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Biomedical subjects

S M Wang

Publications and source records attributed to S M Wang.

At least 19 recordsLinked to original sources
PubMed1992

Specific activation of glucuronide prodrugs by antibody-targeted enzyme conjugates for cancer therapy.

Cancer chemotherapy may be improved by increasing antineoplastic drug specificity for tumor cells. We have synthesized a glucuronide prodrug that can be enzymatically converted to an antineoplastic agent at tumor cells that are able to bind beta-glucuronidase-monoclonal antibody conjugates. The glucuronide prodrug BHAMG, the tetra-n-butyl ammonium salt of (p-di-2-chloroethylaminophenyl-beta-D-glucopyranoside) uronic acid, was 150 times less toxic than the parent drug, N,N-di-(2-chloroethyl)-4-hydroxyaniline, to HepG2 human hepatoma cells and over 1000-fold less toxic than the parent drug to AS-30D rat hepatoma cells in vitro. In the presence of beta-glucuronidase, BHAMG was activated and became as toxic as the parent drug N,N-di-(2-chloroethyl)4-hydroxyaniline. A conjugate (RH1-beta G) was formed by linking beta-glucuronidase to a monoclonal antibody which binds to an antigen expressed on the surface of AS-30D cells. The concentration of BHAMG causing 50% inhibition of AS-30D cellular protein synthesis was reduced over 1000-fold, from greater than 770 microM to less than 0.74 microM after these cells were preincubated with RH1-beta G. Specificity of BHAMG activation at antigen-positive cells was shown by monoclonal antibody RH1 blocking of RH1-beta G conversion of BHAMG to toxic drug and by the inability of BHAMG to be converted to active drug when antigen-negative control cells were preincubated with RH1-beta G. Our results show that the targeted-beta-glucuronidase activation of BHAMG can increase the specificity of chemotherapy for rat hepatoma in vitro and suggest that the targeted activation of glucuronide prodrugs may be useful for cancer therapy.

Aniline Mustard↗
PubMed1992

Two distinct membrane-bound phosphatidylinositol-4-phosphate phosphatases in bovine brain.

Solubilized phosphatidylinositol-4-phosphate 4-phosphatase from bovine brain resolved into two peaks of activity by ion exchange chromatography. Both exhibited substantial detergent binding characteristic of integral membrane proteins, and both appear specific for phosphatidylinositol-4-phosphate, but their pH optima differ: the earlier eluting fraction (peak 1) is optimally active between pH 5.5 and 6, whereas the later eluting fraction (peak 2) is most active around pH 8.5. Detergent inhibition studies suggest that peak 2, but not peak 1, interacts with phosphatidylinositol-4-phosphate in the context of a single mixed micelle. Further characterization of these activities should help shed light on the biological function of polyphosphoinositide phosphatases.

Animals↗
PubMed1992

A PCR-based assay for reporter gene expression.

Transient transfection is a widely used tool for the identification of cis-acting regulatory elements. These elements are detected by their effect on the expression of a reporter gene, which is quantified by measuring the reporter gene product in the form of mRNA, protein (hGH), or enzymes (CAT, luciferase). Measurements of mRNA levels have several advantages over enzyme or protein assays. However, mRNA quantification by RNase protection or S1 mapping has considerably lower signal-to-background ratio than protein assays and is therefore less sensitive. In this paper we report the development of a system that takes advantage of the polymerase chain reaction (PCR) to quantify rabbit beta-globin reporter gene expression. Cells are co-transfected with constructs whose activity is to be tested and a reference plasmid with a small deletion in the second exon of the beta-globin gene. We show that the ratio of the two amplified cDNA signals is a highly reliable measure of test gene expression. The sensitivity of this assay is at least 1000-fold higher than RNase protection.

Animals↗
PubMed1992

[Factors in recurrence after curative resection of rectal cancer].

