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W Ying

Publications and source records attributed to W Ying.

At least 37 records · Page 2Linked to original sources
PubMed1998

Targeted delivery of DNA encoding cytotoxic proteins through high-affinity fibroblast growth factor receptors.

Nonviral DNA delivery strategies for gene therapy have generally been limited by a lack of specificity and efficacy. However, ligand-mediated endocytosis can specifically deliver DNA in vitro to cells bearing the appropriate cognate receptors. Similarly, in order to circumvent problems related to efficacy, DNA must encode proteins with high intrinsic activities. We show here that the ligand basic fibroblast growth factor (FGF2) can target FGF receptor-bearing cells with DNA encoding therapeutic proteins. Delivery of genes encoding saporin, a highly potent ribosomal inactivating protein, or the conditionally cytotoxic herpes simplex virus thymidine kinase, a protein that can kill cells by activating the prodrug ganciclovir, is demonstrated. The saporin gene was codon optimized for mammalian expression and demonstrated to express functional protein in a cell-free assay. FGF2-mediated delivery of saporin DNA or thymidine kinase DNA followed by ganciclovir treatment resulted in a 60 and 75% decrease in cell number, respectively. Specificity of gene delivery was demonstrated in competition assays with free FGF2 or with recombinant soluble FGF receptor. Alternatively, when histone H1, a ligand that binds to cell surface heparan sulfate proteoglycans ("low-affinity" FGF receptors), was used to deliver DNA encoding thymidine kinase, no ganciclovir sensitivity was observed. These findings establish the feasibility of using ligands such as FGF2 to specifically deliver genes encoding molecular chemotherapeutic agents to cells.

Amino Acid Sequence↗
PubMed1998

Deleterious network hypothesis of apoptosis.

Apoptosis or programmed cell death (PCD) plays crucial roles in a number of physiological and pathological processes. Growing evidence has suggested that oxidative damage, altered calcium homeostasis and abnormal mitochondrial functions are three key factors of PCD. Because a number of studies have also indicated that a deleterious network is formed on the basis of the close interactions among these major apoptotic factors, it is proposed that the deleterious network is just the common pathway in PCD. A variety of apoptotic stimuli can trigger the network, leading to the characteristic apoptotic changes. This new theory--the deleterious network hypothesis of apoptosis--appears to unify some major theories of PCD, providing consistent explanations of a significantly larger number of the observations about apoptosis than other hypotheses. Based on this unifying hypothesis, it is suggested that the three major factors of the deleterious network could be targeted for treatment of multiple apoptosis-related disorders.

Animals↗
PubMed1997

Tamoxifen-mediated growth inhibition of human cholangiocarcinoma.

Cholangiocarcinoma represents a challenging primary malignancy of the liver with no effective medical therapy and a poor prognosis. We have investigated the role of tamoxifen and estrogen receptors (ERs) in the regulation of growth of human cholangiocarcinoma. Two human cholangiocarcinoma cell lines, OZ and SK-ChA-1, were grown in the presence of graded concentrations of tamoxifen; the effects on cell growth were determined by cell counting or 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium proliferation assay. The presence of ER protein was tested by indirect immunofluorescence and immunoprecipitation. In addition, cells were grown in estrogen-depleted media supplemented with exogenous 17beta-estradiol. ER mRNA was evaluated by reverse transcription-PCR and Northern blotting. Finally, one cholangiocarcinoma cell line was grown as a xenograft in athymic nude mice; tamoxifen effects on in vivo tumor growth were determined with biweekly caliper measurements. Tamoxifen (5-10 microM) caused dose-dependent in vitro growth inhibition of two human cholangiocarcinoma cell lines. In addition, growth inhibition of one cell line (SK-ChA-1) grown as a xenograft in nude mice by tamoxifen was observed. The presence of ER protein was suggested by 17beta-estradiol stimulation of tumor cell growth in vitro and confirmed by immunoprecipitation. Immunofluorescence microscopy was ineffective at detection of ER protein. Reverse transcription-PCR demonstrated the presence of ER mRNA in both cell lines. Northern blot analysis confirmed the presence of full-length 6.5-kb ER mRNA. No ER deletion mutants were detected. Tamoxifen inhibited the growth of human cholangiocarcinoma in vitro and in vivo. ER protein and mRNA were detected in both cell lines. The mechanism(s) of tamoxifen-mediated growth inhibition is unclear but may occur via ER protein or additional pathways. The ability of tamoxifen to inhibit tumor growth may offer an alternative adjunctive treatment for cholangiocarcinoma.

