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Biomedical subjects

Q Peng

Publications and source records attributed to Q Peng.

At least 91 records · Page 5Linked to original sources
PubMed1997

Effect of surface photorefractive keratectomy and laser in situ keratomileusis on the corneal endothelium.

PURPOSE: To investigate endothelial cell loss in pairs of fresh human autopsy globes following high-diopter myopic photorefractive keratectomy (PRK) or laser in situ keratomileusis (LASIK). SETTING: Center for Research on Ocular Therapeutics and Biodevices and Magill Laser Center for Vision Correction, Storm Eye Institute, Charleston, South Carolina, USA. METHODS: In the first part of the study, 12 globes had either -10 diopters (D) multizone surface PRK or -10 D single-zone LASIK. In the second part, three groups of 5 globes each had -15 D, -20 D, or -25 D multizone-blend LASIK procedures. Fellow globes in both groups were used as untreated controls. Corneoscleral buttons were excised from all globes. Following 7 days in corneal organ culture, the endothelial surface was stained with two vital dyes: calcein-AM and ethidium homodimer. Fluorescence microscopy was used to obtain endothelial cell counts. RESULTS: The mean dead cells per square millimeter (cells/mm2) were 0.94 in the -10 D PRK treated corneas compared with 0.91 in the fellow untreated eyes (P = 0.06(. The mean dead cells/mm2 in the -10 D single-zone LASIK-treated corneas and in the fellow untreated eyes were 0.61 (P = 0.88). The mean dead cells/mm2 in the -15 D, -20 D, and -25 D multizone-blend LASIK-treated corneas were 3.08, 2.33, and 5.55, respectively, compared with 3.49, 1.92, and 5.01 in the fellow untreated eyes (P = 0.276, P = 0.339, and P = 0.427, respectively). Dead cell counts for treated and control paired corneas were highly correlated in all treatment groups. CONCLUSIONS: No significant endothelial cell loss occurred after -10 D PRK or LASIK corrections up to -25 D. Although this study has limitations that prevent direct extrapolation to the clinical situation, it does afford a comparable clinical correlate for endothelial cell toxicity following a typical excimer laser ablations.

Aged↗
PubMed1997

Silicone oil adhesion to intraocular lenses: an experimental study comparing various biomaterials.

PURPOSE: To perform an in vitro experimental study comparing the degree of adherence of silicone oil to various rigid and foldable intraocular lens (IOL) designs and to the human lens capsule. SETTING: Center for Research on Ocular Therapeutics and Biodevices, Department of Ophthalmology, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: Seven IOL styles comprising various biomaterials were studied: fluorine-treated (Fluorlens), heparin-surface-modified (HSM), hydrogel, Memory-Lens, Poly(methyl methacrylate) (PMMA), soft acrylic, and silicone lenses; the human crystalline lens was also studied. Each lens was immersed in silicone oil for 12 hours, than photographed, studied by scanning electron microscopy (except the crystalline lens), and subjected to computer-generated image analysis to determine the silicone oil coverage. RESULTS: Silicone oil coverage of dry silicone lenses was 100% and of lenses immersed in normal saline, 82.5%. The least coverage was on the heparin-surface-modified lens (mean score 9.4%). Coverage of the other four lenses ranged from approximately 15.1% to 33.7%. Mean coverage of the human lens capsule was 10.9%. CONCLUSION: Although a silicone IOL shows maximal adherence to silicone oil, other lens biomaterials are not immune to this complication. Silicone oil coverage was related to the dispersive energy component of the surface charge of the IOL biomaterial. Low dispersive energy materials had less silicone oil coverage, while those with higher dispersive energy had more oil coverage.

Adhesiveness↗
PubMed1997

[Penetration of ciprofloxacin and cefoperazone into human pancreas].

Major pancreatic infection is responsible for more than 80% of deaths in patients with acute pancreatitis. Therefore, the role of antimicrobial drugs in the prevention and treatment of secondary parcreatic infection is very important. The choice of antimicrobial drugs must be based upon the ability of the drug to exceed the therapeutic concentration in pancreas for the common pathogens. The penetration of ciprofloxacin and cefoperazone into pancreas was investigated in ten patients who had undergone pancreatoduodenectomy. The pancreatic juice was temporarily diverted to the exterior via a panoreatic duct catheter. The pancreatic tissue was obtained intraoperatively and pancreatic juice was drained postoperatively. The antimicrobial drug concentrations were determined by high-performance liquid chromatography. The concentrations of ciprofloxacin and cefoperazone in pancreatic juice were 44% and 17%, respectively, of those in serum, and exceeded the in vitro concentration (MIC-90) for most bacteria associated with pancreatic infections. The result indicates that ciprofloxacin and cefoprazone appear to be appropriate for both prophylaxis and therapy of secondary pancreatic infections.

