Role of CD44 in the reaction of vascular smooth muscle cells to arterial wall injury.
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Animals↗
Biomedical subjects
Q He
Publications and source records attributed to Q He.
Immunologic basis of transplant-associated arteriosclerosis.
Although immunosuppressive therapy minimizes the risk of graft failure due to acute rejection, transplant-associated arteriosclerosis of the coronary arteries remains a significant obstacle to the long-term survival of heart transplant recipients. The participation of specific inflammatory cell types in the genesis of this lesion was examined in a mouse model in which carotid arteries were transplanted across multiple histocompatibility barriers into seven mutant strains with immunologic defects. An acquired immune response--with the participation of CD4+ (helper) T cells, humoral antibody, and macrophages--was essential to the development of the concentric neointimal proliferation and luminal narrowing characteristic of transplant arteriosclerosis. CD8+ (cytotoxic) T cells and natural killer cells were not involved in the process. Arteries allografted into mice deficient in both T-cell receptors and humoral antibody showed almost no neointimal proliferation, whereas those grafted into mice deficient only in helper T cells, humoral antibody, or macrophages developed small neointimas. These small neointimas and the large neointimas of arteries grafted into control animals contained a similar number of inflammatory cells; however, smooth muscle cell number and collagen deposition were diminished in the small neointimas. Also, the degree of inflammatory reaction in the adventitia did not correlate with the size of the neointima. Thus, the reduction in neointimal size in arteries allografted into mice deficient in helper T cells, humoral antibody, or macrophages may be accounted for by a decrease in smooth muscle cell migration or proliferation.
Animals↗
Existence of phosphorylated and dephosphorylated forms of cytosolic thymidine kinase (TK1).
In this study we examine whether different TK1 variants of pI 6.9 and 8.3 found by isoelectric focusing gel electrophoresis (IFE) reflect just a phenotype difference due to phosphorylation modifications or have a real phenotypic background. The phosphorylation degree of purified TK1 variants was analyzed by determining the changes in the pI values after treatment with alkaline phosphatase, using IFE. The genetic origin of the two TK1 variants was studied by determining their mol wt. by means of SDS-gelelectrophoresis. Furthermore, the subcellular distribution of the two TK1 variants was also studied. Alkaline phosphatase treatment changed the pI value of purified TK1 from 6.9 to 8.3. No change in the pI value was found when purified TK1 corresponding to pI 8.3 was treated in the same way. Similar results were obtained when treated a cytosolic fraction with alkaline phosphatase. Antibody raised against the C-terminal part of human TK1 only recognized the dephosphorylated TK1 variant corresponding to pI 8.3. There was no difference in the molecular weight between the two TK1 variants. Thus, we concluded that the TK1 variants corresponding to pI 6.9 and 8.3 are of the same genetic origin, but consist of phosphorylated and dephosphorylated forms.
Animals↗
Role of CD44 in the reaction of vascular smooth muscle cells to arterial wall injury.
CD44, the principal receptor for hyaluronic acid, is a widely distributed cell surface proteoglycan involved in cellular activation, proliferation, and migration. These processes are also central to the vascular smooth muscle cell's response to arterial wall injury. We evaluated the expression of CD44 and its isoform, CD44-V6, on vascular smooth muscle cells in vitro and in vivo and assessed the role of CD44 in DNA synthesis. Cultured vascular smooth muscle cells expressed CD44 and CD44-V6 at levels equal to or higher than those of the beta 1 and beta 2 integrins. In a rat carotid artery balloon injury model, CD44 and CD44-V6 mRNAs were unregulated in vascular smooth muscle cells after injury, and CD44 protein expression was greatest at the luminal edge of the growing neointima. CD44-expressing smooth muscle cells proliferated actively, and hyaluronic acid expression increased after injury in a temporal pattern similar to that of CD44. Through binding to hyaluronic acid, CD44 augmented DNA synthesis in cultured human and rat smooth muscle cells by 48 +/- 7.8 and 100 +/- 12.5%, respectively, an effect inhibited by an anti-CD44 antibody that blocked hyaluronate binding. These observations support a role for CD44 in the reaction of vascular smooth muscle cells to arterial wall injury.
Animals↗
Open-angle glaucoma and blood groups. The Barbados Eye Study.
