Biomedical subjects

X Yang

Publications and source records attributed to X Yang.

At least 757 records · Page 42Linked to original sources

PubMed1997

Nitrogen dioxide in indoor ice skating facilities: an international survey.

An international survey of nitrogen dioxide (NO2) levels inside indoor ice skating facilities was conducted. One-week average NO2 concentrations were measured inside and outside of 332 ice rinks located in nine countries. Each rink manager also completed a questionnaire describing the building, the resurfacing machines, and their use patterns. The (arithmetic) mean NO2 level for all rinks in the study was 228 ppb, with a range of 1-2,680 ppb, based on a sample collected at breathing height and adjacent to the ice surface. The mean of the second indoor sample (collected at a spectator's area) was 221 ppb, with a range of 1-3,175 ppb. The ratio of the indoor to outdoor NO2 concentrations was above 1 for 95% of the rinks sampled, indicating the presence of an indoor NO2 source (mean indoor:outdoor ratio = 20). Estimates of short-term NO2 concentrations indicated that as many as 40% of the sampled rinks would have exceeded the World Health Organization 1-hour guideline value of 213 ppb NO2 for indoor air. Statistically significant associations were observed between NO2 levels and the type of fuel used to power the resurfacer, the absence of a catalytic converter on a resurfacer, and the use of an ice edger. There were also indications that decreased use of mechanical ventilation, increased number of resurfacing operations per day, and smaller rink volumes were associated with increased NO2 levels. In rinks where the main resurfacer was powered by propane, the NO2 concentrations were higher than in those with gasoline-powered resurfacers, while the latter had NO2 concentrations higher than in those using diesel. Rinks where the main resurfacer was electric had the lowest indoor NO2 concentrations, similar to the levels measured outdoors.

Air Pollution, Indoor↗

PubMed1997

DNA vaccination with the major outer-membrane protein gene induces acquired immunity to Chlamydia trachomatis (mouse pneumonitis) infection.

The efficacy of DNA vaccination for prevention of Chlamydia trachomatis infection was studied using the murine model of pneumonia induced by the mouse pneumonitis (MoPn) isolate of C. trachomatis. Intramuscular DNA immunization with two chlamydial genes, one that encodes the major outer-membrane protein (MOMP) and one that encodes a cytoplasmic enzyme (cytosine triphosphate [CTP] synthetase) were tested. The MOMP DNA vaccine but not the CTP synthetase DNA vaccine generated significant delayed-type hypersensitivity and serum antibodies to MoPn elementary bodies and reduced the peak growth of MoPn by >100-fold following lung challenge infection. MOMP DNA immunization suggests a new approach to vaccine development for prevention of human chlamydial infection.

Animals↗

PubMed1997

Neutralization of endogenous interferon-beta increases the efficiency of adenoviral vector-mediated gene transduction.

One reason for low transduction efficiency and, hence, the inefficiency of gene therapy using adenoviral vectors may be the natural antiviral defense mechanisms of hosts. In this study, we investigated the effects of endogenous interferon-beta (IFN-beta) on gene transduction by adenoviral vectors. Infection of murine macrophages with Ad5CMV-LacZ produced increased expression of endogenous IFN-beta. Neutralization with anti-IFN-beta antibody (but not control immunoglobulin) during infection with the vector enhanced expression of LacZ. In contrast, IFN-beta gene expression was not detected in readily transduced NIH 3T3 cells, and the transduction efficiency of NIH 3T3 cells was unaffected by the antibody. LacZ gene expression in NIH 3T3 cells was decreased when cocultured with macrophages or in the presence of exogenous IFN-beta. The addition of the anti-IFN-beta antibody reversed this inhibition. These results demonstrate that IFN-beta-mediated cellular antiviral mechanisms are a barrier to gene transduction by adenoviral vectors.

3T3 Cells↗

PubMed1997

Microsatellite instability and loss of heterozygosity in chromosomes 9 and 16 in human breast epithelial cells transformed by chemical carcinogens.

