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Biomedical subjects

R Zhou

Publications and source records attributed to R Zhou.

At least 145 records · Page 8Linked to original sources

Detection of ligands in regions anatomically connected to neurons expressing the Eph receptor Bsk: potential roles in neuron-target interaction.

Neuron-target interaction is a key feature in the establishment of neuronal networks. However, the underlying mechanism remains unclear. We have shown that at the time of target innervation, Bsk, an eph family receptor, is expressed at high levels in several brain regions including the hippocampus, olfactory bulb, and retina. To study whether the ligands are expressed in the target tissues, we investigated the expression of Bsk ligands using a ligand-affinity probe, Bsk-AP, which consisted of the extracellular domain of Bsk fused in frame with a human placental alkaline phosphatase. These analyses showed that the ligands were expressed at high levels in the developing septum, hypothalamus, olfactory neural epithelium, and tectum. In situ hybridization studies revealed that at least three different factors were responsible for the Bsk-AP binding. In the septum, Elf-1, Lerk3 (Eff-2), and AL-1/Lerk7 were transcribed. In the hypothalamus, AL-1/Lerk7 was the ligand detected by Bsk-AP. In the olfactory system, high levels of Lerk3 were detected in the sensory neurons. Both Elf-1 and AL-1/Lerk7 were present in the tectum. These ligand-positive areas are known to be anatomically connected to Bsk-expressing regions. These observations strongly suggest that Bsk and the ligands participate in neuron-target interactions in multiple systems and provide support for their involvement in topographic projection.

Alkaline Phosphatase↗

Chemistry of the pyrrolo[1,2-alpha]benzimidazole antitumor agents: influence of the 7-substituent on the ability to alkylate DNA and inhibit topoisomerase II.

This study addresses the influence the 7-substituent on the cytotoxicity of pyrrolo[1,2-alpha]-benzimidazole quinones possessing a 6-aziridinyl group (PBIs) and a 6-acetamido group (APBIs). Reduction of a PBI to the aziridinyl hydroquinone results in both nucleophile trapping (alkylation) and 1,5-sigmatropic shift reactions. The latter process is essentially an internal redox reaction wherein the hydroquinone causes reductive opening of the aziridinyl ring. The 7-substituent controls the fate of the aziridinyl ring by means of steric and electronic effects. An electron-rich 7-substituent favors the 1,5-sigmatropic shift reaction. If the 7-substituent distorts the 6-aziridinyl group from the conformation required for the 1,5-sigmatropic shift, then nucleophile trapping occurs. The 7-methyl substituent results in significant nucleophilic trapping, and the 7-unsubstituted and 7-methoxy substituents favor the 1,5-sigmatropic reaction. Thus, the 7-methyl PBIs show the most cytotoxicity of the analogues studied. The APBIs are cytotoxic only as quinones, and reduction to the hydroquinone results in loss of activity. Consistent with this observation, the change from 7-methyl to the more electron-rich 7-methoxy results in a substantial loss of APBI cytotoxicity as well as decreased topoisomerase II inhibition. The mechanism of inhibition is thought to involve the interacalation of only electron deficient APBIs into DNA.

Alkylation↗

Regulation of topographic projection in the brain: Elf-1 in the hippocamposeptal system.

The hippocampus and septum play central roles in one of the most important spheres of brain function: learning and memory. Although their topographic connections have been known for two decades and topography may be critical for cognitive functions, the basis for hippocamposeptal topographic projection is unknown. We now report for the first time that Elf-1, a membrane-bound eph family ligand, is a candidate molecular tag for the genesis of the hippocamposeptal topographic projection. Elf-1 is expressed in an increasing gradient from dorsal to ventral septum. Furthermore, Elf-1 selectively allows growth of neurites from topographically appropriate lateral hippocampal neurons, while inhibiting neurite outgrowth by medial hippocampal neurons. Complementary to the expression of Elf-1, an eph family receptor, Bsk, is expressed in the hippocampus in a lateral to medial gradient, consistent with a function as a receptor for Elf-1. Further, Elf-1 specifically bound Bsk, eliciting tyrosine kinase activity. We conclude that the Elf-1/Bsk ligand-receptor pair exhibits traits of a chemoaffinity system for the organization of hippocamposeptal topographic projections.

