Biomedical subjects

T T Chen

Publications and source records attributed to T T Chen.

At least 37 records · Page 2Linked to original sources

PubMed2000

Identifying attitudinal barriers to family management of cancer pain in palliative care in Taiwan.

The purpose of this study was threefold: (1) to examine attitudes held by Taiwanese family caregivers of hospice inpatients with cancer that serve as barriers to cancer pain management; (2) to determine the relationship of attitudinal barriers to family caregiver hesitancy to report pain and to administer analgesics; and (3) to determine the relationship of attitudinal barriers to the adequacy of analgesics used by the patient. A total of 80 pairs of palliative care inpatients and their primary family caregivers participated in this study. Family caregivers completed the Barriers Questionnaire-Taiwan (BQT) form and a demographic questionnaire. The instruments completed by patients consisted of the Brief Pain Inventory-Chinese version and a demographic questionnaire. The data revealed that Taiwanese family caregivers of palliative care patients with advanced cancer had concerns about reporting pain and administering analgesics, particularly as they related to disease progression, side-effects and p.r.n. analgesics. Older and less-educated family caregivers scored significantly higher on the BQT than did their younger, more educated counterparts. Family caregivers' concerns measured by the BQT were related to family caregivers' reluctance to administer analgesics to their patients. Implications for a broader understanding of analgesics in the advancement of pain management in palliative care in Taiwan are discussed.

Adult↗

PubMed2000

The hexapeptide KP-102 (D-ala-D-beta-Nal-ala-trp-D-phe-lys-NH(2)) stimulates growth hormone release in a cichlid fish (Ooreochromis mossambicus).

Abstract Studies in mammals have shown that synthetic Met-enkephalin derivatives, called growth hormone-releasing peptides (GHRPs), stimulate growth hormone (GH) release. The present study was conducted to determine whether the GHRP, KP-102, specifically stimulates GH release in a teleost. Tilapia (Oreochromis mossambicus) were given a single intraperitoneal injection of KP-102 (D-Ala-D-beta;-Nal-Ala-Trp-D-Phe-Lys-NH(2)) or bovine GHRH(1-29)-amide or vehicle and blood was sampled at 1, 6 and 12 h after injection. KP-102 was administered at two doses of 1 ng/g and 10 ng/g body weight, whereas GHRH (positive control) was administered at a single dose of 10 ng/g body weight. Plasma levels of tilapia GH and prolactins (tPRL(177) and tPRL(188)) were determined by radioimmunoassay. As expected, GHRH injection significantly (P<0.001) elevated plasma GH levels (ng/ml) in tilapia at 6 h post-injection. KP-102 also significantly elevated GH levels (at the low dose) at 6 (P<0.05) and 12 (P<0.01) hours post-injection. There were no significant effects on plasma PRL(s) levels, although mean levels of both PRLs were elevated at 6 h post-injection. These results show for the first time that GHRPs stimulate GH release in teleosts and suggest that the GHRP receptor and possibly a "Ghrelin-like" ligand are also present in lower vertebrates.

Analysis of Variance↗

PubMed2000

Cloning and sequence analysis of a cDNA for an inducible 70 kDa heat shock protein (Hsp70) of the American oyster (Crassostrea virginica).

We have been investigating 70 kDa heat shock proteins (Hsp70s) as potential molecular markers for improved breeding and stress management to revitalize stocks of the American oyster, C. virginica. From a C. virginica visceral mass library, a 2.2 kb full-length cDNA was isolated that included a 634 amino acid open reading frame possessing approximately 80% sequence identity with inducible and constitutive Hsp70s of a broad array of animal species. Northern blotting indicated that the cloned cDNA preferentially recognized an mRNA of about 2 kb that was virtually absent from visceral mass under basal conditions but greatly increased after in vivo heat shock of American and Pacific oysters, suggesting that the cDNA codes for an inducible Hsp70.

Amino Acid Sequence↗

PubMed1999

TP53 mutation and haplotype analysis of two large African American families.

