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Biomedical subjects

K S Kang

Publications and source records attributed to K S Kang.

At least 37 records · Page 2Linked to original sources

High susceptibility of a human breast epithelial cell type with stem cell characteristics to telomerase activation and immortalization.

We have recently characterized two types of normal human breast epithelial cells (HBECs) from reduction mammoplasty. Type I cells express estrogen receptor, luminal epithelial cell markers, and stem cell characteristics (i.e., the ability to differentiate into other cell types and to form budding/ductal structures on Matrigel), whereas Type II cells show basal epithelial cell phenotypes. In this study, we have examined whether Type I HBECs are more susceptible to telomerase activation and immortalization after transfection with SV40 large T-antigen. The results show that both types of cells acquire extended life span [(EL); i.e., bypassing senescence] at a comparable frequency. However, they differ significantly in the ability to become immortal in continuous culture, ie., 11 of 11 Type I EL clones became immortal compared with 1 of 10 Type II EL clones. Both parental Type I and Type II cells as well as their transformed EL clones at early passages [approximately 30 cumulative population doubling level (cpdl)] showed a low level of telomerase activity as measured by the telomeric repeat amplification protocol assay. For all 11 of the Type I EL clones and the single Type II EL clone that became immortal, telomerase activities were invariably activated at middle passages (approximately 60 cpdl) or late passages (approximately 100 cpdl). For the four Type II EL clones randomly selected from the nine Type II clones that did not become immortal, the telomerase activities were found to be further diminished at mid-passage, before the end of the life span. Thus, normal HBECs do have a low level of telomerase activity, and Type I HBECs with stem cell characteristics are more susceptible to telomerase activation and immortalization, a basis on which they may be major target cells for breast carcinogenesis.

Breast↗

Inhibitory effect of pentachlorophenol on gap junctional intercellular communication in rat liver epithelial cells in vitro.

To understand the initiating/promoting actions of pentachlorophenol (PCP), a non-mutagenic hepatocarcinogen, and its metabolite, tetrachlorohydroquinone (TCHQ), we investigated the effects of each chemical on gap junctional intercellular communication (GJIC) in rat liver epithelial cells (WB cells) by the scrape-loading and dye transfer method. After treatment with PCP, the GJIC was initially inhibited at 4 h but was restored in 6-8 h, followed by a second phase of inhibition between 16 and 24 h. Both the first and second inhibitions were concentration-dependent and were restored by 2-4 h after removal of PCP. The phosphorylation state of connexin 43 (CX43) and its localization on the plasma membrane were unchanged up to 24 h after treatment; however, this was accompanied by a decrease in the CX43 protein level. No inhibitory effect was apparent on the GJIC of cells treated with TCHQ. These results suggest that PCP may play a critical role of promoting activity via non-mutagenic mechanisms.

Animals↗

Involvement of tyrosine phosphorylation of p185(c-erbB2/neu) in tumorigenicity induced by X-rays and the neu oncogene in human breast epithelial cells.

Ionizing radiation is the exogenous agent best proven to induce breast cancer. c-erbB2/neu amplification and overexpression are known to occur in breast cancer and are correlated with aggressive tumor growth and poor prognosis. We have developed simian virus 40-immortalized cell lines from normal human breast epithelial cells (HBECs) with luminal and stem-cell characteristics. In this study, we examined whether x-rays and a mutated neu oncogene are capable of inducing tumorigenicity in these cells. The results indicated that x-rays were effective in converting immortal non-tumorigenic HBECs to weakly tumorigenic cells that then could be transformed to highly tumorigenic cells by the neu oncogene. The in vitro growth of these tumorigenic cells was significantly faster than that of the parental non-tumorigenic cells in growth factor- and hormone-supplemented or -depleted media. The neu oncogene, however, had no tumorigenic effect on immortal non-tumorigenic cells. The expression of p185(c-erb82/neu) was elevated in neu-transduced immortal or weakly tumorigenic cell lines. However, only in the latter was p185(c-erbB2/neu) found to be phosphorylated at tyrosine residues. Thus, x-rays appear to induce a genetic alteration that confers weak tumorigenicity on immortal HBECs and interacts with p185(c-erbB2/neu) directly or indirectly to give rise to fast-growing tumors.