Animals↗

Higher in vivo protein binding of etoposide in children compared with adult cancer patients.

Etoposide is bound to plasma albumin (94%). Previous studies have revealed altered protein binding of etoposide in cancer patients. This has clinical implications since only the free fraction is considered pharmacologically active. We have studied the etoposide protein binding in 11 children (eight acute lymphocytic leukemia, two malignant histiocytosis, and one oligodendroglioma; age 1-17 years) and 46 adult patients (28 acute myelocytic leukemia, eight lymphoma, one multiple myeloma, and nine small cell lung cancer; age 38-81 years). All patients were treated with etoposide 50-200 mg/m2 i.v. or orally. Plasma from ten healthy volunteers, 26-50 years of age, was spiked with etoposide, 10 micrograms/ml, and the protein binding was compared with that in patient samples. The free etoposide concentration was determined by high performance liquid chromatography (HPLC) after ultrafiltration at room temperature. The free etoposide fraction was lower, 2.5 +/- 0.6% (mean +/- SD), in the children compared with 5.0 +/- 3.6% in adult cancer patients. In plasma from healthy adults it was 3.2 +/- 0.3%. It is concluded that children have significantly lower levels of free etoposide compared with adult patients (P = 0.03) as well as with healthy subjects (P = 0.001), which is likely to affect metabolism and renal clearance as well as cellular uptake of the drug.

Adolescent↗

DNA damage induced by etoposide; a comparison of two different methods for determination of strand breaks in DNA.

Etoposide induces DNA damage to cells by interacting with the nuclear enzyme topoisomerase II. In this investigation the human lymphoblastic leukemia cell line (CEM) was used to study induction of DNA-strand breaks and cellular drug uptake after treatment with etoposide at a concentration of 0.5-2 micrograms/ml. High performance liquid chromatography was used for determination of etoposide concentrations. The alkaline elution assay and the DNA unwinding technique were compared for quantifying strand breaks in DNA induced by etoposide. The concentrations required to increase the level of DNA damage significantly was as follows: the DNA unwinding technique, 0.20 microgram/ml; the alkaline elution assay with proteinase K, 0.45 microgram/ml; the alkaline elution assay without proteinase K, 0.60 microgram/ml. When the half-life was adjusted, considering the efflux time of etoposide from cells, it was found to be only a few minutes. The present data show that the DNA unwinding technique is to be preferred for the screening of DNA damage. This technique is easier and quicker to perform than the alkaline elution technique.

Antineoplastic Agents, Phytogenic↗

The Mos/mitogen-activated protein kinase (MAPK) pathway regulates the size and degradation of the first polar body in maturing mouse oocytes.

Mos is an upstream activator of mitogen-activated protein kinase (MAPK) and, in mouse oocytes, is responsible for metaphase II arrest. This activity has been likened to its function in Xenopus oocytes as a component of cytostatic factor. Thus, Mos-deficient female mice (MOS-/-) are less fertile and oocytes derived from these animals fail to arrest at metaphase II and undergo parthenogenetic activation [Colledge, W. H., Carlton, M. B. L., Udy, C. B. & Evans, M. J. (1994) Nature (London) 370, 65-68 and Hashimoto, N., Watanabe, N., Furuta. Y., Tamemoto, B., Sagata, N., Yokoyama, M., Okazaki, K., Nagayoshi, M., Takeda, N., Ikawa, Y. & Aizawa, S. (1994) Nature (London) 370, 68-71]. Here we show that maturing MOS-/- oocytes fail to activate MAPK throughout meiosis, while p34cdc2 kinase activity is normal until late in metaphase II when it decreases prematurely. Phenotypically, the first meiotic division of MOS-/- oocytes frequently resembles mitotic cleavage or produces an abnormally large polar body. In these oocytes, the spindle shape is altered and the spindle fails to translocate to the cortex, leading to the establishment of an altered cleavage plane. Moreover, the first polar body persists instead of degrading and sometimes undergoes an additional cleavage, thereby providing conditions for parthenogenesis. These studies identify meiotic spindle formation and programmed degradation of the first polar body as new and important roles for the Mos/MAPK pathway.