Animals↗
PubMed1997

Targeted gene delivery to Kaposi's sarcoma cells via the fibroblast growth factor receptor.

Kaposi's sarcoma (KS) is a major AIDS-related malignancy associated with significant morbidity and mortality. Current chemotherapeutic regimens are associated with a dismal prognosis. In an effort to develop a new approach to KS treatment, we devised a gene therapy-based adenovirus retargeting schema that redirects the adenovirus to fibroblast growth factor receptors endogenously present on the cell surface of KS cells. By using a bifunctional conjugate consisting of a blocking antiadenoviral knob Fab linked to basic fibroblast growth factor, FGF2, the gene transduction of KS cells was enhanced 7.7-44 fold; recombinant adenoviruses encoding either the firefly luciferase reporter gene, or the herpes simplex thymidine kinase gene, demonstrated quantitative enhancement of expression in the KS cell lines. In this regard, two KS cell lines that were previously refractory to native adenovirus transduction could be successfully transduced by the addition of the conjugate. This study thus addresses the utility of adenoviral retargeting to the FGF receptor in KS cells that are ordinarily transduction refractory to standardized approaches and allows practical development of gene therapy approaches for the treatment of human KS.

Acquired Immunodeficiency Syndrome↗
PubMed1997

Deleterious network hypothesis of aging.

It is becoming increasingly apparent that free-radical damage, mitochondrial defects, glycation/Maillard reaction and calcium dyshomeostasis play crucial roles in cellular and organism senescence. Based on the evidence indicating close relationships among these four aging-promoting factors, a unifying hypothesis of senescence-the deleterious network hypothesis of aging-is proposed: In living organisms, both endogenous and exogenous detrimental factors produce age-dependent accumulation of triggering of a deleterious network, which is constructed on the basis of the interactions among oxidative impairments, mitochondrial defects, calcium mismetabolism and glycation/Maillard reaction. The age-related triggering of the network leads to numerous senescent alterations. It appears that this novel theory has synthesized multiple hypotheses of aging, and is capable of providing consistent explanations to a larger number of senescent changes than any previous hypotheses. Based on the new theory of senescence, it is proposed that the approaches which can inhibit the initiation of the four key age-promoting factors should be applied combinatively to slow down the aging process and to prevent and treat age-associated illnesses.

Aging↗
PubMed1997

Deleterious network: a testable pathogenetic concept of Alzheimer's disease.

Cumulative evidence has indicated that a deleterious network is formed on the basis of close interactions among abnormal amyloid precursor protein (APP) metabolism, oxidative damage, compromised energy metabolism and impaired calcium homeostasis. A unifying hypothesis-the deleterious network hypothesis of Alzheimer's disease (AD)-proposes that the deleterious network, not any single factor, is the common pathway of AD. Aging and multiple genetic or environmental factors could trigger the network by promoting the occurrence of one or more of the key detrimental factors, leading to a number of pathological changes of the disorder. This new hypothesis appears to unify some major theories of AD, providing a sound basis for consistent explanations to a large variety of the observations about the disorder. In this article upto-date delineation of the novel theory is given. Three types of studies are also proposed for further determining the validity of the new hypothesis. Based on this theory, it is suggested that combinative applications of the approaches which can reduce the incidence of the four key pathological factors could become a new therapeutic strategy of AD.