Two large apparently unrelated African American families with a high incidence of breast cancer and other tumors characteristic of Li-Fraumeni breast sarcoma cancer family syndrome were studied. Mutation screening revealed that in both families the affected members carried a germline mutation of the TP53 gene at codon 133 (ATG--> ACG, M133T). In order to determine whether an ancestral haplotype was shared by these two families, polymorphic markers within and flanking the TP53 gene were studied. Haplotype analysis using five markers revealed an identical haplotype shared by the two families. Loss of heterozygosity at the TP53 locus in the probands' tumor tissues from each family was observed; in each case, the retained allele carried the common haplotype. The frequency of this haplotype in the general African American population is <0.003. This unique haplotype, combined with the rare TP53 mutation, suggests that these African American families share a common ancestry. This finding suggests that other African Americans may be carriers of this mutation and thus may be at risk of early-onset breast cancer or other cancers characteristic of the Li-Fraumeni breast sarcoma cancer family syndrome. The finding of recurring mutations in African Americans may facilitate carrier screening and identification in this population.

Alleles↗

PubMed1999

Early embryonic expression of the growth hormone family protein genes in the developing rainbow trout, Oncorhynchus mykiss.

In fish, growth hormone (GH), prolactin (PRL) and somatolactin (SL) are three major peptide hormones produced in the pituitary gland. Using reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analysis of the amplified products, the expression of GH, PRL, and SL genes were determined during the embryonic development in rainbow trout (Oncorhynchus mykiss). The mRNA for GH, PRL and SL were detected not only in embryos prior to or after organogenesis of the pituitary gland but also in mature oocytes as maternal messages. After hatching, all of these three mRNA species were detected at very high levels. Since the ontogenesis of the pituitary gland takes place on embryonic day (ED) 14, these observations suggest that the GH family protein genes are expressed in the developing embryos prior to the formation of the pituitary gland. Using the same RT-PCR assay, PRL mRNA was detected only in the head part of the fry whereas GH and SL mRNA were detected in both the head and trunk. In adult animals, though high levels of GH mRNA were primarily detected in the pituitary gland, brain, gill, and heart, low levels of GH mRNA were also detected in the kidney, liver, pyloric ceaca, and ovary. Results of the current study and those reported by Yang et al. (1997. Gen Comp Endocrinol 106:271-281) suggest that GH and SL genes are also expressed in extrapituitary tissues even after the organogenesis of the pituitary gland. Furthermore, these results suggest that these hormones may play important roles yet to be identified during embryonic development in fish.

Animals↗

PubMed1999

Quantitation of IGF-I, IGF-II, and multiple insulin receptor family member messenger RNAs during embryonic development in rainbow trout.

The IGF system has been shown to be important for normal embryonic growth in mice. Characterization of the IGF system in lower vertebrates is still in progress. To gain a greater understanding of the IGF system during embryonic development in teleosts, a competitive reverse transcription-polymerase chain reaction (RT-PCR) assay was developed and used to quantitate the levels of IGF-I and IGF-II mRNAs from rainbow trout embryos isolated from a staged series. The absolute number of molecules of IGF-I mRNA/microg total RNA was significantly lower than the absolute number of molecules of IGF-II mRNA/microg total RNA both during early and late embryonic development. The recent identification of multiple IGF type I receptor (rtIGFR) and insulin receptor (rtIR) cDNAs in rainbow trout has provided us with a tool to investigate the expression of these mRNAs. A relative quantitative RT-PCR assay was used to determine the steady state levels of two forms of rtIGFR and three forms of rtIR mRNAs in rainbow trout embryos. The relative levels of rtIGFR mRNAs were greater in embryos compared to adult tissues while the relative levels of rtIR mRNAs were generally lower. In a RT-PCR based assay, a differential ability to detect rtIGFR and rtIR mRNAs was shown, suggesting developmental regulation of polyadenylation. Our results suggest that IGF-II mRNA is the predominant IGF expressed in rainbow trout embryos. Our characterization of IGF ligand and receptor mRNA levels in rainbow trout embryos suggests that a functional IGF system exists during embryonic development in teleosts.