OBJECTIVE: To evaluate the association of open-angle glaucoma (OAG) with ABO, Rh and Duffy blood groups in the population-based Barbados Eye Study. DESIGN: Case-control study. SETTING AND PARTICIPANTS: A subset of black Barbados Eye Study participants, which included 199 OAG cases and 1063 controls. DATA COLLECTION: ABO, Rh and Duffy blood groups were determined as part of a comprehensive study visit, which included assessment for OAG through perimetry, fundus photography, and ophthalmologic examination. OUTCOME MEASURES: Comparison of blood groups between OAG cases and nonOAG controls, expressed as odds ratio and 95% confidence intervals. RESULTS: Associations were found with the Duffy Fya+ group, which is more frequent in white than black populations. In Mantel-Haenszel analyses, OAG was positively associated with Duffy Fya+ in men (odds ratio, 2.67; confidence interval, 1.52 to 4.69) and in persons with intraocular pressure more than 21 mm Hg (odds ratio, 3.32; confidence interval, 1.49 to 7.38). Logistic regression analyses confirmed these findings (interaction of Duffy Fya+ and male gender, P = .01; interaction of Duffy Fya+ and intraocular pressure, P = .04). No associations between OAG and the ABO or Rh blood groups were seen. CONCLUSIONS: The associations with Duffy Fya+, which had not been reported previously in a black population, support the involvement of genetic factors in OAG. However, the lack of association between OAG and blood group markers of African ancestry is inconsistent with a genetic explanation for the differences in OAG prevalence between blacks and whites. Our findings suggest gene-environment interactions in OAG, to be explored by further studies of OAG and Fy markers by racial group and gender.
ABO Blood-Group System↗
Suspension culture of differentiated rat heart myocytes on non-adhesive surfaces.
Cardiac myocytes isolated from adult rat ventricles have been maintained in a stable, differentiated state for prolonged periods by the use of suspension culture on hydrophobic tissue culture inserts or agarose-coated plates. The success of this procedure depends on the use of low-serum media to prevent myocyte-myocyte interaction and proliferation of any residual endothelial cells. Myocytes cultured in this manner retain many of their structural characteristics, suggesting that maintenance of their elongated irregular shape is not dependent on interaction with extracellular matrix. They also exclude trypan blue, can be vitally stained by the uptake and reduction of the tetrazolium dye MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide], synthesize myosin and when returned to adhesive surfaces are capable of attachment and attendant dedifferentiation. Stability of the myocytes in suspension permits their use in co-culture experiments; specifically, myocytes separated from endothelial cells by the hydrophobic membrane of the tissue culture insert stimulated proliferation of the latter cells, suggesting this to be a useful system for studying myocyte-endothelial cell interaction.
Animals↗
Proton detection of choline and lactate in EMT6 tumors by spin-echo-enhanced selective multiple-quantum-coherence transfer.
An extension of the Sel-MQC pulse sequence--SEE-SelMQC (spin-echo-enhanced selective multiple-quantum coherence transfer)--that completely suppresses lipid and water in tissues containing mobile lipid in a single scan and detects 1H resonances of multiple metabolites is described. As in the Sel-MQC lactate-editing experiments [Q. He et al., J Magn. Reson. B 106,203 (1995)], SEE-SelMQC acquires lactate from its ZQ --> DQ coherence-transfer pathway; in addition, the method recovers signal from other metabolites by selective generation of additional spin echoes using extra frequency-selective pulses and gradients. This method introduces no loss of lactate signal intensity beyond the 50% that is lost through the multiple-quantum coherence-transfer process. The spatial distributions of choline and lactate with a phantom and in vivo, in subcutaneously implanted murine EMT6 tumors, have been simultaneously mapped.
Animals↗
Protective role of immunoglobulin G antibodies to filamentous hemagglutinin and pertactin of Bordetella pertussis in Bordetella parapertussis infection.
An outbreak of parapertussis was studied prospectively in 38 first and second grade pupils of an elementary school. Eleven (29%) children were confirmed to be culture positive for Bordetella parapertussis. Serum samples were collected from 31 children for assay of antibodies to filamentous hemagglutinin (FHA), pertactin (PRN), and pertussis toxin of Bordetella pertussis. At the first sampling, ten children were found to have a cough and 21 were asymptomatic. Of the latter, 12 remained asymptomatic and eight developed cough within 11 to 53 days (mean +/- standard deviation, 31 +/- 12 days) after sampling. One child was identified as culture positive for Bordetella pertussis and, thus, not included in the analysis of Bordetella parapertussis infection. The mean levels of IgC antibodies to FHA and PRN were significantly higher in the 12 asymptomatic children than in the eight children who later developed cough or in 20 healthy control children of the same age (for FHA, p = 0.009 and < 0.001, respectively; for PRN, p = 0.002 and 0.002, respectively). These preliminary data suggest that Bordetella parapertussis infection is more prevalent than documented, and that children with high levels of IgG antibodies to FHA and PRN can remain asymptomatic.
