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H Guo

Publications and source records attributed to H Guo.

At least 271 records · Page 15Linked to original sources

Prenatal exposure to methadone delays the development of striatal cholinergic neurons.

The effect of prenatal exposure to methadone via maternal osmotic minipumps on the expression of acetylcholinesterase (AChE) and choline acetyltransferase (ChAT) has been studied by light microscopy in the striatum of male and female rats. At postnatal day 10, rats of both sexes exhibit reduced intensity of ChAT-immunoreactive staining in striatal neurons in the methadone-treated group in comparison to either untreated or water-treated controls. Although the number and distribution of ChAT-immunoreactive neurons appear to be similar across all three groups, the size (cross-sectional area) of these neurons is significantly smaller in the methadone-treated animals. By postnatal day 22, there are no differences in the ChAT immunoreactivity of striatal neurons between the water-treated and methadone-treated animals. The size of these neurons in female animals of both treatment groups is significantly larger than those of male animals in the same groups at postnatal day 22. AChE histochemical staining is qualitatively similar between treatment groups in both age groups. Thus, prenatal exposure to methadone appears to produce a delay in the expression of ChAT in striatal neurons. It remains to be demonstrated whether these differences are a direct effect of methadone exposure or are a consequence of neonatal withdrawal.

Acetylcholine↗

Therapeutic tumor-specific cell cycle block induced by methionine starvation in vivo.

The ability to induce a specific cell cycle block selectively in the tumor could have many uses in chemotherapy. In the present study we have achieved this goal of inducing a tumor-specific cell cycle block in vivo by depriving Yoshida sarcoma-bearing nude mice of dietary methionine. Further, we demonstrate that methionine depletion also causes the tumor to eventually regress. The antitumor effect of methionine depletion resulted in the extended survival of the tumor-bearing mice. The mice on the methionine-deprived diets maintained their body weight for the time period studied, indicating that tumor regression was not a function of body weight loss. The data reported here support future experiments utilizing methionine depletion as a target for tumor-selective cell cycle-dependent therapy.

Animals↗

Mechanism of DmS-II-mediated pause suppression by Drosophila RNA polymerase II.

Transcription elongation factor S-II mediates nascent transcript cleavage by RNA polymerase II (Reines, D. (1992) J. Biol. Chem. 267, 3795-3800). We have examined the mechanism of action of the Drosophila S-II analog, DmS-II, in a defined transcription system. Our results show that DmS-II is necessary and sufficient to activate nascent transcript cleavage by RNA polymerase II during transcription of a dC-tailed template. The pattern of transcripts resulting from prolonged action by DmS-II indicates that there are kinetic barriers to transcript shortening. During the cleavage reaction, the polymerase remains in register with the template strand and generates mainly nucleotide dimers. The ability of DmS-II to mediate transcript shortening resides in the carboxyl-terminal half of the protein. Our results support a model for pause suppression in which DmS-II binds to the paused polymerase, causes one cleavage event and is then released from the complex. Elongation by the polymerase then allows a second encounter with the pause site and a second chance of passing the site. Complete pause suppression may require multiple transcript shortening events for some polymerase molecules.

Animals↗

Phosphorylation and activation of protamine kinase by two forms of a myelin basic protein kinase from extracts of bovine kidney cortex.

Two myelin basic protein kinases designated MBPK-1 and MBPK-2 were purified to apparent homogeneity from extracts of bovine kidney cortex. The purified preparations exhibited an apparent M(r) approximately 40,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and approximately 42,000 (MBPK-1) and 45,000 (MBPK-2) by gel permeation chromatography. Up to 0.4 and 1.8 mol of phosphoryl groups were incorporated per mol of MBPK-1 and MBPK-2, respectively, on threonines following incubation with ATP. Autophosphorylation, incubation with protein phosphatase 2A2 (PP2A2), CD45, or T-cell protein tyrosine phosphatase did not affect MBPK-1 activity. Autophosphorylation increased by about 3-fold MBPK-2 activity. This autophosphorylation and activation was reversed by PP2A2 but not by CD45 or T-cell protein tyrosine phosphatase. MBPK-1 and MBPK-2 displayed a positive reaction with an antibody to mitogen-activated protein kinase. Purified preparations of protamine kinase were activated by about 1.5-6-fold and, after inactivation with PP2A2, were reactivated by about 30% by MBPK-1 and MBPK-2. Activation and reactivation correlated with the incorporation, respectively, of 0.1-0.5 and 0.5 mol of phosphoryl groups/mol of the protamine kinase on serines. The results show that MBPK-1 and MBPK-2 are protamine kinase-activating kinases and suggest that MBPK-1 and MBPK-2 may be related to mitogen-activated protein kinase.

