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Biomedical subjects

T L Lee

Publications and source records attributed to T L Lee.

At least 19 recordsLinked to original sources
PubMed2002

Teaching first-year medical students physiology: does the human patient simulator allow for more effective teaching?

BACKGROUND: Although the Human Patient Simulator (HPS) is an effective teaching tool in many medical fields, literature supporting its use in the teaching of physiology to medical students is lacking. This study investigated the effectiveness of HPS-based teaching of cardiovascular physiology to first-year medical students. METHODS: Two hundred and ten first-year medical students were scheduled to our HPS laboratory with the purpose of demonstrating "physiology in action". Students were divided into groups of 19-25 each, and attended a lecture followed by a HPS session. Using a theatre-type simulator complete with mannequin, anaesthesia machine and monitors (METI, Sarasota FL), the scenarios of hypovolaemia, sepsis, and cardiac failure were run to demonstrate the physiological changes that occur with changes in preload, afterload, and cardiac contractility. Each student was given a true/false test before, and again after the HPS session, followed by a survey of their learning experience. RESULTS: There was marked improvement in test scores after the HPS session (82.1% vs. 64.6%, P < 0.001). Most of the students felt that HPS was a better teaching tool (94.5%) and raised more questions (76.5%) than lectures. They wanted more topics to be taught this way (96%), as they could apply and re-enforce textbook knowledge, and visualise real-time changes. However, they felt that their experience could have been enhanced with more time and smaller groups. DISCUSSION: HPS is an excellent teaching tool as it stimulates student curiosity and makes knowledge acquisition and understanding easier. It is highly desirable to be incorporated into the teaching of physiology.

Anesthesiology↗
PubMed2001

Alterations of frizzled (FzE3) and secreted frizzled related protein (hsFRP) expression in gastric cancer.

Wnt signaling pathway is important for development and carcinogenesis. Alterations of this pathway, such as mutations in adenomatous polyposis coli (APC) gene and activation mutations of beta-catenin, would result in stabilization of beta-catenin and subsequent translocation to nucleus where genes are transcribed. Recently, a receptor of Wnt, FzE3 was found to be up-regulated in esophageal carcinoma while a non-receptor antagonist of Wnt, secreted frizzled related protein (hsFRP) was found to be down-regulated in some cancer. These findings suggested that FzE3 is a potential oncogene while hsFRP is a potential tumor suppressor gene. We aimed to investigate whether FzE3 and hsFRP were altered in gastric cancer. Twelve cases of gastric cancer, including 7 cases of intestinal type, 4 cases of diffuse type and I case of mixed type, were studied. FzE3 and hsFRP mRNAs were expressed in most of the paired normal gastric tissues. FzE3 was over-expressed in 9 cases (75%) of gastric carcinoma tissues while hsFRP was down-regulated in 2 cases (16%). Beta-catenin nuclear staining was identified in 3 cases (27%) and cyclin D1 was expressed in 5 cases (41%) of cancer samples. All these cases were associated with either up-regulation of FzE3 or down-regulation of hsFRP. Our results suggested that alterations of FzE3 or hsFRP were frequent in gastric cancer. These provide alternative mechanisms leading to activation of Wnt signaling pathway in gastric carcinogenesis.

Adenocarcinoma↗
PubMed2001

Inverse association between cyclooxygenase-2 overexpression and microsatellite instability in gastric cancer.

This study examined the association between cyclooxygenase-2 (COX-2) overexpression and microsatellite instability (MSI) in gastric cancer. COX-2 expression was assessed by immunohistochemistry and scored in a semi-quantitative manner whereas MSI status was characterized by nine microsatellite markers. The clinicopathological features of cancers including survival data were analyzed. Of the 109 gastric cancers studied, COX-2 overexpression and high level of MSI (MSI-H) was detected in 64.2 and 22.0% cases respectively. Gastric tumors with MSI-H phenotypes had significantly lower level of COX-2 expression levels when compared to MSI-L and MSS tumors (P=0.002). Moreover, COX-2 overexpression was associated with tumor invasion beyond submucosa (P=0.045) and there was a trend favoring better survival in gastric cancers without COX-2 overexpression (P=0.07). The results from this study suggest that gastric cancer with microsatellite instability or COX-2 overexpression present with diverse clinicopathological features.

