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Biomedical subjects

S L Chen

Publications and source records attributed to S L Chen.

At least 109 records · Page 6Linked to original sources
PubMed1996

[The study on the in-vitro stability of Tc(V)-99m dimercaptosuccinic acid].

Tc(V)-99m DMS, developed by Yokoyama et al. in 1981, has been recognized to be advantageous for the scintigraphic diagnosis of various malignant tumors and their metastasis, the aim of this study is to assess the in-vitro stability of Tc(V)-99m DMS. Thin-layer chromatography, including paper chromatography and silica gel thin layer chromatography, is performed to determine the change of radiochemical species presented in the reconstituted solution of Tc(V)-99m DMS prepared from the DMS kit (Institute of Nuclear Energy Research, Atomic Energy Council R.O.C.) and the commercial DMSA kit (Nephroscint, IRE CELLTARG Radiopharmaceuticals Japan). The bioscan imaging scanner is used to measure the Rf value and labeling efficiency of radiochemical species on the chromatographic strip. The in-vitro stability of Tc(V)-99m DMS prepared from the DMS kit and the commercial DMSA kit is studied by examining various parameters which include temperature(degree C) and time(hr) after reconstitution. The results show that the in-vitro stability of Tc(V)-99m DMS prepared from the DMS kit is actually better than that from the commercial DMSA kit. The one-step labeling method of DMS kit is much simpler than the two-step labeling method of DMSA kit.

Drug Stability↗
PubMed1996

Respiratory distress and selective muscle involvement in central core disease: report of a case.

A 10-year-old girl who had suffered life-threatening respiratory distress in late infancy and mild muscle weakness in the extremities was found to have central core disease (CCD). Her developmental milestones were markedly delayed and she became ambulant at the age of 6 years. On physical examination, she had a high-arched palate, nasal tone vocalization, a positive Gowers' sign, waddling gait, and decreased deep tendon reflexes. On magnetic resonance imaging (MRI) examination, the thigh, calf, arm, and paravertebral muscles were selectively involved, especially in the paravertebral, biceps brachii, gluteus maximus and medius, vastus lateralis and medialis, sartorius, adductor magnus, tibialis anterior, soleus, lateral head of gastrocnemius, and peroneus longus muscles. Two biopsy specimens from the left biceps brachii and rectus femoris muscles showed the common histochemical findings of CCD, including type 1 fiber atrophy and predominance, and core structures. Proliferation of adipose tissue was only present in the biceps brachii muscle. The patient improved clinically in muscle strength after three year follow-up. Muscle MRI may provide a non-invasive evaluation of gross muscle involvement in CCD.

Arm↗
PubMed1996

Cloning of the tryptophan operon of Brevibacterium divaricatum and its expression in E. coli.

A genomic bank from Brevibacterium divaricatum has been prepared using lambda EMBL3 as a vector. The genomic bank's titers are 2.2 x 10(6) pfu/micrograms. Through screening by plaque hybridization, a 9.6 kb NcoI fragment which contains the entire trp operon has been isolated. Polymerase chain reaction amplification and restriction endonuclease analysis of PCR fragments indicated that there is homology between the coryneform bacteria; however, some genetic diversity among the species still exists. By complementation tests using subcloning of the 9.6 kb NcoI fragments and various E. coli tryptophan auxotrophs, this fragment was found to contain a gene cluster composed of trpE, trpD, trpC, trpB and trpA in this order. This revealed that the tryptophan biosynthesis genes in B. divaricatum may be an operon.

Bacterial Proteins↗
PubMed1996

Progesterone stimulates in vitro release of prolactin and thyrotropin involving cAMP production in rat pituitary.

