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Biomedical subjects

C H Lee

Publications and source records attributed to C H Lee.

At least 19 recordsLinked to original sources

Disruption of gastrulation movements in Xenopus by a dominant-negative mutant for C-cadherin.

Gastrulation in Xenopus laevis transforms a ball of undifferentiated cells into a well-organized elongated embryo with a distinct body axis, as a result of extensive cell rearrangements and movements. Since cell adhesion is thought to play an important role during tissue morphogenesis, we investigated the potential role for C-cadherin, a primary cell-cell adhesion molecule in Xenopus, in gastrulation movements. A deletion mutant consisting of the extracellular and transmembrane domains but lacking the cytoplasmic tail (Ctrunc) was constructed to act as a dominant-negative inhibitor of the endogenous protein. Injection of in vitro transcribed Ctrunc mRNA into the prospective dorsal involuting marginal zone, the region that undergoes the most extensive movements, caused a reproducible defect in gastrulation. The defect, an inability to complete involution and close the blastopore, was cadherin-specific, because it could be rescued by expressing full length C-cadherin. Ctrune also seems to inhibit morphogenetic movements of animal cap explants isolated from injected embryos. Expression of Ctrunc did not appear to interfere with inductive processes in the embryos that developed gastrulation defects, because they formed complete axial structures, including a complete head and notochord. These results demonstrate that C-cadherin plays a critical role in Xenopus laevis gastrulation movements, and they support the hypothesis that cadherins can mediate cellular rearrangements during tissue morphogenesis.

Animals

Assessment of skull base involvement in nasopharyngeal carcinoma: comparisons of single-photon emission tomography with planar bone scintigraphy and X-ray computed tomography.

The diagnostic contribution of single-photon emission tomography (SPET) to the detection of bone lesions of the skull base was explored in 200 patients with nasopharyngeal carcinoma (NPC). Comparison of SPET with planar bone scintigraphy showed that SPET improved the contrast and better defined the lesions in 107 out of the 200 patients. Comparison of SPET with X-ray computed tomography (CT) showed that SPET did not miss the lesions detected by CT while CT missed 49% of the lesions detected by SPET. The only false-positive lesion with SPET was detected in the mastoid bone. SPET detected skull base lesions in all of the 35 patients with cranial nerve involvement, while CT missed eight and planar bone scintigraphy missed four. The findings suggest that SPET should be included in the routine check-up examinations of patients with NPC.

Female

Retinoic acid induction of mouse cellular retinoic acid-binding protein-I gene expression is enhanced by sphinganine.

Cellular retinoic acid-binding protein-I (CRABP-I) gene expression is induced in mouse embryonal carcinoma P19 cells specifically by retinoic acid (RA) and the induction is enhanced by sphinganine. The effects of retinoic acid and sphinganine on CRABP-I gene expression can be accounted for by a stimulation of its transcription rate. Using a lacZ reporter system, it was determined that a DNA fragment containing a putative AP-1 binding site in the promoter region of CRABP-I gene is required for the up-regulation of CRABP-I gene transcription.

Animals

Correlation between presence of clonal rearrangements of immunoglobulin heavy chain genes and B-cell antigen expression in Hodgkin's disease.

Southern blot analysis of Hodgkin's disease (HD), although often compromised by the small number of abnormal cells present in the tissue, have tended to favor a B-cell derivation of the Hodgkin's and Reed-Sternberg (HRS) cells in cases of nodular sclerosis (NS) and mixed cellularity (MC) Hodgkin's disease. Eighteen frozen and 29 paraffin-embedded sections of lymph node specimens from 29 patients with pretreatment HD (22 NSHD and 7 MCHD) were studied by molecular analysis and immunohistochemistry to determine the phenotype of HRS cells. All cases were reviewed and showed typical morphology and CD45-, CD30+, CD15+, BLA.36+ HRS cells. In 11 of 29 (38%) cases, HRS cells were reactive with at least one B-cell marker (CD20, CD79a, MB2), 7 of 29 (24%) cases showed reactivity with the T-cell marker CD3, and 11 of 29 (38%) cases displayed a "null" phenotype. By using a polymerase chain reaction (PCR) and consensus primers for the V and J regions of the immunoglobulin heavy chain (IgH) gene, the authors were able to detect B-cell clonality in 9 of 18 (50%) frozen samples of HD analyzed. IgH gene rearrangement was present in 8 of 15 (53%) NSHD and in 1 of 3 (33%) MCHD. In five of nine (56%) of these cases, HRS cells were reactive with at least one B-cell marker, whereas one case expressed the T-cell marker CD3. The other three cases with IgH gene rearrangement showed a "null" immunophenotype. IgH gene analysis was negative in all remaining CD3+ cases and in two other cases that expressed B-cell markers by immunohistology. Southern blotting failed to detect rearrangement of the T-cell receptor beta-chain gene and immunoglobulin heavy and light genes in any of these cases. The results show that PCR represents a specific and sensitive technique for the detection of IgH gene rearrangements in cases of Hodgkin's disease. The results also suggest a lymphoid B-cell derivation of HRS cells in a high proportion of the cases.

