Biomedical subjects

T W Chan

Publications and source records attributed to T W Chan.

At least 19 recordsLinked to original sources

PubMed2001

Tetrapyrrole derivatives substituted with ferrocenylethynyl moieties. synthesis and electrochemical studies.

Up to eight redox-active ferrocenyl units have been incorporated, through the unsaturated ethynyl linkers, on the periphery of a series of cyclic tetrapyrrole derivatives including zinc(II) phthalocyanine and 2,3-naphthalocyanine, and nickel(II) meso-diphenylporphyrin. The synthesis of the former two macrocycles 4 and 7 involves the Sonogashira coupling reaction of ferrocenylethyne with 4,5-dichlorophthalonitrile (1) or 6,7-dibromonaphthalonitrile (5), respectively, followed by a base-promoted cyclization. The meso-bis(ferrocenylethynyl)porphyrin 11 has been prepared from the dibromo analogue 10 also by a palladium-catalyzed coupling reaction. These novel macrocyclic compounds have been spectroscopically and electrochemically characterized. As revealed by cyclic voltammetry, the ferrocenyl moieties appear to be electrochemically independent in these complexes and there is no significant electronic coupling among the iron(II) centers.

Journal Article↗

PubMed2001

Induction of UO-44 gene expression by tamoxifen in the rat uterus and ovary.

A complementary DNA, uterine-ovarian-specific gene 44 (UO-44), has been isolated from tamoxifen-induced rat uterine complementary DNA library using differential display techniques. UO-44 transcripts are found to be abundant in the uterus and ovary. UO-44 gene expression in the uterus is strictly regulated by estrogens, tamoxifen, and GH, whereas the pure antiestrogen ICI 182780 is inhibitory. Treatment of ovariectomized rats and hypophysectomized rats with tamoxifen and GH, respectively, resulted in up-regulation of UO-44 expression in a dose-dependent manner. In situ hybridization revealed that UO-44 gene expression was restricted to the luminal and glandular epithelial cells of the uterus and to granulosa cells of medium-size ovarian follicles. Transfection studies showed that UO-44 was a membrane-associated protein. Because estrogens, tamoxifen, and GH are stimulators of uterine luminal epithelial cell growth in vivo, UO-44 protein may serve as a mediator of the effect of these compounds in inducing epithelial proliferation and differentiation in these tissues.

Animals↗

PubMed2001

Cloning and characterization of a novel pregnancy-induced growth inhibitor in mammary gland.

Growth factors and growth inhibitors play crucial roles in the growth regulation and differentiation of mammary epithelial cells. Studies have shown that during pregnancy, with the onset of terminal differentiation, there is a dramatic decrease in the proliferation of the mammary epithelial cells. Here we report the cloning and characterization of a novel pregnancy-induced cDNA, OKL38, from a human ovarian cDNA library. This cDNA encodes for a protein of approximately 34.5 kDa. Tissue distribution studies through Northern analyses revealed the ubiquitous nature of OKL38 transcripts in most tissues, with the highest levels observed in the ovary, kidney, and liver. The onset and advancement of pregnancy also gave rise to a concomitant increase in OKL38 gene expression. In situ hybridization revealed that OKL38 mRNA was further detected in mammary secretory epithelial cells. However, low levels of OKL38 transcripts were observed in the various human breast cancer cell lines studied and were barely detectable in all dimethylbenz(A)anthracene-induced mammary tumors examined. Transfection studies with OKL38 cDNA with MCF-7 cells resulted in growth inhibition in vitro and reduction in tumor formation in vivo. These observations led to speculation that OKL38 may play a vital role in the growth regulation and differentiation of breast epithelial cells during pregnancy and its implications in tumorigenesis.

Amino Acid Sequence↗

PubMed2001

Antioxidative glucosides from the fruits of Ligustrum lucidum.

The ethanol extract of the fruits of Ligustrum lucidum was shown to have inhibitory effects on the hemolysis of red blood cells induced by 2,2'-azo-bis-(2-amidinopropane) dihydrochloride. Bioassay-guided analysis led to the isolation of ten secoiridoid glucosides. Two of them were new, lucidumosides C and D. Their structures were elucidated by spectroscopic methods. The other eight compounds were identified as oleoside dimethyl ester, ligustroside, oleuropein, nuezhenide, isonuezhenide, neonuezhenide, lucidumoside A and lucidumoside B. Five compounds, oleoside dimethyl ester, oleuropein, neonuezhenide, lucidumoside B and lucidumoside C, exhibited strong antioxidant effect against hemolysis of red blood cells induced by free radicals.

