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Biomedical subjects

B Xu

Publications and source records attributed to B Xu.

At least 325 records · Page 18Linked to original sources
PubMed1993

Identification of the metal ligands and characterization of a putative zinc finger in methionyl-tRNA synthetase.

A truncated form of the methionyl-tRNA synthetase (delta MTS), which has been cloned, overproduced, and characterized, was used in an attempt to better understand the role of the enzyme-bound zinc in the amino-acylation process. Apo-, Zn(2+)-, Co(2+)-, and 113Cd(2+)-substituted delta MTS proteins were prepared in vivo and purified to homogeneity. Apo-delta MTS was devoid of enzymatic activity in the aminoacylation of tRNA(fMet) and in the methionine-dependent ATP-pyrophosphate exchange reactions. Kinetic constants in both the aminoacylation and ATP-pyrophosphate exchange reactions for the Co(2+)- and 113Cd(2+)-substituted delta MTS proteins were found to be identical with those of the native Zn2+ protein. The low energy absorption spectrum of Co(2+)-substituted delta MTS resembles the d-d transition bands characteristic of tetrahedrally coordinated Co(2+)-substituted proteins. A strong S-->Co2+ charge transfer absorption at 350 nm was clearly evident having a molar absorptivity consistent with four thiolate ligands. The environment of the metal center was further probed by measuring the 113Cd chemical shift of 113Cd(2+)-substituted delta MTS. A single resonance at 759.6 ppm was observed. This chemical shift is consistent with Cd2+ coordinated to four thiolate ligands. The Escherichia coli methionyl-tRNA synthetase contains a potential metal binding sequence Cys-X2-Cys-X9-Cys-X2-Cys in a connecting polypeptide within the nucleotide fold. Titration of a 21-amino acid peptide corresponding to this putative metal binding site, Cys145-Cys161, was shown to bind Co2+ with a Kd of 120 +/- 11 microM. These results demonstrate that the isolated zinc finger binding domain is capable of specifically forming a stoichiometric complex with the divalent cation. Taken together, our studies identify the 4 cysteine residues in the zinc finger-like domain as the metal binding ligands in the E. coli methionyl-tRNA synthetase. The role of the enzyme-bound metal appears to be structural and not directly involved in catalysis.

Amino Acid Sequence↗
PubMed1993

Cytotoxic effect of interferon on primary malignant tumour cells. Studies in various malignancies.

It is a well established fact that interferon (IFN) can inhibit cell growth, but only recently has it been found that IFN can exert a direct cytotoxic effect on primary tumour cells. This was shown in malignant cells from patients with multiple myeloma. In this study the influence of IFN on the viability of primary malignant cells from patients with different malignancies was studied. As previously described a direct cytotoxic effect of IFN on multiple myeloma cells was observed. No major effects on cell viability could be found in malignant cells from patients with lymphoma, chronic lymphocytic leukaemia, hairy cell leukaemia, chronic myelogenous leukaemia and carcinoma. This indicates that the direct cytotoxic effect of IFN in multiple myeloma may be relatively specific for this malignancy. It could be due to a specific differentiation stage in the myeloma cells, specific genetic alterations and/or abrogation of an autocrine/paracrine loop.

Cell Count↗
PubMed1993

Arginine vasotocin gene expression and hormone synthesis during ontogeny of the chicken embryo and the newborn chick.

Chicken embryos at different developmental stages (embryonal day (E) 6 to 21) and chicks at posthatch day 1 (D1) were monitored for the development of their hypothalamo-neurohypophysial system as indicated by the kinetics of arginine vasotocin (AVT) gene expression via mRNA concentration and brain AVT content. Our data concerning the onset of gene expression support previous results from our laboratory and others about an early activation of the AVT gene transcriptional and translational activity around E6. We could detect measurable amounts of AVT in chicken embryo brains at E6 and an exponential increase during further development until D1. Dot blots of hypothalamic RNA extracts indicated that AVT gene transcript concentrations rose between E12 and E17 and slightly dropped thereafter. Northern hybridization showed that this drop was caused by a decrease of full length message and an increase of smaller transcripts during late embryonal and D1 stages, probably an AVT mRNA specific degradation phenomenon. The dissociation between the increase of AVT concentration and AVT mRNA concentration visible at the D1 stage might be due to accumulation and storage of AVT in the magnocellular neurons, preferentially in their axon terminals in the neurohypophysis. Blood samples taken from E14 onwards revealed a constant increase in plasma osmolality and plasma AVT concentration. Our data suggest that, in the chicken, AVT seems to be required early during embryonal development, either for osmoregulatory or further unknown functions.

Animals↗
PubMed1993

A human mitochondrial transcriptional activator can functionally replace a yeast mitochondrial HMG-box protein both in vivo and in vitro.