Animals↗

Stimulation of cell proliferation and inhibition of gap junctional intercellular communication by linoleic acid.

The effect of linoleic acid (LA) on gap-junction permeability, connexin 43 mRNA level, protein level, and phosphorylation, and the numbers of gap-junctional membrane plaques were studied in the rat liver epithelial cell line WB-F344 to determine whether changes in these parameters correlated with the enhanced cell growth and the inhibition of gap-junction function. When cultured in a medium with low serum (1%), these cells exhibited a slower growth rate than in the high serum medium (7%). Addition of linoleic acid (0.01-3 mg/ml) to the low serum medium increased the growth rate and inhibited gap junctional intercellular communication (GJIC) in a dose-dependent manner. In a comparison of short-term and long-term treatments with LA, GJIC in short-term treated (1 h) WB cells was inhibited at 3 mg/ml LA but readily recovered by washing and removing LA from cells, whereas GJIC in long-term treated (6 days) WB cells did not recover by washing and removing LA from WB cells. Western blot analysis of connexin 43 showed that a short-term incubation with linoleic acid increased the relative amount of unphosphorylated connexin 43 protein, but a long-term incubation with linoleic acid decreased the amount of unphosphorylated connexin 43 protein and increased the relative amount of hyperphosphorylated connexin 43 protein. Connexin 43 and p53 mRNA levels decreased in a time- and dose-dependent manner in linoleic acid-treated cells. These results suggest that growth stimulation and gap junctional intercellular communication inhibition of rat liver epithelial cells by linoleic acid may be mediated in part through modulation of p53 expression and function.

Animals↗

Hydrogen peroxide inhibits gap junctional intercellular communication in glutathione sufficient but not glutathione deficient cells.

Cell to cell communication via gap junctions is essential in the maintenance of the homeostatic balance of multicellular organisms. Aberrant intercellular gap junctional communication (GJIC) has been implicated in tumor promotion, neuropathy and teratogenesis. Oxidative stress has also been implicated in similar pathologies such as cancer. We report a potential link between oxidative stress and GJIC. Hydrogen peroxide, a known tumor promoter, inhibited GJIC in WB-F344 rat liver epithelial cells with an I50 value of 200 microM. Inhibition of GJIC by H2O2 was reversible as indicated by the complete recovery of GJIC with the removal of H2O2 via a change of fresh media. Free radical scavengers, such as t-butyl alcohol, propylgallate, and Trolox, did not prevent the inhibition of GJIC by H2O2, which indicated that the effects of H2O2 on GJIC was probably not a consequence of aqueous free radical damage. The depletion of intracellular GSH reversed the inhibitory effect of H2O2 on GJIC. The treatment of glutathione-sufficient cells with H2O2 resulted in the hyperphosphorylation of connexin43, which is the basic subunit of the hexameric gap junction protein, as determined by Western blot analysis. TPA, a well-known tumor promoter, also inhibits GJIC via hyperphosphorylation of GJIC, which is a result of protein kinase-C activation. However, H2O2 also induced hyperphosphorylation in GSH-deficient cells that had normal rates of GJIC. Therefore, the mechanism of GJIC inhibition must be different from the TPA-pathway and involves GSH.

Animals↗

Expression of estrogen receptors in a normal human breast epithelial cell type with luminal and stem cell characteristics and its neoplastically transformed cell lines.

Although approximately two-thirds of breast cancers are estrogen receptor (ER)-positive, only a small proportion of epithelial cells in the mammary gland express the ER. The origin of the ER-positive breast cancers is unknown. Recently, we have developed a culture method to grow two morphologically and antigenically distinguishable types of normal human breast epithelial cells (HBEC) derived from reduction mammoplasty. In this report, we studied the expression of ER in these two types of cells and their transformed cell lines. The results indicate that Type I HBEC with luminal and stem cell characteristics expressed a variant ER (approximately 48 kd) by Western blot analysis. This variant ER contains a deletion in the DNA binding domain (exon 2) as revealed by RT-PCR analysis. The lack of the DNA-binding domain of the variant ER was also confirmed by the ER-estrogen responsive element binding assay, as well as by the immunofluorescence staining of the ER using anti-ER antibodies which recognize either the C-terminal or N-terminal region. In contrast, Type II HBEC with basal epithelial phenotype are ER-negative. Simian virus 40 (SV40) transformed Type I and Type II HBEC lines also expressed the variant ER. Tumors formed in athymic nude mice by in vitro transformed tumorigenic Type I cell lines, however, expressed a high level of wild type ER which was undetectable in these cells grown in vitro before and after tumor formation. Thus, there appears to be a differential ER mRNA splicing between the in vitro and in vivo mileu.