Animals↗

PubMed1999

Characterization of teleost insulin receptor family members.

Insulin from mammals and fish has been used to determine insulin-binding affinities and receptor numbers with remarkable similarities between these two vertebrates, suggesting functional conservation. Yet, the nature and structure of teleost insulin receptors are not known. Therefore, the cloning and mRNA characterization of rainbow trout insulin receptors were undertaken. Three insulin receptor cDNAs were isolated by screening a cDNA library, confirmed as separate genes by genomic Southern hybridization, and designated as rainbow trout insulin receptor a (rtIR a), rainbow trout insulin receptor b (rtIR b), and rainbow trout insulin receptor c (rtIR c). A high degree of amino acid identity was observed between rainbow trout insulin receptors (rtIRs) and their human homolog, confirming the structural similarities between mammalian and fish insulin receptors. Reverse transcription-polymerase chain reaction from total RNA using either oligo(dT) or random hexamer primers resulted in a diminished ability to detect rtIR a and rtIR b mRNA when oligo(dT) was used, suggesting developmental and tissue-specific polyadenylation. The highest steady-state levels of rtIR mRNAs were consistently detected in juvenile and adult pyloric caeca (which also contained adipose and pancreatic tissue), while the lowest levels were consistently found in muscle. A high level of rtIR b and rtIR c mRNA was also found in ovary, while a high level of rtIR a was found in adult brain. Significant differences were also found between steady-state rtIR mRNA levels in corresponding juvenile and adult tissues. These results suggest a complex expression pattern of insulin receptor mRNAs in partial tetraploid fish.

Aging↗

PubMed1999

Characterization of teleost insulin receptor family members. II. Developmental expression of insulin-like growth factor type I receptor messenger RNAs in rainbow trout.

The insulin-like growth factor (IGF) system in teleosts consists of two ligands, IGF-I and IGF-II, multiple binding proteins, and high-affinity transmembrane receptors. There exists a large gap in our knowledge of the structure and expression of receptors mediating the biological effects of the IGFs in teleosts. For example, nucleotide sequence data other than those from the kinase domain, evidence of multiple genes, mRNA expression pattern and polyadenylation status in multiple tissues at different developmental stages, and quantitation of mRNA levels in multiple tissues are not known for any teleost. In the study described here, two rainbow trout IGF type I receptor cDNAs (rtIGFR Ia and rtIGFR Ib) were isolated by a 5' rapid amplification of cDNA ends method and confirmed as separate genes by genomic Southern blot hybridization. The predicted amino acid sequences are 85% identical to each other in the tyrosine kinase domain. Both cDNAs are more homologous to mammalian IGF type I receptors than to insulin receptors. Reverse transcription-polymerase chain reaction from total RNA using either oligo(dT) or random hexamers as primers resulted in a diminished ability to detect IGF receptor mRNAs when oligo(dT) was used, suggesting developmental and tissue-specific polyadenylation. The highest steady-state mRNA levels of rtIGFR Ia were found in juvenile gill and adult heart, while the highest levels of rtIGFR Ib were found in adult pyloric caeca, which also contained diffuse pancreatic and adipose tissue. The lowest steady-state mRNA levels of both rtIGFR Ia and rtIGFR Ib were found in juvenile heart, liver, muscle, and spleen, and adult liver. Significant differences in steady-state mRNA levels were also found between juveniles and adults. These results suggest a complex expression pattern of IGF type I receptor mRNAs in partial tetraploid fish.

Amino Acid Sequence↗

PubMed1999

Regression of endometrial explants in a rat model of endometriosis treated with the immune modulators loxoribine and levamisole.