Alzheimer Disease↗
PubMed1996

Deleterious network hypothesis of Alzheimer's disease.

Numerous studies indicate that aberrant amyloid precursor protein metabolism, elevated peroxidative damage, depressed energy metabolism and altered calcium homeostasis are four pivotal deleterious factors in the pathogenesis of Alzheimer's disease. Cumulative evidence further suggests that these four factors are intimately interrelated, forming a deleterious network. Based on this new concept of 'deleterious network', a unifying hypothesis-the deleterious network hypothesis of Alzheimer's disease-is proposed. The main ideas of the hypothesis are delineated as follows: increases in free radical damage, alterations in amyloid precursor protein metabolism, impairment of energy metabolism and abnormalities of calcium homeostasis are four cornerstones of a deleterious network. Various risk factors of Alzheimer's disease can triger the network by promoting the occurrence of one of these key components, resulting in the biological abnormalities of Alzheimer's disease. Based on this new theory, a majority of the important observations about Alzheimer's disease can be explained consistently and succinctly.

Alzheimer Disease↗
PubMed1996

A new hypothesis of neurodegenerative diseases: the deleterious network hypothesis.

Growing evidence has indicated the existence of deleterious networks in the brains of neurodegenerative disorders, including Alzheimer's disease. Parkinson's disease, amyotrophic lateral sclerosis and Huntington's disease. The deleterious networks are formed on the basis of the intimate interactions among the key pathogenic factors, including oxidative damage, aberrant calcium homeostasis, metabolic compromise and, under certain circumstances, amyloid precursor protein mismetabolism. Based on the novel concept, deleterious network, a unifying hypothesis, the deleterious network hypothesis of neurodegenerative diseases, is proposed. This new theory stresses that the deleterious network is just the common pathway of the degenerative disorders, triggering of which by aging, certain genetic or environmental factors leads to a cascade of pathological alterations of the illnesses. It appears that this new theory has synthesized some most appealing hypotheses about neurodegenerative illnesses, providing consistent explanations to a larger number of observations about those diseases than other hypotheses. Because the disorders appear to result from the interactions among the key detrimental factors, it is suggested that the patients of the neurodegenerative diseases should be treated by combinative application of the drugs which can diminish peroxidative damage, calcium mismetabolism, and metabolic compromise.

Alzheimer Disease↗
PubMed1995

Saporin toxins directed to basic fibroblast growth factor receptors effectively target human ovarian teratocarcinoma in an animal model.

BACKGROUND: The antitumor activity of the chemical conjugate and recombinant forms of the mitotoxin basic fibroblast growth factor (bFGF) saporin (SAP) and the bFGF receptor-directed immunotoxin 11A8-SAP against human ovarian teratocarcinoma PA-1 was examined in athymic nude mice. Alternative administration schedules to prolong therapeutic efficacy were explored. METHODS: Intravenous dosing (0.01-125 micrograms/kg) of chemical conjugate and recombinant bFGF-SAP or 11A8-SAP beginning 5 days after subcutaneous implantation of PA-1 cells was administered by i) weekly injection for 4 weeks, ii) continuous infusion for one week, or iii) daily injection five times a week for 4 weeks. RESULTS: Weekly injections (31.25 micrograms/kg) of chemical conjugate bFGF-SAP or 11A8-SAP, the latter of which is 25% the molarity of the former, resulted in mean tumor volumes that were, respectively, 35% or 52% of controls (day 30) and 52% or 76% of controls (day 60). Chemical conjugate or recombinant bFGF-SAP administered weekly resulted in mean tumor volumes that were, respectively, 51% or 77% (0.5 microgram/kg) and 42% or 31% (50 micrograms/kg) of controls (day 30). A mean tumor volume of 35% of controls (day 30) and of 49% of controls (day 60) were observed in animals treated by constant infusion of chemical conjugate bFGF-SAP (125 micrograms/kg, total dose). Alternatively, tumors of animals receiving daily injections (125 micrograms/kg, total dose) exhibited a mean volume of 21% of controls (day 30) and prolonged growth inhibition as demonstrated by a mean tumor volume of 22% of controls (day 60). CONCLUSIONS: These studies suggest a therapeutic potential for bFGF-receptor-directed saporin toxins in the treatment of ovarian teratocarcinoma and the importance of frequency of administration in achieving optimal tumor responses.