Adult↗

Inhibition of gap junctional intercellular communication in heptachlor- and heptachlor epoxide-treated normal human breast epithelial cells.

Based on the concern of organochlorides in the environment and in human tissue, this study was designed to determine whether various noncytotoxic levels of heptachlor and heptachlor epoxide could inhibit, reversibly, gap junctional intercellular communication in human breast epithelial cells (HBEC). Cytotoxicity and gap junctional intercellular communication (GJIC) were evaluated by lactate dehydrogenase assay and fluorescence redistribution after photobleaching analysis, respectively. Both heptachlor and heptachlor epoxide were noncytotoxic up to 10 microg/ml. At this concentration, heptachlor and heptachlor epoxide inhibited GJIC of normal human breast epithelial cells after 1 h treatment. Within a 24 h treatment with heptachlor and heptachlor epoxide at 10 microg/ml, recovery of GJIC had not returned. GJIC completely recovered after a 12 h treatment of 1 microg/ml heptachlor epoxide, but it did not recover after a 24 h treatment of 1 microg/ml heptachlor. RT-PCR and Western blots were analyzed to determine whether the heptachlor or heptachlor epoxide might have altered the steady-state levels of gap junction mRNA and/or connexin protein levels or phosphorylation state. No significant difference in the level of connexin 43 (Cx43) message between control and heptachlor-treated cells was observed. Western blot analyses showed hypophosphorylation patterns in cells treated with 10 microg/ml heptachlor and heptachlor epoxide for 1 h with no recovery within 24 h. Immunostaining of Cx43 protein in normal HBEC indicated that heptachlor and heptachlor epoxide caused a loss of Cx43 from the cell membranes at noncytotoxic dose levels. Taken together, these results suggest that heptachlor and heptachlor epoxide can alter GJIC at the post-translational level, and that, under the conditions of exceeding a threshold concentration in the breast tissue containing 'initiated' cells for a long time and not being counteracted by anti-tumor-promoting chemicals, they could act as breast tumor promoters.

Base Sequence↗

Lossless medical image compression by multilevel decomposition.

Lossless image coding is important for medical image compression because any information loss or error caused by the image compression process could affect clinical diagnostic decisions. This paper proposes a lossless compression algorithm for application to medical images that have high spatial correlation. The proposed image compression algorithm uses a multi-level decomposition scheme in conjunction with prediction and classification. In this algorithm, an image is divided into four subimages by subsampling. One subimage is used as a reference to predict the other three subimages. The prediction errors of the three subimages are classified into two or three groups by the characteristics of the reference subimage, and the classified prediction errors are encoded by entropy coding with corresponding code words. These subsampling and classified entropy coding procedures are repeated on the reference subimage in each level, and the reference subimage in the last repetition is encoded by conventional differential pulse code modulation and entropy coding. To verify this proposed algorithm, it was applied to several chest radiographs and computed tomography and magnetic resonance images, and the results were compared with those from well-known lossless compression algorithms.

Algorithms↗

Inhibition of gap junctional intercellular communication in normal human breast epithelial cells after treatment with pesticides, PCBs, and PBBs, alone or in mixtures.

