wild-type p53. Presence of wild-type p53 was found only in cell lines derived from either a low-grade, papillary tumor (RT4) or fetal bladder (FHs 738Bl). The T24 cell line was found to contain a novel p53 mutant having an in-frame deletion of tyrosine 126. This p53 mutant does not bind SV40 large T antigen, yet is expressed at low levels, comparable to cell lines containing wild-type p53 alleles. Our findings characterize p53 mutations in a panel of bladder carcinoma cell lines, and provide a model for testing the role of wild-type or mutant p53 cDNA to suppress or induce tumorigenic properties.

Animals↗
PubMed1994

[Comparative study of organ storage solutions for rat liver preservation: effects of HX and Collins I solution on sinusoidal lining cells and hepatic functions].

The liver sinusoidal lining cell injury and biochemical function changes were studied with isolated perfusion after rat livers had been preserved in HX solution or in Collins II solution. Seventy inbred Wistar rats were randomly divided into control group (n = 10), group H (preserved with HX solution, n = 30), and group C (preserved with Collins II solution, n = 30). The effects of the storage solutions were assessed by measuring the sinusoidal lining cell mortality (SLCM), Krebs-Henseleit perfusate ketone body ratio (PKBR), the hepatic sugar release (SL), and the number of livers secreting bile during isolated perfusion. The results suggest that the HX solution be much superior to Collins II solution in preventing sinusoidal lining cell injury and maintaining normal hepatic functions.

Animals↗
PubMed1994

[Evaluation of MRI of knee by arthroscopy].

Between August 1989 and April 1993, 61 patients (68 knees) suspected of having intraarticular knee pathology underwent magnetic resonance imaging (MRI) before arthroscopy. In order to evaluate the diagnostic value of the MRI of knee joints, a retrospective study was undertaken. The analysis of the correlation between the arthroscopic findings and the results of MRI indicated the high sensitivity and specificity of MRI in diagnosing disorders of the knee joint. Also the characteristics of MRI of knee and the affecting factors have been discussed in this paper.

Adult↗
PubMed1994

Isolation, mapping, DNA sequence and RFLPs studies of random single-copy DNA segments on human X chromosome.

Using the total human/mouse DNA as the probe, screening has been carried out three times with in situ plaque hybridization to obtain the single-copy DNA sequence from the human X chromosome genomic library. The effective rate of screening is 1.45%. DNAs from clones containing single-copy inserts have been analyzed by a panel of hybrid cells with or without human X chromosome. Three segments, designated by DXFD52,73,75, are mapped to the X chromosome. DXFD52 has been precisely localized on Xq12-q13 with in situ chromosomal hybridization. DXFD52 has been partially sequenced. The results indicate that DXFD52 is a new isolated single-copy segment on the X chromosome. Great progress in the RFLPs study with DXFD52 has been achieved in the population of Chongqing, Sichuan Province. The results show that the DXFD52 can be used to detect the RFLP with Hind III, Bgl II, and Hinf I. DXFD52 will be a potential "landmark" for the construction of the complete linkage map of human genome and the analysis of genomic sequence. And also, the results provide the experience in the development of a useful DNA probe to be applied to diagnosing X-linked disorders.

Base Sequence↗
PubMed1993

Synthesis, cloning and expression of a synthetic gene for high potential iron protein from Chromatium vinosum.

A synthetic gene encoding the peptide sequence for the low molecular weight (M(r) approximately 9600 Da) high-potential iron protein (HiPIP) from the photosynthetic bacterium Chromatium vinosum has been constructed by shotgun ligation of twelve complimentary oligonucleotides varying in size from 42-mers to 48-mers. After cloning the gene into a pET-21d(+) vector, expression of holoprotein in yields of 35 mg/liter of culture was obtained following induction with isopropyl-beta-D-thiogalactoside (IPTG). The recombinant protein was characterized by electronic absorption, 1H NMR, electrochemistry, N-terminal sequencing and amino acid analysis. This is the first example of the expression of a high potential ferredoxin containing a fully constituted [Fe4S4] cluster.

