Search PubMed⌕ Search

Biomedical subjects

C Zhang

Publications and source records attributed to C Zhang.

785 records · Page 44Linked to original sources
PubMedDate not supplied

Genetic engineering strategies for purification of recombinant proteins from canola by anion exchange chromatography: an example of beta-glucuronidase.

The elution behavior of native canola proteins from different anion-exchange resins was determined. The elution profiles showed the potential for simplified recovery of acidic recombinant proteins from canola. When Q-sepharose fast flow was used, there were three optimal salt elution points at which a recombinant protein would have minimal contamination with native proteins. The feasibility of exploiting this advantage was examined for recovery of the acidic protein beta-glucuronidase (GUS/GUSD0 from the Escherichia coli gene) along with three polyaspartate fusions to the wild-type GUS. The fusions contained 5 (GUSD5), 10 (GUSD10), or 15 (GUSD15) aspartic acids fused to the C-terminus and were chosen to extend the elution time. The three fusions and the wild-type enzyme were produced in E. coli, purified, and added to canola extracts before chromatography. The equivalence of this spiking experiment to that of extracting a recombinant protein from transgenic canola was determined in a control experiment using transgenic canola expressing the wild-type enzyme. Behavior in the transgenic and spiked experiments was equivalent. GUSD0 eluted at the earliest optimal elution point; the addition of polyaspartate tails resulted in longer retention times and better selective recovery. If one assumes binding through a single fusion (the protein is a tetramer), there is a nearly linear shift in elution within the salt gradient of 17 mM per added charge up to 10, with a reduced increment from 10 to 15. The fusions and their enzymatic activity proved very stable in the canola extracts through 7 days in cold storage, providing flexibility in process scheduling.

Anion Exchange Resins↗
PubMedDate not supplied

New flavonol glycosides and new xanthone from Polygala japonica.

Three new flavonol glycosides and a new xanthone were isolated from Polygala japonica HOUTT. with eight known compounds. Their structures were identified as 1,7-dihydroxy-3,4-dimethoxy-xanthone (1), kaempferol-7,4'-dimethyl ether (2), physcion (3), guazijinxanthone (4), rhamnetin (5), polygalin A (6), 3,5,7-trihydroxy-4'-methoxy-flavone-3-O-beta-d-galactopyranoside (7), 3,5,3'-trihydoxy-7,4'-dimethoxy-flavone-3-O-beta-d-galactopyranoside (8), 3,5,3',4'-tetrahydroxy-7-methoxy-flavone-3-O-beta-d-galactopyranoside (9), 3,5,3',4'-tetrahydroxy-7-methoxy-flavone-3-O-beta-d-glucopyranoside (10), polygalin B (11), polygalin C (12). Among them, compound 4 is a new xanthone, and 6, 11 and 12 are new flavonol glycosides. Compounds 1, 4, 7 and 8 were tested for cytotoxic activity with MTT assays on five human tumor cell lines, K562, A549, PC-3M, HCT-8 and SHG-44. Compound 4 showed cytotoxic activity against all the five cell lines.

Antineoplastic Agents, Phytogenic↗
PubMedDate not supplied

Unusual selective immunoglobulin deficiency in an Arabian foal.

A 10-month-old Arabian foal was evaluated for a suspected immunoglobulin (Ig) M deficiency. Decreased to nondetectable concentrations of IgM, IgA, and IgG (T), and a normal concentration of IgG, were present. Results of in vitro testing of the blood lymphocyte blastogenesis showed a weak response to the B-cell mitogen, lipopolysaccharide (LPS), but normal responses to T-cell mitogens. Results of postmortem examination showed synovitis of the left tibiotarsal and both scapulohumeral joints. Atrophy and edema of the lymph nodes and lymphocyte depletion in the thymus and spleen were seen. A subacute inflammatory infiltrate was observed in the kidney, synovium, liver, and brain. Etiologic agents were not identified. This case represents a previously unreported form of immunodeficiency disease in the horse.

Animals↗
PubMedDate not supplied

Intravascular leukostasis and systemic aspergillosis in a horse with subleukemic acute myelomonocytic leukemia.

Leukemia is a neoplastic disease of one or more of the cell types of the hemopoietic system and is rarely diagnosed in the horse. This report describes a case of subleukemic acute myelomonocytic leukemia in an 11-year-old gelding. Preliminary cytological diagnosis was supported by two types of laboratory investigations. Cytochemical characterization of blood and bone marrow neoplastic cells was consistent with a myelomonocytic origin. Neoplastic blast cells in peripheral blood were labeled by monoclonal antibodies specific for cell surface molecules of horse granulocytes, but they were not labeled by antibodies to T- or B-lymphocytes or macrophages. Treatment was attempted but was unsuccessful. At necropsy, intravascular leukostasis was present in all tissues examined. Fungal hyphae were also found in lung interstitium and colonic submucosa, suggesting the presence of a systemic mycosis. Nucleated cells were isolated from peripheral blood and cultured in vitro; they survived for up to 2 weeks and had evidence of cell division that was not sustained. Frozen-thawed cells stored in liquid nitrogen were also successfully cultured in vitro, but no permanent cell lines could be established.