Data on 409 Patients who had a curative resection for rectal cancer at the National Taiwan University Hospital between 1977 and 1989 were analysed to determine the independent effect on recurrence. In our series, the total operative mortality rate was 1.7% and the resectability rate was 88.1%. For these cases who received curative resection, the overall 5-year survival rate was 62.6%. The 5-year survival rate varied according to the Dukes' stage: stage A, 96.4%; stage B, 73.3% and stage C, 39.6%. The total recurrence rate after curative resection was 35.9%, including local recurrence 18.3%, systemic recurrence 12.0%, and combined recurrence 5.6%. According to Dukes' staging system, the recurrence rate for stage A, B and C were 0%, 22.4% and 63.8%, respectively. We used Cox's regression model to analyse the patients characteristics and pathological variables on recurrence and to assess the independent influence of each when all other factors were held constant. The pathological stage of the cancer had the strongest association. Other variables found to have an independent yet significant importance, were CEA level and tumor size. The identification of the patient group, at a high risk of recurrence, might promote a more judicious selection for surgical procedure and trials of adjuvant therapy.

Adult↗
PubMed1992

The role of endothelin-1 during ischemia-reperfusion injury.

In order to investigate the role of endothelin-1 during ischemia-reperfusion injury, 80 adult male Wistar rats were subjected to three hours of ischemia and one hour of reperfusion. Animals were evenly divided into eight groups. The rats in group 1 served as the normal control group, while the rats in group 2 received an intravenous infusion of endothelin-1 in a dosage of 0.5 ng/kg/min, group 3 in a dosage of 5 ng/kg/min, and group 4 in a dosage of 50 ng/kg/min. The rats in group 5 were infused with angiotensin II (10 ng/kg/min). The rats in group 6 received an intravenous infusion of 10,000 units of superoxide dismutase and 10,000 units of catalase. Group 7 rats were infused with endothelin-1 (50 ng/kg/min), superoxide dismutase (10,000 units), and catalase (10,000 units). Group 8 rats received an infusion of angiotensin II (10 ng/kg/min), superoxide dismutase (10,000 units) and catalase (10,000 units). The infusions were given during the reperfusion period. After one hour of reperfusion, the gastrocnemius and soleus muscles of the experimental animals were excised and assayed for ischemia-reperfusion injury by measuring triphenyltetrazolium chloride (TTC) reduction. The results showed that the limb activity of the ischemic extremity was 40.33 +/- 2.75% in group 1, 41.62 +/- 4.08% in group 2, 14.42 +/- 3.14% in group 3, 4.43 +/- 1.05% in group 4, 23.81 +/- 3.51% in group 5, 57.23 +/- 4.52% in group 6, 31.79 42- 3.63% in group 7, and 27.39 +/- 3.95% in group 8. Endothelin-1 reduced the limb activity in a dose-dependent manner.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Studies on the interaction between titin and myosin.

This study examines the interaction of titin and myosin. In order to analyze the domains of myosin contributing to the binding for titin, we conducted a solid phase binding assay. Different portions of myosin (heavy chains, light chains and myosin fragments) were coated on the microtiter wells and reacted with biotinylated titin. Then the binding of biotinylated titin to these polypeptides was detected by using the avidinbiotin-peroxidase method. The results demonstrated that light meromyosin and subfragment 1 were the major domains of myosin interacting with titin. Titin fragments obtained by trypsin digestion were allowed to react with myosin in an affinity column, and the bound fragments were isolated by an acidic elution. Immunoblot analysis of myosin-bound titin fragments revealed that an A-band domain of titin was responsible for the binding of myosin. In addition, biotinylated titin labelled the outer A-bands and Z-bands in intact myofibrils, thus confirming the in situ binding of titin to myosin.

Animals↗
PubMed1992

Effects of light deprivation on prolactin cells in golden hamsters: an immunoelectron microscopic study.

In the golden hamster light deprivation has been shown to induce gonadal regression and reduction of pituitary and plasma levels of prolactin (PRL). In the present study we examined changes in morphology and population ratios of three types of PRL cells 8 weeks after light deprivation, by means of blinding or exposure of hamsters to continuous darkness. In the pituitary of intact hamsters of either sex, which were entrained to a 14-h light: 10-h dark cycle, Type C cells with large secretory granules were the most numerous and Type A with smaller granules the least. After light deprivation the pituitary was found to contain remarkably atrophic PRL cells and showed a profound change in population ratio of PRL cell types, i.e., Type A cells prevailed over the other two types. Pituitary glands from light-deprived and concurrently pinealectomized hamsters exhibited structures and a population ratio of three types of PRL cells similar to those from intact animals. It is suggested that small-granule-containing PRL cells represent an inactive stage of PRL cells, whereas medium- and large-granule-containing cells are functionally active cells. The atrophy of PRL cells can account for the decreased pituitary level of PRL in light-deprived hamsters reported previously.