Animals↗

AtJ1, a mitochondrial homologue of the Escherichia coli DnaJ protein.

The nucleotide sequence of a cDNA clone from Arabidopsis thaliana ecotype Columbia was determined, and the corresponding amino sequence deduced. The open reading frame encodes a protein, AtJ1, of 368 residues with a molecular mass of 41 471 Da and an isoelectric point of 9.2. The predicted sequence contains regions homologous to the J- and cysteine-rich domains of Escherichia coli DnaJ, but the glycine/phenylalanine-rich region is not present. Based upon Southern analysis, Arabidopsis appears to have a single atJ1 structural gene. A single species of mRNA, of 1.5 kb, was detected when Arabidopsis poly(A)+ RNA was hybridized with the atJ1 cDNA. The function of atJ1 was tested by complementation of a dnaJ deletion mutant of E. coli, allowing growth in minimal medium at 44 degrees C. The AtJ1 protein was expressed in E. coli as a fusion with the maltose binding protein. This fusion protein was purified by amylose affinity chromatography, then cleaved by digestion with the activated factor X protease. The recombinant AtJ1 protein was purified to electrophoretic homogeneity. In vitro, recombinant AtJ1 stimulated the ATPase activity of both E. coli DnaK and maize endosperm cytoplasmic Stress70. The deduced amino acid sequence of AtJ1 contains a potential mitochondrial targeting sequence at the N-terminus. Radioactive recombinant AtJ1 was synthesized in E. coli and purified. When the labeled protein was incubated with intact pea cotyledon mitochondria, it was imported and proteolytically processed in a reaction that depended upon an energized mitochondrial membrane.

Adenosine Triphosphatases↗

[Comparative study on the long-term effect of permanent embolization of hepatic artery with Bletilla striata in patients with primary liver cancer].

From May, 1990 to Sept., 1993 106 cases of primary liver cancer were treated by temporary or permanent hepatic arterial embolization with Bletilla striata powders (permanent, 56 cases) or gelfoam powders (temporary, 50 cases) under controlled technical conditions, in term of degree of tumor necrosis, revascularization of tumor, and long-term effect, the results of Bletilla striata group were better as compared with those of gelfoam group. The 1, 2 and 3 year survival rates were 44.9%, 33.6% and 33.6% in Beltilla striata group while the rates were 48.9%, 31.1% and 16.0% in gelfoam group, suggesting that Bletilla striata is superior to gelfoam as an embolizing agent for hepatic carcinoma.

Antineoplastic Agents, Phytogenic↗

A multiple wavelength algorithm in color image analysis and its applications in stain decomposition in microscopy images.

Stains have been used in optical microscopy to visualize the distribution and intensity of substances to which they are attached. Quantitative measures of optical density in the microscopic images can in principle be used to determine the amount of the stain. When multiple dyes are used to simultaneously visualize several substances to which they are specifically attached, quantification of each stain cannot be made using any single wavelength because attenuation from the several stain components contributes to the total optical density. Although various dyes used as optical stains are perceived as specific colors, they, in fact, have complex attenuation spectra. In this paper, we present a technique for multiple wavelength image acquisition and spectral decomposition based upon the Lambert-Beer absorption law. This algorithm is implemented based on the different spectral properties of the various stain components. By using images captured at N wavelengths, N components with different colors can be separated. This algorithm is applied to microscopy images of doubly and triply labeled prostate tissue sections. Possible applications are discussed.

Algorithms↗

[The inhibitory effects of serum of patients with acute promyelocytic leukemia on the formation of megakaryocytopoiesis colony unit].