Animals↗
PubMed1994

Identification of soluble forms of the fibroblast growth factor receptor in blood.

We have purified three acidic (FGF-1) and basic (FGF-2) fibroblast growth factor binding proteins (FGF-BP1, FGF-BP2, and FGF-BP3) from human plasma and calf serum and demonstrate the presence of these circulating FGF-BPs in blood. Each are truncated forms of the high-affinity FGF receptor (FGFR-1). FGF-BP1 and FGF-BP2 have estimated molecular masses of 70-85 kDa and 55-60 kDa, respectively, and are detected by using 125I-labeled FGF-2 ligand blotting. Immunoblotting with four distinct antibodies to FGFR-1 reveals that FGF-BP1 and FGF-BP2 are immunologically and biochemically related to the extracellular domain of FGFR-1. Reverse-phase HPLC chromatography resolves FGF-BP2 into two proteins with estimated molecular masses of 55 kDa and 60 kDa. Protein sequencing of the amino terminus of FGF-BP2 and FGF-BP3 reveals identity with the extracellular domain of the two-IgG-loop form of human FGFR-1. The FGF-BPs do not require heparin to bind FGF-2 on affinity columns, but heparin does enhance their recovery from blood. These FGF-BPs may play an important physiological role in regulating the biological activity of FGF and the other members of the FGF family of growth factors.

Amino Acid Sequence↗
PubMed1994

Anti-B16-F10 melanoma activity of a basic fibroblast growth factor-saporin mitotoxin.

BACKGROUND: The authors attached basic fibroblast growth factor (FGF-2), a growth factor for numerous tumors and normal cell types, to saporin (SAP), a ribosome-inactivating protein isolated from the plant Saponaria officinalis. The conjugate (FGF-SAP) then was tested for antitumor activity using B16-F10 melanoma cells. This rapidly growing murine melanoma cell line has been used classically as a model to screen antitumor agents. METHODS: B16-F10 cells in culture were used for in vitro experiments or introduced into C57BL/6 mice to demonstrate the in vivo antitumor activities of FGF-SAP. RESULTS: FGF-SAP was found to be an extremely effective cytocidal agent in vitro with an ED50 of 30-60 pM. The effects were specific for FGF-2 receptors, as shown by the ability of FGF-2 to block FGF-SAP action. In the in vivo models, FGF-SAP was found to increase survival time, inhibit tumor growth, and decrease metastases. CONCLUSIONS: The authors conclude that this mitotoxin has potent in vitro and in vivo effects on B16-F10 cells, supporting the hypothesis that ligand-mediated cytotoxicity can control tumor growth.

Animals↗
PubMed1994

Expression and activities of a recombinant basic fibroblast growth factor-saporin fusion protein.

A fusion protein containing the full-length sequences of the mitogen, basic fibroblast growth factor (FGF-2), and the ribosome-inactivating protein, saporin (SAP), has been expressed in E. coli. As expected, it binds with high affinity to heparin-Sepharose like FGF-2 and can displace the binding of radiolabeled FGF-2 to its high affinity receptor. In contrast, the fusion protein only has much lower ribosome-inactivating activity than free saporin, although full ribosome-inactivating protein activity can be generated by proteolytic removal of the FGF-2 moiety. Cytotoxicity experiments with B16-F10 mouse melanoma cells establish that the fusion protein is active as a chemical conjugate against these intact cells. Presumably these cells have the ability to activate the SAP component of the fusion protein through an intra-cellular metabolism of the fusion protein. Because we also show the fusion protein has tumor growth inhibition properties and antimetastatic activity in in vivo models of melanoma, the findings support the hypothesis that FGF-based ligand-mediated cytotoxicity can serve to target cytotoxic agents in vivo.