Amino Acid Sequence↗
PubMed1993

A complex response element in intron 1 of the androgen-regulated 20-kDa protein gene displays cell type-dependent androgen receptor specificity.

The androgen-regulated 20-kDa protein gene consists of four exons that code for a major secretory protein of rat ventral prostate. Analysis of its potential cis-acting transcriptional regulatory elements revealed that a large intron 1 region (In-1) had stronger androgen response element (ARE) activity than did the 5'-flanking DNA. In cotransfected CV1 cells, In-1 and its most active subfragment In-1c functioned as AREs but not glucocorticoid response elements (GRE). Nevertheless several ARE/GRE-like partial palindromic sequences are present in In-1c, and it bound both androgen receptors and glucocorticoid receptors in mobility shift assays. A cluster of three ARE/GRE-like sequences contained within a 39-base pair sequence of In-1c had both ARE and GRE activities when analyzed as an isolated oligonucleotide, suggesting that other elements within In-1c determined its ARE specificity. In addition to ARE/GRE-like sequences, In-1c contains putative response elements for the transcription factors AP1, CREB, AP2, OCT-1, C/EBP, and a number of inverted and direct repeats. The ARE specificity of In-1c observed in CV1 cells was diminished in PC3 and HeLa cells transiently cotransfected with an androgen receptor or glucocorticoid receptor expression vector together with an In-1c reporter vector; however, the ARE activity of In-1c was greater than its GRE activity in these cell lines. Interestingly, a 131-base pair subfragment of In-1c retained ARE specificity in all three cell lines.

Amino Acid Sequence↗
PubMed1993

Regulatory elements necessary for termination of transcription within the Ig heavy chain gene locus.

Previous experiments have shown that the extent of delta gene transcription during B cell development is regulated primarily at the transcriptional level. We have shown that deletion of a sequence located between the mu and delta coding regions in the Ig heavy chain locus where transcriptional termination has been previously mapped abrogates the termination. Restoration of termination requires reintroduction of this segment as well as sequence elements within the microM poly (A) site which cannot be substituted by the microS poly (A) site. Recognition of the termination site by non-lymphoid cells suggests that initiation of delta transcription in mature B lymphocytes requires the activation of an anti-termination mechanism not yet developed in early B cells.

Animals↗
PubMed1993

Binding of the ligand [3H]MK-801 to the MK-801 binding site of the N-methyl-D-aspartate receptor during experimental encephalopathy from acute liver failure and from acute hyperammonemia in the rabbit.

Binding of the ligand [3H]MK-801 to the MK-801 binding site of the N-methyl-D-aspartate (NMDA) receptor population on brain homogenates in rabbits was studied during experimental encephalopathy from acute liver failure and from acute hyperammonemia in the rabbit. Homogenates were prepared from brain cortex, hippocampus and striatum. Hepatic encephalopathy was induced by a two-stage liver devascularization procedure and acute hyperammonemia by a prolonged ammonium-acetate infusion; rabbits receiving a sodium-potassium-acetate infusion served as controls. In these animal models extracellular brain glutamate levels are known to be elevated. However no significant alterations in the number nor the affinity of the MK-801 binding sites of the NMDA receptors were found during acute liver failure and acute hyperammonemia. These findings suggest that the NMDA receptor population remains unaltered in experimental encephalopathy from acute liver failure and acute hyperammonemia, despite alterations in extracellular brain glutamate levels.

Acute Disease↗
PubMed1993

Treatment of chronic severe alopecia areata with topical diphenylcyclopropenone and 5% minoxidil: a clinical and immunopathologic evaluation.

