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Y Tan

Publications and source records attributed to Y Tan.

At least 109 records · Page 6Linked to original sources
PubMed1998

Origins and antiquity of X-linked triallelic color vision systems in New World monkeys.

It is known that the squirrel monkey, marmoset, and other related New World (NW) monkeys possess three high-frequency alleles at the single X-linked photopigment locus, and that the spectral sensitivity peaks of these alleles are within those delimited by the human red and green pigment genes. The three alleles in the squirrel monkey and marmoset have been sequenced previously. In this study, the three alleles were found and sequenced in the saki monkey, capuchin, and tamarin. Although the capuchin and tamarin belong to the same family as the squirrel monkey and marmoset, the saki monkey belongs to a different family and is one of the species that is most divergent from the squirrel monkey and marmoset, suggesting the presence of the triallelic system in many NW monkeys. The nucleotide sequences of these alleles from the five species studied indicate that gene conversion occurs frequently and has partially or completely homogenized intronic and exonic regions of the alleles in each species, making it appear that a triallelic system arose independently in each of the five species studied. Nevertheless, a detailed analysis suggests that the triallelic system arose only once in the NW monkey lineage, from a middle wavelength (green) opsin gene, and that the amino acid differences at functionally critical sites among alleles have been maintained by natural selection in NW monkeys for >20 million years. Moreover, the two X-linked opsin genes of howler monkeys (a NW monkey genus) were evidently derived from the incorporation of a middle (green) and a long wavelength (red) allele into one chromosome; these two genes together with the (autosomal) blue opsin gene would immediately enable even a male monkey to have trichromatic vision.

Alleles↗
PubMed1998

Widespread skeletal metastatic potential of human lung cancer revealed by green fluorescent protein expression.

To understand the skeletal metastatic pattern of non-small cell lung cancer, we developed a stable high-expression green fluorescent protein (GFP) transductant of human lung cancer cell line H460 (H460-GFP). The GFP-expressing lung cancer was visualized to metastasize widely throughout the skeleton when implanted orthotopically in nude mice. H460 was transduced with the pLEIN retroviral expression vector containing the enhanced GFP and the neomycin (G418) resistance gene. A stable high GFP-expressing clone was selected in vitro using 800 microg/ml G418. Stable high-level expression of GFP was maintained in s.c.-growing tumors formed after injecting H460-GFP cells in nude mice. To use H460-GFP for visualization of metastasis, fragments of s.c.-growing H460-GFP tumors were implanted by surgical orthotopic implantation in the left lung of nude mice. Subsequent micrometastases were visualized by GFP fluorescence in the contralateral lung, plural membrane, and widely throughout the skeletal system including the skull, vertebra, femur, tibia, pelvis, and bone marrow of the femur and tibia. The use of GFP-expressing H460 cells transplanted by surgical orthotopic implantation revealed the extensive metastatic potential of lung cancer in particular to widely disseminated sites throughout the skeleton. This new metastatic model can play a critical role in the study of the mechanism of skeletal and other metastasis in lung cancer and in screening of therapeutics that prevent or reverse this process.

3T3 Cells↗
PubMed1998

Modeling pH effects on microbial growth: a statistical thermodynamic approach.

This paper applies a statistical thermodynamic approach to the kinetics of microbial growth influenced by pH. A general equation is developed and shown to provide a good theoretical basis for the existing pH models that have been widely used to describe the effects of pH on microbial growth kinetics. Four experimental data sets are used to test the general equation developed. The four data sets exhibited a variety of functional curve shapes, for example, symmetrical and asymmetrical bell-shaped, when the specific growth rate of microorganisms is plotted as a function of pH. All four data sets are found to be well represented by the general equation. The existing pH model was, however, found to represent only one out of four data sets, i.e., the symmetrical case.

Bacteria↗
PubMed1998

Presenilin-1 mutations associated with familial Alzheimer's disease do not disrupt protein transport from the endoplasmic reticulum to the Golgi apparatus.