Cohort Studies↗
PubMed2001

Concurrent hypermethylation of multiple tumor-related genes in gastric carcinoma and adjacent normal tissues.

BACKGROUND: Transcriptional silencing by CpG-island hypermethylation now is believed to be an important mechanism of tumorigenesis. To date, studies on CpG-island hypermethylation in gastric carcinoma and adjacent normal tissues are few. METHODS: The authors examined 5 gastric carcinoma cell lines, 26 frozen gastric carcinoma tissues and their adjacent nontumor area for concurrent CpG-island hypermethylation in 6 tumor-related genes (p15, p16, E-cadherin, GST-pi, hMLH1, and VHL) by methylation-specific polymerase chain reaction. Nontumorous gastric tissues from 10 gastritis patients were used as controls. RESULTS: Hypermethylation was not detected in any tissue taken from gastritis patients but was identified in all 5 cell lines and in 24 (92.3%) gastric carcinoma patients. CpG-island methylation in tumor-related genes also was detected in 7 out of the 25 adjacent normal tissues from cancer patients. Hypermethylation of E-cadherin, p15, and p16 were detected more frequently than GST-pi and hMLH1, whereas aberrant methylation of VHL was not detected. Concurrent hypermethylation in 2 or more tumor-related genes was detected in 3 out of the 5 gastric carcinoma cell lines, 22 (84.6%) tumor samples, and 5 (20%) adjacent gastric tissues. Eighteen (69.2%) tumor samples showed hypermethylation in >or= 3 genes. CONCLUSIONS: The current study showed that concurrent hypermethylation of multiple tumor-related genes is detected frequently in gastric carcinoma and adjacent normal tissues. Study findings suggested that a mechanism that leads to dysregulation in CpG-island methylation is likely to be involved in the early gastric carcinogenesis process.

Adaptor Proteins, Signal Transducing↗
PubMed2001

Somatic beta-catenin mutation in gastric carcinoma--an infrequent event that is not specific for microsatellite instability.

We screened 90 cases of gastric carcinoma (GCA) samples for beta-catenin exon 3 mutation and assessed its possible relationship with microsatellite instability (MSI). Three mutations were detected in two samples, including a single mutation in an intestinal type and double mutations in a diffuse type GCA. One of the mutations found in the diffuse type GCA sample was a non-sense mutation at codon 68 (CAG-->TAG). This novel mutation was predicted to disrupt the binding of beta-catenin to alpha-catenin and may be related to the diffuse type morphology. The other two mutations were missense mutations involved or related to the GSK-3beta phosphorylation site, which have been reported previously. No MSI can be demonstrated in the two cases with beta-catenin mutation. Our results suggested that beta-catenin mutation was infrequent in GCA and appeared not specific for MSI.

Adult↗
PubMed2001

Association between cyclo-oxygenase-2 overexpression and missense p53 mutations in gastric cancer.

Wild-type p53 competitively binds to the promoter region of COX-2 in vitro and inhibits its transcription. We examined the association between p53 mutation and COX-2 expression in gastric cancer. COX-2 over-expression was seen in 19 (48.7%) cases. These tumours had more lymph-node metastasis (P = 0.048) and tended to have a poorer survival (P = 0.07). Missense mutations of p53 were detected in 20 (51.3%) patients and had a significantly stronger COX-2 expression than tumours without p53 mutation (P = 0.016). Our results suggest a link between p53 mutation and COX-2 overexpression in gastric cancer.

Adult↗
PubMed2001

IRF-1-mediated CAS expression enhances interferon-gamma-induced apoptosis of HT-29 colon adenocarcinoma cells.