Antigens, Neoplasm

The effect of dietary protein levels on the responses of the taste nerve to sodium chloride in spontaneously hypertensive rats (SHRs).

The present study was undertaken to clarify the effect of dietary protein level on the response of the taste nerve to NaCl solutions in spontaneously hypertensive rats (SHRs). The results showed that the taste sensitivity to NaCl in SHRs fed a 5% whole-egg protein diet for 3 weeks was significantly lower than in those fed a 15% protein diet. This observation suggests that chronic feeding of a low-protein diet causes an impairment of salt-taste reception or subsequent transduction.

Animals

Early repair of traumatic ventricular septal defect and mitral valve regurgitation.

Traumatic ventricular septal defect with valvular injury is an uncommon blunt trauma. It may develop either immediately or be delayed, but it should be corrected electively. With hemodynamic instability and cardiopulmonary deterioration, however, early repair is necessary as a lifesaving procedure. Two-dimensional echocardiography and Doppler color flow mapping are very important for rapid detection in patients who are critically injured. This is a case report of the successful repair of ventricular septal defect and posterior leaflet disruption of mitral valve right after blunt trauma.

Adult

Antisense gene suppression against human ICAM-1, ELAM-1, and VCAM-1 in cultured human umbilical vein endothelial cells.

Antisense gene suppression has been carried out for human ICAM-1, ELAM-1, and VCAM-1 in cultured human umbilical vein endothelial cells (HUVEC) stimulated by lipopolysaccharide, tumor necrosis factor alpha, or interleukin-1 beta. A panel of antisense phosphorothioate oligodeoxyribonucleotides (PS-ODN), complementary to mRNA or pre-mRNA of these molecules, were tested for their gene suppression activity monitored by radioimmunoassay of the respective cell surface adhesion molecules. Sequences targeted by effective antisense PS-ODNs were located throughout the mRNA and pre-mRNA. "Hot spots" of gene suppression sites for each region were observed. Shift of the PS-ODN hybridizing site upstream or downstream by a few bases resulted in drastic change of gene suppression efficiency. In addition to translation arrest and RNase H activity, a third mechanism was proposed for antisense gene suppression, involving multiple binding sites for PS-ODN and the activities of RNase H and RNases other than RNase H. Suppression of ICAM-1, ELAM-1, or VCAM-1 in HUVEC by their antisense PS-ODNs resulted in the reduction of adhesion of monocytes and U937 to HUVEC. This may suggest cooperativity among the adhesion molecule pairs in endothelial-leukocyte adhesion, since decrease of a single adhesion molecule on EC surface significantly reduced cell-cell adherence.

Base Sequence

The sodium lauryl sulfate model: an overview.

Irritant contact dermatitis is a complex entity with several clinical forms (acute, cumulative, etc.) and multiple mechanisms. Sodium lauryl sulfate (SLS), the most widely utilized model for studying acute and cumulative irritation, has proved highly practical and informative for such studies. This article summarizes several decades of investigation, and provides details of dosing, application method (closed versus open), and biologic endpoints (visual grading, transepidermal water loss) that may be utilized in future studies.

Acute Disease

Extreme fitness differences in mammalian and insect hosts after continuous replication of vesicular stomatitis virus in sandfly cells.

Continuous, persistent replication of a wild-type strain of vesicular stomatitis virus in cultured sandfly cells for 10 months profoundly decreased virus replicative fitness in mammalian cells and greatly increased fitness in sandfly cells. After persistent infection of sandfly cells, fitness was over 2,000,000-fold greater than that in mammalian cells, indicating extreme selective differences in the environmental conditions provided by insect and mammalian cells. The sandfly-adapted virus also showed extremely low fitness in mouse brain cells (comparable to that in mammalian cell cultures). It also showed an attenuated phenotype, requiring a nearly millionfold higher intracranial dose than that of its parent clone to kill mice. A single passage of this adapted virus in BHK-21 cells at 37 degrees C restored fitness to near neutrality and also restored mouse neurovirulence. These results clearly illustrate the enormous capacity of RNA viruses to adapt to changing selective environments.