OBJECTIVE: To investigate the effects of the immune modulators levamisole and loxoribine in a rat model of endometriosis. DESIGN: Prospective, placebo-controlled study. SETTING: Hospital-based research facility. ANIMAL(S): Nineteen rats with experimentally induced endometriosis. INTERVENTION(S): Rats were treated with three weekly intraperitoneal injections of levamisole (2 mg per rat; n = 6), loxoribine (1 mg per rat; n = 6), or saline (control; n = 7) and killed 8 weeks after treatment. MAIN OUTCOME MEASURE(S): Histologic and immunohistochemical analysis of endometriotic explants. RESULT(S): The loxoribine-treated group showed marked regression of both epithelial and stromal components. Epithelial regression was noted in the control group, but the epithelium was strikingly preserved in the levamisole group. There were significantly greater numbers of dendritic cells in the explants of animals treated with loxoribine and levamisole. The number of natural killer cells was significantly reduced in loxoribine-treated explants. CONCLUSION(S): Loxoribine, a potent immunomodulatory drug, appeared to cause regression in both stromal and epithelium components in a rat model of endometriosis. Further, specific cell-mediated immune responses in this model of endometriosis were elucidated.

Adjuvants, Immunologic↗

PubMed1999

Application of rapid random stimulation (RRS) to visual evoked potentials in children.

The present study was performed to examine the effects of regular (1 Hz) and modified rapid random stimulation (RRS) (6 and 12 Hz) on visual evoked potentials (VEPs), by simultaneously recording negative waves around 100 ms, wave IV-latency, positive waves around 60 ms, wave III-latency, and amplitudes calculated from peak to peak, without causing impairment of visual acuity, in 44 patients aged 5-17 years. The wave IV-latencies of VEPs obtained by 6 and 12 Hz RRS were easily determined, and the latencies were not significantly changed compared to those obtained by previous 1 Hz regular stimulation. On the other hand, the amplitudes decreased in a frequency-dependent manner (1 < 6 < 12 Hz). These results were found to be similar in both preschool and school children. The examination time of VEPs determined by RRS is one-tenth shorter than that of 1 Hz regular stimulation. Thus, this method has the benefit of shortening the examination time, which decreases fatigue and inattention of the subjects, suggesting that modified RRS is a practically useful method for children.

Adolescent↗

PubMed1999

Interaction between estrogen receptor and Pit-1 protein is influenced by estrogen in pituitary cells.

The estrogen responsiveness of the rat prolactin gene expression requires the presence of both the estrogen receptor (ER) and the tissue-specific transcription factor, Pit-1 protein. We performed protein interaction assays using anti-rat Pit-1 antiserum (a-rPit-1) to investigate the physical interactions which occur between ER and Pit-1 proteins following estrogen treatment. After fusing maltose binding protein (MBP) and Pit-1 protein, we used the resulting MBP Pit-1 fusion protein to prepare a-rPit-1. Our results show that the estrogen receptor readily co-precipitated with the Pit-1 protein drawn from the lysates of two prolactin-expressing pituitary cell lines GH3 and PR1. The rate of precipitation appears to be both estrogen- and time-dependent. Cellular levels of estrogen receptors and Pit-1 proteins did not show significant changes during the time of estrogen treatment. We therefore suggest that an estrogen-dependent physical interaction between ER and Pit-1 protein exists in vivo, and that this interaction may play an important role in the regulation of prolactin gene expression.

Animals↗

PubMed1999

Evolution of a cytokine using DNA family shuffling.

DNA shuffling of a family of over 20 human interferon-alpha (Hu-IFN-alpha) genes was used to derive variants with increased antiviral and antiproliferation activities in murine cells. A clone with 135,000-fold improved specific activity over Hu-IFN-alpha2a was obtained in the first cycle of shuffling. After a second cycle of selective shuffling, the most active clone was improved 285,000-fold relative to Hu-IFN-alpha2a and 185-fold relative to Hu-IFN-alpha1. Remarkably, the three most active clones were more active than the native murine IFN-alphas. These chimeras are derived from up to five parental genes but contained no random point mutations. These results demonstrate that diverse cytokine gene families can be used as starting material to rapidly evolve cytokines that are more active, or have superior selectivity profiles, than native cytokine genes.