The expression of CAS is reported to be upregulated in a variety of human tumor cells, and such expression correlates with the development of tumors. CAS also plays a role in apoptosis. We investigated whether CAS expression affects the susceptibility of tumor cells to IFN-gamma-induced apoptosis. Our data show that IFN-gamma treatment induces CAS expression in HT-29 tumor cells. IFN-gamma-induced gene expression is primarily mediated by the transcriptional factor, IRF-1. Our data show that IRF-1 mediates IFN-gamma-induced CAS expression. Transfection of HT-29 cells with CAS expression vector did not induce apoptosis of cells; nevertheless, CAS overexpression greatly enhanced IFN-gamma-induced apoptosis of cells. CPP32 is regarded as one of the central apoptosis executioner molecules. CAS overexpression enhances IFN-gamma-induced CPP32 expression. These results indicate that tumor cells highly expressing CAS may be more susceptible to apoptosis induced by reagents that are capable of inducing CAS expression. Thus, CAS may be a target for the elimination of tumors.

Apoptosis↗
PubMed2001

Up-regulation of cyclooxygenase-1 and -2 in human gastric ulcer.

BACKGROUND: The expression of cyclooxygenase (COX) in human gastric ulcers is unknown. AIM: To study the expression and cellular localization of cyclooxygenase in human gastric ulcers. METHODS: A total of 38 surgical gastric ulcer specimens were studied; 20 were Helicobacter pylori-positive and 18 were associated with NSAID use. Twenty non-ulcerated, histologically normal gastric specimens were used as controls. The cellular localization of COX-1 and COX-2 were determined by immunohistochemistry and double immunofluorescence. Cyclooxygenase messenger RNA (mRNA) was measured by reverse transcription-polymerase chain reaction and localized by in situ hybridization. RESULTS: In control specimens, COX-1 was detected in stromal cells in the lamina propria. There was focal and weak immunostaining for COX-2 in the foveolar epithelium. At the ulcer edge, COX-1 was significantly increased in lamina propria cells whereas COX-2 was strongly expressed in the hyperplastic foveolar epithelium in H. pylori- and non-steroidal anti-inflammatory drugs (NSAID)-associated ulcers. At the ulcer base, there was strong expression of COX-1 and COX-2 in myofibroblasts, macrophages and endothelial cells in the granulation tissue, irrespective of H. pylori status or NSAID use. Messenger RNA of COX-1 and COX-2 were demonstrated by reverse transcription-polymerase chain reaction. Double immunofluorescence and in situ hybridization confirmed the cellular localization of cyclooxygenase at protein and mRNA levels, respectively. CONCLUSION: Both COX-1 and COX-2 are up-regulated in human gastric ulcers.

Adult↗
PubMed2001

Expression and cellular localization of COX-1 and -2 in Helicobacter pylori gastritis.

BACKGROUND: There are conflicting reports on the expression of cyclooxygenase in Helicobacter pylori infection. AIM: To evaluate the expression of COX-1 and COX-2 in H. pylori gastritis at messenger RNA (mRNA) and protein levels. METHODS: Endoscopic gastric biopsies were obtained from patients with non-ulcer dyspepsia. The levels of COX-1 and COX-2 mRNA were compared between H. pylori-infected and uninfected specimens using reverse transcription-polymerase chain reaction. The immunohistochemical findings were correlated with the cellular localization of cyclooxygenase mRNA using in situ hybridization. RESULTS: A total of 40 H. pylori-infected and 40 uninfected specimens were studied. mRNA of COX-2 but not COX-1 was elevated in H. pylori-infected mucosa. COX-1 was localized to the mononuclear inflammatory, endothelial and smooth muscle cells in the lamina propria. COX-2 was barely detectable in uninfected mucosa but was strongly expressed in the foveolar and glandular epithelia in H. pylori gastritis. CONCLUSION: Cyclooxygenase-1 is expressed in the mononuclear inflammatory, endothelial and smooth muscle cells in the lamina propria irrespective of the H. pylori status. By contrast, H. pylori induces COX-2 expression in the foveolar and glandular epithelia.

Adult↗
PubMed2001

Detection of microchimerism by PCR is a function of amplification strategy.