Adult↗
PubMed1997

[Studies on expansion ex vivo of murine bone marrow cells and its hematopoietic reconstitution capacity].

OBJECTIVE: To investigate the effects of stem cell factor (SCF) in combination with interleukin-1 (IL-1) or/and interleukin-3 (IL-3) on ex vivo expansion of 5FU treated bone marrow cells and hematopoietic recovery in lethally irradiated mice transplanted with the expanded cells. METHODS: 5FU treated bone marrow cells (d3-5FU-BMC) were cultured in a cytokines-containing medium, and the net increments of CFU-GM and high proliferative potential colony forming cell (HPP-CFC) were evaluated. RESULTS: CFU-GM increased by 33.7 +/- 18.1- or 18.1 +/- 6.3- fold, and HPP-CFC by 17.8 +/- 10.5- or 12.7 +/- 9.1- fold, respectively, in cultures containing SCF with IL- or IL-3, as compared with that in control; while SCF alone had little effect. Compared with fresh d3-5FU-BMC, transplantation of the expanded bone marrow cells accelerated the recovery of recipients' peripheral blood cell counts by 1 approximately 3 days and increased the survival rate of the transplanted animals (d3-5FU-BMC group 50% vs expansion group 8U approximately 100%). CONCLUSION: SCF in combination with IL-1 or IL-3 synergetically ex vivo expands hematopoietic cells. Transplantation of the expanded bone marrow cells accelerates the recipient's hematopoietic reconstitution.

Animals↗
PubMed1997

[Molfig, a software for displaying the structure and vibration of molecule].

A Molfig software has been developed for displaying the structure and vibrational mode of molecule in our lab. Various functions and a friendly interface are equipped in the software. The testing results showed that the software may enhance our understanding of the relationship between the vibrational behavior and the structure of molecules.

English Abstract↗
PubMed1997

[Curvefit, an overlapping bands resolving software for IR spectra].

A Curvefit software has been developed for resolving the overlapping band of the vibartional spectra in our lab . Various functions and a friendly interface are equipped in the software. The testing results showed that both the speed of the calculation and the reliability of the final results are satisfactory.

English Abstract↗
PubMed1996

Characterization of the protein product encoded by a splicing variant of the Marek's disease virus Eco-Q gene (Meq).

In the present study, we report the characterization of a 212-amino-acid polypeptide encoded by a splicing variant of the Marek's disease virus Eco-Q gene (Meq). This protein, referred to as Meq-sp, contains the N-terminal 100 amino acids of Meq, which include part of Meq's DNA binding/dimerization domain, but lacks the transactivation domain of Meq. Thus, Meq-sp was examined for its ability to bind to DNA and act as a transactivator. Results indicated that while Meq and Meq-sp could both bind to the AP-1 binding site, the 110 C-terminal amino acid residues of Meq-sp lacked the ability to function as a transactivator when fused to the GAL4 (1-147) DNA binding motif. To investigate whether Meq-sp can interact with Meq or with c-jun, protein-protein and protein-DNA interactions in vitro were examined. Results showed that Meq-sp can associate with both Meq and c-jun and bind to the AP-1 site with a higher affinity as a heterodimer with c-jun. These results suggest that Meq-sp could compete with Meq for heterodimer formation with c-jun and dimer binding to DNA and possibly act as a transdominant negative regulator of Meq activity in vivo.

Alternative Splicing↗
PubMed1996

Isolation and characterization of Marek's disease virus (MDV) cDNAs from a MDV-transformed lymphoblastoid cell line: identification of an open reading frame antisense to the MDV Eco-Q protein (Meq).

Two Marek's disease virus (MDV) cDNAs of 852 and 1168 bp, which map to the right end of the BamHI-I2 fragment of the MDV genome, were isolated from a cDNA library derived from the MDV transformed lymphoblastoid cell line MKT-1. These cDNAs hybridized to relatively abundant leftward mRNA transcripts in MKT-1 cells and cells lytically infected with MDV. The transcriptional initiation site for these transcripts was located in the adjacent BamHI-Q2 fragment, as determined by RNase protection and primer extension assays. A computer search for the presence of leftward open reading frames (ORFs) revealed two ORFs encoding 135- and 195-amino-acid polypeptides. A polyclonal antibody raised against a protein sequence in the N-terminus of the latter ORF detected a 23-kDa protein in the nuclear fraction of MDV-transformed lymphoblastoid cells. Furthermore, this ORF was antisense to part of the MDV Eco-Q protein (Meq) sequence.