Antioxidants↗

PubMed2001

Co-administration of finasteride and the pure anti-oestrogen ICI 182,780 act synergistically in modulating the IGF system in rat prostate.

Prostate cancer is the most diagnosed invasive malignancy in males. Androgens and oestrogens have been implicated in the pathogenesis of prostate cancer. We report herein that the pure anti-oestrogen ICI 182,780 (ICI) reduces Ki-67 labelling index and IGF-I receptor levels in rat prostate. Increase of IGF-I mRNA and IGF-binding protein 3 (IGFBP-3) accumulation occur without any effect on prostate weight. Finasteride significantly decreases prostate weight and inhibits IGF-I gene expression. IGFBP-3 mRNA, Akt and phospho-Akt are not affected by finasteride. Co-administration of ICI plus finasteride reduces prostate weight by approximately 50% and causes acinar dilation with decreased luminal epithelial cell thickness. The acinar epithelial cells became atrophic and inactive with minimal cytoplasm. We also demonstrate a synergistic effect of ICI and finasteride on induction of IGFBP-3 accumulation and inhibition of Akt phosphorylation. Because the IGF and IGFBP-3 system plays an important role in prostate epithelial cell proliferation, apoptosis and tumour progression, the inhibitory effects of finasteride and ICI on IGF system may contribute to their anti-proliferative activity. These observations support a potential use of ICI in conjunction with finasteride in the prevention and/or treatment of prostate cancer.

5-alpha Reductase Inhibitors↗

PubMed2001

Induction of mammary epithelial cell differentiation and inhibition of dimethylbenz(A)anthracene-induced mammary tumour by co-administration of a pure antiestrogen ICI 182,780 and testosterone enanthate.

Epidemiological studies have shown that early first pregnancy is associated with a life-long reduction in breast cancer risk. The terminal differentiation associated with pregnancy and lactation has been proposed as a mechanism underlying the protective effect of pregnancy. We report that treatment of rats with ICI 182,780 (ICI) caused a marked reduction in epithelial cells and Ki-67 labelling index as compared to controls and testosterone enanthate-treated (TE) mammary glands. TE increased the Ki-67 labelling index, stimulated lobuloalveolar and ductal growth, as well as the secretory activity of acinar cells. Co-administration of TE and ICI resulted in a reduction in Ki-67 labelling index. Mammary epithelial cells became differentiated, resembling that observed at the end of pregnancy and during lactation as indicated by marked increase in secretory activity, lipid accumulation and presence of basal nuclei. The expression of differentiation markers such as whey acidic protein, mammary derived growth inhibitor, alpha-casein and beta-casein was detected only in TE plus ICI treated mammary tissues. Unlike TE, ICI caused a significant reduction in DMBA-induced tumour incidence, number of tumour bearing and tumour size. Tumour incidence was reduced to 8% when both ICI and TE were co-administered. Our data provide the novel molecular interactions between the estrogen and androgen in regulation of mammary growth and differentiation. These observations may give insight into novel actions of ICI and TE on breast differentiation and protection against carcinogenesis which may be useful in designing novel strategies for cancer prevention and/or treatment based on maximizing mammary epithelial cell differentiation.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed2001

Inhibition of insulin-like growth factor signaling pathways in mammary gland by pure antiestrogen ICI 182,780.

The antiestrogens ICI 182,780 (ICI) and tamoxifen are clinically useful in the treatment of estrogen receptor-positive breast tumors. We assessed the in vivo effects of ICI, tamoxifen, and estradiol on the insulin-like growth factor (IGF) signaling pathway in the rat mammary gland. ICI significantly decreased the size of the lobular structures, Ki-67 labeling index, and insulin-like growth factor binding protein (IGFBP)-2 and IGFBP-5 gene expression. Treatment of rats with 1, 1.5, and 2 mg of ICI/kg body weight/week resulted in a 2-, 7-, and 8-fold increase in IGFBP-3 transcripts. High doses of ICI increased mammary IGF-1 gene expression by 2-fold (P < 0.01) but decreased IGF-1R and its autophosphorylation to approximately 30% of the control mammary gland. IRS-1, IRS-2, and c-Raf-1 levels in the ICI-treated mammary glands were approximately 30, 15, and 40% of controls, respectively. Basal phosphorylation of IRS-1, Akt-1, and the p85 subunit of phosphatidylinositol 3-kinase (PI-3K) were low but detectable after ICI treatment. Despite a significant reduction in levels of IGF-1R, IRS-1, and IRS-2 phosphorylation, phospho p42/p44 MAPK levels were only slightly decreased. Tamoxifen-induced growth inhibition was associated with slight stimulation of IGFBP-3 gene expression and reduction in IRS-2 levels. Basal phosphorylation of IGF-1R, IRS-1, and p85 subunit of PI-3K was decreased by tamoxifen. Estradiol-induced epithelial cell proliferation was associated with inhibition of IGFBP-3 gene expression, stimulation of IGFBP-2 gene expression, and increases in IGF-1R, IRS-1, IRS-2, and c-Raf-1 levels. Although basal phosphorylation of IGF-1R, IRS-1, IRS-2, Akt-1, and the p85 subunit of PI-3K was significantly increased by estradiol, basal phospho p44/42 MAPK was significantly reduced. The data indicate that in addition to their classic actions, antiestrogens have major effects on IGF signaling pathways.