Animals↗
PubMed1994

Photoaffinity-labeling peptide substrates for farnesyl-protein transferase and the intersubunit location of the active site.

--CAAX motif peptides, which are substrates for isoprenylation, were synthetically derivatized with the light-sensitive benzophenone (Bz) group in order to determine their potential use as catalytic site-directed covalent photocross-linking ligands for one of the enzymes catalyzing protein isoprenylation, farnesyl-protein transferase (FPTase). Bz-peptides could be synthesized with [3H]benzophenone and possessed eiter one or two benzophenone groups located at or near the peptide's NH2 terminus (e.g. the mono-Bz probes Bz-ACVIM and Bz-LPCVVM, and the di-Bz derivatized probe Bz-GY-(Bz)PCVVM, referred to as Bz2-GYPCVVM). Each type of derivatized peptide behaved as a substrate for farnesylation in vitro without irradiation, while under 366-nm irradiation each demonstrated covalent cross-linking ability as a catalytic site-directed photoaffinity ligand with tissue-purified or enriched but impure fractions from rat and bovine brain FPTase, as well as with a recombinant human FPTase variant, FPTase (beta alpha t) expressed in Escherichia coli. Without photoactivation, Bz-ACVIM yielded a Kd of 37 nM for the cloned variant of human FPTase. Pseudo first-order photolytic inhibition of FPTase preparations with Bz-peptides, as well as protection from photoinactivation by unmodified -CAAX motif peptides, supported the capacity of these Bz-peptides to serve as co-substrates and their specificity for seeking the catalytic site of the enzyme. SDS-polyacrylamide gel electrophoresis analysis subsequent to photolysis indicated that the mono-Bz-derivatized peptides (e.g. [3H]Bz-LPCVVM or 3H]Bz-ACVIM) became covalently cross-linked preferentially to the approximately 49-kDa beta subunit of the alpha beta dimeric FPTase. The farnesyl-PP cosubstrate bound equally well to unmodified and Bz-ACVIM-labeled enzyme. The di-Bz derivative, [3H]Bz2-GYPCVVM, in contrast, revealed exclusive photocovalent cross-linking with a species of molecular mass approximately 95-97 kDa, indicating that both FPTase subunits were tethered together covalently by the di-Bz probe. Similar differential SDS-polyacrylamide gel electrophoresis cross-linking patterns were obtained with homogeneous FPTases as well as with partially purified rat or bovine brain enzyme preparations. The absence of nonspecific photolabeling of any proteins in the partially purified rat or bovine brain enzyme preparations other than FPTase independently attested to the high efficiency of photocross-linking of the FTPase, and the selective catalytic site-seeking ability of these Bz-derivatized peptide substrates, verifying their potential as structural probes for the active site domain on the enzyme.(ABSTRACT TRUNCATED AT 400 WORDS)

Affinity Labels↗
PubMed1994

Fibroblast growth factor in the hypothalamic-pituitary axis: differential expression of fibroblast growth factor-2 and a high affinity receptor.

In situ hybridization and immunohistochemistry were used to map gene expression and protein distribution of basic fibroblast growth factor (FGF-2) in the hypothalamic-pituitary system. Although the expression of FGF-2 mRNA in the pituitary is low, the protein is widely distributed in both its neural and anterior lobes. In the anterior lobe, immunoreactive (ir-) FGF-2 localizes to basement membranes and select endocrine cells. In the neural lobe, ir-FGF-2 is detected in basement membranes, pituicytes, and Herring bodies. Analyses of FGF high affinity receptor (FGFR) immunoreactivity in the anterior pituitary establishes a distribution of FGFR similar to that of FGF-2. In the neural lobe, ir-FGFR is associated with nerve fibers, pituicytes, and Herring bodies. Unlike FGF-2, the distribution of FGFR1 mRNA correlates well with the presence of the immunoreactive receptor. In the hypothalamus, magnocellular neurons of paraventricular and supraoptic nuclei contain ir-FGF-2 and ir-FGFR. In the median eminence, ir-FGF-2 and ir-FGFR is associated with fibers, glial, and endothelial cells. Ependymal and subependymal cells lining the third ventricle also show high levels of ir-FGF-2 and ir-FGFR and mRNAs. Overall, there is a specific and selective distribution of FGF-2 and its high affinity receptor(s) in the hypothalamo-pituitary axis. This localization lead us to postulate a role in neurohypophyseal functions, possibly water balance.