Animals↗
PubMedDate not supplied

Diagnostic imaging of breast cancer using fluorescence-enhanced optical tomography: phantom studies.

Molecular targeting with exogenous near-infrared excitable fluorescent agents using time-dependent imaging techniques may enable diagnostic imaging of breast cancer and prognostic imaging of sentinel lymph nodes within the breast. However, prior to the administration of unproven contrast agents, phantom studies on clinically relevant volumes are essential to assess the benefits of fluorescence-enhanced optical imaging in humans. Diagnostic 3-D fluorescence-enhanced optical tomography is demonstrated using 0.5 to 1 cm(3) single and multiple targets differentiated from their surroundings by indocyanine green (micromolar) in a breast-shaped phantom (10-cm diameter). Fluorescence measurements of referenced ac intensity and phase shift were acquired in response to point illumination measurement geometry using a homodyned intensified charge-coupled device system modulated at 100 MHz. Bayesian reconstructions show artifact-free 3-D images (3857 unknowns) from 3-D boundary surface measurements (126 to 439). In a reflectance geometry appropriate for prognostic imaging of lymph node involvement, fluorescence measurements were likewise acquired from the surface of a semi-infinite phantom (8x8x8 cm(3)) in response to area illumination (12 cm(2)) by excitation light. Tomographic 3-D reconstructions (24,123 unknowns) were recovered from 2-D boundary surface measurements (3194) using the modified truncated Newton's method. These studies represent the first 3-D tomographic images from physiologically relevant geometries for breast imaging.

Algorithms↗
PubMedDate not supplied

Metabolism of 3-(13)C-malate in primary cultures of mouse astrocytes.

Malate, specifically labeled with carbon 13 on C(3), was synthesized by chemical means and used to study malate metabolism by primary cultures of mouse cortical astrocytes. 3-(13)C-Malate in combination with glucose as well as 3-(13)C-malate alone were used as substrates; the effect of 3-nitropropionic acid, an inhibitor of succinate dehydrogenase and fumarase was also examined. The consumption of malate was only 0.26 micromol/mg of protein, approx. 25-fold lower than the consumption of glucose. Besides lactate, glutamine and fumarate were the two major metabolites released to the medium. Very low and similar levels of isotopic enrichment were detected on C(2) and C(3) of lactate; glutamine was labeled on C(2) and C(3) to a similar extent as well and labeling on C(4) was only detected when glucose was not added. These labeling studies suggest that cytosolic malic enzyme is not active in primary astrocytes and support the occurrence of pyruvate recycling in astrocytes.

Animals↗
PubMedDate not supplied

A novel hydroxamic acid compound, BMD188, demonstrates anti-prostate cancer effects by inducing apoptosis. II: In vivo efficacy and pharmacokinetic studies.

BACKGROUND: In the preceding paper, we demonstrated that, BMD188 [cis-1-hydroxy-4-(1-naphthyl)-6-octylpiperidine-2-one], a newly synthesized cyclic hydroxamic acid compound, induces potent apoptotic death of prostate cancer cells in vitro. In this project, we studied the in vivo pharmacokinetic behavior and anti-tumor efficacy of this novel compound. MATERIALS AND METHODS: A bioavailability/elimination study was first performed using radiolabeled BMD188 administered to rats through intraperitoneal (i.p.), intravenous (i.v). or oral (p.o.) routes. Based on these pharmacokinetic data as well as pilot experiments on in vivo toxicity, two sets of efficacy studies, with i.p. administered BMD188, were performed in SCID mice or athymic nude mice which had been orthotopically transplanted with Du145 human prostate cancer cells. Tumor growth rate was measured and the final tumor weights and sizes determined. Subsequently, histopathological data were obtained and tumor tissue sections were used for apoptosis (i.e., TUNEL) staining. RESULTS: The pharmacokinetic studies revealed low (approximately 8%) absorption through the p.o. route and high (approximately 70%) absorption through the i.p. route. The average plasma half life (T1/2) of BMD188 was approximately 50 h. Post-absorption, plasma elimination of radioactivity was similar to that in animals given [3H]-188 intravenously. The in vivo efficacy results indicate that i.p. administered BMD188 significantly inhibited the primary growth and local invasion of Du145 prostate cancer cells orthotopically implanted into SCID or athymic nude mice. The tumor-inhibitory effect of BMD188 was due to apoptosis induction in vivo, as revealed by histological analysis as well as TUNEL staining of the tumor tissue sections. CONCLUSION: Collectively, the preceding in vitro and the current in vivo studies suggest that BMD188 and its analogs may find clinical applications in the treatment of prostate cancer patients by inducing apoptotic death of prostate cancer cells.

