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Biomedical subjects

W Hong

Publications and source records attributed to W Hong.

At least 145 records · Page 8Linked to original sources

Retention of a type II surface membrane protein in the endoplasmic reticulum by the Lys-Asp-Glu-Leu sequence.

Soluble luminal proteins of the endoplasmic reticulum (ER) are known to be retained by a tetrapeptide retention signal, KDEL. We report in this communication that the KDEL sequence when appended to the carboxy terminus of a cell surface membrane protein, dipeptidyl peptidase IV (DPPIV), resulted in its retention in the endoplasmic reticulum of transfected Madin-Darby canine kidney cells as assessed by indirect immunofluorescence. Selective surface biotinylation revealed that about 90-95% of the expressed DPPIV was retained in the ER. Appendance of the sequence KDEV did not, however, result in ER retention, illustrating the functional specificity of the retention signal. The ER retention was not due to misfolding of the mutant protein, as the mutant proteins remained enzymatically active. Our data suggest that the KDEL receptor is able to recognize and recycle type II membrane proteins containing a carboxyl-terminal KDEL sequence and postulates the existence of such yet to be identified endogenous proteins.

Amino Acid Sequence↗

The 17-residue transmembrane domain of beta-galactoside alpha 2,6-sialyltransferase is sufficient for Golgi retention.

beta-Galactoside alpha 2,6-sialyltransferase (ST) is a type II integral membrane protein of the Golgi apparatus involved in the sialylation of N-linked glycans. A series of experiments has shown that the 17-residue transmembrane domain of ST is sufficient to confer localization to the Golgi apparatus when transferred to the corresponding region of a cell surface type II integral membrane protein. Lectin affinity chromatography of chimeric proteins bearing this 17-residue sequence suggests that these chimeric proteins are localized in the trans-Golgi cisternae and/or trans-Golgi network. Further experiments suggest that this 17-residue sequence functions as a retention signal for the Golgi apparatus.

Amino Acid Sequence↗

Selective inhibition of protein targeting to the apical domain of MDCK cells by brefeldin A.

Dipeptidyl peptidase IV (DPPIV) is mainly vectorially targeted to the apical surface in MDCK cells. BFA was found to abolish the apical targeting of DPPIV. This BFA effect could be achieved under conditions where the ER to Golgi transport and the total surface expression of DPPIV were essentially unaffected. BFA executed its effect during the transport from the trans-Golgi network (TGN) to the surface. The inhibition of apical targeting resulted in enhanced mis-targeting to the basolateral surface. The mistargeted DPPIV was transcytosed back to the apical domain only after BFA withdrawal. In contrast, the basolateral targeting of uvomorulin was unaffected by BFA. These results established that the apical targeting of DPPIV was selectively abolished by BFA.

Animals↗

[Eclampsia complicated with cerebrovascular disease].

Ten eclamptic patients complicated by acute cerebrovascular disease were admitted during the past eight years from Feb. 1982 to Feb. 1990. There were six cases of superior sagittal sinus thrombosis, two cases of cerebral hemorrhage, one cases of subarachnoid hemorrhage and the other one of cerebral infarction. Among them, eight patients were cured. Our analysis demonstrates that it is advisable to terminate pregnancy by cesarean section as soon as possible. A comprehensive management proposed by good cooperation between neurologist, neurosurgeon and obstetrician is the key to get a good way to cure these patients.

Adult↗

Immobilization of nucleoside on silica gel and specific separation of oligonucleotides.

Deoxyadenosine was immobilized on silica gel, in order to use as HPLC resins for selective separation of oligonucleotides. The longest retention time was observed for the complementary pd(T)4, and increased with decrease of temperature. This fact suggested that the main separation factor was the base pairing between complementary nucleic acid bases. These resins may be useful for separation of components of nucleic acids and polynucleotides as a specific separation system.

Chromatography, High Pressure Liquid↗

Age distribution and turnover of physicians in nonmetropolitan counties of the United States.

Using data for 1975 to 1985 from the Area Resource File (ARF), net number changes in different age groups of physicians practicing in nonmetropolitan counties are examined. Small net increases are seen in the higher age groups in most county size categories but the most striking change is large increases in the age groups up to age 44. Family practitioners show a net decline in all county size categories for ages 45-54, but these are offset by increases in medical specialists. In counties of less than 25,000 population, the rate of turnover of physicians over the 1983-1988 period was 25 percent; physicians exiting these counties had a mean age of 52.3 years. The data indicate that aging of physicians in rural counties should not affect maintenance of current supply in the short run, but that increasing the physician-to-population ratio in areas with less adequate supply will be difficult, particularly if the rate of increase of younger physicians in nonmetropolitan counties does not continue.

Adult↗

Inhibition by brefeldin A of protein secretion from the apical cell surface of Madin-Darby canine kidney cells.

The effect of brefeldin A (BFA) on total and polarized protein secretion was examined in MDCK cells. Increasing concentrations of BFA have increasingly inhibitory effects on total protein secretion. The total protein secretion was essentially unaffected by BFA at 0.5 microgram/ml. When the BFA concentration was increased to 10 and 30 micrograms/ml, the total protein secretion was reduced to about 70 and 25%, respectively, of the control level. Consistent with this effect on total protein secretion, the Golgi structure as revealed by C6-NBD-ceramide (a fluorescent ceramide analog) staining was essentially unaltered by 0.5 microgram/ml BFA, while 10 and 30 micrograms/ml BFA significantly dispersed the Golgi apparatus. When the polarity of protein secretion was examined, it was found that the ratio of proteins secreted from the apical to those from the basolateral surface was reduced from 1.5-2.0 to 0.4-0.7 by all three BFA concentrations. Furthermore, several proteins which are preferentially released from the apical surface were found to be released without apparent surface polarity, while several other proteins which were preferentially released from the basolateral surface were unaffected. This study suggests that BFA, at 0.5 microgram/ml, can selectively inhibit protein secretion from the apical surface without affecting total protein secretion. The inhibition of apical secretion results in enhanced protein secretion from the basolateral surface.