Effects of progesterone administration in vivo or in vitro on the release of pituitary prolactin (PRL) and thyroid stimulating hormone (TSH) in vitro, as well as the generation of pituitary cyclic adenosine 3', 5'-monophosphate (cAMP) were studied in ovariectomized (Ovx) rats. Ovx rats were either decapitated or injected subcutaneously with progesterone, or oil for 3 days before decapitation. The anterior pituitary glands (APs) of progesterone treated Ovx rats were incubated with Locke's medium containing thyrotropin-releasing hormone (TRH, 10 nM) and/or 3-isobutyl-1-methylxanthine (IBMX, 1 mM) at 37 degrees C for 30 min. The APs of untreated Ovx rats were incubated in vitro with IBMX and progesterone (0, 0.1, 1, 10 nM). Progesterone increased spontaneous and TRH-induced release of PRL and TSH in vitro. Administration of progesterone, either in vivo or in vitro, increased the production of pituitary cAMP following incubation of IBMX. These results suggest that progesterone increases the release of PRL and TSH through a mechanism associated with an increase of cAMP production in rat APs.

Animals↗
PubMed1995

Human papillomavirus 11 E5a delays the growth restriction induced by temperature shift in temperature-sensitive simian virus 40 T antigen-immortalized keratinocytes.

The transfection with human papillomavirus type 11 E5a oncoprotein can cause longer and more active proliferation in the human keratinocytes previously transfected stably with temperature sensitive SV40 T antigen at 39 degrees C. Also, after the E5a transfection in parental temperature sensitive SV40 T antigen gene transfected cells, we observe a delay in the accumulation of p21 gene at 39 degrees C. Moreover, T antigen degradation did not occur at 39 degrees C in the E5a transfected cells as it did in the parental cells. We draw from these observations that E5a may transiently stabilize temperature sensitive T antigen and/or repress p21 synthesis.

Antigens, Polyomavirus Transforming↗
PubMed1995

Differential regulation of cyclin A, cyclin B and p21 concentrations in a growth-restricted human fibroblast cell line.

When the culture temperature was shifted from 35 degrees C to 39 degrees C, human fibroblasts immortalized by the temperature-sensitive simian virus 40 T antigen became larger and acquired the morphological characteristics of senescent fibroblasts. After culture at 39 degrees C for 48 h, most cells had ceased to proliferate. A rapid depletion of cells with S-phase DNA content was observed after the temperature shift. To elucidate the mechanism governing this rapid arrest of proliferation, we studied the expression of genes involved in the regulation of cell cycle progression. Cyclin A, cyclin B and p34cdc2 concentrations were not changed during growth restriction, whereas p21 was rapidly induced in these growth-restricted cells. Transient expression of exogenous p21 in cells cultured at 35 degrees C led to growth restriction and morphological changes characteristic of senescence. Furthermore, we studied the reversibility of growth restriction induced by the temperature increase. The results showed that senescent morphology and growth arrest were not reversible. In these cells the p21 concentration remained high and p34cdc2 remained undetectable. This indicates that p21 accumulation might be responsible for the maintenance of senescence. Our findings provide information on the use of growth restriction of immortalized fibroblasts induced by a temperature shift as a model system to study senescence.

CDC2 Protein Kinase↗
PubMed1995

Protein kinase C-and ras-dependent activation of c-jun gene by human papillomavirus type 11 E5a oncoprotein.

E5a of HPV-11 is a transforming oncogene. Previously, we have shown that E5a constitutively activates the expression of protooncogene c-jun by transcriptional regulation through the AP-1 binding site in the c-jun promoter. In the present study, we used two different types of cells: the E5a transfected NIH 3T3 cells and human epidermal keratinocytes, and selectively inhibited different signal transduction pathways to investigate effects of E5a on c-jun expression. We find that protein kinase C and ras-dependent pathways are important for the c-jun induction by E5a, but not the cAMP-dependent pathway.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1995

Trace element concentration and arsenic speciation in the well water of a Taiwan area with endemic blackfoot disease.