Mutations in genes encoding presenilin-1 (PS1) and presenilin-2 (PS2) have been linked to familial forms of Alzheimer's disease (AD). Cells expressing mutant presenilins produce elevated levels of Abeta42, the major amyloid peptide found in AD plaques. The mechanism whereby this occurs remains unknown, but the localization of presenilins to endoplasmic reticulum (ER) and Golgi compartments has suggested that they may function in intracellular trafficking pathways involved in processing beta-amyloid precursor proteins (APP). To test this possibility, we coexpressed PS1(wt), PS1(M146L), or PS1(L286V) in HEK293 cells together with the LDL receptor, a classic glycoprotein marker that undergoes post-translational O-glycosylation in the Golgi compartment. Pulse-chase analysis of the receptor indicated that mutant presenilins had no effect on ER-->Golgi transport. Similar results were obtained when the studies were carried out with cells expressing the Swedish variant of APP (SWAPP751) instead of the LDL receptor. Moreover, secretion of the soluble exodomain polypeptide fragments of SWAPP751 that arise from alpha-secretase and beta-secretase cleavage was not markedly affected by the PS1 mutants. Despite the lack of discernible effect of the PS1 mutants on trafficking of proteins through the Golgi apparatus, they caused a substantial increase in the proportion of Abeta42 relative to total Abeta in the culture medium. The results suggest that mutant forms of PS1 cause elevated production of Abeta42 by a mechanism that is independent of a major disruption of exocytic trafficking of APP.

Alzheimer Disease↗
PubMed1998

Solid-phase extraction of 18beta-glycyrrhetinic acid from plasma and subsequent analysis by high-performance liquid chromatography.

A new method is described for the solid-phase extraction of 18beta-glycyrrhetinic acid from plasma or serum, with subsequent analysis by HPLC. New aspects of the method include the use of commercially available 18alpha-glycyrrhetinic acid as the internal standard and the use of a Bond Elut C2 (ethyl) extraction column, to avoid the need to use large volumes of organic solvent to elute the isolates from the columns. Separation was achieved on a Shandon Hypersil BDS C18 analytical column, with a mobile phase consisting of acetonitrile-0.02 M phosphate buffer, pH 5.7 (55:45, v/v). The column effluent was monitored at 248 nm. Compared with previous methods, the procedure is much easier to carry out, whereas the sensitivity (limit of detection, 10 ng/ml, and limit of quantitation, 50 ng/ml), the precision (0.3-6.2%) and the accuracy (97.2-101.9%) are of the same order of magnitude.

Calibration↗
PubMed1998

Polyethylene glycol conjugation of recombinant methioninase for cancer therapy.

Recombinant methioninase (rMETase) is a homotetrameric pyridoxal 5'-phosphate enzyme of 172-kda molecular mass derived from Pseudomonas putida and cloned in Escherichia coli. rMETase has been found previously to be an effective, anti-tumor agent in vitro and in vivo. The enzyme targets the elevated minimal methionine requirement seen in all tumor types. In order to prevent immunological reactions which might be produced by multiple dosing of rMETase and to prolong the serum half-life of rMETase, the N-hydroxysuccinimidyl ester of methoxypolyethylene glycol propionic acid (M-SPA-PEG 5000) has been coupled to rMETase. Molar ratios of M-SPA-PEG-5000 (PEG) to rMETase from 10 to 40 were used for PEGylation of rMETase. PEGylation reactions were run at 20 degrees C for 30 to 60 min in reaction buffer (20 mM sodium phosphate buffer, pH 8.3). The PEGylated molecules (PEG-rMETase) were purified from unreacted PEG with Amicon 30 K centriprep concentrators or by Sephacryl S-300 HR gel-filtration chromatography. Unreacted rMETase was removed by DEAE Sepharose FF anion-exchange chromatography. The resulting PEG-rMETase subunit, from a PEG/rMETase ratio of 30/1 in the synthetic reaction, had a molecular mass of approximately 53 kda determined by matrix-assisted laser desorption/ionization mass spectrometry, indicating the conjugation of two PEG molecules per subunit of rMETase and eight per tetramer. PEG-rMETase molecules obtained from reacting ratios of PEG /rMETase of 30/1 had enzyme activities of 70% of unmodified rMETase. PEGylation of rMETase increased the serum half-life of the enzyme in rats to approximately 160 min compared to 80 min for unmodified rMETase. PEG-rMETase could deplete serum methionine levels to less than 0.1 microM for approximately 8 h compared to 2 h for rMETase in rats. Efficacy studies of PEG-rMETase on human lung cancer and kidney cancer cells in vitro demonstrated a 50% inhibitory concentration (IC50) of 0.04 and 0.06 units/ml, respectively. These IC50 values were almost identical to unmodified rMETase, thus indicating maintenance of antitumor efficacy in the PEGylated enzyme. PEG-rMETase had an IC50 for normal lung and kidney cells of 0.8 and 1.5 units/ml, respectively, similar to rMETase. The efficacy data indicated that PEG-rMETase maintained the high level tumor selectivity of rMETase. PEG-rMETase injected intravenously in mice demonstrated a tumor/blood retention ratio of approximately 1/6 compared to 1/10 of unmodified enzyme, indicating that PEG-rMETase distributes to the tumor at least as effectively as rMETase.