Antibodies, Bacterial↗
Incidence and progression of nuclear opacities in the Longitudinal Study of Cataract.
PURPOSE: To estimate incidence and progression rates of nuclear opacities in the Longitudinal Study of Cataract, an epidemiologic study of the natural history of all types of lens opacities. METHODS: The Lens Opacities Classification System III was used to assess longitudinal changes between baseline and follow-up lens photographs for the 764 Longitudinal Study of Cataract participants. Baseline data, collected until December 1988 as part of a case-control study, included color slit, retroillumination, and Scheimpflug photographs. The same data were collected by the longitudinal Study of Cataract at four subsequent visits at yearly intervals. RESULTS: Among patients free of nuclear opacities at baseline, the incidence of new opacities was 6% after 2 years and 8% after 5 years of follow-up. The progression of pre-existing nuclear opacities was much higher. After 2 years, nuclear opacities had progressed in more than one third of the patients with pre-existing opacities; after 5 years, almost half had progressed. Older age was significantly related to higher incidence of new nuclear opacities, but not to progression of pre-existing opacities. Patients with other opacity types had higher nuclear incidence and progression rates. CONCLUSIONS: In this clinic-based, older-patient population, new nuclear opacities developed in less than one tenth of the patients after 5 years of follow-up. In contrast, almost one half of the patients with pre-existing opacities had worsened after 5 years. These estimated rates can be used to plan intervention or other studies of nuclear changes in similar populations.
Aged↗
Impact of polymerase chain reaction on clinical pertussis research: Finnish and Swiss experiences.
Since April 1993 in Finland and March 1994 in Switzerland, polymerase chain reaction (PCR) has been used routinely nationwide for the diagnosis of pertussis. Nasopharyngeal specimens from 3794 patients suspected of having pertussis and 1125 controls were tested. Finnish and Swiss assays found 23% and 36% of clinical specimens positive, respectively. PCR showed a higher incidence of pertussis infection among 1- to 6-year-old children in Switzerland than in Finland (P < .001). This difference may be due to the booster dose of vaccine given at 2 years of age in Finland but not in Switzerland. In Finland, PCR-confirmed asymptomatic cases were more common among children <7 years old than in older children (P < .001), whereas older children tended to have symptomatic infection. The use of PCR markedly improves the diagnosis of pertussis and opens new perspectives for epidemiologic and vaccine efficacy studies.
Adolescent↗
Detection of Bordetella pertussis by polymerase chain reaction and culture in the nasopharynx of erythromycin-treated infants with pertussis.
BACKGROUND: Pertussis is a highly contagious respiratory disease and the most serious effects occur in young infants. Recently it has been shown that rapid and highly specific PCR can be a useful diagnostic tool for detection of pertussis infection. To our knowledge there are no previous studies concerning the disappearance of Bordetella pertussis DNA from the nasopharynx during antimicrobial treatment. METHODS: We studied prospectively how rapidly live B. pertussis organisms and DNA of these bacteria disappear from the nasopharynx during erythromycin therapy in unvaccinated infants. Eighty-five nasopharyngeal swabs obtained from nine erythromycin-treated infants with pertussis on consecutive days during hospitalization were tested by PCR and culture. The PCR products were further analyzed by Southern hybridization. RESULTS: On the fourth day of treatment 56% of the samples were positive by culture and 89% by PCR, whereas after 7 days the rates were 0 and 56%, respectively. In seven of nine patients PCR remained positive for 1 to 7 days longer than culture. The follow-up study also showed the semiquantitative nature of the PCR assay. The intensity of the PCR products in agarose gel usually weakened with time during erythromycin therapy. CONCLUSIONS: The results of this study show that PCR assay can achieve the specific diagnosis of pertussis infection in a large proportion of infants even when antimicrobial treatment has killed the organisms and culture is no longer positive.