Blackfoot disease is a peripheral vascular disease resulting in gangrene of the lower extremities. Although extensive epidemiological study has implicated high arsenic content in artesian well water in the endemic area, there is more to learn about the etiology of the disease. In this study, effort is paid on multielement determination and arsenic speciation in order to find out whether the trace element concentration pattern in well water in the Blackfoot disease endemic area is different from those of two control areas. Experimental results indicate that the concentrations of Fe, P, Na, and Ba in well water in the Blackfoot disease endemic area are found to be significantly higher than those of the controls, but they are still below the drinking water standard. The total arsenic in well water in the endemic area (671 +/- 149 ppb) is much higher than that of one normal control area of Hsin-Chu (< 0.7 ppb), but is a similar level as that of other control areas of I-Lan (653 +/- 71 ppb) where no Blackfoot disease has ever been found. It was also found that the insoluble arsenic in the endemic area (21.9 ppb) is much higher than that in two control areas (< or = 1.8 ppb), and the concentration ratio between As(III) and As(V) species in the endemic area (2.6) is much lower than that in one of the control areas, where the total arsenic is also high (14.7). The possible connection of Blackfoot disease with trace elements, arsenic species, and possibly other as yet undefined environmental factors in the artesian well water, is discussed.

Arsenic↗
PubMed1995

Myocardial metabolism of 123I-BMIPP in patients with hypertrophic cardiomyopathy: assessment by radial long-axis SPET.

To elucidate the metabolism of free fatty acid in the myocardium of hypertrophic cardiomyopathy (HCM), SPET was performed with 123I-15-(p-iodophenyl)-3-R,S-methylpentadecanoic acid (BMIPP), an analogue of free fatty acid, and with 201Tl in nine patients with HCM and eight healthy volunteers who acted as controls. For quantitative analysis of the apical area, a radial long-axis tomogram was reconstructed every 6 degrees after short-axis tomography. The relative regional uptake (RRU, %) in each segment was obtained by normalizing to the maximal value among all segments. The 3-h washout rate was calculated both for BMIPP and for 201Tl. The ratio of the RRU of BMIPP to that of 201Tl was significantly lower in the apical and antero-septal regions compared with other walls (P < 0.01) in the patients with HCM, whereas uptake of both BMIPP and 201Tl in the controls was homogeneous and there was a significant correlation between them. The coefficient of variation (CV) in all segments in the HCM patients was significantly higher (P < 0.01) than that of the controls both BMIPP and 201Tl, indicating inhomogeneous uptake of BMIPP in the HCM patients. The washout rate of BMIPP over 3 h was significantly higher in the patients with HCM (mean +/- S.D. 26.3 +/- 7.3%) than the controls (18.3 +/- 7.5%, P < 0.05). The ratio of segmental washout rate of BMIPP to that of 201Tl in the HCM patients was increased, especially in the septum and apex.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1995

Differential induction and regulation of c-jun, junB, junD and c-fos by human papillomavirus type 11 E5a oncoprotein.

The E5a gene of human papillomavirus type 11 (HPV-11) is a transforming oncogene. In this study, we investigated the mechanism of E5a induced transformation. Our results show that the expression of c-jun and junB, but not junD, was activated by HPV-11 E5a in NIH 3T3 cells and human epidermal keratinocytes. However, the expression of c-fos was activated by E5a in NIH 3T3 cells, but not in keratinocytes. We further investigated the mechanism of c-jun and junB induction by E5a. The amount of c-jun and junB RNAs correlated with the amount of E5a RNA in the heavy metal inducible system. E5a constitutively activated the expression of c-jun and junB at the initiation of transcription level. In addition, analyses of the effect of serum on c-jun expression in E5a transformed human epidermal keratinocytes show that EGF might have a stimulatory effect on c-jun gene expression in E5a expressing keratinocytes.

3T3 Cells↗
PubMed1995

Synthesis, DNA binding, and sequence specificity of DNA alkylation by some novel cyclic peptide-chlorambucil conjugates.