Animals↗

PubMed2000

Differentiation and authentication of Panax ginseng, Panax quinquefolius, and ginseng products by using HPLC/MS.

An LC/MS-based method is established for the differentiation and authentication of specimens and commercial samples of Panax ginseng (Oriental ginseng) and Panax quinquefolius (American ginseng). This method is based on the separation of ginsenosides present in the ginseng methanolic extracts using high-performance liquid chromatography (HPLC), followed by detection with electrospray mass spectrometry. Differentiation of ginsenosides is achieved through simultaneous detection of intact ginsenoside molecular ions and the ions of their characteristic thermal degradation products. An important parameter used for differentiating P. ginseng and P. quinquefolius is the presence of ginsenoside Rf and 24-(R)-pseudoginsenoside F11 in the RICs of Oriental and American ginsengs, respectively. It is important to stress that ginsenoside Rf and 24(R)-pseudoginsenoside F11, which possess the same molecular weight and were found to have similar retention times under most LC conditions, can be unambiguously distinguished in the present HPLC/MS method. The method developed is robust, reliable, reproducible, and highly sensitive down to the nanogram level.

Chromatography, High Pressure Liquid↗

PubMed2000

Differentiation of enantiomers using matrix-assisted laser desorption/ionization mass spectrometry.

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has successfully been used to differentiate pseudo-enantiomeric (isotopically labelled) amino acids by using cyclodextrin as complexing host. By using different pseudo-enantiomeric mixtures (i.e. R(Dn) + S; and R + S(Dn)), it has been demonstrated that the preference of cyclodextrin for S-enantiomers is not due to the size differences caused by the hydrogen/deuterium substitution. It is postulated that this method can be extended to differentiate enantiomers (and determine enantiomeric excess) by using a pair of enantiomeric hosts, as demonstrated previously using other ionization techniques, but with much higher sensitivity.

Amino Acids↗

PubMed2000

Effect of sample preparation methods on the analysis of dispersed polysaccharides by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Effect of sample preparation methods on the matrix-assisted laser desorption/ionization (MALDI) analysis of dispersed polysaccharides is reported. By using the conventional drop-drying method, the measured number-averaged molecular weights were found to differ significantly from the values obtained from gel-permeation chromatography (GPC). These discrepancies were found to increase as the average masses of the polysaccharides increased. To understand the impact of the sample preparation method on the MALDI measurement, a dispersed dextran sample was separated into ten narrow-distribution fractions. Mixtures of different mass fractions were prepared and analyzed by using different mixing and preparation methods. By using the ratio of the signal intensity for the low-mass fraction to that of the high-mass fraction as an indicator, the impact of sample preparation conditions on the enhancement of low-mass components (or suppression of high-mass components) was determined. From the results obtained, it is postulated that the difference in solubilities between the low-mass and high-mass components for dispersed polymers might be large enough to induce an enhancement of low-mass components (or suppression of high-mass components) at the surface of the crystals during sample crystallization in the drop-drying method.

Algorithms↗

PubMed2000

Modulation of transforming growth factor beta1 gene expression in the mammary gland by insulin-like growth factor I and octreotide.