Bacteriological Techniques↗
Effects of nifedipine, metronidazole, and nitric oxide donors on spore germination and cell culture infection of the microsporidia Encephalitozoon hellem and Encephalitozoon intestinalis.
Two species of microsporidia, Encephalitozoon hellem and Encephalitozoon intestinalis, were isolated from AIDS patients and cultured in green monkey kidney cells. A spore germination assay and a cultured-cell infection assay were used to test the efficacy of candidate antiparasitic agents. The calcium channel blocker nifedipine, metronidazole, and two nitric oxide (NO) donors, S-nitroso-N-acetylpenicillamine and sodium nitroprusside, were tested in the two assays. Nifedipine (10(-8) M) significantly inhibited E. hellem spore germination in three of four germination media. Metronidazole (10(-5) M) inhibited germination weakly and significantly inhibited E. intestinalis germination in a single germination medium. The inhibitory effect of nifedipine and metronidazole used together was greater than the sum of the effects of the drugs used alone in all E. hellem germination assays. The NO donors also inhibited spore germination. The inhibitory effect of nifedipine and metronidazole could be reversed by washing the spores, while that of the NO donors was not reversible. In early cultured-cell infections, both nifedipine (10(-8) M) and metronidazole (10(-5) M) significantly reduced the number of cells being infected. As the infection spread, these agents were less effective. Some inhibition of the spread of the infection was also demonstrated with the NO donors at a concentration (10(-5) M) not obviously toxic to the cultured cells. These data suggest that combination drug therapy targeting spore germination and intracellular parasite development is promising.
Acquired Immunodeficiency Syndrome↗
High-frequency oscillations in membrane potentials of medullary inspiratory and expiratory neurons (including laryngeal motoneurons).
1. In midcollicular decerebrate, unanesthetized, paralyzed cats ventilated with a cycle-triggered pump system, the properties of high-frequency oscillations (HFOs, 50-100 Hz) in membrane potentials (MPs) of medullary inspiratory (I) and expiratory (E) cells were studied. Simultaneous recordings were taken from bilateral phrenic and recurrent laryngeal (RL) nerves and from cells in the intermediate ventral respiratory group (intVRG, 0-1 mm rostral to the obex) or the caudal ventral respiratory group (cVRG, 2-4 mm caudal to the obex). 2. Spectral coherence analyses were used to detect the presence of HFOs during I in I and E cell MPs. Cross-correlation histograms (CCHs) between the cell and phrenic signals were used to ascertain cell-nerve HFO phase relations and to identify cells as RL motoneurons. Of the 103 cells that had significant HFOs (cell-phrenic coherences > or = 0.1), measurable HFO peak lags in the CCH were seen in 53 cells: 1) RL cells (9 I cells and 7 E cells); and 2) other types of cell (8 intVRG I cells, 18 intVRG E cells, and 11 cVRG E cells). These cells had high HFO correlations; the cell-phrenic coherence range was 0.35-0.94, with a mean HFO frequency of 58 Hz. 3. The cell-phrenic HFO lag (in ms) was measured in the CCH as the lag of the primary peak (peak located nearest to 0 lag). The phase lag was defined as (lag of primary peak in ms)/(HFO period in ms). The phase lags differed markedly between two subsets of cells: 1) RL I cells had HFO depolarization peaks that lagged the phrenic HFO peaks (average cell-phrenic phase lag = -0.18); and 2) the non-RL cells, regardless of location (intVRG or cVRG) and type (I or E), had HFO depolarization peaks leading (preceding) the phrenic HFO peaks (average cell-phrenic phase lag = 0.28). In addition, the cVRG E cells had significantly shorter cell-phrenic phase lags than the intVRG E cells (0.23 vs. 0.31, respectively). 4. These lags can be compared with the (I unit)-phrenic phase lags (average approximately 0.3) found in earlier extracellular studies. 1) There is a transmission delay of about one half HFO cycle from excitatory I cells to RL I cells. 2) Because a depolarization peak in the MP of an E cell corresponds to the start of a hyperpolarizing wave, the excitatory bulbospinal pathways from I cells have transmission times comparable with those of the inhibitory intramedullary pathways from I cells to E cells. 5. These results indicate that study of HFO phase relations can furnish useful information on functional connectivity of medullary respiratory neurons during the I phase.
Animals↗
Analyses of reported family history of glaucoma: a preliminary investigation. The Barbados Eye Study Group.