Human mitochondrial transcription factor A is a 25-kDa protein that binds immediately upstream of the two major mitochondrial promoters, thereby leading to correct and efficient initiation of transcription. Although the nature of yeast mitochondrial promoters is significantly different from that of human promoters, a potential functional homolog of the human transcriptional activator protein has been previously identified in yeast mitochondria. The importance of the yeast protein in yeast mitochondrial DNA function has been shown by inactivation of its nuclear gene (ABF2) in Saccharomyces cerevisiae cells resulting in loss of mitochondrial DNA. We report here that the nuclear gene for human mitochondrial transcription factor A can be stably expressed in yeast cells devoid of the yeast homolog protein. The human protein is imported efficiently into yeast mitochondria, is processed correctly, and rescues the loss-of-mitochondrial DNA phenotype in a yeast abf2 strain, thus functionally substituting for the yeast protein. Both human and yeast proteins affect yeast mitochondrial transcription initiation in vitro, suggesting that the two proteins may have a common role in this fundamental process.

Amino Acid Sequence↗
PubMed1993

Role of neuropeptide-Y in episodic luteinizing hormone release in ovariectomized rats: an excitatory component and opioid involvement.

We tested the hypothesis that hypothalamic neuropeptide-Y (NPY) is an excitatory signal in the episodic secretion of LH in ovariectomized (ovx) rats and that the suppression of LH secretion that consistently follows intracerebroventricular administration of NPY is due to concurrent release of opioids or CRH, both previously shown to readily inhibit LH release. In the first experiment, ovx rats received continuous intraventricular infusion of either serum containing NPY antibodies (NPY-Ab) or normal rabbit serum (control) at dilutions of 1:5 or 1:1. NPY-Ab infusion at a 1:5 dilution significantly decreased mean plasma LH levels and LH pulse amplitude without affecting LH pulse frequency over a 3-h period of observation. However, infusion of relatively more concentrated NPY-Ab (1:1) markedly decreased not only mean plasma LH levels and LH pulse amplitude, but also the frequency of LH episodes. In the next experiment, we observed that intraventricular administration of NPY (0.2 nmol) suppressed LH release for 60 min. However, blockade of opiate receptors with iv infusion of naloxone (2 mg/h) before and after NPY injection completely counteracted the NPY-induced inhibition of LH release. On the other hand, prior blockade of the CRH receptors with alpha-helical CRH-(9-41) (25 or 100 micrograms/rat) was ineffective in reversing the inhibitory LH response of NPY (0.125 nmol). These results together with our previous demonstration of morphological communication between NPY and beta-endorphin neurons, show that suppression of LH by exogenous NPY in ovx rats may result from concurrent stimulation of opioids, primarily beta-endorphin. However, diminution of all parameters of episodic LH secretion by NPY-Ab affirms the notion that the NPY network is a physiologically important excitatory component of the hypothalamic pulse generator circuitry that regulates episodic LH secretion in rats.

Animals↗
PubMed1993

Increased pressor responsiveness of femoral arteries to exogenous norepinephrine in renal hypertensive dogs mediated through alpha 2 adrenoceptors.

Changes of perfusion pressure induced by femoral arterial perfusion of norepinephrine (NE) were studied in renal hypertensive dogs made with the method of wrapping both kidneys. The NE threshold dose which induced perfusion pressure increase was lower in hypertensive dogs than that in normotensive dogs. The NE pressor response in low concentration (1, 2, 10 ng.kg-1) was increased in hypertensive dogs, after alpha 2 adrenoceptors antagonized by idazoxan this increased response had been eliminated. These suggested that increased vasoconstriction by exogenous NE in hypertensive dogs was mediated through the increased reactivity of postsynaptic alpha 2 adrenoceptors to NE.

Adrenergic alpha-Antagonists↗
PubMed1993

Inhibition of phosphorylation of histone H1 and H3 induced by 10-hydroxycamptothecin, DNA topoisomerase I inhibitor, in murine ascites hepatoma cells.

Hydroxycamptothecin (HCPT), isolated from Camptotheca acuminata, is a powerful antitumor alkaloid. Previous studies indicated that the molecular target of this agent was DNA topoisomerase I. The present results demonstrated that in vitro treatment of murine ascites hepatoma cells with HCPT resulted in a marked reduction in DNA syntheses and the inhibition of phosphorylation in histone was in a time-dependent manner. Gel electrophoresis found that HCPT had a selectively inhibitory effect on the phosphorylation of histone H1 and H3, but less effect on the other kinds of histones. In vivo, HCPT also exhibited a suppressive effect on histone H1 and H3 phosphorylation. These data suggested that HCPT-induced cell killing may be, at least in part, associated with the suppression of histone H1 and H3 phosphorylation.

Animals↗
PubMed1993

Transformation of Escherichia coli with foreign DNA by electroporation.