Amino Acid Sequence↗

Purification and characterization of an autophosphorylation-activated protein serine threonine kinase that phosphorylates and inactivates protein phosphatase 2A.

Protein phosphatase 2A2 is inactivated by phosphorylation following incubation with purified preparations of an autophosphorylation-activated protein kinase (Hong Guo and Zahi Damuni (1992) Proc. Natl. Acad. Sci. U.S.A. 90, 2500-2504). This protein kinase was purified about 250,000-fold from extracts of bovine kidney to apparent homogeneity. The purified preparations exhibited a single polypeptide of apparent M(r) approximately 36,000. Up to 1 mol of phosphoryl groups was incorporated per mol of the purified kinase following incubation with ATP. This autophosphorylation reaction (t1/2 approximately 0.5-1 min) was accompanied by a approximately 10-fold activation of the kinase. Autophosphorylation and activation were reversed by protein phosphatase 2A2 or the catalytic subunit of protein phosphatase 1. Phosphoamino acid analysis indicated that the kinase underwent autophosphorylation on threonines. The rate of autophosphorylation was independent of the concentration of the enzyme and a slope of 0.97 (gamma = 0.998) was obtained by van't Hoff's plot indicating that autoposphorylation was intramolecular. Relative to myelin basic protein, the enzyme exhibited about 8, 62, 130, 33, 5, and < 0.1% activity with histones H1, H2A, H2B, H3, and H4 and with glycogen synthase alpha, respectively. Heparin inhibited the activity of the enzyme half-maximally at about 20 micrograms/ml. The results indicate that this autophosphorylation-activated kinase is a new protein kinase.

Adenosine Triphosphate↗

Autophosphorylation-activated protein kinase phosphorylates and inactivates protein phosphatase 2A.

Purified preparations of a distinct autophosphorylation-activated protein kinase from bovine kidney phosphorylated and inactivated purified preparations of protein phosphatase 2A2 (PP2A2) by about 80% with the autophosphorylation-activated protein kinase, protamine kinase, and 32P-labeled myelin basic protein as substrates. Analysis of incubations performed in the presence of 0.2 mM [gamma-32P]ATP by autoradiography following SDS/PAGE and by FPLC gel permeation chromatography on Superose 12 demonstrated that the catalytic subunit of PP2A2 was phosphorylated in the incubation mixtures containing the kinase and phosphatase. Up to 0.3 mol of phosphate groups was incorporated per mol of the catalytic subunit of PP2A2 following incubation with the kinase. This phosphorylation was enhanced about 5-fold in the presence of 0.4 microM microcystin-LR. In addition, up to 1 mol of phosphate groups was incorporated per mol of the PP2A2 subunit of apparent M(r) approximately 60,000 when microcystin-LR was included. Analysis by thin-layer chromatography indicated that PP2A2 catalyzed an autodephosphorylation reaction which was inhibited by microcystin-LR. Phospho amino acid analysis showed that the catalytic subunit of PP2A2 was phosphorylated on threonine residues by the autophosphorylation-activated protein kinase. Together with previous observations, the results suggest that inactivation of PP2A by phosphorylation catalyzed by the autophosphorylation-activated protein kinase could contribute to the marked increase in the phosphorylation of cellular proteins in response to insulin and other mitogens.

Adenosine Triphosphate↗

Hypothermia blunts acetylcholine increase in CSF of traumatically brain injured rats.

Activation of muscarinic acetylcholine (ACh) receptors contributes to the pathophysiological consequences of moderate experimental traumatic brain injury (TBI). Hypothermia (30 degrees C) provides protection in experimental TBI. We measured ACh levels in CSF and plasma 5 min after moderate fluid percussion TBI under normothermic or hypothermic conditions, because ACh in the CSF has been correlated with the severity of behavioral deficits after TBI. Three groups were examined: TBI with hypothermic brain (30 degrees C), TBI with normothermic brain (37 degrees C), or sham TBI with normothermic brain (37 degrees C). ACh concentrations in CSF were significantly higher in 37 degrees C TBI rats, but not in 30 degrees C TBI rats compared to shams. ACh concentrations in plasma did not differ between groups. These results suggest that a contributing factor to the neuroprotective effects of moderate hypothermia in TBI may be related to the reduction of excessive ACh levels in the central nervous system following injury.

Acetylcholine↗

[The study of the RFLP of D14S13 site of human genome].

The single-locus probe pCE 1.2 recognizes the D14S13 site in human genome. Allele-frequency of HaeIII digested D14S13 site is drawn out from 269 unrelated individuals. It displayed that there were at least 66 alleles and more than 93% heterogeneity. No mutation in 93 individuals of 8 families was observed. This RFLP technique has been applied in the identification and parentage testing in forensic science, and the accuracy of identity was calculated. Some further applications were discussed.

Alleles↗