OBJECTIVES: To determine if the sera of patients with acute promyelocytic leukemia (APL) and all-trans retinoic acid (ATRA) inhibit the formation of colony-forming-unit of megakaryocyte (CFU-Meg). METHODS: By cellular culture assay, the sera of 22 APL patients treated with ATRA and 10(-6)' mol/L ATRA were kinetically used to perform the investigation. Meanwhile, serum tumor necrosis factor (TNF) activity and bone marrow CFU-L were also detected. RESULTS: Before treatment, there was an inhibitory activity on CFU-Meg in the serum of APL patients (29.6% +/- 2.4%). The activity reduced gradually with the treatment and was stimulus when bone marrow remission obtained (-18.7% +/- 4.1%, P < 0.001). It was positively correlated with TNF activity in the serum and the number of CFU-L however negatively correlated with the number of megakaryocyte in bone marrow and PLT. Serum TNF activity and the number of CFU-L before treatment were higher than those after treatment (P < 0.05 and < 0.001, respectively). ATRA could stimulate the growth of CFU-Meg. CONCLUSIONS: There was an inhibitory activity in the serum of APL patients which could be used to monitor the efficacy of the therapy. ATRA stimulates the growth of CFU-Meg.

Adolescent↗

[New use of Bletilla striata as embolizing agent in the intervention treatment of hepatic carcinoma].

A Chinese herb, Bletilla striata, was used as embolizing agent in order to improve the therapeutic results of intervention treatment of liver cancer. From October 1991 to January 1995, 56 cases of hepatic carcinoma were treated with Bletilla striata by hepatic artery embolization, with conventional gelform embolization in 50 cases as control. Patients were followed-up for 10-48 months. Embolization with Bletilla striata led to extensive and permanent vascular obstruction, accompanied with marked shrinkage of tumor size and significant decrease in serum AFP levels. Collaterals were few in number and collateral circulation was established late so that the treatment intervals could be prolonged, with an average of 7 months. The 1-, 2-, and 3-year survival rate was 81.9%, 44.9% and 33.6%, respectively, with a median survival time of 19.8 months. All the clinical parameters were better than those treated with conventional gelform embolization. The results indicate that Bletilla striata is an ideal vascular embolizing agent.

Adult↗

[Study on local huangjiabang delivery device].

This article reports the pharmacological functions of local Huangjiabang delivery device which mainly includes medicinal herbs. The experimental study showed that Huangjiabang could inhibit the predominant cultivable organisms in periodontal disease significantly, and could also inhibit inflammatory swelling caused by xylene. Jiaochacaijiao, egg serum and filter-paper-granuloma significantly. The preliminary clinical observation showed that GI, PLI, SBI and PD were reduced remarkably in Huangjiabang administered pockets after two days and seven days of treatment. It is suggested that this new direct drug delivery system with medicinal herbs has wide prospect in the treatment of periodontal disease.

Administration, Topical↗

Correlation of survival with quantitative tissue staining of prostate specific acid phosphatase in patients with prostate carcinoma by using microscopic image analysis: a preliminary report of correlative studies on RTOG protocols 75-06, 77-06, and 83-07.

PURPOSE: We have previously shown that the intensity (graded semiquantitatively as 1-4+) of tissue prostate-specific acid phosphatase (PSAP) staining determined immunocytochemically in a cohort of prostate carcinoma patients from Radiation Therapy Oncology Group Protocols (RTOG) protocols 75-06 and 77-06 correlated with survival. The extent of this staining was heterogeneous and was estimated. The extent of staining was not found to be significantly associated with survival. We undertook the present quantitative study to see if the improved precision and reliability of measurement of the intensity and extent of prostate specific acid phosphatase staining would confirm and extend our previous observations. METHODS AND MATERIALS: Patient cohorts representative of the entire group were obtained from RTOG 75-06 plus 77-06 and 83-07. The RTOG 77-06 plus 75-06 patients (No-Hormone population) did not receive preradiation hormonal therapy. RTOG 83-07 patients (Prehormone population) received one of two types of preradiation chemical androgen ablation. In this study, histologic slides of tumors were immunocytochemically stained for PSAP by the peroxidase-antiperoxidase (PAP) technique using diaminobenezidene (DAB) as a substrate and hematoxylin as a nuclear counterstain. The intensity and extent of immunocytochemical PSAP staining (IPSAP stain) was quantified using our dual wavelength and batch mode image process technique. RESULTS: Our study of 151 cases confirmed that overall survival of patients in both populations was positively correlated with the intensity and extent of IPSAP stain. Results of the two studies were similar. The statistical significance of the relationship of both extent and intensity was greater in the cohort from protocol 83-07, which was the patient group receiving pretreatment with hormones. In a Cox multiple regression analysis including clinical stage, Gleason and M. D. Anderson grades, and the cohort of patients (Prehormone or No-Hormone group) as covariables, both the intensity and extent of the IPSAP stain significantly correlated with survival along with M. D. Anderson grade of the tumor. CONCLUSION: Quantitative image analysis of the IPSAP stain predicts survival in patients treated with external beam radiotherapy with and without prior hormonal therapy.