Animals↗
PubMed1991

Location of the C-terminus of titin at the Z-line region in the sarcomere.

Limited proteolysis of titin with trypsin yielded a number of polypeptides which were electrophoresed and transferred to a nitrocellulose membrane. Proteolytic removal of the C-terminal residues on the nitrocellulose-bound polypeptides was achieved by using carboxypeptidase Y. The species of the polypeptides left after the digestion was quantified by immunoblotting with two distinct monoclonal anti-titin antibodies A2 and A12 of which the epitopes were located at 0.74 micron and 0.69 micron away from the center of an A-band, respectively. Two polypeptides (266 kd and 84 kd) reactive to both antibodies were identified in the control group. Fifteen minutes after the digestion, the immunoreactivities of A2 on 266 kd and 84 kd polypeptides were disappeared, while those of A12 on these polypeptides were not affected. The results indicate that the C-terminal end of titin is located near the Z-line region and the N-terminal end at the M-line region in the sarcomere.

Animals↗
PubMed1991

Chicken cardiac myofibrillogenesis studied with antibodies specific for titin and the muscle and nonmuscle isoforms of actin and tropomyosin.

Myofibrillogenesis was studied in cultured chick cardiomyocytes using indirect immunofluorescence microscopy and antibodies against alpha- and gamma-actin, muscle and nonmuscle tropomyosin, muscle myosin, and titin. Initially, cardiomyocytes, devoid of myofibrils, developed variable numbers of stress fiber-like structures with uniform staining for anti-muscle and nonmuscle actin and tropomyosin, and diffuse, weak staining with anti-titin. Anti-myosin labeled bundles of filaments that exhibited variable degrees of association with the stress fiber-like structures. Myofibrillogenesis occurred with a progressive, and generally simultaneous, longitudinal reorganization of stress fiber-like structures to form primitive sarcomeric units. Titin appeared to attain its mature pattern before the other major contractile proteins. Changes in the staining patterns of actin, tropomyosin, and myosin as myofibrils matured were interpreted as due to longitudinal filament alignment occurring before ordering in the axial direction. Non-muscle actin and tropomyosin were found with sarcomeric periodicity in the initial stages of sarcomere myofibrillogenesis, although their staining patterns were not identical. The localization of the "sarcomeric" proteins alpha-actin and muscle tropomyosin in stress fiber-like structures and the incorporation of non-muscle proteins in the initial stages of sarcomere organization bring into question the meaning of "sarcomeric" proteins in regard to myofibrillogenesis.

Actins↗
PubMed1991

Some dicopper complexes of benzimidazole-containing ligands.

Dicopper complexes of the following benzimidazole-containing ligands have been studied as possible models for the active site of hemocyanin: EDTB (N,N,N',N'-tetrakis-(2-benzimidazolylmethyl)-1,2-ethanediamine), EGTB (1,1,10,10-tetrakis-(2-benzimidazolylmethyl)-1,10-diaza-4,7- dioxadecane), and MEGTB (1,1,10,10-tetrakis-(1-methylbenzimidazol-2-y lmethyl)-1,10-diaza-4,7-dioxadecane). The initial oxygenation product of Cu2(EDTB)(ClO4)2 in Me2SO gives optical absorption maxima at 315 nm (epsilon = 3750 M-1 cm-1) and 690 nm (epsilon = 100 M-1 cm-1). The fluorescence emission intensities of Cu2(EDTB)(ClO4)2 at 400 and 700 nm (excitation at 350 nm) decreases rapidly on exposure to air. This suggests oxidation of Cu2(I) to Cu2(II). The x-ray absorption edge spectra suggest that both coppers in the oxygenation product, analyzed as Cu2(EDTB)(ClO4)2(O).3H2O, are Cu(II). From spectrophotometric titration of Cu2(MEGTB)Cl4 with azide, formation constant of the Cu2(MEGTB)N3Cl3 complex has been obtained. Data from cyclic voltammetry experiments suggest that in the presence of azide, Cu(II)(N3)Cu(II) species is present.