Chemical pollutants in the Great Lakes have found their way through the food chain into humans because of their environmental persistence and lipophilicity. Some epidemiological studies have claimed an association between metabolites of 2,2-bis(p-chlorophenyl)-1,1,1-trichloroethane (DDT), polychlorinated biphenyls (PCBs), and polybrominated biphenyls (PBBs) and breast cancer, but others have reported no such association. We examined various halogenated hydrocarbons for their capacity to inhibit gap junctional intercellular communication (GJIC) in normal human breast epithelial cells (HBEC) when given as single compounds or as mixtures. The scrape-loading/dye transfer and fluorescent redistribution after photobleaching techniques were used to measure GJIC; immunostaining and Western and Northern analyses were performed on connexin 43 (Cx43) gap junction protein and message to determine how halogenated hydrocarbons might affect GJIC. DDT, dieldrin, and toxaphene inhibited GJIC in a dose-responsive manner after 90 min treatments. Dieldrin suppressed GJIC within 30 min with no recovery after 24 hr. Inhibition of GJIC by DDT and toxaphene was partially restored after 12 hr and fully restored after 24 hr. Several PCB and PBB congeners inhibited GJIC in a dose-responsive and time-dependent manner, but GJIC was almost restored to control values 24 hr after exposure. The highest concentrations of the individual chemicals that did not inhibit GJIC was determined, and mixtures containing two of these chemicals were tested for their ability to inhibit GJIC. Significant inhibition of GJIC was observed when cells were treated with a mixture of DDT and 2,4,5-hexachlorobiphenyl (2,4,5-HCB), dieldrin and 2,4,5-HCB, or dieldrin and 2,4,5-hexabromobiphenyl (2,4,5-HBB). These results indicate that halogenated hydrocarbons, alone or in specific combinations, can alter GJIC at the post-translational level. These results are consistent with the hypothesis that DDT, dieldrin, toxaphene, 2,3,4-HCB, 2,4,5-HCB, and 2,4,5-HBB could have tumor-promoting potential in human breast tissue.

Breast↗

Assessment of bone mineral density in postmenopausal and senile osteoporosis using quantitative CT.

The authors analyzed bone mineral density in 213 patients from January 1988 to September 1989. Bone mineral density of the vertebral body in the osteoporosis group of patients was compared with that in the normal group to investigate the correlation between bone mineral density and age distribution, and to estimate fracture threshold in the osteoporosis group. It was found that men, by linear regression, lose an average of 0.91% of bone per year, and women, by cubic regression, lose an average of 1.14% per year, accelerating at menopause. In the osteoporotic group, bone mineral density of men decreased an average of 29.7% compared with the non-osteoporotic group; that of women decreased an average of 29.9%. There was no difference between sexes. The fracture threshold of the osteoporotic patient was estimated to be about 90 mg/cm3.

Adult↗

[A study on the effect of cold applications using a sponge bath in healthy adults].

This study was a quasi-experimental research study to test the characteristics of temperature regulation according to sponge bath methods of cold application. Thirteen volunteers were selected from among nursing college students according to an established criteria using a purposive sampling technique. Four different cold application methods were used: (1) tepid water sponge bath at 28 degrees C, (2) 20% alcohol sponge bath at 28 degrees C, (3) 40% alcohol sponge bath at 28 degrees C and (4) tepid water sponge bath at 28 degrees C plus an ice bag to the head. Changes in rectal temperature, mean skin temperature, mean body temperature, heat content change and thermal discomfort during the cold application were measured at 5 minute intervals over a 120 minute period. The data collection period was from Dec. 20, 1988 to Feb. 3, 1989. The data were analyzed using descriptive statistics, simple regression, ANOVA, Duncan's multiple range test and Pearson correlation coefficient using the SPSS-X Program. The results of the study are summarized as follows. Five general hypothesis were tested. Hypothesis 1 that "Change in heat content will be decreased for each cold application method according to the cold application time" was rejected. (tepid water sponge bath: after 10 minutes of cold application, 20% alcohol sponge bath: after 25 minutes of cold application: 40% alcohol sponge bath: after 45 minutes of cold application, tepid water sponge bath plus an ice bag to the head: after 80 minutes of cold application). Hypothesis 2 that "Thermal discomfort will be changed for each cold application method according to the cold application time" was rejected after 5 minutes of cold application. Hypothesis 3 that "Change in heat content will differ among the cold application methods" was accepted except 0-5, 0-10, 0-65, 0-105 and 0-120 minute. This difference showed significance only between sponge bath methods and tepid water sponge bath plus an ice bag to the head. Hypothesis 4 that "Thermal discomfort will differ among the cold application methods" was accepted at 15, 20, 35, 45, 75, 80, 90, 95, 100, 105, 110, 115 and 120 minute of cold application time. This difference showed significance only between sponge bath methods and tepid water sponge bath plus an ice bag to the head. Hypothesis 5 that "The higher the change in heat content, the higher the thermal discomfort during the cold application time" was accepted for between 10-60 and 75 minute of cold application.(ABSTRACT TRUNCATED AT 400 WORDS)

Acclimatization↗

Genetic and physical analyses of a cluster of genes essential for xanthan gum biosynthesis in Xanthomonas campestris.