BACKGROUND: Suitable detection methods are needed to support larger studies of microchimerism and the allogeneic exposures that may be etiologically related to it. STUDY DESIGN AND METHODS: A twotier PCR strategy for microchimerism detection was developed on the basis of the observation that assay sensitivity for the detection of microchimerism depends on the specificity with which primer pairs recognize sequences unique to the minor population. First, specimens are tested to determine the host HLA class II genotype by using a locus-specific PCR strategy with low sensitivity for microchimerism. Then, a sequence-specific PCR analysis having high sensitivity for detection of microchimerism is applied to detect and quantitate the minor population. Locus-specific, group-specific, and sequence-specific amplification strategies for the detection of distinct minor WBC populations prepared ex vivo were compared. In addition, 39 clinical samples from patients with known transfusion-associated microchimerism and 20 umbilical cord blood (CB) specimens containing maternal WBCs were studied. RESULTS: Locus-specific amplification detected 17 (94%) of 18 cases in which microchimerism was present at 10 percent, but only 1 of 51 cases with microchimerism < or = 1 percent. Group-specific amplification detected all 63 cases with minor populations present at > or = 0.10 percent, but only 16 of 21 cases at the 0.01 percent level. Sequence-specific amplification detected 100 percent of cases down to the 0.01 percent level. When applied to clinical samples, locus-specific amplification reliably identified the major population but proved insensitive to low-level minor populations. CONCLUSIONS: For the detection of microchimerism, assay sensitivity is a function of amplification strategy. These results suggest a simple approach to population screening for microchimerism: the background population of WBCs is typed by a locus-specific method, while minor population(s) can then be sought by using one or several sequence-specific amplifications.

Alleles↗
PubMed2001

Adenomatous polyposis coli gene mutation and decreased wild-type p53 protein expression in oral submucous fibrosis: a preliminary investigation.

OBJECTIVE: The purpose of this study was to identify the adenomatous polyposis coli (APC) tumor suppressor gene mutation and level of wild-type p53 protein expression in patients with oral submucous fibrosis (OSF). STUDY DESIGN: Cells from OSF and control subjects were cultured in Dulbecco modified Eagle medium with 10% fetal bovine serum at 37 degrees C. Genomic DNA was extracted from cultured cells and used as a template for polymerase chain reaction amplification of the APC tumor suppressor gene. The presence of wild-type p53 protein in cell lysates of cultured cells was analyzed by Western blot. Data were analyzed by the sign test for nonparametric samples and by analysis of variance. RESULTS: The results showed that the APC gene of explant cultured cells from OSF patients (8/8) had a CGA-to-GGA transition mutation at codon 498 that resulted in an Arg-to-Gly missense mutation (P <.01). All (8/8) normal HGF cultures revealed expression of the wild-type APC protein. Cells cultured from 7 of 8 OSF patients were also found to have a single nucleotide deletion at nucleotide 1494 that resulted in creating a stop codon (TGA) at codon 504 (P <.01). This created a premature signal for the endpoint of translation and thus resulted in the generation of a truncated protein product that encodes a polypeptide of 503 amino acid residue. It was found that wild- type p53 protein in human gingival fibroblast cell cultures was significantly higher than in OSF cells (P <.01). CONCLUSION: Alterations of the APC and wild-type p53 tumor suppressor genes in OSF may imply a risk for progression to oral cancer.

Adenomatous Polyposis Coli↗
PubMed2001

Two novel quantitative trait loci on mouse chromosomes 6 and 4 independently and synergistically regulate plasma apoB levels.