Benzimidazoles↗
PubMed1991

Reduced DNA repair in progeria cells and effects of gamma-ray irradiation on UV-induced unscheduled DNA synthesis in normal and progeria cells.

A reduction in the amount of UV-induced unscheduled DNA synthesis (UDS), and reduced cell survival and host-cell reactivation against UV exposure in Hutchinson-Gilford progeria syndrome cell strains were shown. UV-induced UDS in 4 progeria cell strains was 33-50% of the normal level. A similar reduction in the UV-induced UDS in normal cells was caused by gamma-ray irradiation to the cells before UV irradiation. The dose of gamma-rays required to cause a reduction in UDS of normal cells to the level of progeria cells was 40 Gy and the reduction was reversible after 2 days. In progeria cells, gamma-ray irradiation further reduced UDS with a lower gamma-ray dose required than in normal cells, and the reduction was also reversible but with less relative recovery than in normal cells. The presence of a 'built-in' defect in progeria cells responsible for the reduced DNA-repair capacity was suggested, and such defect may share a common mechanism with the reduction of UV-induced UDS in normal cells caused by gamma-ray irradiation.

Adolescent↗
PubMed1991

Primary malignant tumor of the small intestine.

The records of 101 patients with primary small intestinal malignant tumor at NTUH, collected from 1960 to 1989, were reviewed. These patients represented 1.2% of the patients with gastrointestinal cancer at NTUH over the same period. Fourty-two (41.6%) of the cancer patients had lymphomas, 30(29.7%) had adenocarcinomas, 26 (25.7%) had leiomyosarcomas, and 3(3.0%) had carcinoid tumors. The average age at cancer presentation was 47.5 years (range from 3 to 96). The lymphoma patients had an average age of 35.1 years, while adenocarcinoma patients averaged 60.4 years of age. Leiomyosarcoma and carcinoid tumors averaged 51.2 years and 59 years, respectively. There were 65 male patients and 36 female patients, and there was a male predominance in all groups except for the leiomyosarcoma group which had an equal sex ratio. Generally speaking, the incidence rate for the areas involved were similar in the duodenum, jejunum and ileum. However, adenocarcinomas were more common in the duodenum (53%) and 45% of lymphomas were found in the ileum, as were the carcinoid tumors (66%). The most common presenting symptom was abdominal pain (62%), with bleeding second (32%). Obstruction and palpable mass together were present in 29% of the cases. Body weight loss was found in 25% of patients, and 14% of the patients presented with acute abdomen due to intestinal perforation. Laparotomy was the most common diagnostic procedure (60%). Preoperative diagnoses were possible in cases of duodenal and upper intestinal malignancies, but were rarely possible in patients with lower intestinal malignancies. Sixty-eight patients (68%) underwent tumor resection for palliation or cure. The operation mortality was 4%.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1991

Factors affecting local or regional recurrence in breast cancer.

Between 1975 and 1989, 535 patients with infiltrating ductal carcinoma of the breast in various clinical stages were treated by modified radical mastectomy and axillary node clearance, followed by chemotherapy, endocrine therapy or radiotherapy. Median follow-up was 3 years (5 months to 15 years). Fifty-eight local or regional recurrences occurred. The 5-year and 10-year actuarial risks of breast recurrences were 14% +/- 2% and 23% +/- 4%, respectively. Using multivariate analysis with the Cox regression model, the most important multivariate-adjusted independent contributing factors to local or regional breast recurrences were age less than or equal to 40 (p less than 10(-4), relative risk = 2.94), positive lymph nodes greater than 3 (p less than 10(-3), relative risk = 2.57), and multicentricity (p = 0.044, relative risk = 2.73). The overall 5-year survival rate of recurrent patients was 64 +/- 7%, and was significantly worse when breast recurrence occurred in the first 18 months after operation. The multivariate-adjusted relative risk of death for early recurrence was 4.35 (p = 0.04). The unadjusted 5-year survival rate for early recurrence was 58% +/- 1%, and 85% +/- 8% for late recurrence. This study confirms the relationship between young age and low breast control rates. It emphasizes the adverse prognosis of early breast recurrences as compared to the relatively favorable outcome of late recurrences.

Adult↗
PubMed1991

Tumoricidal activity of interferon-r activated peripheral monocytes in colorectal cancer patients.