Xanthomonas campestris produces copious amounts of a complex exopolysaccharide, xanthan gum. Nonmucoid mutants, defective in synthesis of xanthan polysaccharide, were isolated after nitrosoguanidine mutagenesis. To isolate genes essential for xanthan polysaccharide synthesis (xps), a genomic library of X. campestris DNA, partially digested with SalI and ligated into the broad-host-range cloning vector pRK293, was constructed in Escherichia coli. The pooled clone bank was conjugated en masse from E. coli into three nonmucoid mutants by using pRK2013, which provides plasmid transfer functions. Kanamycin-resistant exconjugants were then screened for the ability to form mucoid colonies. Analysis of plasmids from several mucoid exconjugants indicated that overlapping segments of DNA had been cloned. These plasmids were tested for complementation of eight additional nonmucoid mutants. A 22-kilobase (kb) region of DNA was defined physically by restriction enzyme analysis and genetically by ability to restore mucoid phenotype to 10 of the 11 nonmucoid mutants tested. This region was further defined by subcloning and by transposon mutagenesis with mini-Mu(Tetr), with subsequent analysis of genetic complementation of nonmucoid mutants. A region of 13.5 kb of DNA was determined to contain at least five complementation groups. The effect of plasmids containing cloned xps genes on xanthan gum synthesis was evaluated. One plasmid, pCHC3, containing a 12.4-kb insert and at least four linked xanthan biosynthetic genes, increased the production of xanthan gum by 10% and increased the extent of pyruvylation of the xanthan side chains by about 45%. This indicates that a gene affecting pyruvylation of xanthan gum is linked to this cluster of xps genes.

Chromosome Mapping↗

Agar-like polysaccharide produced by a pseudomonas species: production and basic properties.

A new species of Pseudomonas was isolated that produced copious amounts of an exocellular heteropolysaccharide (PS-60) after incubation for 3 days at 30 degrees C in media containing 3% glucose as a carbon source. The polysaccharide was composed of approximately 46% glucose and 30% rhamnose and, in addition, contained 21% uronic acid and 3% O-acetyl. Upon deacetylation by a mild alkaline treatment, PS-60 produced a brittle, firm, and optically clear gel. This gelling property was thermoreversible. The PS-60 gel exhibited excellent heat stability that withstood autoclaving (i.e., 121 degrees C for 15 min) for several cycles. The gel strength, melting point, and setting point of the polysaccharide were controlled primarily by the concentration of cations. PS-60 was not affected by a variety of enzymes. The results of tests involving various culture media and biochemical test media indicate that PS-60 is an excellent alternative gelling agent to agar.

Journal Article↗

Large bodies of Mycoplasma and L-form organisms.

The large bodies of various Mycoplasma and L-form organisms were studied by ultraviolet fluorescence microscopy of preparations stained with various fluorochromes. Primuline and Thioflavine S specifically stained the outer portion or rim of the large bodies, and the fluorescence characteristics of the stained bodies differed from those for other microorganisms and for spheroplasts and protoplasts. Small granular structures similar in size and morphology to minimal reproductive units were observed within some of the large bodies by phase microscopy and by fluorescence microscopy with acridine orange or Coriphosphine O. Micromanipulation probing of the large bodies revealed their elastic nature; many of the large bodies could be subdivided into two or more smaller circular bodies, each retaining the fluorescence staining properties of the parent body. Under these conditions, however, a few of the large bodies were ruptured, leaving the stainable outer boundary area as a stable residual structure. The large bodies were somewhat resistant to various rigorous treatments normally employed to eliminate viability of Mycoplasma and L-form cultures. Structures similr to large bodies were observed in various natural tissues, and structures resembling large bodies in size, morphology, fluorescence staining characteristics, and reaction to micromanipulation probing were reconstructed from an acetone extract of egg yolk. Overall, the large bodies of Mycoplasma and L-form organisms appeared to be structures resulting from accumulations of metabolic by-products and medium components within or on which minimal reproductive units had become entrapped, although it could not be ruled out that they might be defined structures specifically formed during culture as protective lipoidal sacs for the minimal reproductive units.

Bacteria↗