Animals↗
PubMed1996

Autoimmune lpr/lpr mice deficient in CD40 ligand: spontaneous Ig class switching with dichotomy of autoantibody responses.

Fas-deficient MRL/Mp-lpr/lpr mice develop a syndrome that resembles human systemic lupus erythematosus, including production of IgG autoantibodies against small nuclear ribonucleoproteins (snRNPs), dsDNA, and self IgG (rheumatoid factor). To investigate the necessity for T-B cell contact in MRL autoimmunity, mice deficient in CD40 ligand (CD40L) were backcrossed onto this background, and Ab synthesis was assessed. In comparison to their CD40L-intact lpr/lpr counterparts, CD40L-deficient lpr/lpr mice had elevated levels of serum IgM and lower levels of IgG; however, a subset of animals had IgG2a, and to a lesser extent, IgG2b levels similar to those found in wild-type lpr/lpr mice. Levels of both isotypes in CD40L-deficient lpr/lpr mice were significantly greater than those found in nonautoimmune CD40L-deficient animals. IgG autoantibodies, including those directed against small nuclear ribonucleoproteins, also arose in CD40L-deficient lpr/lpr mice; however, they did not develop IgG rheumatoid factors or anti-dsDNA, and lacked histologic evidence of overt glomerulonephritis at age 3 mo, in contrast to CD40L-intact lpr/lpr animals. These results indicate that isotype switching occurs in lpr/lpr mice deficient in CD40L, and that production of IgG autoantibodies to ribonucleoproteins is at least partially preserved. They also suggest that different mechanisms may be responsible for eliciting autoantibody responses in lpr/lpr mice.

Animals↗
PubMed1996

Germinal center formation, immunoglobulin class switching, and autoantibody production driven by "non alpha/beta" T cells.

The production of class-switched antibodies, particularly immunoglobulin (Ig) G1 and IgE, occurs efficiently in T cell receptor (TCR) alpha-/- mice that are congenitally devoid of alpha/beta T cells. This finding runs counter to a wealth of data indicating that IgG1 and IgE synthesis are largely dependent on the collaboration between B and alpha/beta T cells. Furthermore, many of the antibodies synthesized in TCR alpha-/- mice are reactive to a similar spectrum of self-antigens as that targeted by autoantibodies characterizing human systemic lupus erythematosus (SLE). SLE, too, is most commonly regarded as an alpha/beta T cell-mediated condition. To distinguish whether the development of autoantibodies in TCR alpha-/- mice is due to an intrinsic de-regulation of B cells, or to a heretofore poorly characterized collaboration between B and "non-alpha/beta T" cells, the phenotype has been reconstituted by transfer of various populations of B and non-alpha/beta T cells including cloned gamma/delta T cells derived from TCR alpha-/- mice, to severe combined immunodeficient (SCID) mice. The results establish that the reproducible production of IgG1 (including autoantibodies) is a product of non-alpha/beta T cell help that can be provided by gamma/delta T cells. This type of B-T collaboration sustains the production of germinal centers, lymphoid follicles that ordinarily are anatomical signatures of alpha/beta T-B cell collaboration. Thus, non-alpha/beta T cell help may drive Ig synthesis and autoreactivity under various circumstances, especially in cases of alpha/beta T cell immunodeficiency.

Animals↗
PubMed1996

Influenza A virus RNA-dependent RNA polymerase cleaves influenza mRNA in vitro.

We have investigated the endonuclease activity of the influenza A virus RNA polymerase in an in vitro assay with an artificial influenza-like mRNA containing a cap structure at its 5' terminus, followed by a 10 nt beta-globin mRNA sequence, and the 5' and 3' conserved termini of a truncated nucleoprotein (NP) cRNA influenza sequence. Results showed that partially purified virion ribonucleoprotein complexes (RNPs) and micrococcal nuclease treated RNPs cleaved the artificial influenza-like mRNA substrate specifically at positions near the 5' terminus to generate capped 14 and 15 nucleotide long RNA fragments which subsequently served as primers to initiate transcription. The endonuclease activity was completely blocked by addition of cap analog and competitively inhibited by added globin mRNA. Furthermore, an in vitro reconstituted influenza RNA transcription reaction containing a truncated NP vRNA as template, micrococcal nuclease treated RNPs and globin mRNA as primer, synthesized capped and uncapped full length (+) sense products. Enzyme kinetics showed that capped RNA was made earlier in the reaction; it reached a peak at 120 min and then declined. However, uncapped cRNA synthesis appeared later and remained as the dominant product later in the reaction. The nature of these products was confirmed by ribonuclease protection assays and by primer extension.