Acid Phosphatase↗

Mutagenic analysis of functional domains of the mos proto-oncogene and identification of the sites important for MAPK activation and DNA binding.

We constructed in-frame deletion/replacement mutations in the Xenopus mos proto-oncogene that lie within conserved Mos-specific codons, but outside of the regions that are conserved among the src kinase family of genes. All gene products were assayed in vitro for kinase activity and in vivo for their ability to induce oocyte maturation, embryonic cleavage arrest and cellular transformation. Most mutations in Mos eliminated both kinase and biological activity. However, a mutation in Mos that removed two basic amino acid residues (R94 and K97) downstream from the lysine at the ATP binding site (K90) markedly enhanced autophosphorylation activity. Moreover, this mutant displayed markedly reduced biological activity, lacked transforming activity, and failed to activate mitogen activated protein kinase (MAPK). A second mutant Mos product, lacking amino acids R45-A54, displayed a five-fold increase in cellular transforming activity. This Mos mutant specifically localized to the cytoplasm; in contrast to wild-type (wt) Mos that localized to both the nucleus and the cytoplasm. These data indicate that Mos transforming activity is mediated via signalling exerted in the cytoplasm, presumably through MAPK, and that nuclear localization of the oncogene product interferes with transforming activity. We also show that amino acids R45-A54 are important for Mos DNA binding activity.

3T3 Cells↗

Full-matrix refinement of the protein crambin at 0.83 A and 130 K.

This paper describes the first successful full-matrix least-squares (FMLS) refinement of a protein structure. The example used is crambin which is a small hydrophobic protein (4.7 kDa, 46 residues). It proves the feasibility of refining such large molecules by this classic method, routinely applied to small molecules. The final structure with 381 non-H protein atoms (54 protein atoms in alternative positions), 367 H atoms, 162 water molecules (combined occupancy 93) and one disordered ethanol molecule converged to a standard unweighted crystallographic R-factor of R = 9.0% when refined against F with reflections stronger than F > 2sigma(F) and R = 9.5% when refined against F(2). The programs RFINE [Finger & Prince (1975). Natl Bur. Stand. (US) Tech. Note 854. A System of Fortran IV Computer Programs for Crystal Structure Computations] and SHELXL93 [Sheldrick (1993). SHELXL93. Program for Crystal Structure Refinement, Univ. of Göttingen, Germany] were used for FMLS refinement with the high-resolution low-temperature (0.83 A, 130 K) data set of a mixed-sequence form of crambin. A detailed analysis of the models obtained in FMLS and PROLSQ [restrained least squares or RLS; Teeter, Roe & Heo (1993). J. Mol. Biol. 230, 292-311] refinements with the same data set is presented. The differences between the models obtained by both FMLS and RLS refinements are systematic but negligible and advantages and shortcomings of both methods are discussed. The final structure has very good geometry, fully comparable to the geometry of other structures in this resolution range. Ideal values used in PROLSQ and those by Engh & Huber [Engh & Huber (1991). Acta Cryst. A47, 392-400] differ significantly from this refinement and we recommend a new standard. FMLS refinement constitutes a sensitive tool to detect and model disorder in highly refined protein structures. We describe the modeling of temperature factors by the TLS method [Schomaker & Trueblood (1968). Acta Cryst. B24, 63-76]. Rigid body-TLS refinements led to a better understanding of different modes of vibrations of the molecule. Refinements using F(2) or F protocols converged and reached slightly different minima. Despite theoretical support for F(2)-based refinement, we recommend refinement on structure factors.

Journal Article↗