Electroporation has been widely applied in molecular biology in recent years. In this study, successful transformation of plasmid DNA and transfection of phage DNA into E. coli were described by using intense electrical field of exponential decay waveform (electroporation) generated by a Gene Pulsar LN-101 (made in Tianjin, Tianjin Institute of Technology and Nankai University). We have obtained 10(9)-10(10) transformants/micrograms with strain DH5 alpha and plasmid pUC18, by a single voltage pulse at 1.0-2.5 kV (initial electric field strength = 2.8-16 kV/cm) with 5-20 microF capacitor. The efficiency of electroporation depends on various parameters: the electric field strength, capacitance, the pulse length (RC time constant), etc. The frequency of transformation was a linear function of the DNA concentration, as well as the density of recipient cell. The incubation time of before or after electroporation has no significant effect on transformation. The high transfection efficiency was also achieved with strain JM109 and M13mp19RF by electroporation. The method was easier, more time-saving and more efficient than Ca(2+)-dependent transformation (transfection) method. The results obtained by Gene-Pulsar LN-101 was similar to that by the Bio-Rad Gene-Pulsar apparatus.

Calcium↗
PubMed1993

[Studies on the establishment of malarial animal model of short-term relapse. IV. Short-term relapse in rhesus monkeys infected with sporozoites of Plasmodium cynomolgi].

The present paper reports that the short-term relapse could be artificially made by the application of the experimental method, and thus we established the monkey model of the short-term relapse. According to the experimental design, when the parasitemia was detected in rhesus monkeys infected with sporozoites of Plasmodium cynomolgi, a combined therapy of pyronaridine 6 mg/kg body weight, artemether 10 mg/kg and chloroquine 20 mg/kg once daily for 3 days was carried out to clear the erythrocytic parasite and then the short-term relapse was observed in the animal follow-up for 100 days. The combined therapy was given again when relapse occurred. One onset of relapse occurred on 47 days after therapy in monkey M194 infected with sporozoites 11 x 10(3). In M195 infected with sporozoites 55 x 10(4), relapses occurred for 3 times on 30, 37 and 51 days respectively after medication, but during a follow-up period of 200 days, no relapse was shown in M192 and M193 infected with sporozoites 50 and 55 x 10(2), respectively. The results showed that the frequency of the short-term relapse was relative to the sporozoites inoculation, and sporozoites less than 11 x 10(3) were not suitable for making short-term relapse in animal model. No long term relapse could be seen in all the 4 monkeys until 400 days. Under existing conditions that the relapse was generally explained with hypnozoite assumption, the establishment of animal model of short-term relapse could be tenable.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

An interactive image processing system for the quantification of cardiac function in the embryonic chick.

This paper describes an interactive computer-based method for quantifying embryonic chick heart blood pool volumes that have been recorded as digitized image sequences. The primary motivation for the development of such a system is to be able to extract measurement data that is useful for evaluating potential drug-induced effects on intracardiac blood flow while minimizing variabilities in the measurements that typically occur when human evaluators extract the measurements visually from photographs. The processing that is used to achieve this goal can be divided into four basic steps: enhancing the contrast of the digitized embryonic chick heart image, extracting the shape of the blood pool in the embryonic chick heart ventricle from the enhanced image, quantifying the axial dimensions of the blood pool and calculating the required cardiac blood flow data. The technical details and the basic theory motivating each processing step are provided as well as an analysis of the results obtained by applying the method to actual image data.

Animals↗
PubMed1993

[Distribution of 14C labeled at dioxopiperazine or methyl morpholine group of probimane by whole body autoradiography].

Probimane (AT-2153) is a new anticancer compound. It was first developed in this Institute. It is effective against mouse tumors S37, S180, Lewis lung carcinoma, L1210 and human pulmonary adenocarcinoma heterotransplanted into nude mice. In the present work, 14C was labeled at central dioxopiperazine or methyl morpholine group of probimane 120 mg.kg-1 was injected iv in mice bearing Lewis lung carcinoma by whole body autoradiography. The results showed that probimane was broken into at least two parts: a central part and a methyl morpholine group. The central part of compound hardly penetrated through the blood-brain barrier, but accumulated in the urinary bladder. The methyl morpholine group showed a high affinity to tumor tissue and accumulated in spleen, bone and liver.

Animals↗
PubMed1993

[A study on classification of Xanthomonas by isoelectric focusing].

Studying Xanthomonas strains of 4 species and 29 pathovars by isoelectric focusing, it was shown that the protein patterns were very different among the species and pathovars. Through cluster analysis of IEF date, it was found that the protein pattern differences among certain pathovars were no fewer than that among the species. It revealed that some pathovars could become to be species. There were few differences of 6 protein bands among X. campestris pv. cerealis, X. campestris pv. undulosa and X. campestris pv. hordei which cause bacterial black cheff, so the classification of the three pathovars need to be re-considered. It also showed that isoelectric focusing could be used to classify Xanthomonas below species.

Cluster Analysis↗