Transforming growth factor beta1 (TGF-beta1) has been shown to exhibit anti-proliferative activity for mammary gland epithelial cells and for human breast cancer cells. Insulin-like growth factor I (IGF-I), in contrast, is a well-characterized mitogenic and anti-apoptotic factor involved in mammary gland physiology. In order to examine the hypothesis that IGF-I suppresses TGF-beta1 expression in the mammary gland, we studied the effect of various manipulations of the growth hormone - IGF-I axis on TGF-beta1 mRNA abundance. Administration of IGF-I to intact animal suppressed TGF-beta1 mRNA levels in a dose-dependent manner to approximately 20% of control levels. Administration of the somatostatin analogue octreotide in a manner previously shown to acutely suppress the growth hormone - IGF-I axis increased mammary gland TGF-beta1 expression approximately 3-fold. Transgenic mice overexpressing growth hormone expressed TGF-beta1 in the mammary gland at only approximately 12% of the level of control animals, while mice IGF-I deficient due to the mutation expressed TGF-beta1 at slightly higher levels than control animals. The large differences in TGF-beta1 expression between control and GH-transgenic animals were correlated with major differences in architecture of the mammary gland, while the appearance of mammary glands of normal and animals was similar. These data document a previously unrecognized relationship between TGF-beta1 and IGF-I physiology in the mammary gland, and suggest a novel mechanism by which somatostatin analogues influence the proliferative behaviour of breast epithelial cells.

Animals↗

PubMed1999

A novel nonmetallized tip for electrospray mass spectrometry at nanoliter flow rate.

We report the fabrication of a durable nonmetallized nanospray tip. This nanospray tip does not require complex preparation procedures such as chemical treatment, deposition of gold or SiOx vapor. It was fabricated by pulling a heated glass capillary of 1.1 mm internal diameter to produce a fine tip with an orifice of 10-15 microns in diameter. A 10 microns gold-plated tungsten wire was inserted through the capillary tip. This tungsten wire played a central role in the operation of this durable nanospray tip by providing electrical contact. This type of nanospray tip could withstand electrical discharges and sustained spraying of solution at nanoliter flow rate for more than 3 h. Using insulin (35 microM) and myoglobin (1 microM) solutions, useful mass spectrum could be acquired with low fmol sensitivity.

Animals↗

PubMed1998

Formulation of matrix solutions for use in matrix-assisted laser desorption/ionization of biomolecules.

We report a simple method for converting solid matrices into useful matrix solutions for matrix-assisted laser desorption/ionization (MALDI). This method is based on the dissolution of the solid matrix in a liquid support of low volatility such as glycerol. An appropriate solubilizing reagent was added to promote the dissolution of the matrix materials into the liquid support. Selection of the solubilizing reagent is empirically related to an acid-base relationship, i.e., an acidic solid matrix requires a basic organic compound to form a stable matrix solution in the liquid support and vice versa. A tenfold increase in the solubility can be obtained for many solid matrices when appropriate solubilizing reagents are added into the glycerol support. This solubility enhancement is tentatively attributed to the ion-pair formation in a polar nonvolatile liquid support. In addition, the hydrophobicity of the solid matrix seems to play an important role in the efficiency of the resulting matrix solution. By using glycerol as liquid support, a hydrophilic matrix, such as 2,5-dihydroxybenzoic acid (DHB), showed a substantial "peripheral effect," in which good analyte ion signals could only be recorded at the peripheral region of the sample droplet. More hydrophobic matrices, such as alpha-cyano-4-hydroxycinnamic acid (alpha-CCA), exhibit better and more homogeneous responses at different regions of the droplets. The performance of these matrix solutions was evaluated in terms of the durability, reproducibility, sensitivity, high mass capability, and generality. A typical sample droplet can afford more than an hour of repeated sampling with excellent shot-to-shot reproducibility. A low picomole sensitivity was demonstrated using a luteinizing hormone releasing hormone (LHRH) in a Fourier transform ion cyclotron resonance mass spectrometer with a homemade external MALDI ion source. By using a commercial MALDI time-of-flight mass spectrometer, proteins with masses as high as 66,000 Da were successfully analyzed by using these matrix solutions.

Cyclotrons↗

PubMed1998

Interpretive algorithms for the symptom-limited exercise test: assessing dyspnea in Persian Gulf war veterans.