Globins↗
PubMed1996

Apoptosis and necrosis induced with light and 5-aminolaevulinic acid-derived protoporphyrin IX.

The mode of cell death induced by photodynamic treatment (PDT) was studied in two cell lines cultured in monolayer, V79 Chinese hamster fibroblasts and WiDr human colon adenocarcinoma cells. The cells were incubated with 5-aminolaevulinic acid (5-ALA) as a precursor for the endogenously synthesised protoporphyrin IX, which was activated by light. Free DNA ends, owing to internucleosomal DNA cleavage in apoptotic cells, were stained specifically with a fluorescent dye in the terminal deoxynucleotidyl transferase (TdT) assay. The free DNA ends were measured by flow cytometry and the fractions of apoptotic cells determined. Total cell death was measured in a cell survival assay to determine the necrotic fraction after subtraction of the apoptotic fraction. V79 cells did undergo apoptosis while WiDr cells were killed only through necrosis. With time, the apoptotic fraction of V79 cells increased until a maximum was reached about 3-4 h after ALA-PDT treatment. For increasing ALA-PDT doses, a maximal apoptotic fraction 75-85% of the cells was measured at about 85% of total cell death. The flow cytometric assay of apoptosis was confirmed by the typical ladder of oligonucleosomal DNA fragments obtained from agarose gel electrophoresis, by fluorescence micrographs visualising the induced free DNA ends and by electron micrographs showing the typical morphology of apoptotic cells.

Adenocarcinoma↗
PubMed1996

Influenza A virus RNA-dependent RNA polymerase: analysis of RNA synthesis in vitro.

Influenza A virus RNA-dependent RNA polymerase, purified from virion ribonucleoprotein particles and from which endogenous genomic RNA (vRNA) has been depleted by treatment with micrococcal nuclease, was used to study transcription initiation, elongation, and termination in vitro. Templates that contained either minus- or plus-sense influenza virus nucleoprotein minigenes with conserved 5' and 3' termini and the uridylate tract were constructed. The dinucleotide ApG and alfalfa mosaic virus RNA4 (AlMV4) were used as primers. ApG primed the synthesis of full-length positive-strand or cRNA products and shorter transcripts, depending upon the molar ratio between the nucleoprotein and the vRNA template. Sequence analysis of the ends of these transcripts demonstrated that the 5' termini of both transcripts and the 3' terminus of the full-length product were complementary to the 3' and 5' termini of the vRNA template, respectively, whereas the 3' terminus of the incomplete product corresponded to a sequence located 40 bases downstream from the 5' terminus of the template and was about 20 nucleotides downstream from the uridylate tract, which is the putative signal for polyadenylation. Binding of the cap structure of AlMV4 by the polymerase activated RNA synthesis by ligation-elongation of small genomic RNA fragments which were likely derived from a genome segment protected by the polymerase from micrococcal nuclease digestion. The sequence of these fragments mapped to a region 14 to 28 nucleotides upstream of the 3' terminus of the viral genome. Polymerase subunit involvement in transcription initiation with ApG or AlMV4 was characterized by studying the effect of purified polyclonal antisubunit immunoglobulins of the G class (IgGs) in transcription assays. These results showed that anti-PB2 IgG inhibited transcription initiation in both ApG- and AlMV4-primed reactions, whereas anti-PB1 antibodies also blocked transcription initiated with AlMV4. The differences observed in product size, product sequence, and differential inhibition by antisubunit IgGs are discussed. These observations would support the notion that the influenza virus RNA-dependent RNA polymerase undergoes a conformational change after the binding of the cap structure of host cell heterogeneous nuclear RNA by PB2, which then usually leads to endonucleolytic cleavage of the capped primer 13 nucleotides downstream from the cap.

Animals↗
PubMed1995

Isolation and characterization of Marek's disease virus (MDV) cDNAs mapping to the BamHI-I2, BamHI-Q2, and BamHI-L fragments of the MDV genome from lymphoblastoid cells transformed and persistently infected with MDV.

