Biomedical subjects

S M Kuo

Publications and source records attributed to S M Kuo.

At least 19 recordsLinked to original sources

PubMed2006

Preparation of nano-sized particles from collagen II by a high-voltage electrostatic field system.

The pilot study describes a novel method for preparing nano-sized particles from collagen II using a high-voltage electrostatic field system. Observations from transmission electron microscopy showed that, in one of the cases, the nano-sized collagen II particles exhibited good sphericity, and the particles were in the range of 23.3+/-1.7 nm in diameter at the experimental setting of 3 kV cm(-1), for a 3 h treatment period and at 25 degrees C (with a collagen concentration of 0.2 mg ml(-1)). When the treatment temperature increased to 30 degrees C, the collagen II began to lose the tendency to form individually separated spherically shaped nano-particles. Moreover, a fibrous structure of collagen II was formed instead of a nano-particle shape at the temperature of 37 degrees C. This result is probably contributed to by an entropy-driven process that is termed fibrillogenesis, a larger force causing the collagen molecules to self-assemble and then form collagen fibrils. It is interesting to note that this is practically the first attempt to produce nano-particles directly from collagen II solution under the treatment of a high-voltage electrostatic field, together with a set of working parameters for the collagen concentration and low-temperature setting.

Biocompatible Materials↗

PubMed2001

Dihydropyridine calcium channel blockers inhibit ascorbic acid accumulation in human intestinal Caco-2 cells.

We investigated the effect of commonly used medications on the accumulation of ascorbic acid in human intestinal Caco-2 cells. Although ascorbic acid is negatively charged at physiological pH, anionic compounds including drugs and metabolites had little effect on its accumulation. On the other hand, hydrophobic 1,4-dihydropyridine compounds (nifedipine and nicardipine), but not other structurally unrelated calcium channel blockers, were found to be potent inhibitors. They inhibited both Na+-dependent and Na+-independent (K+ substituting Na+) accumulation of ascorbic acid. The inhibition was non-competitive with a Ki of 108 microM and 9 microM for nifedipine and nicardipine, respectively. The efflux of ascorbic acid from cells was not affected. Previously, we reported a similar inhibition of ascorbic acid accumulation by estrogens. When nifedipine and estrogens were included in the buffer together, the combined inhibitory effect was less than additive implying that they may act through the same mechanism. The potential clinical significance of dihydropyridine usage on ascorbic acid status in human needs to be considered.

Ascorbic Acid↗

PubMed2001

Quercetin cumulatively enhances copper induction of metallothionein in intestinal cells.

Wilson's disease, a genetic copper-overload condition, is currently treated with zinc because of the ability of zinc to induce metallothionein. We are interested in nonmetal chemicals that may alter intestinal copper metabolism and thus help to alleviate copper toxicity. Previously, we have shown that quercetin, a dietary flavonoid, can chelate copper. This study further examined the interaction of quercetin and copper in intestinal epithelial cells. We found that quercetin enhanced metallothoinein induction by copper and the effect was dose dependent. Quercetin also exerted a cumulative effect after repeated exposure. Repeated low-dose treatment (3-10 microM) of cells with quercetin can lead to the same effect on metallothoinein as one higher concentration treatment (100 microM). This property of quercetin is distinct from its chemical interaction with copper, but both can contribute to a reduction of copper toxicity. Among other flavonoids tested, two other copper chelators, catechin and rutin, did not increase copper induction of metallothionein, whereas genistein, an isoflavone that does not interact with copper chemically, increased copper induction of metallothionein. The effect of quercetin on copper metabolism is unique. Quercetin decreased zinc-stimulated metallothionein expression and had no effect on the cadmium induction of metallothionein. The clinical application of our observation needs to be explored.

Cadmium↗

PubMed2000

Determination of 14 elements in Taiwanese bones.