This study investigated the self-reported family history of open-angle glaucoma (OAG) among 4,314 black participants in the Barbados Eye Study (BES), which was based on a random sample of Barbados-born citizens between 40 and 84 years of age. Data collection included Humphrey perimetry, fundus photography, various ophthalmic and other measurements and a comprehensive interview, including family history. Results showed that participants with OAG and previous OAG treatment reported more family history; maternal history was reported twice as often as paternal history. In persons without previous OAG treatment, those with newly diagnosed OAG reported more sibling history (Odds Ratio = 4.5). The Statistical Analysis for Genetic Epidemiology (S.A.G.E.) system was used to test the transmission models for OAG in a subset of 1,048 families (5,806 individuals) with the most complete self-reported family information. The S.A.G.E. results are consistent with the existence of a major dominant allele for OAG. These results should be viewed as promising, but preliminary, since they are based on self-reported data. More definitive information is currently being collected by the Barbados Family Study of Open-angle Glaucoma.
Adult↗
[Experimental study on "three-dimensional" culture of scar-derived fibroblasts].
Based on the monolayer cell culture system, we resuspended the scar-derived fibroblasts in a solution of polymerizing collagen to develop a "three-dimensional" cell culture system. In this system, cells, extracellular matrices and various modulating factors were associated to form an organic entity, in which the fibroblasts express their biologic characteristics similar to that in living body. With the three-dimensional system, we could study not only the morphology, proliferation and metabolism of fibroblasts, but also their cytophysiology, such as the role on connective tissue contraction. We recommend this culture system as a better model in the research of wound healing and scar formation.
Adolescent↗
[Experimental study on antibacterial activity of subdermal vascular network island skin flap].
A comparison study was carried out on the antibacterial activity between two kinds of pig buttock flaps: the conventional island skin flap and the subdermal vascular network island skin flap which was created by thinning the distal half of a conventional island flap, preserving its subdermal vascular network. Antibacterial activity was evaluated by bacterial counts, phagocytic index and intracellular bacterial killing ratio of leukocytes, skin temperature, laser Doppler, ink perfusion, microangiography and transparent specimen technique. The results indicated that antibacterial activity of the subdermal vascular network island skin flap was much lower than that of the conventional island skin flap. The reduced blood supply and decreased leukocyte function were responsible for the low antibacterial activity after the conventional island flap was thinned.
Animals↗
Secretory expression of a single-chain insulin precursor in yeast and its conversion into human insulin.
A synthetic single-chain porcine insulin precursor (PIP) gene and an alpha-mating factor leader sequence (alpha MFL) gene obtained by the PCR method are inserted between the promoter and 3'-terminating sequence of the alcohol dehydrogenase gene ADH1 in plasmid pVT102-U to form plasmid pVT102-U/alpha MFL-PIP. The single-chain insulin precursor is expressed and secreted to the culture medium by Saccharomyces cerevisiae transformed by pVT102-U/alpha MFL-PIP. The precursor is purified and converted into human insulin by tryptic transpeptidation. The purified human insulin is fully active and can be crystallized. The overall yield of human insulin is 25 mg per liter of culture medium.
Base Sequence↗
Characterization of a peptide antibody against a C-terminal part of human and mouse cytosolic thymidine kinase, which is a marker for cell proliferation.
An affinity-purified anti-TK1 antibody (pAb1) raised against a synthetic peptide (amino acids K211PGEAVAARKLFAPQ225) corresponding to part of the C-terminus of human cytosolic thymidine kinase (TK1) was produced and characterized by enzyme-linked immunosorbent assay, Western immunoblotting and immunoprecipitation as well as by immunostaining of intact cells. pAb1 recognized a single 25 kDa TK1 polypeptide in extracts of human and rodent cells. The protein was localized to the cytoplasm, as studied by immunohistochemistry and there was no staining in G1/G0 cells or mutant cells lacking TK1 activity, while it was high in S-phase and G2 cells. When series of peptides were tested for antibody binding in which alanine was replacing each of the other amino acids one by one, lysines 211 and 220, proline 212 and glutamic acid 214 were found to be important for antibody reactivity. These results indicate that amino acids 211-214, which may form a turn region, constitute a major recognition site for pAb1, and this structure may also be involved in the cell cycle-dependent modification of TK1, pAb1 is a very useful tool for studies of the cell cycle regulation of TK1, and it may be used to identify and quantify rapidly proliferating cells such as tumor cells.
Amino Acid Sequence↗