We have isolated and sequenced two cDNAs of sizes 2674 and 677 bp from a cDNA library derived from MKT-1, a lymphoblastoid cell line transformed and latently infected with Marek's disease virus (MDV) using probes corresponding to the right-hand end of the BamHI-I2 fragment of the MDV genome. The larger cDNA clone represents an abundant transcript, which extends from the right-hand end of BamHI-I2 to the adjacent BamHI-Q2 and BamHI-L fragments of the MDV genome and contains the Meq (MDV Eco-Q) open reading frame. The smaller cDNA clone represents a spliced transcript containing the putative DNA binding domain of Meq as well as sequences in the BamHI-L region. We prepared a polyclonal antibody against part of the protein sequence of Meq and detected a 44-kDa protein in MKT-1 cells and in cells lytically infected with MDV. In addition, riboprobes corresponding to sequences specific to each cDNA as well as shared sequences between cDNAs detected a number of transcripts in cells either lytically or latently infected with MDV. Our results indicate that the Meq transcriptional unit extends to the BamHI-L fragment and that the transcripts mapping to the right-hand end of the BamHI-I2 and adjacent BamHI-Q2 and BamHI-L fragments are not preferentially expressed during latency.

Amino Acid Sequence↗
PubMed1995

Uptake, localization, and photodynamic effect of meso-tetra(hydroxyphenyl)porphine and its corresponding chlorin in normal and tumor tissues of mice bearing mammary carcinoma.

By using a chemical extraction assay and confocal laser scanning fluorescence microscopy, the kinetic patterns of uptake, elimination, and localization of meso-tetra(hydroxyphenyl)porphine (m-THPP) and its corresponding chlorin (m-THPC) in tumors and various normal tissues of female C3D2/F1 mice bearing CaD2 mammary carcinoma were studied after an i.p. injection of either 5 mg/kg body weight of m-THPP or 1 mg/kg body weight of m-THPC. Moreover, the histological and ultrastructural alterations of the tumors were evaluated after photodynamic therapy (PDT) with m-THPP or m-THPC. The PDT efficacy with m-THPP and m-THPC was also compared. Both m-THPP and m-THPC had a similar kinetic pattern of distribution in the tumors and most normal tissues examined. The concentrations of the dyes in the tissues peaked at 24-48 h after injection. The peak values of the uptake of m-THPP by the tissues were found to decrease in the following order: spleen > urinary tract > kidney > liver > lung > tumor > heart > skin > muscle > brain. However, higher concentrations of m-THPC were taken up by the tumors than by most of the normal tissues studied except for the liver, urinary tract, and skin. m-THPP was mainly localized in the stroma of the tumors, whereas m-THPC was distributed in both vascular interstitium and neoplastic cells of the tumors. Morphological studies showed that PDT with m-THPP resulted in destructive changes in the microvasculature of the tumors, whereas m-THPC-based PDT destroyed both vascular walls and tumor cells of the tumors. The m-THPP-PDT of the tumors was much less efficient than m-THPC-PDT of the tumors, although the dose of m-THPP used was five times higher than that of m-THPC. m-THPP and m-THPC have different efficiency of sensitizing tumors to photodestruction, although they are similar with respect to hydrophobicity. This is likely due to the differences in their intratumoral localization patterns and in their absorption spectra.

Animals↗
PubMed1995

The haem b558 component of the cytochrome bd quinol oxidase complex from Escherichia coli has histidine-methionine axial ligation.

The cytochrome bd ubiquinol oxidase from Escherichia coli is induced when the bacteria are cultured under microaerophilic or low-aeration conditions. This membrane-bound respiratory oxidase catalyses the two-electron oxidation of ubiquinol and the four-electron reduction of dioxygen to water. The oxidase contains three haem prosthetic groups: haem b558, haem b595 and haem d. Haem d is the oxygen binding site, and it is likely that haem d and b595 form a bimetallic site in the enzyme. Haem b558 has been previously characterized spectroscopically as being low spin and has been shown to be located within subunit I (CydA) of this two-subunit enzyme. It is likely that haem b558 is associated with the quinol oxidation site, which has also been shown to be within subunit I. In a previous effort to locate the specific amino acids axially ligated to haem b558, all six histidines within subunit I were altered by site-directed mutagenesis. Only one, histidine-186, was identified as a likely ligand to haem b558. Hence it was suggested that haem b558 could not have bis(histidine) ligation. In the current work, a combination of low-temperature near-infrared magnetic circular dichroism (NIR-MCD) and EPR spectroscopies have been employed to identify the nature of the haem b558 axial ligands. The NIR-MCD spectrum at cryogenic temperatures is dominated by the low-spin haem b558 component of the complex, and the low-energy band near 1800 nm is strong evidence for histidine-methionine ligation. It is concluded that haem b558 is ligated to histidine-186 plus one of the methionines located within subunit I of the oxidase.

Bacterial Proteins↗