The objective of this study was to measure the concentration of 14 elements in human bone and investigate the affecting factors. This study aims to establish a database of elements in Taiwanese bone. Seventy-seven bone samples were taken from 70 subjects who were undertaking various bone surgeries from a medical center in central Taiwan. Bone samples were pretreated using microwave digestion and then, after determining the optimal conditions for analysis, fourteen elements were analyzed using Inductively Coupled Plasma-Atomic Emission Spectroscopy (ICP-AES). Results showed that Ca had the highest concentration, followed by Mg and Zn. The lowest concentrations were of Mn, followed by Cd and Co. There was a positive correlation between the frequency of seafood intake and presence of all the 14 elements in human bone. There were several elements that had positive inter-correlation: As with Co; Al, Ag and Ca; Ni with Co, Mn and Al; Co with Al, Ag and Ca; Mn with Cr and Cu; Cr with Cu; Mg with Ca; Al with Ag and Ca; Ag with Ca; and also Zn with Mg. There were no significant differences found between concentrations of elements and etiology of bone disease and age. The authors suggest that further studies be conducted to establish a normal value of elements in human bone in Taiwan. This will greatly benefit the understanding of the relationship between disease pattern and elements in human bone.

Adult↗

PubMed1999

Expression of antioxidant proteins in human intestinal Caco-2 cells treated with dietary flavonoids.

Dietary flavonoids are known to scavenge free radicals but little information is available on their roles in antioxidant protein gene expression. The goal of this paper is to investigate the effect of flavonoid treatment on the antioxidant protein expression in human intestinal Caco-2 cells. The antioxidant proteins of interest were metallothionein (MT), catalase (CAT), and superoxide dismutase (SOD). Treatment of Caco-2 cells with 100 microM genistein, biochanin A, daidzein or kaempferol significantly increased MT mRNA up to 15 fold. On the contrary, CAT mRNA level was not affected by various flavonoids. We also developed gel activity assays to determine the specific activities of CAT and Cu/Zn SOD in flavonoid-treated Caco-2 cells. Compared to the conventional spectrophotometric assays, the gel assays allow a separation of antioxidant activities of the enzymes from that of the flavonoids. CAT and Cu/Zn SOD were found not to be affected by 48-h treatment of 100 microM dietary flavonoids (genistein, biochanin A, daidzein, flavone, quercetin, or kaempferol). In conclusion, the effects of flavonoids on antioxidant protein expression are structure- and gene-specific. When evaluating antioxidant capacity of flavonoids, their ability to modulate antioxidant protein expression should also be taken into consideration.

Caco-2 Cells↗

PubMed1999

Genistein increases metallothionein expression in human intestinal cells, Caco-2.

Flavonoids found in common vegetables, fruits, and legumes have been shown to possess antioxidant property. This study is the first to demonstrate that one member of the flavonoid family, genistein, can induce the expression of metallothionein (a metal-binding protein with antioxidant property). We found the effect of genistein to be time- and dose-dependent (10-100 microM). The effect can be observed at both protein and mRNA levels and was synergistic to that of 30 microM zinc. Genistein was shown previously to interact with the estrogen receptor and induce gene expression similar to estrogens at a lower affinity. We thus tested the hypothesis that the effect of genistein on metallothionein expression was mediated through the steroid hormone pathway. We found that various glucocorticoids do not affect metallothionein expression in Caco-2 cells. 17Beta-estradiol at 10-100 microM (concentrations much higher than needed to activate the estrogen response element) induced metallothionein expression in Caco-2 cells. However, a synthetic estrogen, diethylstilbestrol, did not increase metallothionein level at 10 microM. 17Beta-estradiol also did not act synergistically with zinc. Thus, genistein may enhance metallothionein expression through an uncharacterized mechanism. Further studies are needed to delineate the molecular mechanism and to determine whether the expression of other genes is also affected by genistein.

Caco-2 Cells↗

PubMed1999

Amination of polycarbonate surface and its application for cell attachment.

The polycarbonate sheet was modified with ammonia gaseous plasma and characterized by the contact angle measurement and ESCA analysis. The contact angles decreased significantly from 77 degrees to about 20 degrees-40 degrees, indicate that the polycarbonate sheet become more hydrophilic after plasma treatment. The ESCA analysis results showed that the hydrophilicity was mainly derived from the amino groups on the modified surface. In this study, a flow-chamber system was also constructed to evaluate the 3T3 fibroblast cells attachment phenomena on these modified sheets. Before the experimental run, the parameters of inoculated cell number and cell passage were examined previously. The results revealed that these two parameters are independent in shear experiment. And besides, 3-hours plating time has the better adhering fraction. The experimental results showed that the 3T3 fibroblast cells adhesion strength increased significantly on the plasma modified sheet.