Animals↗
PubMedDate not supplied

Improved safety, pharmacokinetics and therapeutic efficacy profiles of a novel liposomal formulation of mitoxantrone.

We describe here the preclinical studies of a novel formulation of liposome-entrapped mitoxantrone (LEM). The liposome entrapment efficiency of mitoxantrone was 93.4 +/- 2.8%. In vitro cytotoxicity studies in HL60 cells comparing LEM with conventional mitoxantrone (MTO) showed IC50 values of 0.31 +/- 0.05 ng/ml and 0.48 +/- 0.06 ng/ml for LEM and MTO, respectively. In CD2F1 mice, LEM was significantly less toxic as compared with MTO. A single intravenous (i.v.) dose of 15 mg/kg MTO produced 100% mortality in CD2F1 mice by Day 10, whereas a single i.v. dose as high as 35 mg/kg LEM caused no mortality for at least up to Day 60 post-treatment. Multiple doses of MTO (i.v., 5.0 mg/kg, 1x daily, x5) caused 100% mortality by Day 10, whereas a similar dose regimen of LEM caused no mortality in CD2F1 mice. Clinical and histopathology evaluations indicated long-term normal tissue protection in mice treated with relatively high single dose (i.v., 35 mg/kg) or multiple doses of LEM (i.v., 5.0 mg/kg, 1x daily, x5). LEM also demonstrated favourable pharmacokinetic profiles. CD2F1 mice injected with 5 mg/kg i.v. dose of LEM showed plasma levels 51-fold higher than with an equivalent dose of MTO. The area under the plasma concentration-time curve was 200-fold greater with LEM as compared to MTO. The plasma half-lives were 0.96 hours and 0.11 hours for LEM and MTO, respectively. An altered tissue distribution was observed with LEM; cardiac tissue demonstrating at least 2.6-fold lower levels of mitoxantrone with LEM vs. MTO. LEM exhibited significant anti-tumor activity against murine ascitic L1210 leukemia in CD2F1 mice. Treatment with a single dose of 20.0 mg/kg LEM resulted in 100% long-term survivors. LEM 2.5 mg/kg (i.v., x4) had antitumor activity against a human hormone-independent prostate carcinoma (PC-3) grown in athymic mice, while a comparable dose of MTO was too toxic. A significant decrease in toxicity, altered pharmacokinetics, and enhanced efficacy of LEM suggest that LEM may provide a viable alternative to the clinical use of conventional mitoxantrone.

Animals↗
PubMedDate not supplied

Inhibitory action of ganglioside GM3 on murine neuroblastoma cell proliferation: modulating effect of fetal calf serum.

Exogenous gangliosides have been shown to exert a regulatory influence on the proliferation and differentiation of several cell lines in tissue culture. The effect of ganglioside GM3 on C-1300 murine neuroblastoma (MNB) cell proliferation and the modulating action of fetal calf serum (FCS) concentration in the culture media have been investigated. MNB cells were cultured in DMEM containing 1, 2.5, 5 or 10% FCS, and incubated with GM3 at concentrations ranging from 1.95 to 500 microM. Cell proliferation was assayed 4 days after the addition of GM3 using the CellTiter 96 Non-Radioactive Cell Proliferation Assay. GM3 inhibited MNB cell proliferation in a dose-dependent manner regardless of the FCS concentration in the culture media. However, the magnitude of this inhibitory effect was inversely proportional to the FCS concentration in the culture media. The addition of albumin to MNB cells cultured in DMEM containing 1% FCS exerted no effect on the antiproliferative action of GM3. FACS cell cycle analyses demonstrated that MNB cultured in DMEM containing 1% FCS had a higher proportion of cells in the G0/G1 compartment when compared to those cultured in 10% FCS. The enhanced response of MNB cells to GM3 observed in 1% FCS, may be due to a preferential action on cells in the G0/G1 stage of the cell cycle. These studies have demonstrated that the ganglioside GM3 inhibited MNB cell proliferation in tissue culture and this effect was modulated by FCS concentration in the culture media. Since protein-binding of GM3 by FCS does not appear to be the primary mechanism by which FCS exerts its antagonistic effects, we hypothesize that this may be due to the opposing action of stimulatory growth factors present in FCS.

Animals↗
PubMedDate not supplied

Physical and psychosocial side effects of antepartum hospital bed rest.

The purpose of this study was to describe the physical and psychosocial effects of antepartum hospital bed rest on pregnant women and to determine whether the symptoms exhibited differed from those exhibited by pregnant women not on bed rest. Women on complete bed rest (n = 10) had greater gastrocnemius muscle dysfunction, weight loss, and dysphoria than women on partial bed rest (n = 7) or no bed rest (n = 18). Separation from family was the greatest hospital stressor. Postpartum recovery from the side effects of bed rest was prolonged and included symptoms of muscular and cardiovascular deconditioning. The severity of side effects appeared to be directly related to the degree of severity restriction.

Activities of Daily Living↗