Animals↗
PubMed1994

Establishment of a new cell line with the characteristics of a multipotential progenitor from a patient with chronic myelogenous leukemia in early erythroblastic crisis.

A novel cell line (KH88) was established from a patient with chronic myelogenous leukemia in blastic crisis. The leukemic blasts had the features of undifferentiated blasts with basophilic agranular cytoplasm and they were focally positive for acid phosphatase and alpha-naphthyl acetate esterase. CD36, CD33, HLADR, and CD71 were expressed on the surfaces of the blast cells. Most blasts were positive for platelet peroxidase activity, and some of them had granules containing aggregates of ferritin molecules. These findings were compatible with those of 'early' erythroblastic leukemia, this established cell line (KH88) having similar characteristics, and actually producing hemoglobin A and hemoglobin F. Although the KH88 cells were negative for megakaryocytic markers, they were induced to express CD41 by phorbol ester. Further, a few KH88 cells were positive for myeloperoxidase. This cell line was thus revealed to have the capacity to differentiate into three lineages, providing a useful model for studying the differentiation of multipotential stem cells. Moreover, a subline of KH88 had a peculiar chromosome abnormality, del(3)(q21q25); it would be useful to study the significance of this chromosomal abnormality.

Aged↗
PubMed1983

Effect of vasectomy on the steroid profile of human seminal plasma.

The effect of vasectomy on the steroid profile of seminal plasma was studied in 19 healthy men at 4-monthly intervals during a 2-year post-vasectomy period. The steroids analyzed by radioimmunoassays were: androstenedione (A), testosterone (T), dihydrotestosterone (DHT), oestradiol (E2) and the sulphoconjugated forms of pregnenolone (delta 5P-S), dehydroepiandrosterone (DHEA-S), testosterone (T-S), dihydrotestosterone (DHT-S), oestrone (E1-S) and oestradiol (E2-S). The quantitatively most important steroids present in the seminal plasma were DHEA-S, delta 5P-S and DHT-S. Vasectomy resulted in a significant but non-progressive reduction in DHT (40%) and T (23%) levels. The levels of E2, delta 5P-S and DHT-S also exhibited a smaller reduction following vasectomy. No significant changes were observed in the levels of the other steroids studied. The results are interpreted as suggesting that under normal conditions the bulk of the steroids present in the ejaculate is contributed by the accessory reproductive organs. In addition, a significant part of the DHT and T reach the ejaculate together with the sperm directly from the testis and/or epididymis. It is suggested that the normal steroid levels in seminal plasma established in this investigation could serve as reference values in the hormonal evaluation of male infertility.

Adult↗
PubMed1980

An evaluation of the CODABAR symbol in blood-banking automation.

The CODABAR system has been used in blood banking automation since 1976 when it was introduced by 16 blood centers and transfusion services throughout the United States in a test program coordinated by the American Blood Commission Committee for Commonality in Blood Banking Automation. Since then CODABAR has been used in sample identification on Groupamatic and Technicon automated blood-typing equipment, in the labeling of blood products and in their distribution. It is widely used by a number of blood centers in the United States and has been adopted by the 14 regional blood centers in the United Kingdom, as well as blood centers in Switzerland, the Netherlands and Japan. Some properties of the CODABAR symbol that are relevant to the high accuracy requirements of blood banking are described and preliminary test data are presented.

Autoanalysis↗