Animals↗
PubMed1998

High expression, purification, and properties of recombinant homocysteine alpha, gamma-lyase.

Homocysteine alpha,gamma-lyase from the anaerobic protozoan parasite Trichomonas vaginalis has been cloned from genomic DNA using PCR methods and expressed in Escherichia coli with a vector containing the T7 promoter. The recombinant homocysteine alpha,gamma-lyase (rHCYase) is expressed as the major protein in the host E. coli cells. The enzyme was purified to approximately 90% purity using heat treatment at 50 degreesC, precipitation steps with polyethyleneimine, polyethylene glycol 8000, and high sodium chloride, DEAE-Sepharose FF chromatography, and phenyl-Sepharose 6 FF chromatography. The final yield was greater than 50%, which encompassed an approximate 18-fold purification. The enzyme is a homotetramer with a monomer molecular weight of 43K and contains pyridoxal phosphate. The Trichomonas rHCYase is selective for homocysteine with respect to very low cysteinase activity in contrast to the alpha,gamma-lyase from Pseudomonas putida, which has very high cysteinase activity with respect to homocysteine. The T. vaginalis and P. putida alpha,gamma-lyases readily separate on a phenyl-Sepharose 6 FF column with the T. vaginalis enzyme eluting first. rHCYase is stable up to 50 degreesC and active over a pH range of 6-8. These properties of high recombinant expression in E. coli, a simple and effective high-yield purification procedure and high relative specificity for homocysteine with respect to cysteine, make rHCYase a promising candidate to use for the diagnosis of hyperhomocystenemia, which has been demonstrated to be a major risk factor for the onset and mortality of cardiovascular disease of all types.

Animals↗
PubMed1998

A gross and microscopic study of cerebral injuries accompanying maxillofacial high-velocity projectile wounding in dogs.

PURPOSE: This study investigated the pathologic characteristics and some related factors of cerebral damage associated with maxillofacial high-velocity missile wounds. MATERIALS AND METHODS: Sixty dogs, divided into two groups, were wounded randomly by steel spheres weighting 1.03 g at impact velocities of 1,400 m/s (46 dogs) and 800 m/s (14 dogs). Six dogs served as controls. The maxillofacial wounds and cerebral injuries were examined grossly. The distance between the center of wound and the base of skull and the largest diameter of the wound were measured, and the incidence of the brain injury in the two groups was compared. The cerebral specimens, dissected at 1 and 6 hours after trauma, were observed by light and electron microscopy. RESULTS: Cerebral hyperemia, contusion, spotty hemorrhage, and intracerebral hematoma were found in some of the dogs. In the 1,400 m/s velocity missile wound group, the incidence of the brain injury was 71.7% (33 of 46), and in the 800 m/s group it was 7.1% (1 of 14) on macroscopic examination. Microscopic observation showed intracerebral microhemotomas and degeneration and necrosis of the nervous cells. CONCLUSION: High-velocity projectile maxillofacial wounds can induce associated brain injury.

Animals↗
PubMed1998

The distribution and translocation of the G protein ADP-ribosylation factor 1 in live cells is determined by its GTPase activity.

ADP-ribosylation factors (ARF) are small G proteins that play key roles in vesicular transport processes. We have studied the distribution of ARF1 in live cells using chimeras of ARF1 mutants (wild type (wt) ARF1; Q71L-ARF1 (reduced GTPase); T31N (low affinity for GTP); and (Delta)Nwt (deletion of amino acids 2-18)) with green fluorescent protein (GFP). Confocal microscopy studies showed that the wt and Q71L proteins were localized in the Golgi and cytoplasm. The (Delta)Nwt and the T31N mutants were exclusively cytoplasmic. The behavior of the wt and Q71L proteins was studied in detail. About 15% of wt-ARF1-GFP was bound to the Golgi. Bound wt-ARF1-GFP dissociated rapidly after addition of Brefeldin A (BFA). This process did not appear to be a consequence of BFA-induced disappearance of the Golgi. Photobleaching recovery showed that essentially all the ARF-GFP was mobile, although it diffused very slowly. In contrast, about 40-50% of the Q71L mutant was found in the Golgi, and its rate of dissociation in the presence of BFA was slow and biphasic. Q71L-ARF1-GFP diffused more slowly than the wt. We conclude that ARF1 proteins exist in a dynamic equilibrium between Golgi-bound and cytosolic pools, and that the translocation of ARF in live cells requires the hydrolysis of GTP by the Golgi-bound protein.