3T3 Cells↗

PubMed1998

17beta-estradiol inhibition of ascorbic acid accumulation in human intestinal Caco-2 cells.

We investigated the effect of estrogen on the accumulation of ascorbic acid by human intestinal Caco-2 cells. 17beta-estradiol, synthetic estrogen diethylstilbestrol, and partial agonist tamoxifen were found to inhibit ascorbic acid accumulation in a dose-dependent fashion. The inhibitory effect of estrogens can be observed at as short as 5 min of incubation. An additive effect was observed when they were used in combination. Similar to dietary flavonoids, inhibition was also observed in two other intestinal cell lines, HT-29 and IEC-6. These chemicals affected both Na+ -dependent and Na+ -independent(K+ substituting Na+) accumulation of ascorbic acid and did not affect the efflux of accumulated ascorbic acid. Kinetic analysis of diethylstilbestrol showed a non-competitive inhibition with an apparent Ki of 23 microM. The hormone-ascorbic acid interaction in the intestinal cell could help to explain the known reduction in blood ascorbic acid level among oral contraceptive users and female guinea pigs given contraceptive hormones.

Ascorbic Acid↗

PubMed1998

The protein thiol metallothionein as an antioxidant and protectant against antineoplastic drugs.

Metallothioneins (MTs) are major zinc-binding protein thiols that are readily inducible and whose functions remain unclear. Recent evidence supports a role for MT as an antioxidant. Mechanisms underlying this function may include direct interception of free radicals, complexation of redox sensitive transition metals, altered zinc homeostasis or interaction with glutathione (GSH). MT overexpression after direct gene transfer in cultured cells, decreases cytotoxicity, to partially reduce reactive oxygen and nitrogen species and markedly attenuates intracellular oxidation of reporter molecules including dichlorofluorescein and cis-parinaric acid. Conversely, enhanced intracellular oxidation is seen in cells derived from mice lacking both functional MTI and MTII genes. GSH levels are unaffected in MT null cells relative to wildtype, suggesting the antioxidant function of MT is independent of GSH. In tumor cells there is at least a 400-fold range in MT levels and a 10-fold difference in the ratio of nuclear to cytoplasmic distribution. No correlation exists between MT levels and GSH levels demonstrating the autonomous regulation of intracellular thiol pools. This may be important for cancer chemotherapies since MT overexpression is seen in human tumor cells with acquired drug resistance. The authors found no evidence for altered MT isoform profiles in drug resistant cells that overexpress MT. Recent evidence suggests MT subcellular location may dictate functionality and MT may help determine the threshold for apoptosis. Thus, MT is a stress-inducible protein with antioxidant attributes that may participate independently or in conjunction with GSH to protect cells against injurious agents.

Animals↗

PubMed1998

Dietary flavonoids interact with trace metals and affect metallothionein level in human intestinal cells.

Flavonoids are natural compounds found in food items of plant origin. The study examined systematically the interaction of structurally diverse dietary flavonoids with trace metal ions and the potential impact of dietary flavonoids on the function of intestinal cells. Spectrum analysis was first performed to determine flavonoid-metal interaction in the buffer. Among the flavonoids tested, genistein, biochanin-A, naringin, and naringenin did not interact with any metal ions tested. Members of the flavonol family, quercetin, rutin, kaempferol, flavanol, and catechin, were found to interact with Cu(II) and Fe(III). On prolonged exposure, quercetin also interacted with Mn(II). Quercetin at 1:1 ratio to Cu(II) completely blocked the Cu-dependent color formation from hematoxylin. When quercetin was added to the growth medium of cultured human intestinal cells, Caco-2, the level of metal binding antioxidant protein, metallothionein, decreased. The effect of quercetin on metallothionein was dose- and time-dependent. Genistein and biochanin A, on the contrary, increased the level of metallothionein. The interaction between dietary flavonoids and trace minerals and the effect of flavonoids on metallothionein level imply that flavonoids may affect metal homeostasis and cellular oxidative status in a structure-specific fashion.

Cadmium↗

PubMed1998

Transepithelial transport and accumulation of flavone in human intestinal Caco-2 cells.

