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Biomedical subjects

J Q Zhou

Publications and source records attributed to J Q Zhou.

15 recordsLinked to original sources

Quasi-monochromatic fiber depolarizer and its application to polarization-dependent loss measurement.

We theoretically derive the relationship between the degrees of polarization (DOPs) of input and output for an optical component with polarization-dependent loss (PDL) and birefringence. Based on the theoretical result, we propose a novel depolarizer for quasi-monochromatic light that can depolarize a fully polarized light with a 50 MHz linewidth to less than 0.2% in the DOP The depolarized light is then used to measure PDL in a single-mode optical fiber link. To the best of our knowledge, our new PDL measurement method is significantly faster than all known methods. Experimental results show excellent agreement with other methods.

Journal Article↗

An evaluation of para toluene-sulfonamide metabolism and effect with regard to CYP isoforms, P-glycoprotein, and drug interactions.

Aim of this study was to investigate liver metabolism of with regard to para toluene-sulfonamide (PTS), CYP isoforms, P-glycoprotein (P-gp), and drug interactions. Known substrates, inducers and inhibitors of CYP and inhibitor of P-gp were employed and metabolites were determined with HPLC. Male Wistar rats were pretreated with ip phenobarbital (PB), ketoconazole (Ket), or verapamil (Ver) for 3 days and in situ liver perfusion of PTS was conducted in a recirculation system. Rats were also pretreated with ip PTS (33 mg x kg(-1) x d(-1) or PTS 99 mg x kg(-1) x d(-1)) for 4 days before liver perfusions with dextromethorphan (Dex) and phenacetin (Phe) preparations were conducted. Microsome incubation was used to investigate PTS effect on five CYP isoforms and PTS-drug interactions probability with phyllotoxin and 5-fluorouracil (5-FU) in vitro. PTS at 60 min perfusates had areas of 61.4% and 133.6% of the blank control in PB group and Ket group, respectively. The result that PTS metabolism was enhanced by PB and inhibited by Ket treatments suggested liver CYP was attributed to PTS metabolism. PTS 99 mg x kg(-1) x d(-1) pretreatment slowed down the metabolism of Dex and Phe while in vitro incubations did not show a PTS (0-160 micromol/L) effect on CYP activities. PTS metabolite formation when co-incubated with phyllotoxin was 50.7% of the negative control. The potent inhibitory ability of phyllotoxin to PTS requires further clinical investigation regarding in concomitant administration.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

The Saccharomyces Pif1p DNA helicase and the highly related Rrm3p have opposite effects on replication fork progression in ribosomal DNA.

Replication of Saccharomyces ribosomal DNA (rDNA) proceeds bidirectionally from origins in a subset of the approximately 150 tandem repeats, but the leftward-moving fork stops when it encounters the replication fork barrier (RFB). The Pif1p helicase and the highly related Rrm3p were rDNA associated in vivo. Both proteins affected rDNA replication but had opposing effects on fork progression. Pif1p helped maintain the RFB. Rrm3p appears to be the replicative helicase for rDNA as it acted catalytically to promote fork progression throughout the rDNA. Loss of Rrm3p increased rDNA breakage and accumulation of rDNA circles, whereas breakage and circles were less common in pif1 cells. These data support a model in which replication fork pausing causes breakage and recombination in the rDNA.

Amino Acid Sequence↗

The small subunit is required for functional interaction of DNA polymerase delta with the proliferating cell nuclear antigen.

DNA polymerase delta is usually isolated as a heterodimer composed of a 125 kDa catalytic subunit and a 50 kDa small subunit of unknown function. The enzyme is distributive by itself and requires an accessory protein, the proliferating cell nuclear antigen (PCNA), for highly processive DNA synthesis. We have recently demonstrated that the catalytic subunit of human DNA polymerase delta (p125) expressed in baculovirus-infected insect cells, in contrast to the native heterodimeric calf thymus DNA polymerase delta, is not responsive to stimulation by PCNA. To determine whether the lack of response to PCNA of the recombinant catalytic subunit is due to the absence of the small subunit or to differences in post-translational modification in insect cells versus mammalian cells, we have co-expressed the two subunits of human DNA polymerase delta in insect cells. We have demonstrated that co-expression of the catalytic and small subunits of human DNA polymerase delta results in formation of a stable, fully functional heterodimer, that the recombinant heterodimer, similar to native heterodimer, is markedly stimulated (40- to 50-fold) by PCNA and that the increase in activity seen in the presence of PCNA is the result of an increase in processivity. These data establish that the 50 kDa subunit is essential for functional interaction of DNA polymerase delta with PCNA and for highly processive DNA synthesis.

Animals↗

Purification and characterization of the catalytic subunit of human DNA polymerase delta expressed in baculovirus-infected insect cells.