In an effort to investigate the potential of cyclic peptides as carriers for cytotoxic agents, we synthesized four cyclic peptide-chlorambucil conjugates: cyclo[Lys(CHB)-Lys(CHB)-Gaba-] (peptide Y), cyclo[Lys(CHB)-Gly-Lys(CHB)-Gaba-] (peptide A), cyclo[Lys(CHB)-beta-Ala-Lys(CHB)-Gaba-] (peptide B) and cyclo[Lys(CHB)-Gaba-Lys(CHB)-Gaba-] (peptide C). The cyclic peptides were synthesized by coupling protected amino acid residues in solution and the subsequent cyclization was performed by the pentafluorophenyl ester method as described previously (Sheh et al., 1990, 1993a,b). After deblocking the lysyl-carbobenzyloxy protecting group (Z), the conjugation was achieved by reaction with the pentafluorophenyl ester of chlorambucil (CHB). These cyclic peptides differ from one another in ring size and are disubstituted with CHB via the epsilon-amino group of the lysyl residue. The various conjugates were designed to study the effect of ring size on the mode of DNA binding and alkylation. A DNA-binding assay using lambda-DNA with ethidium bromide showed that whereas peptide Y and CHB have no observable binding affinity, the apparent binding constants for peptide A, peptide B and peptide C on lambda-DNA were determined to be 2.36 x 10(5), 1.27 x 10(5) and 3.50 x 10(5), respectively. Thus, it is suggested that cyclic peptides bearing aliphatic side chains attached to a ring larger in size than 14 members would be more favourable as regards augmenting the binding affinity. DNase I footprinting showed that no footprinting patterns were observed for the 253-mer fragment and 117-mer fragment with peptide A, but two new bands corresponding to G69 and G80 were observed for the 117-mer fragment. DNA alkylation studies using a piperidine cleavage assay on the 117-mer DNA fragment showed that the sequence selectivity, judged by reaction intensity observed with peptide A, peptide B and peptide C, was similar to that seen with CHB alone. The selectivity of alkylation for both CHB and its peptide derivatives appears to be: 3'-Pur-G-Pyr-5' > 3'-Pyr-G-Pyr-5' > 3'-Pyr-G-Pur-5' = 3'Pur-G-Pur-5'. However, there are apparent differences in the intensity of alkylation by peptides A, B, C and CHB at certain guanine residues.

Alkylation↗
PubMed1994

Retinoic acid represses the gene expression of topoisomerase II in HEP3B cells.

All-trans-retinoic acid (RA) affects cell growth and regulates gene expression. We examined the expression of topoisomerase 11 gene in Hep3B cells treated with RA. At low RA concentration which did not significantly affect the growth rate of Hep3B cells, RA inhibited the synthesis of topoisomerase II mRNA as revealed by Northern analysis and nuclear run-on analysis. These results indicated that the repression of topoisomerase II gene expression could be directly induced by RA rather than was a secondary event which occurred after cell growth was inhibited by RA. An unexpected finding is that after up to 72 h continuous exposure to RA, the topoisomerase II protein concentration remained unchanged.

Blotting, Northern↗
PubMed1994

Antisense oligodeoxynucleotides to c-jun inhibits proliferation of transformed NIH 3T3 cells induced by E5a of HPV-11.

E5a of HPV-11 is a transforming oncogene. Previously, we had shown that the E5a gene is required only for the initiation of transformation; c-jun might be involved in the maintenance of transformation. In this study, we exposed E5a transformed NIH 3T3 cells to antisense oligodeoxynucleotides complementary to the 24 nucleotides corresponding to the translation initiation site of the c-jun gene, and examined the effects of this treatment on cell proliferation. Results show that antisense c-jun oligodeoxynucleotides could repress c-jun production and inhibit cell proliferation in E5a transformed cells.