BACKGROUND: Topical diphenylcyclopropenone (DPCP) and minoxidil have been used in the treatment of alopecia areata with variable results. OBJECTIVE: This study was designed to evaluate the efficacy of DPCP alone or in combination with topical 5% minoxidil for the treatment of chronic severe alopecia areata. The effect of therapy on cutaneous T-cell and Langerhans cell subpopulations and intercellular adhesion molecule-1 (ICAM-1) expression was also examined. METHODS: Fifteen patients with chronic (more than 2 years), severe (more than 50% scalp involvement) alopecia areata participated in a 24-week trial. Half of the scalp was treated with DPCP once weekly and with either 5% minoxidil solution or a vehicle solution twice daily in a randomized double-blind design. Skin biopsy specimens from each half of the scalp were obtained before therapy and after 12 and 24 weeks of therapy for histologic and immunophenotypic analysis. RESULTS: Thirteen patients completed the study. Five of 13 patients (38%) showed marked regrowth of coarse terminal hair after 24 weeks of treatment with DPCP. The addition of topical 5% minoxidil did not produce any significant clinical benefit in this 24-week trial. Immunophenotypic analysis showed no differences between responders and nonresponders at baseline. During treatment, Leu-4, Leu-2, Leu-3, and keratinocyte ICAM-1 expression were significantly reduced in biopsy specimens of responders versus nonresponders. CONCLUSION: DPCP treatment showed a 38% success rate in producing cosmetically acceptable regrowth in patients with chronic severe alopecia areata.

Administration, Cutaneous↗
PubMed1993

Sympathetic skin response and R-R interval variation in rheumatoid arthritis. Two simple tests for the assessment of autonomic function.

Autonomic nervous system involvement with peripheral neuropathy is a well known complication of rheumatoid arthritis (RA). In the assessment of autonomic function of patients with rheumatoid arthritis, we performed sympathetic skin response (SSR) and R-R interval variation (RRIV) tests in 30 patients and in 30 normal controls. Of 30 patients, 5 had complaints of clinical dysautonomic symptoms. SSRs were abnormal in 6 of 30 patients, whereas 8 showed abnormal RRIVs during deep breathing. Nine of 30 patients also showed electrophysiologic evidence of peripheral neuropathy. All 5 of the patients with clinical dysautonomic symptoms showed abnormal SSR and RRIV test results. No patients with normal results on both tests had clinical dysautonomia. This study showed that there are frequent abnormalities in SSR and RRIV tests in patients with rheumatoid arthritis whether there is a clinical symptom of autonomic dysfunction or not.

Action Potentials↗
PubMed1993

[The role of hemorheologic disturbance in experimental acute pancreatitis].

The role of hemorheologic disturbance in experimental acute pancreatitis was studied in Wistar male rats. It was found that pancreatic duct obstruction with secretin stimulation could induce acute interstitial pancreatitis (AIP), but when high molecular dextran intravenous injection being added, acute necrotizing pancreatitis (ANP) was induced. The authors suggest that the high molecular dextran can increase blood viscosity, aggregation of erythrocytes and decrease erythrocyte deformability, which results in pancreatic microcirculatory disturbances and promotes progressive pancreatic necrosis. Hemorheologic disturbance is one of the key factors in the pathogenesis of ANP.

Acute Disease↗
PubMed1993

[Morphometric evaluation on the pathologic changes in ovariectomized endometria influenced by estrogen use].

Thirty SD female rats were divided into the following groups: (1) sham operation; (2) bilateral ovariectomized (OVX); (3) OVX and estrogen-treated (E). Computerized Image Analyser was used to evaluate the pathologic changes of endometria in these animals. The results showed the endometria of OVX group was in the state of atrophia, while the endometria of E group was in the state of early proliferative stage. The heights of endometria, endometrial epithelia cells and gland epithelia cells, the volume fraction and numerical density of endometrial glands, and the cavity diameter of the glands could effectively express the proliferative changes of the endometria. It is suggested that this method be used to study the pathology of the different endometric diseases.

Animals↗