Amino Acid Sequence↗

PubMed1999

Recombinant E-peptides of pro-IGF-I have mitogenic activity.

In mammals, the post-translational proteolytic modification of many pro-hormones generates multiple peptides with similar or distinct biological activities. The production of mature insulin-like growth factor I (IGF-I) involves the cleavage of an E-peptide from pro-IGF-I. Although the IGF-I E-peptides are conserved among vertebrate species, their fate and biological roles have not been identified. To test whether the E-peptides possess any biological activity, three recombinant E-peptides of pro-IGF-I, namely Ea-2-, Ea-3- and Ea-4-peptides of rainbow trout, Oncorhynchus mykiss, were produced in vitro with a His-tag expression system and partially purified with an affinity Ni++ column. The mitogenic activity of each E-peptide was determined by (1) the stimulation of total DNA content increase as measured by a fluorometric method and/or (2) stimulation of [3H]-thymidine incorporation into DNA. Recombinant Ea-4-peptide elicited a dose-related increase in both mitogenic assays in NIH3T3 cells. To further test the specificity of the mitogenic activity of Ea-4-peptide, three other cell lines were used: retroviral transformed human embryonic kidney cells (293GP), human mammary gland tumor cells (MCF-7) and caprine mammary epithelial cells (CMEC). Similar levels of mitogenic activity were observed in all cell lines tested for Ea-4-peptide. Mitogenic activity was also observed with recombinant Ea-2- and Ea-3-peptides when assayed in NIH3T3 cells. These results suggest that E-peptides of rainbow trout pro-IGF-I possess mitogenic activity in heterologous systems.

Animals↗

PubMed1998

Optimal flexible designs in phase II clinical trials.

In the conduct of a phase II cancer clinical trial, patients usually enter in two stages. If the response rate from the first stage is low, then the study terminates. Within various two-stage designs, Simon proposed the optimal and minimax criteria. In the co-operative group setting, practical considerations make it difficult to arrive at the planned sample size exactly. Green and Dahlberg proposed and compared several flexible designs. In this paper, we explicitly define a flexible design as a collection of two-stage designs where the first stage size is in a set of consecutive values (n1, ..., nk) and the second stage size is also in another set of consecutive values (N1, ..., Nk), and each of k2 possible designs has the same probability of occurrence. We apply Simon's optimal and minimax criteria to flexible designs for phase II trials in order to minimize the number of patients tested on an ineffective drug.

Clinical Trials, Phase II as Topic↗

PubMed1998

Inhibition of DNA synthesis by RB: effects on G1/S transition and S-phase progression.

The retinoblastoma tumor suppressor protein, RB, is a negative regulator of cell proliferation. Growth inhibitory activity of RB is attenuated by phosphorylation. Mutation of a combination of phosphorylation sites leads to a constitutively active RB. In Rat-1 cells, the phosphorylation-site-mutated (PSM)-RB, but not wild-type RB, can inhibit S-phase entry. In PSM-RB-arrested G1 cells, normal levels of cyclin E and cyclin E-associated kinase activity were detected, but the expression of cyclin A was inhibited. The ectopic expression of cyclin E restored cyclin A expression and drove the PSM-RB expressing cells into S phase. Interestingly, Rat-1 cells coexpressing cyclin E and PSM-RB could not complete DNA replication. Microinjection of cells that have passed through the G1 restriction point with plasmids expressing PSM-RB also led to the inhibition of DNA synthesis. The S-phase inhibitory activity of PSM-RB could be attenuated by the coinjection of SV40 T-antigen, adenovirus E1A, or a high level of E2F-1 expression plasmids. However, the S-phase inhibitory activity of PSM-RB could not be overcome by the coinjection of cyclin E or cyclin A expression plasmids. These results reveal a novel role for RB in the inhibition of S-phase progression that is distinct from the inhibition of the G1/S transition, and suggest that continued phosphorylation of RB beyond G1/S is required for the completion of DNA replication.