Flavonoids are found in many food items of plant origin. Intake of flavonoids has been linked to the prevention of human diseases including cancer. However, little is known about the intestinal absorption of flavonoids in the cellular level. This study was designed to study the absorption of dietary flavonoids using cultured human intestinal epithelial cell monolayer as a model system and 14C-flavone as a model compound. 14C-flavone at 10 microM was found to move across the cell monolayer rapidly both from the luminal to basolateral direction and from the basolateral to luminal direction. The rate of transport from the luminal to basolateral direction was 5 times of the rate for phenylalanine, an aromatic amino acid. Flavone also accumulated substantially in the cells. Replacing sodium in the transport buffer with potassium did not affect the transport but reducing the incubation temperature significantly decreased the initial rate of transport. The presence of protein in the transport buffer reduced the initial rate of transport to half. Other flavonoids and hydrophobic chemicals at 100 microM had no effects on the transport. Together with the evidence from microscopic observation (Cancer Letts. 110: 41-48, 1996), this study supports that rapid diffusional transport may be the main route for flavonoid absorption. The ability of intestinal cells to accumulate flavone is consistent with the role of flavonoids in colon cancer prevention.

Biological Transport↗

PubMed1997

Effect of antiproliferative flavonoids on ascorbic acid accumulation in human colon adenocarcinoma cells.

Dietary flavonoids were found to be antiproliferative for human colon cancer cells, Caco-2 and HT-29, and rat nontransformed intestinal crypt cells, IEC-6. The antiproliferative potency was found to be structure-dependent. We report here a correlation between the antiproliferative potency of these flavonoids and their ability to inhibit cellular accumulation of ascorbic acid (vitamin C). Caco-2, HT-29 and IEC-6 cells were found to accumulate ascorbic acid in a sodium-dependent fashion although some ascorbic acid may also enter the cells through sodium-independent mechanisms. Flavonoids that have been found to be antiproliferative, quercetin and genistein, inhibited the accumulation of ascorbic acid. The inhibition was dose-dependent and could be observed after as short as 10-min of incubation. The degree of inhibition of accumulation was more during rapid cell division as compared to post-confluency Caco-2 cells. Flavonoids that were found to show little antiproliferative effect, naringenin and catechin, also had little effect on ascorbic acid accumulation. The antiproliferative property of flavonoids could be linked to their ascorbic acid deprivation property.

Animals↗

PubMed1997

Effect of PVA-AA on dentine bonding of HEMA.

PVA-AA, an esterification product of poly(vinyl alcohol) and acryloyl chloride, was synthesized and tested for its dentine bonding ability as an additive to 2-hydroxyethyl methacrylate (HEMA). The dentine bonding strength increased significantly by increasing the concentration of PVA-AA in HEMA. The dentine tensile bonding strength attained by the mixture of 10 wt% PVA-AA in HEMA is about 38% higher than that of HEMA or Gluma bonding agent. The dentine shear bonding strength also increased by increasing the acrylate content of the PVA-AA up to about 30%. Test results of cell culturing indicate that no toxic substance is released from PVA-AA to inhibit the cell growth.

Acrylates↗

PubMed1997

Dietary flavonoid and cancer prevention: evidence and potential mechanism.

Dietary flavonoids represent a family of polyphenol compounds found in common food items derived from plants. Depending on structural features, flavonoids can be further subdivided into flavones, flavonols, isoflavones, flavanes, and flavanols. The biological activities of flavonoids are structure dependent and epidemiological studies support their role in human cancer prevention. Several flavonoids inhibit cancer development in animal models of chemical and UV carcinogenesis. However, at high dose some flavonoids themselves may also increase cancer incidence. Although flavonoids have been shown to inhibit cancer cell growth in vitro, the ability of flavonoids to limit cancer progression is limited in animal studies. A potential application is the possible synergisticaction of flavonoids with chemotherapy agents. Molecularly, flavonoids have antioxidant properties and can further enhance the antioxidant protein activities in cells and in animals. Isoflavones and some other flavonoids have weak affinity for the estrogen receptor. Neonatal exposure of animals to isoflavonoids affects the development of reproductive organs, an observation that opens the possibility of using isoflavonoids in the prevention of cancers of the reproductive system. Some growth-inhibiting flavonoids also bind to the low-affinity type II estrogen binding sites, but the biochemical identity of type II sites is unknown.

Animals↗

PubMed1997

Plasma-modified nylon meshes as supports for cell culturing.