Animals

Multiple brushings with immediate Riu's stain via flexible fibreoptic bronchoscopy without fluoroscopic guidance in the diagnosis of peripheral pulmonary tumours.

BACKGROUND: Accurate diagnosis of peripheral pulmonary lesions usually relies on fluoroscopic guided procedures. As fluoroscopy is not routinely available in many respiratory units, an approach not using fluoroscopy but with a high diagnostic yield is highly desirable. METHODS: Immediate cytological examination of multiple brushings using Riu's stain, a modified Wright's stain, was performed in 38 patients with peripheral pulmonary lesions not visible at bronchoscopy. The results were compared with the final diagnoses determined by histological examination or subsequent Papanicolaou staining of cytological specimens and clinical course. RESULTS: Of the 38 patients 29 were subsequently confirmed to have a malignant tumour. Our method provided a diagnosis of malignancy in 86% of these lesions. The accuracy (91%) and sensitivity (88%) were higher for lesions > 3 cm in diameter than for those of diameter < or = 3 cm (87% and 83%). There were no false positive results. The 29 lesions correctly diagnosed as malignant by Riu's stain required significantly fewer brushings (mean (SD) 3 (2)) than the nine benign lesions (5 (4)). CONCLUSIONS: This technique provides a high diagnostic yield, avoids the need for fluoroscopy, and is probably safer than percutaneous biopsy.

Adenocarcinoma, Bronchiolo-Alveolar

CD34 immunohistochemical staining of bone marrow biopsies in myelodysplastic syndromes.

Although it has been shown that the percentage of bone marrow blasts in myelodysplastic syndrome (MDS) constitute the only independent determinant of survival and progression to acute leukemia, the great variability in survival among patients with MDS of similar percentage of blasts has prompted us to investigate new objective, independent prognostic parameters for the selection of high-risk patients. It was suggested that CD34 antigen expression adversely affected the prognosis of acute myelogenous leukemia. However, no study has been published so far on clinical and prognostic significance of CD34 antigen expression in MDS. Bone marrow biopsies from 58 patients diagnosed as primary MDS were studied using QBEND/10, a monoclonal antibody which recognized the human progenitor CD34 antigen on routine aldehyde-fixed, paraffin-embedded samples. The high percentage of CD34-positive cells (above 3% of total bone marrow nucleated cells) was predominantly observed in cases with RAEB-T, CMML, and to a lesser degree in RAEB. But neither age, hemograms, bone marrow findings including percentage of blasts, ALIP, nor leukemic transformation correlated with the percentage of CD34-positive cells. The median actuarial survival time in the high positive group was significantly shorter (12.0 months) than that of the low group (30.0 months; p = 0.028). The high CD34 aggregate (> or = 3) was selectively found in cases with RAEB, RAEB-T, and CMML. The percentage of bone marrow blasts (p = 0.007) and ALIP (p = 0.030) significantly correlated with number of CD34 aggregates.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Studies of cloning, chromosomal mapping, and embryonic expression of the mouse Rab geranylgeranyl transferase beta subunit.

The mouse Rab geranylgeranyl transferase beta subunit has been cloned from a mouse E8.5 embryonic cDNA library. Sequence comparison reveals 97.4% sequence identity at the amino acid level to the rat clone isolated from an adult rat brain cDNA library. This gene, given a gene symbol of Rabggtb, is mapped in the distal region of mouse chromosome 3. It is ubiquitously expressed in adult animals but displays an interesting pattern of expression during a specific time of embryonic development. The expression of this gene can be detected in the whole embryos during early embryonic stages and is specifically concentrated in the developing brain, heart, and liver between gestation stages of E11.5 and E13.5. In addition, the expression of this gene is induced by retinoic acid in a mouse embryonal carcinoma cell line, P19.

Animals

Emergency surgical intervention for severe corrosive injuries of the upper digestive tract.