ADP-Ribosylation Factor 1↗
PubMed1998

Basic fibroblast growth factor-mediated lymphangiogenesis of lymphatic endothelial cells isolated from dog thoracic ducts: effects of heparin.

We have attempted to evaluate whether, similar to the angiogenesis of blood vessels, cultures of lymphatic endothelial cells (LEC) isolated from dog thoracic ducts have an ability to induce lymphangiogenesis in response to basic fibroblast growth factor (bFGF), then to examine the effects of heparin on the bFGF-mediated morphogenesis. The effects of bFGF and/or heparin on the proliferation and migration of the LEC were evaluated by changing the number of the subconfluent cells and by wound migration assay, respectively. The effects of the agents on invasion and tube formation of the LEC into a three-dimensional collagen gel and on collagen gel induced tube formation of the LEC were also investigated by a phase-contrast microscope and an electron microscope. The bFGF (10 ng/ml) caused a significant induction of proliferation and migration of the LEC, the induction of which was augmented dose-dependently by an additional treatment with heparin ranging from 1 to 100 microg/ml. The bFGF produced invasion and tube formation of the LEC into a three-dimensional collagen gel. The bFGF also facilitated to form capillary-like tubes of the LEC between two layers of collagen gels. Heparin (10 microg/ml) accelerated both processes of bFGF-mediated lymphangiogenesis of the LEC. These findings suggest that the cultured LEC isolated from dog thoracic ducts have an ability to form lymphatic capillary-like tubes in response to bFGF and that heparin accelerates dose-dependently the process of the bFGF-mediated neovascularization of lymph vessels.

Animals↗
PubMed1998

[A study on the relationship between Lp(a) and arteriosclerotic cerebral infarction].

Using a 1:1 matched case control study, we analysed the data of 105 arteriosclerotic cerebral infarction cases and controls to detect the levels of TC, TG, HDL-C, LDL-C and Lp(a). The results showed that the average level of Lp(a) in cases was (238.38 mg/L), significantly higher than that of controls (149.98 mg/L t = 3.96, P < 0.001). Comparing the levels of different age groups and sex, no significant difference, except the group over 70 years, which was higher than that in other groups was found. Dose-response analysis also showed that the OR value was positively correlated with the Lp(a). Multi-variable logistic regression model indicated that Lp(a) was an independent risk factor with OR = 1.93. Another risk factor was TG. However, LDL-C, HDL-C were found to be the protective factors of arteriosclerotic cerebral infarction, with an OR = 0.32.

Aged↗
PubMed1998

The effect of thyrotropin receptor antibodies on the proliferation of FRTL-5 cells and the expression of protooncogene c-fos mRNA.

OBJECTIVE: Hyperthyroidism and a diffuse goiter are the main symptoms of Graves' disease (GD) associated with autoantibodies to thyroid-stimulating hormone (TSH) receptor (TRAb). The present study was conducted to evaluate effects of autoantibodies in patients with GD (TRAb-IgG) on induction of the proliferation and c-fos mRNA expression in FRTL-5 cells (Fisher rat thyroid cell line). METHODS: Highly purified IgG fractions were isolated from 11 patients with GD, TRAb-IgG and 15 normal individuals (normal controls) with Protein A Sepharose CL-4B affinity column chromatograph. FRTL-5 cells, which had been grown to subconfluency and deprived of TSH for a few days. Then, these cells were used for measuring cAMP content, 3H-thymidine incorporation in cells and the expression of c-fos mRNA respectively. RESULTS: After stimulation of TRAb-IgG, the cAMP production and 3H-thymidine incorporation in FRTL-5 cells were much higher than those from normal controls (P < 0.05 respectively). Using 32P labelled v-fos probe by the Northern Blot method, the expression of c-fos mRNA could be induced by IgGs from patients with GD. CONCLUSIONS: These data suggest that the stimulation of TRAb-IgG followed by cAMP production and 3H-thymidine incorporation is related to the induction of c-fos mRNA and, thus, to the growth of FRTL-5 cells.