Interpretation of symptom-limited exercise testing requires analysis of a large body of simultaneously recorded cardiopulmonary data. Karlman Wasserman has recommended an algorithmic approach to interpretation (WA) that leads to a dichotomous choice between pulmonary and cardiovascular impairment. An alternative algorithm published by William Eschenbacher (EA) provides for concurrent assessment of cardiovascular and pulmonary exercise impairment. We analyzed a group of 29 individuals referred to the Pulmonary Physiology Laboratory at the Washington Veterans Affairs Medical Center for evaluation of dyspnea following service in the Persian Gulf War to assess the concordance of the two algorithms in determining the cause of dyspnea and exercise impairment in these individuals. They each performed a progressive, ramped, symptom-limited exercise test on a bike for a minimum of 6 min. Exercise measurements were analyzed by both interpretive algorithms. Concordance was found in 28% of tests. The greatest discordance occurred in identifying pulmonary limitation. Eleven had pulmonary limitation by EA; of these, WA found 1 to have pulmonary limitation, 5 to be normal, 4 indeterminate, and 1 musculoskeletal limitation. Of the 11 with pulmonary limitation by EA, but not by WA, 5 had abnormal resting pulmonary function measurements. Analysis of the differences between these two interpretive approaches is given. The EA algorithm may be more sensitive for detecting exercise impairment of pulmonary origin, but its specificity remains to be determined.

Adult↗

PubMed1996

Positive-ion electrospray-fourier transform ion cyclotron resonance mass spectra of polypeptides and proteins.

We have determined the mass spectra of three polypeptides which have been designed as an effective reagent to be used in the diagnosis of a Chinese strain of Hepatitis E Virus (HEV). The spectra were determined by electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometer. These are peptide 30, peptide 33 and peptide 42, where the guanidine amino group of arginine in each polypeptide was protected. The molecular weights were respectively measured as 3575, 4094 and 5390 Dalton, against the molecular weights of bovine serum albumin (66430 Da) and bovine insulin (5732.77 Da).

Amino Acid Sequence↗

PubMed1996

Use of ammonium halides as co-matrices for matrix-assisted laser desorption/ionization studies of oligonucleotides.

Four ammonium salts, NH4F, NH4Cl, NH4Br and NH4I were tested as co-matrices for the matrix-assisted laser desorption/ionization (MALDI) analysis of a number of DNA homopolymers, including phosphorylated d(T)8, d(A)4, d(C)4 and non-phosphorylated d(G)4, using 2-amino-5-nitropyridine (ANP) matrix. In the present study, all the ammonium halides displayed significant enhancement effects on the signal intensities of the intact molecular ions of the DNA homopolymers. Among the halides used, NH4F was found to exhibit the greatest enhancement effects. By comparison with results obtained using the corresponding isomorphous potassium halides as co-matrices, it is postulated that both the cationic and anionic portions of the co-matrix molecule play important roles in the desorption/ionization of the oligonucleotides under typical MALDI conditions. It was also demonstrated that the ANP matrix exhibits a strong inhibitory effect on the formation of alkali-metal adducts in oligonucleotide analysis.

DNA, Single-Stranded↗

PubMed1996

Induction of seminal vesicle morphogenesis from dog epididymal epithelium.

A number of carnivores, including the dog, do not have seminal vesicles (SVs). They have been induced experimentally by heterotypic recombination of adult dog epididymal epithelium with mesenchyme of neonatal rat SV and allowed to grow for 4 weeks under the renal capsule of male athymic nude mice. The induced SVs bore a striking similarity to rat SV both histologically and ultrastructurally. However, they secreted proteins which were different from those in the rat SV and reacted negatively to antibodies specific to rat SV secretory proteins and dog prostate secretory proteins. The results strongly suggested that the glands induced from dog epididymal epithelium were likely to be "dog" SVs.

Animals↗

PubMed1995

Hepatic MR imaging with ferumoxides: a multicenter clinical trial of the safety and efficacy in the detection of focal hepatic lesions.

PURPOSE: To assess the safety and diagnostic efficacy of intravenous ferumoxides, a superparamagnetic iron oxide, for depiction of focal hepatic lesions on magnetic resonance (MR) images. MATERIALS AND METHODS: This open-label study included 208 patients with known or suspected focal hepatic lesions. MR images were obtained before and 45 minutes to 4 hours after intravenous infusion of ferumoxides (10 mumol/kg). The effect of ferumoxides on signal intensity of the liver was assessed with quantitative analysis. Safety was evaluated with patient monitoring and laboratory measurements. RESULTS: Mean lesion-to-liver contrast-to-noise ratio on T2-weighted images was 9.1 on unenhanced images and 12.7 on enhanced images. Signal intensity of normal liver on enhanced images decreased to 37% of that on unenhanced images. In blinded image evaluations, additional lesions were identified on 27% of enhanced images. No serious adverse events occurred. CONCLUSION: Ferumoxides is a safe and efficacious contrast agent for the detection of focal liver lesions on T2-weighted images.

Contrast Media↗