4-Chloro-7-nitrobenzofurazan↗

Evaluation of second-look laparotomy for ovarian cancer: second-look vs. serum CA125.

Thirty patients with ovarian cancers underwent second-look operations in our hospital between 1981 and 1989, and the results were compared with serum CA125 values. The over-all positive rate determined by second-look laparotomy in the present series was 56.7% (17/30)-40% as gross and 17% an microscopic tumors. The result of the second-look was related to the staging and size of the residual tumor after the previous operation: 1/3 of stage I and II lesions and 16/27 of stage III and recurrent lesions were positive on second-look. The positive rates of those patients who were free from residual tumor and those with residual tumors less than or equal to 2 cm or greater than 2 cm were 28.6% (2/7), 60% (9/15) and 75% (6/8), respectively. Among the 23 patients assayed as to serum CA125 level before the second-look, 18 (78%) were within normal limits (less than 65 U/ml), including 12 patients (66.6%) who were second-look positive and 6 who were negative. Five patients with microscopic tumors and 3 patients with gross tumors less than or equal to 2 cm all had normal CA125 levels before the second-look. Among the 5 of 9 patients with gross tumors greater than 2 cm who had elevated CA125 levels, not one was found to be negative on second-look. This may imply that the elevated CA125 levels indicate recurrence, though a normal CA125 assay certainly does not preclude recurrence.

Adult↗

[Practical anatomy of the frontal and facial veins].

The frontal and facial veins of 15 adult cadavers were dissected. The main branch(es) of the frontal vein may be single (13.3%), dual (80.0%), or triple (6.7%). Blood from the veins around the orbit is collected by the venous sinus in the inner canthus, and the transverse dorsal nasal vein is a bridge between the venous sinuses of both sides. Therefore, veins around the orbit on both sides can be radiographed by venous puncture on either side. The facial vein is S-shaped and composed of 4 segments. Projective locations of the veins are described, with suggestions for practical clinical applications.

Face↗

cDNA cloning for a bile canaliculus domain-specific membrane glycoprotein of rat hepatocytes.

Hepatocytes are polarized cells with distinct sinusoidal, bile canalicular, and basolateral plasma membrane domains. Each domain contains proteins that are specific for it. We have isolated three cDNA clones encoding a rat liver bile canaliculus domain-specific glycoprotein with Mr 110,000 (gp110) by immunologically screening a rat kidney lambda gt11 cDNA library with a rabbit polyclonal antiserum directed against purified gp110. The authenticity of these clones was verified as follows. (i) The antiserum recognizes specifically isopropyl beta-D-thiogalactoside-induced fusion proteins on electrophoretic transfer blots of total lysogen lysates containing these cDNA clones. (ii) Antibodies epitope-selected by these clones are able to interact with gp110 on electrophoretic transfer blots. (iii) The amino acid sequencing derived from the DNA sequence was confirmed by amino acid sequencing of a tryptic peptide of gp110. Rescreening of the same library with the cDNA clones identified a full-length cDNA clone for this glycoprotein. Sequence analysis indicates that the N-linked carbohydrate chains are concentrated on the N-terminal part of this highly glycosylated protein.

Amino Acid Sequence↗

Identification and characterization of a murine receptor for galactose-terminated glycoproteins.

The asialoglycoprotein receptor, the hepatic binding lectin for galactose-terminated glycoproteins, has been isolated and characterized from human, rabbit and rat liver. Several recent studies have shown the existence of the same receptor in murine liver. However, the biochemical structure of the receptor in murine liver has not been resolved. In this paper, we describe the identification and purification of the receptor for asialoglycoproteins from murine liver. The purified receptor has three polypeptides with apparent molecular weights of 42,000, 45,000 and 51,000, respectively, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Furthermore, our studies suggest that the receptor from murine liver is very similar to its counterpart in rat liver, although some potential interesting differences have also been observed. Initial studies indicate that this receptor is well conserved in different mouse strains.

Animals↗

Effects of NH4Cl and nocodazole on polarized fibronectin secretion vary amongst different epithelial cell types.

The extracellular matrix protein fibronectin was found to be secreted by three polarized epithelial cell lines Madin-Darby canine kidney (MDCK), Caco-2 and LLC-PK1. About 54 and 46% of fibronectin was secreted from the apical and basolateral cell surfaces, respectively, in MDCK cells. In Caco-2 and LLC-PK1 cells, the majority (about 92-93%) of fibronectin secretion occurs from the basolateral cell surface, with the remaining 7-8% from the apical surface. In all three cell types, NH4Cl was found to inhibit basolateral secretion (resulting in enhanced apical secretion), while total fibronectin secretion was not significantly affected (although a delay in secretion was observed). Nocodazole reduced total fibronectin secretion to about 70% of control levels in MDCK and Caco-2 cells, with significant inhibition on secretion from both surfaces. In contrast, total fibronectin secretion was enhanced by nocodazole in LLC-PK1 cells. Furthermore, the majority of fibronectin secretion was redirected to the apical cell surface in LLC-PK1 cells. These observations demonstrate that the nature as well as the extent of the effects of NH4-Cl and nocodazole on polarized fibronectin secretion varies amongst different epithelial cell types.

Ammonium Chloride↗