An elevated plasma apolipoprotein B (apoB) level is a strong predictor of atherosclerosis and coronary heart disease. Epidemiologic and family linkage studies have suggested a genetic basis for the wide variations of plasma apoB levels in the general population. Using a human apoB transgenic (HuBTg) mouse model, we have previously shown that hepatic apoB-100 secretion is a major determinant of the high and low plasma human apoB levels in HuBTg mice of the C57BL/6 (B6) and 129/Sv (129) strains, respectively. In the present article, we present the identification of two novel quantitative trait loci (QTL) as major regulators of plasma human apoB levels in the F(2) and N(2) (backcrossed) offspring (n = 572) derived from crosses between the B6 and 129 mouse strains. These loci were designated ApoB regulator genes (Abrg), because the gene products are likely to be involved in the regulation of plasma apoB levels either directly or indirectly. The first locus, designated Abrg1, was mapped to chromosome 6 in 8-week-old male and female mice with a combined logarithm of odds ratio (LOD) score of 14 at the D6Mit55 marker ( approximately 45.9 cM). Abrg1 contributed approximately 35% of the genetic variance. The second locus, designated Abrg2, was mapped to chromosome 4 with an LOD score of 8.6 in 8-week-old male mice but an LOD score of only 2.0 in 8-week-old female mice at the D4Mit27 marker ( approximately 35 cM). Abrg2 contributed approximately 26% of the genetic variance. Epistasis between Abrg1 and Abrg2 was detected and accounted for approximately 12% of the genetic variance. The combination of these two QTL has major effects (>70%) on the regulation of plasma human apoB levels in the tested population. In summary, we have identified two novel loci that have a major role in the regulation of plasma apoB levels and are likely to regulate the secretory pathway of apoB. The human orthologs for the Abrg loci are strong candidates for human disorders characterized by altered plasma apoB levels, such as FCHL and familial hypobetalipoproteinemia.

Analysis of Variance↗
PubMed2000

One-codon alternative splicing of the CpG MTase Dnmt1 transcript in mouse somatic cells.

The genomic methylation patterns in the mammalian somatic cells are presumably maintained by a single enzyme, dnmt1. In mouse, this DNA (cytosine-5)-methyltransferase, or CpG MTase, is encoded by the Dnmt1 gene. We now present evidence that in different tissues and cell types, the primary transcript of mouse dnmt1 is alternatively spliced to generate two poly-(A) RNAs of approximately similar abundance. This alternative splicing most likely originates from the existence of two tandemly arranged acceptor sites separated by only 3 nt. The two Dnmt1 mRNAs thus encode two CpG MTases differing by two amino acids. We discuss the implications of the discovery of two dnmt1 isozymes, instead of one enzyme as previously thought, in the somatic cells of both mouse and human.

Alternative Splicing↗
PubMed2000

Incidence of and breed-related risk factors for gastric dilatation-volvulus in dogs.

OBJECTIVE: To compare incidence of and breed-related risk factors for gastric dilatation-volvulus (GDV) among 11 dog breeds (Akita, Bloodhound, Collie, Great Dane, Irish Setter, Irish Wolfhound, Newfoundland, Rottweiler, Saint Bernard, Standard Poodle, and Weimaraner). DESIGN: Prospective cohort study. ANIMALS: 1,914 dogs. PROCEDURE: Owners of dogs that did not have a history of GDV were recruited at dog shows, and the dog's length and height and depth and width of the thorax and abdomen were measured. Information concerning the dogs' medical history, genetic background, personality, and diet was obtained from owners, and owners were contacted by mail and telephone at approximately 1-year intervals to determine whether dogs had developed GDV or died. Incidence of GDV based on the number of dog-years at risk was calculated for each breed, and breed-related risk factors were identified. RESULTS AND CLINICAL RELEVANCE: Incidence of GDV for the 7 large (23 to 45 kg [50 to 99 lb]) and 4 giant (> 45 kg [> 99 lb]) breeds was 23 and 26 cases/1,000 dog-years at risk, respectively. Of the 105 dogs that developed GDV, 30 (28.6%) died. Incidence of GDV increased with increasing age. Cumulative incidence of GDV was 5.7% for all breeds. The only breed-specific characteristic significantly associated with a decreased incidence of GDV was an owner-perceived personality trait of happiness.

Age Factors↗
PubMed2000

Molecular cloning, characterization, and expression of a cDNA coding Copper/Zinc superoxide dismutase from black porgy.