The catalytic subunit of human DNA polymerase delta has been overexpressed in insect cells by a recombinant baculovirus. The recombinant protein has a Mr = approximately 125,000 and is recognized by polyclonal antisera against N-terminal and C-terminal peptides of the catalytic subunit of human DNA polymerase delta. The recombinant protein was purified to near homogeneity (approximately 1200-fold) from insect cells by chromatography on DEAE-cellulose, phosphocellulose, heparin-agarose, and single-stranded DNA-cellulose. The purified protein had both DNA polymerase and 3'-5' exonuclease activities. The properties of the recombinant catalytic subunit were compared with those of the native heterodimeric DNA polymerase delta isolated from fetal calf thymus, and the enzymes were found to differ in several respects. Although the native heterodimer is equally active with either Mn2+ or Mg2+ as divalent cation activator, the recombinant catalytic subunit is approximately 5-fold more active in Mn2+ than in Mg2+. The most striking difference between the two proteins is the response to the proliferating cell nuclear antigen (PCNA). The activity and processivity of native DNA polymerase delta are markedly stimulated by PCNA whereas it has no effect on the recombinant catalytic subunit. These results suggest that the small subunit of DNA polymerase delta is essential for functional interaction with PCNA.

Animals↗

Endurance-training induced changes in skeletal muscle phosphoglycerate kinase of old Wistar rats.

Sufficiently intense, long-term, endurance training has been shown in several studies to induce a variety of adaptations in skeletal muscle, including a substantial restoration of the activities of several muscle enzymes which are known to be modified during biological aging. This activity-restoration may reflect either an increase in the amounts of enzyme proteins or an enhancement of the specific activities of these molecules. The present study examined the effect of long-term endurance training on the status of phosphoglycerate kinase in skeletal muscle of old rats, as compared with the enzyme isolated either from non-trained old or young animals. The kinetics of heat inactivation, which differ markedly between young and old forms of phosphoglycerate kinase, were used as a sensitive probe for the status of the enzyme. The results reveal a remarkable similarity between the heat inactivation patterns of phosphoglycerate kinase from the muscle of old, exercise-trained rats and enzyme purified from young animals, while enzyme samples isolated from sedentary old animals are significantly more heat-stable. Adaptation to endurance-training is thus evident at the molecular level, and maintains phosphoglycerate kinase in its young form. The aging of this enzyme has been previously shown to involve only conformational changes, which develop following a reversible partial oxidation of reactive cysteine residues. Whether the adaptation of the enzyme to endurance-training results from enhancement in its turnover rate (i.e., dwell time in the cell becoming too short for modifications to develop) or is due to increased protection against oxidation (being the first step in the enzyme's aging) remains to be studied.

Aging↗

Exposure of rat muscle phosphoglycerate kinase to a nonenzymatic MFO system generates the old form of the enzyme.

The occurrence of age-related modifications in enzymes is a well-established symptom of aging that has been explained by several possible mechanisms including the oxidation of amino acid residues by mixed-function oxidation (MFO) systems. In the present study native old phosphoglycerate kinase was compared with young enzyme which had been modified by oxidation with ascorbate: FeCl3 followed by reduction. The comparison was done by monitoring the rates of heat denaturation of these enzyme forms, as well as their inactivation by trypsin. A remarkable similarity between the old and treated young enzyme was revealed, while native young phosphoglycerate kinase was inactivated with a different rate. Extensive unfolding followed by refolding converted both old and MFO-treated young phosphoglycerate kinase to species which greatly resemble the native young enzyme in their heat inactivation kinetics. These results demonstrate that the exposure of phosphoglycerate kinase to the mixed-function oxidation system introduces some modifications which are not reversed by subsequent enzyme reduction and which resemble those found in the native old enzyme. This mechanism, therefore, may account for the aging of phosphoglycerate kinase in vivo.

Aging↗

[Effect of expression of exogenous PDGF-A chain on growth and transformation of CHO cells].

CHO cells were transfected with plasmid pSV2-PDGF-A (containing human PDGF-A cDNA) by calcium phosphate method. Twenty transfected cell lines were obtained after G418 selection. The selected 2 cell lines At1 and Aot7), with prominent changes in morphology and growth behaviour, showed transcription of PDGF-A chain mRNA much higher than CHO cells, strong fluorescent PDGF-specific reaction, appearing that PDGF-like proteins were synthesized in cytoplasm of these cells. At1 and Aot7 cells not only had increased growth rate, but also formed large colonies in soft agar and grew into fibrosarcomas in nude mice. These results suggested that the expression of exogenous PDGF-A gene might cause the uncontrolled growth and malignant transformation of CHO cells.

Animals↗

Earthworm muscle preparation for studying several potent compounds--the possible existence of a "Soman receptor.".

A new model of earthworm (Eisenia foedida) dorsal muscle preparation was studied. It is a specific preparation for testing GABA and its agonist muscimol. The response of muscle could be recorded through mechanical functional changes instead of potential variations. Thus, it is a direct and simple means for measuring GABA activity. ChE-deactivated preparation was suggested for this purpose. By mild membrane modifications of earthworm muscle preparation, it was possible to remove or inactivate GABA, glutamic acid or Soman combining protein. In conjunction with superfusion of Ach-receptor antagonist (alpha-Bungaratoxin) or GABA antagonist (picrotoxin, isopropylbicyclophosphate), we could differentiate Soman receptor from Ach and GABA receptor. The possible existence of a Soman receptor in the earthworm muscle was therefore proposed.

Animals↗