Adenovirus E1A Proteins↗

PubMed1998

Characterization of a breast cancer cell line derived from a germ-line BRCA1 mutation carrier.

A tumor cell line, HCC1937, was established from a primary breast carcinoma from a 24-year-old patient with a germ-line BRCA1 mutation. A corresponding B-lymphoblastoid cell line was established from the patient's peripheral blood lymphocytes. BRCA1 analysis revealed that the tumor cell line is homozygous for the BRCA1 5382insC mutation, whereas the patient's lymphocyte DNA is heterozygous for the same mutation, as are at least two other family members' lymphocyte DNA. The tumor cell line is marked by multiple additional genetic changes including a high degree of aneuploidy, an acquired mutation of TP53 with wild-type allele loss, an acquired homozygous deletion of the PTEN gene, and loss of heterozygosity at multiple loci known to be involved in the pathogenesis of breast cancer. Comparison of the primary tumor with the cell line revealed the same BRCA1 mutation and an identical pattern of allele loss at multiple loci, indicating that the cell line had maintained many of the properties of the original tumor. This breast tumor-derived cell line may provide a useful model system for the study of familial breast cancer pathogenesis and for elucidating BRCA1 function and localization.

Adult↗

PubMed1998

CPAN, a human nuclease regulated by the caspase-sensitive inhibitor DFF45.

Induction of apoptosis by death receptors such as Fas or tumour necrosis factor (TNF) R1 leads to distinct changes in cell morphology, activation of the caspase protease cascade, and the degradation of nuclear chromatin by activated nucleases. Here, we describe the purification and cDNA cloning of a novel 40 kDa endonuclease from Jurkat cells that is activated by caspases. This protein, designated caspase-activated nuclease (CPAN), is sufficient to degrade naked DNA and to induce apoptotic morphology and DNA fragmentation in naive nuclei. CPAN is highly homologous to a recently described mouse nuclease, CAD [1], and may represent the human homologue. Our data on the human cDNA as well as additional data on the mouse homologue suggest that a 30 amino-acid portion of the recently published mouse sequence [1] is incorrect. We show that the activity of human CPAN is regulated by DFF45 [2], an inhibitor necessary for CPAN expression and stabilization in an inactive state in living cells. Proteolytic cleavage of DFF45 by caspases in vitro leads to dissociation of DFF45 fragments from CPAN and activation of CPAN as an endonuclease. CPAN is a tightly regulated endonuclease with unique characteristics that might represent a distinctive family of endonucleases.

Amino Acid Sequence↗

PubMed1998

Interaction of 2,2',4,4',5,5'-hexachlorobiphenyl with hepatic cytochrome P-4501A in rainbow trout (Oncorhynchus mykiss).

Di-ortho polychlorinated biphenyls (PCBs) are prominent environmental contaminants and their biological activity in fish may be more significant than previously thought. Four weeks after intraperitoneal (i.p.) injection with 50 or 250 microg 2,2',4,4',5,5'-hexachlorobiphenyl (2HxCB)/g fish, rainbow trout livers were removed and frozen at -80 degrees C or microsomes were prepared. Microsomal ethoxyresorufin O-deethylase (EROD) activity was approximately one and two orders of magnitude greater than controls in fish treated with 50 and 250 microg 2HxCB/g fish, respectively. Cytochrome P4501A (CYP1A) Western immunoblot relative optical density increased with 2HxCB dose. Hepatic CYP1A1 mRNA levels were approximately threefold greater in fish treated with 250 microg 2HxCB/g fish than in controls, while hepatic CYP1A1 mRNA levels in fish treated with 50 microg 2HxCB/g fish were not significantly induced. There was no increase of CYP1A3 mRNA in 2HxCB-treated fish. The study showed 2HxCB induced hepatic EROD activity, CYP1A protein, and CYP1A1 mRNA content in rainbow trout.

Animals↗