BACKGROUND: In treating severe corrosive injury of the esophagus and stomach, prompt diagnosis, adequate fluid resuscitation and warranted surgical intervention are the most important factors in rescue of critically ill patients. The purpose of this study was to evaluate the need for, and the advantages of, a surgical approach to treatment of such corrosive injuries to the upper gastrointestinal (UGI) tract, as well as to select the most suitable technique to achieve a good survival rate. METHODS: From January 1983 to December 1991, 220 patients were treated for caustic ingestion injury to the UGI tract. A retrospective review of their records allowed targeting of 27 patients with severe corrosive injury that surgical intervention was required. In this study, peritoneal sign was taken as the key indicator for early emergency operation. The age, sex, elapsed time from injury to operation, the sort and quantity of caustic agent used, injury mechanism, clinical manifestations, alternative surgical treatment methods and causes of death were also reviewed and analyzed in this study. RESULTS: The patients included 13 men and 14 women, of whom the majority were adults (96.3%) who had attempted suicide (85.2%). All of them had taken liquid corrosive agents, usually hydrochloric acid (63%). Eighteen underwent emergency operations; the other nine received only supportive treatment, given their terminal status. The mortality rates for patients with surgery and supportive treatment were 66.7% and 100%, respectively. Four patients died after undergoing esophagectomy with resection of the stomach using the thoracoabdominal method. Only three of the eight patients who received esophageal stripping combined with resection of the stomach through the abdomen died (37.5%). CONCLUSIONS: The time elapsed between injury and development of peritoneal sign is a good indicator of the severity and extent of the injury. When peritoneal sign manifests at a very early stage, it is an indicator that the corrosive injury is very advanced in its progress and that, no matter what procedures were performed, the outcome would be the same. Yet if there were a six-hour gap then aggressive surgical management can rescue some patients. It is recommended based on experience here, that when using the surgical approach, resection of the stomach with stripping of the esophagus is superior to the thoracoabdominal method.

Adolescent

Crystal structures of peptide complexes of the amino-terminal SH2 domain of the Syp tyrosine phosphatase.

BACKGROUND: Src homology 2 (SH2) domains bind to phosphotyrosine residues in a sequence-specific manner, and thereby couple tyrosine phosphorylation to changes in the localization or catalytic activity of signal transducing molecules. Current understanding of SH2 specificity is based on the structures of SH2-peptide complexes of the closely-related Src and Lck tyrosine kinases. The tyrosine phosphatase Syp contains two SH2 domains that are relatively divergent from those of the tyrosine kinases, with distinct target specificities, and is thus well suited for structural studies aimed at extending our understanding of SH2 specificity. RESULTS: Crystal structures of the amino-terminal SH2 domain of Syp in separate complexes with two high-affinity peptides, in complex with a non-specific peptide and in the uncomplexed form have been determined at between 2 A and 3 A resolution. The structure of the SH2 domain and the mode of high-affinity peptide binding is essentially similar to that seen in the Src and Lck structures. However, the binding interface is more extensive in Syp. CONCLUSIONS: Most SH2 targets have hydrophobic residues at the third position following the phosphotyrosine, and the Syp structure confirms that the peptide is anchored to the SH2 surface by this residue and by the phosphotyrosine. In addition, the Syp structure has revealed that sequence specificity can extend across the five residues following the phosphotyrosine, and has shown how the SH2 domain's surface topography can be altered with resulting changes in specificity, while conserving the structure of the central core of the domain.

Amino Acid Sequence

Demethylation in the 5'-flanking region of mouse cellular retinoic acid binding protein-I gene is associated with its high level of expression in mouse embryos and facilitates its induction by retinoic acid in P19 embryonal carcinoma cells.

The mouse cellular retinoic acid binding protein-I (CRABP-I) gene is specifically up-regulated by retinoic acid (RA) in P19 mouse embryonal carcinoma cells, and its expression in animals is spatially and temporally restricted to RA-sensitive tissues during embryonic development. This study demonstrates that, in adult mouse tissues and P19 cells where the expression of CRABP-I is detected at the basal level, the 5'-flanking region of the CRABP-I gene is hypermethylated at the C residues of all the Hpa II sites. Conversely, in mouse embryos during early stages of development when the expression of CRABP-I gene is detected at a much higher level, this region is demethylated at these Hpa II sites. In P19, enhancement on the RA-induced up-regulation of CRABP-I can be observed in cells treated with 5-azacytidine (5-AzaC) in conjunction with RA, where partial demethylation in the 5'-flanking region of CRABP-I gene is observed. Nuclear run-on experiments indicate that increased message levels of CRABP-I in P19 cells can be accounted for, at least partially, by increases in its transcription rates. The induction of retinoic acid receptor (RAR) beta by RA can also be enhanced by 5-AzaC, but to a much lesser degree. In contrast, all the Hpa II sites in the structural gene portion, at least in the first two exons, are fully demethylated at the C residues.

Animals