Animals↗
PubMed1998

Experimental study on firearm wound in maxillofacial region.

OBJECTIVE: To make clear the range of firearm wound in the maxillofacial region, the optical repair time and the characteristics of accompanied indirect brain damage, and to offer the principle of emergency treatment and the early repair of war wound. METHODS: With the aid of the standard Sweden model, 200 dogs were used in the experiment. Varies tissues around the primary canal were harvested chronologically, in different zone and different tissue, for histopathological examination. RESULTS: The necrotic range of various tissues in the maxillofacial region was less than that in the extremities. In the maxillofacial region, there was a significant temporary cavity following the passing of bullet, which caused indirect brain damages. CONCLUSION: These findings are helpful to the treatment of war wound in the maxillofacial region. Early bone transplantation using microvascular anastomosis in the treatment of gunshot wound in the maxillofacial region is recommendable.

Adenosine Triphosphatases↗
PubMed1998

[The method of removing methamidophos from contaminated vegetables].

Since the massive food poisoning outbreak in 1987, imported vegetables contaminated with methamidophos continued to cause sporadic food poisoning outbreaks in Hong Kong. Despite various administrative measures to lower the risk of importing vegetables, it is evident that the occurrence of sporadic food poisoning outbreaks cannot be completely stemmed out. The education of the public on the effective ways to remove methamidophos from the contaminated vegetables was reckoned to the another preventive measure against food poisoning. A study was carried out to evaluate the effectiveness of various ways of treating the vegetables before consumption. In this study, the removal of methamidophos by simply washing with water at near room temperature was found to be a slow process. The concentration of methamidophos was reduced by about 65% after washing in water for an hour. Further washing did not improve the situation. The addition of detergents and various washing aids including potassium permanganate, hydrogen peroxide, sodium bicarbonate and vinegar did not greatly enhance the removal effectiveness. Among the various treatment procedures, soaking in hot water was the most effective way to remove methamidophos from vegetables. Less than 10% of the pesticide remained in the vegetables after soaking in boiling water for 1 minute.

Food Contamination↗
PubMed1998

[Mutation analysis of the p15 gene exon 2 in human primary hepatocarcinoma].

To investigate the role p15 gene plays in the pathogenesis of human primary hepatocarcinoma, 35 human hepatocarcinomas, 35 cases of adjacent non-cancerous liver cirrhosis and the blood cells of 10 normal human were analyzed for somatic mutation in p15 gene with PCR-SSCP. One case of adjacent non-cancerous liver cirrhosis showed abnormal migration single strand. In the hepatocarcinomas and in the other cases of adjacent non-cancerous liver cirrhosis, no mutation was found. Cloning and sequencing of the amplified abnormal migration single strand DNA revealed that it contained a wild type exon 2 of p15 gene in 345 bp length. The results indicate that the inaction of p15 gene by point mutation is a very uncommon event in human hepatocarcinoma.

Carcinoma, Hepatocellular↗
PubMed1998

[Neuroendocrine differentiation in ovarian mucinous tumors].

OBJECTIVE: To investigate the relationship between neuroendocrine differentiation of ovarian mucinous tumors and its genesis. METHODS: A morphologic study on 73 cases of ovarian mucinous tumors (32 benign, 20 borderline, 21 malignant) using immunohistochemical and immunohistochemical/histochemical double staining techniques. RESULTS: The incidences of CgA positive cell in tumors of benign, borderline and malignant were 62.5%, 75%, 76% and that of 5-HT positive cells were 31.3%, 40%, 39% respectively. But these NEC were not evenly distributed. The interesting point was that in four cases of the benign tumors the number of CgA positive cell was more than 30 per cent localizing between the glandular basement membrane and the mucinous epithelial cells. There were also many intermediate cells containing both CgA and PAS positive granules. CONCLUSIONS: The positive rate of CgA was increased corresponding to the lowering down of differentiation grades. The four cases of the benign tumors might be originating from a common stem cell, such as so-called amphocrine cell. The relationship between these four tumors and neuroendocrine differentiation in ovarian mucinous tumors remain further clarified.

Cell Differentiation↗