Peripheral blood monocytes obtained from 8 colorectal cancer patients and 6 normal controls were incubated in vitro with interferon-r (IFN-r) in the presence of bacterial lipopolysaccharide (LPS). The cytotoxic properties of the monocyte were determined subsequent to the interaction with radiolabeled autologous, allogeneic, as well as cultured colorectal cancer cells. Monocytes from normal controls and all colorectal cancer patients were activated in vitro to become tumoricidal; monocytes lysed tumorigenic cells but not nontumorigenic cells. Activators of protein kinase C (e.g. phorbol esters, PMA) and Ca2+ ionophores (A23187) when added alone did not effect the activation state of the monocyte. Whereas, PMA and A23187 cooperatively reproduced the ability of IFN-r to prime monocytes for tumoricidal activity. In the presence of PMA, A23187, and EGTA, the addition of excessive Ca2+ was sufficient for priming, whereas the addition of excessive Mg2+ was much less efficient. Priming by IFN-r, however, was not blocked by EGTA. An efflux of Ca2+ from preloaded monocytes was significantly increased by A23187 and by IFN-r. Quin-2/AM, an intracellular chelator of Ca2+, blocked priming by IFN-r. The results suggest that priming of monocytes for tumoricidal function by IFN-r may be involved in the activation of protein kinase C and mobilization of intracellular Ca2+.

Aged↗
PubMed1991

An immunocytochemical study of effects of light deprivation on prolactin cells in the adenohypophysis of the golden hamster.

Population ratio and morphology of prolactin cells were studied by employing immunohistochemical methods in the adenohypophysis of normal and experimental golden hamsters of both sexes at 16 weeks of age. Prolactin cells occupied 29% of the total adenohypophyseal cells in the intact males exposed to 14/10 h light/dark schedule. After stimulation of the pineal activity by binding or exposure of males to continuous darkness for eight weeks, prolactin cells became atrophic and were reduced in population to 17% and 13%, respectively. Pinealectomy prevented to some extent the effects of the above treatments; thus, prolactin cells constituted 27% in the pinealectomized and blinded hamsters, and 19% in the pinealectomized and darkness-treated group; and their morphology was comparable with that of the intact controls. Prolactin cells in the normal females were apparently larger in size and more numerous as compared with those of the normal males, comprising 47% of cell population in the anterior pituitary. In response to light deprivation, prolactin cells were atrophic with a diminished cytoplasm and decreased in cell number as reflected in the population ratio of 27% in the blinded and 21% in the darkness-treated groups. In pinealectomized females combined with binding or darkness-treatment, prolactin cells contained an abundance of secretory granules in the cytoplasm and maintained the population ratio comparable to that in the intact females. The present study revealed that light deprivation exerts an inhibitory effect on the secretory activity of prolactin cells and also causes hypoplasia of prolactin cells in the hamster adenohypophysis, the effects being mediated by the pineal gland.

Animals↗
PubMed1990

An immunoelectron microscopic study of corticotrophs in the golden hamster.

The ultrastructure of corticotrophs in the pituitary of golden hamsters was studied by immunocytochemistry. Corticotrophs were classified into three types according to the different size of the secretory granules. Type A and B cells were oval or polygonal in shape containing small (158 +/- 38 nm) and medium-sized (250 +/- 53 nm) secretory granules, respectively. Type C cells were usually pyramidal or irregular in shape, and contained large secretory granules (380 +/- 78 nm). The cytoplasmic organelles of type B and C cells were fairly well developed. In all types of corticotrophs, the secretory granules varied in electron density, and were either arranged in a single row along the cell membrane, or concentrated in the vascular pole of the cytoplasm. Many of the corticotrophs sent processes to encircle neighboring somatotrophs. In the female, the proportions of type A and B cells were higher than those in the male, whereas that of the type C cells was lower. All three types of corticotrophs were observed in the early postnatal stage. The population of type A cells decreased with the advancement of postnatal development, with a concomitant increase of that of the type C cells. Thus, type A cells might represent the immature type of corticotrophs which would evolve to become the type C cells. One to 2 weeks after adrenalectomy, the number of type C cells was drastically increased. This was accompanied by their prominent changes in ultrastructure as in the type B cells. The cytoplasm became hypertrophic with extensive dilated cisternae of rough endoplasmic reticulum.(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenalectomy↗