The polymeric surfaces of three commercially available nylon films with mesh openings of 5 microns, 10 microns and 20 microns were treated with anhydrous ammonia gaseous plasma. Cells cultured on the plasma-treated nylon films have higher proliferation rate and assume morphology distinct from those cultured on the unmodified films. Of the three plasma-modified membranes, the one with 5 microns mesh openings supported a largest population of cell growth. The plasma-treated nylon meshes provided a stronger anchorage for the collagen matrices formed within the mesh openings. Application of this collagen/nylon meshes for cell culturing is demonstrated.

3T3 Cells↗

PubMed1996

Antiproliferative potency of structurally distinct dietary flavonoids on human colon cancer cells.

Dietary flavonoids are known to be antiproliferative and may play an important role in cancer chemoprevention, especially cancers of the gastrointestinal tract, because of a direct contact with food. This study was designed to compare the antiproliferative potency of several structurally distinct dietary flavonoids in colon cancer cells, Caco-2 and HT-29, and in rat non-transformed intestinal crypt cells, IEC-6. Flavonoids varied significantly in their antiproliferative potency depending on the structural features but the observations were consistent among the three cell lines studied. Of the two most potent flavonoids, quercetin and genistein, the effect was found to be dose-dependent and chromatin condensation, an indication of apoptosis, was noticed. Quercetin was found to distribute throughout the cell with higher amounts in the perinuclear and nucleoli areas. The lack of specific cell membrane enrichment by quercetin was consistent with its lack of effect on the transepithelial resistance. While several flavonoids including quercetin were found to be unstable, the chemical instability did not correlate with the antiproliferative potency, although it may contribute to the antiproliferative effect.

Animals↗

PubMed1996

Pathways for oxalate transport in rabbit renal microvillus membrane vesicles.

Recent evidence suggests that apical membrane Cl--oxalate exchange plays a major role in mediating Cl- absorption in the renal proximal tubule. To sustain steady-state Cl- absorption by a mechanism of exchange for intracellular oxalate requires the presence of one or more pathways for recycling oxalate from lumen to cell. Accordingly, we evaluated the mechanisms of oxalate transport in luminal membrane vesicles isolated from the rabbit renal cortex. We found that transport of oxalate by Na+ cotransport is negligible compared to the transport of sulfate. In contrast, we demonstrated that oxalate shares the electroneutral pathway mediating Na+-independent sulfate-carbonate exchange. We also demonstrated the presence of OH--oxalate exchange (indistinguishable from H+-oxalate cotransport). The process of OH--oxalate exchange was electrogenic and partially inhibited by Cl-, indicating that it occurs, at least in part, as a mode of the Cl--oxalate exchanger described previously. An additional component of OH--oxalate exchange was insensitive to inhibition by either Cl- or sulfate, suggesting that it takes place by neither the Cl--oxalate exchanger nor the sulfate-carbonate exchanger. We conclude that multiple anion exchange mechanisms exist by which oxalate can recycle from lumen to cell to sustain Cl- absorption occurring via apical membrane Cl--oxalate exchange in the renal proximal tubule.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

PubMed1995

Metallothionein localization and cisplatin resistance in human hormone-independent prostatic tumor cell lines.

Metallothioneins (MT) are major cysteine-rich proteins with poorly characterized functions. We have examined the MT amount, isotype expression, and subcellular distribution in 4 human hormone-independent prostatic carcinoma cell lines. Both PC-3 and DU-145 cells were thiol-rich cells with similar MT and glutathione levels, while HPC36M and PC-3 MA2 were thiol-poor cells with lower MT and glutathione levels. All 4 prostatic cell lines expressed the MTIIA isoform at a basal level; DU-145 cells also constitutively expressed MTIE mRNA. Using antibodies for both total MT and MTIIA, we defined MT to cytoplasmic and nuclear domains in PC-3 cells, to perinuclear and nuclear domains in HPC36M cells, and to prominent nonnucleolar nuclear domains in DU-145 and PC-3 MA2 cells. These results indicate that the subcellular distribution is cell type specific and not reflective of the total MT content or MT isoform. Resistance to cadmium in all 4 cell lines was correlated with total MT levels, while resistance to the anticancer agent cisplatin correlated best with nuclear MT content. We suggest that the subcellular localization of MT is functionally important in cellular protection against the anticancer agent cisplatin in human prostatic cancer cells.

Blotting, Northern↗