A full-length complementary DNA (cDNA) clone encoding a putative copper/zinc superoxide dismutase (Cu/Zn-SOD) was amplified by a Polymerase Chain Reaction (PCR) based technique from cDNA synthesized from black porgy, Acanthopagrus schlegeli, mRNA. Nucleotide sequence analysis of this cDNA clone revealed that it comprised a complete open reading frame coding for 154 amino acid residues. The deduced amino acid sequence showed slightly higher identity (72.8-78.1%) with shark and swordfish Cu/Zn-SOD than with Cu/Zn-SOD from mammalian (68.1-70.7%) and plant (55.5-56.5%) sources. The residues required for coordinating copper and zinc are conserved as they are among all reported Cu/Zn-SOD sequences. The deduced amino acid sequence lacks mitochondria targeting sequence, which suggests that the black porgy cDNA clone encodes a cytosolic Cu/Zn-SOD. The coding region of Cu/Zn-SOD from black porgy was introduced into an expression vector, pET-20b(+), and transformed into Escherichia coli AD494(DE3)pLysS. A predominant achromatic zone was detected by activity staining of native PAGE. This indicates that the Cu/Zn-SOD cDNA clone can express active Cu/Zn-SOD enzyme in E. coli.

Amino Acid Sequence↗
PubMed2000

Interaction of Helicobacter pylori eradication and non-steroidal anti-inflammatory drugs on gastric epithelial apoptosis and proliferation: implications on ulcerogenesis.

BACKGROUND: Apoptosis is associated with loss of gastric mucosal integrity and may play an important role in ulcer development. AIM: To examine how Helicobacter pylori and NSAIDs interact to effect apoptosis and proliferation of the gastric mucosa. METHODS: Patients presenting with musculoskeletal pain requiring NSAID treatment and without previous exposure to NSAID or pre-existing ulcers were recruited. Patients were divided into three groups: (A) H. pylori-infected; (B) H. pylori-eradicated; and (C) non-infected patients. They were given naproxen for 8 weeks. Patients with non-ulcer dyspepsia and H. pylori infection who were given anti-Helicobacter therapy were recruited as controls (D). Endoscopy was performed at baseline and 8-weeks after receiving naproxen. Gastric antral biopsies were obtained to assess apoptosis by terminal uridine deoxynucleotidyl nick end-labelling (TUNEL) and proliferation by Ki67 immunostaining. RESULTS: A total of 55 patients were studied. H. pylori-positive patients had a higher apoptosis and proliferation index at baseline than non-infected patients (P < 0.0001), and eradication of H. pylori resulted in a significant reduction in these parameters. The NSAID induced apoptosis in non-infected subjects (P=0.03) whilst apoptosis was reduced in H. pylori-positive patients receiving NSAID (P=0.02). After 8 weeks of NSAID, post-treatment apoptosis was significantly higher in patients with persistent H. pylori infection than in non-infected patients (P=0.01). CONCLUSIONS: Eradication of H. pylori prior to NSAID therapy significantly reduces the level of apoptosis in the gastric mucosa, which may contribute to maintaining mucosa integrity and preventing ulcer development.

Adult↗
PubMed2000

An assessment of the effects on coagulation of midtrimester and final-trimester amniotic fluid on whole blood by Thrombelastograph analysis.

UNLABELLED: The Thrombelastograph((R)) test (TEG; Haemoscope Corporation, Skokie, IL) was used to assess the effects of midtrimester and final-trimester amniotic fluid (AF) on whole blood coagulation. Different volumes of midtrimester and final-trimester AF were added to whole blood from nonpregnant volunteers in a series of TEG tests. The addition of both midtrimester and final-trimester AF resulted in significant decreases in reaction time (P < 0.001) and time from reaction to a fixed level of clot firmness (P < 0.05) and significant increases in angle (P < 0.05) and coagulation index (P < 0.05) values. This reflects accelerated clot initiation and propagation. There was no significant change in the maximal amplitude or % lysis at 30 and 60 min with the addition of either midtrimester or final-trimester AF. There was no significant difference between the effects of midtrimester and final-trimester AF on whole blood TEG. TEG may be an additional useful tool in the treatment of coagulopathy in AF embolism. IMPLICATIONS: We used the Thrombelastograph((R)) test (Haemoscope Corporation, Skokie, IL) to assess the effects of midtrimester and final-trimester amniotic fluid (AF) on whole blood coagulation. Results demonstrate that AF accelerates clot initiation and propagation. The Thrombelastograph((R)) test may be useful in assessing coagulopathy in patients with AF embolism.

Adult↗