3T3 Cells↗
PubMed1994

Responses of NOD congenic mice to a glutamic acid decarboxylase-derived peptide.

Type 1 diabetes in man and the NOD (H-2g7) mouse is frequently associated with an autoimmune response to two isoforms of glutamic acid decarboxylase (GAD), GAD65 and GAD67. GAD-specific autoantibodies produced by B cells and GAD-specific T cells have been observed in both species. In the current study, the response to a GAD65-derived peptide, GAD65 524-543, previously reported to be an epitope recognized by spleen cells obtained from 3-week-old NOD mice, was assessed in NOD MHC and non-MHC congenic strains. Although spontaneous reactivity to GAD65 524-543 was not observed in NOD mice, the peptide was immunogenic in NOD mice as well as in two NOD congenic strains which are both H-2g7, B10.H-2g7 and NOD.B6Il2-Tshb. This was surprising since the B10.H-2g7 strain does not develop diabetes or insulitis and fewer than 3% of NOD.B6Il2-Tshb mice develop diabetes. The response to GAD65 524-543 was shown to be controlled by the MHC since neither the B10 nor the NOD.H-2b strain, both of which are H-2b, responded to the peptide. This study demonstrates that T cell responsiveness to GAD-derived peptides can be elicited in strains of mice that are resistant to the development of spontaneous diabetes, suggesting that peripheral tolerance to GAD is not associated with protection from diabetes.

Amino Acid Sequence↗
PubMed1994

Effects of E5a and E7 genes of human papillomavirus type 11 on immortalized human epidermal keratinocytes and NIH 3T3 cells.

The E5a gene of HPV-11 expressed in NIH 3T3 cells led to tumorigenesis in nude mice; whereas when expressed in keratinocytes, E5a induced anchorage independent growth, but was nontumorigenic in nude mice. The E7 gene of HPV-11 expressed NIH 3T3 cells led to tumors in nude mice and morphological transformation, but not anchorage independent growth. Keratinocytes expressing the E7 gene induced colony formation in soft agarose, but not tumorigenesis in nude mice. Comparison of transforming activities of HPV-11 E5a and E7 genes of HPV-11 showed that the efficiency of cell transformation by E7 was weaker than that of E5a.

3T3 Cells↗
PubMed1994

Genital human papillomavirus infections in Taiwan.

OBJECTIVES: Identification and typing of HPV infections in genital condyloma and normal cytological cervix. METHODS: Cervical cells from 289 Pap cases with normal cytological findings were examined for HPV infection by slot blot hybridization. Fifteen condyloma biopsy specimens were studied by Southern blot hybridization. RESULTS: Thirty-six cases (12.5%) with normal cervical cytology were HPV positive. The predominant HPV type in women with normal cytology is HPV-16. Risk factors for HPV infection in women with normal cytology depend on age and history of pregnancies. Twelve cases (80%) of condyloma contained HPV-6 or -11 sequences. The predominant HPV type in genital condyloma is HPV-11. CONCLUSIONS: HPV detection in population-based screening programs for cervical neoplasia can be an important tool in identifying women who are at risk of developing dysplasia and cervical cancer.

Adult↗
PubMed1994

Epitope mapping of human ETS1 monoclonal antibody.

The epitope for E44 monoclonal antibody (mAb) was mapped using mutated ETS1 proteins lacking different carboxy-terminal regions and by the employment of synthetic oligopeptides spanning the epitope region. This epitope lies around Arg211 of the human ETS1 protein since substitution of Arg211 by Gln211 in the epitope region results in the loss of recognition of the mouse ETS1 protein by E44 mAb. Substitution of Leu214 by valine214 in the epitope region (as is found in the chicken ETS1 and viral Ets proteins) does not alter the capacity of the E44 mAb to recognize this antigen. Taken together, these results suggest that a specific ionic interaction is able to play a pivotal role in the recognition of the ETS1 protein by the E44 mAb.

Amino Acid Sequence↗