Microsatellite instability (MSI) and loss of heterozygosity (LOH) in chromosomes 9 and 16 have been reported in human breast cancers. In order to determine whether changes in these chromosomes play a role in the initiation and progression of this disease, we performed microsatellite polymorphism analyses in human breast epithelial cells (HBEC) transformed by chemical carcinogens, an in vitro system that recapitulates various stages of neoplastic transformation. In this experimental system we studied the mortal HBEC MCF-10M, immortal MCF-10F cells, derived from MCF-10M cells, and clones derived from MCF-10F cells treated with benzo[a]pyrene (B[a]P) (BP1 and BP1E) and 7,12-dimethylbenz[a]anthracene (DMBA) (D3 and D3-1). The four clones of transformed cells were injected into severe combined immunodeficient (SCID) mice. Only BP1-E cells induced the formation of tumors, designated BP1E-Tp cells. These cells originated six additional tumors, designated BP1E-Tf no. 1 through Tf no. 6. Microsatellite analyses were carried out using five markers for chromosome 9 and 20 for chromosome 16. There was no evidence of MSI or LOH in clones BP1 and BP1E when compared with the MCF-10M and MCF-10F cells, whereas BP1E-Tp cells and Bp1E-Tf no. 1-Tf no. 6 tumors exhibited MSI at loci p23 and p21, and LOH at p21-22 of chromosome 9. They also exhibited MSI and LOH at multiple loci of both the short and long arms of chromosome 16, i.e. p13.13, p13.3, p12, q12.1, q12.2, q23 and q24, to which putative tumor suppressor genes have been localized. Clones D3 and D3-1 exhibited no genomic changes in chromosome 9, but did show MSI at locus q12.1 of chromosome 16 using marker D16S285. Although the cells treated with DMBA expressed early phenotypes of neoplastic transformation, they were not tumorigenic, and also manifested fewer changes than the tumorigenic BP1E-Tp cells and the tumors BP1E-Tf. The changes in chromosomes 9 and 16 observed in these latter ones indicated an association with the expression of tumorigenesis, which represents a late event in the progression of the neoplastic transformation of HBEC. Of interest was the observation that HBEC transformed by chemical carcinogens in vitro express genomic changes similar to those found in spontaneous breast carcinomas.

Animals↗

PubMed1997

Dynamics of maturation-promoting factor and its constituent proteins during in vitro maturation of bovine oocytes.

Maturation-promoting factor (MPF) is known to be a key regulator of both mitotic and meiotic cell cycles. MPF is a complex of a B cyclin and the cyclin-dependent kinase cdkl (p34cdc2). Oocyte maturation and its arrest at metaphase of meiosis II (MII) are regulated by changes in MPF activity. In this study, experiments were conducted to examine the dynamics of MPF activity and its constituent proteins during in vitro maturation of bovine oocytes. Bovine oocytes displayed relatively low levels of MPF (histone H1 kinase) activity at the germinal vesicle stage during the first 8 h of maturation. MPF activity increased gradually thereafter, and its first peak of activity occurred at 12-14 h of maturation (presumptive metaphase I), which was followed by an abrupt reduction in activity at 16-18 h, during presumptive anaphase and telophase. MPF activity then increased, reaching a plateau at 20-24 h of maturation (MII stage). This high level of MPF activity was maintained for several hours but decreased gradually after 30 h of maturation and became barely detectable by 48 h of in vitro maturation (IVM) culture. At each time point, there was a significant variation among individual oocytes in histone H1 kinase activity, which was probably due to asynchronous maturation. Abundance of cdk1 increased gradually during the first 8 h and then remained relatively constant except for an apparent reduction at 18-22 h of IVM. The level of cyclin B2 increased quickly during the initial 2 h of culture, and this high level was maintained until 16 h, after which a significant reduction was observed between 18 and 22 h of IVM. The de novo synthesis of cyclin B2, however, exhibited a biphasic oscillation during maturation, with peaks before the onset of MI and of MII. These results have defined the profiles of MPF activity and its individual components during bovine oocyte maturation in vitro. We conclude that active MPF regulates bovine oocyte maturation and that de novo synthesis of cyclin B2 occurs during the process of maturation.

Animals↗

PubMed1997

Expression of Mos proto-oncoprotein in bovine oocytes during maturation in vitro.

The c-mos proto-oncogene product Mos is believed to be an active component of the cytostatic factor that stabilizes and sustains the activity of maturation-promoting factor. Mos has been found to be responsible for the metaphase arrest of oocytes at the second meiotic division in both Xenopus and the mouse. In this study, we have demonstrated, by Western blot and immunoprecipitation analysis, that an approximately 39-kDa protein, identified as Mos, was present in in vitro-matured (metaphase II stage) bovine oocytes but disappeared in parthenogenetically activated oocytes. The oocytes actively synthesized p39mos at the metaphase II stage (between 22 and 26 h of in vitro maturation [IVM]), whereas little p39mos synthesis was detected during the first 4 h of IVM and it was nondetectable during aging at 44-48 h of IVM, when oocytes lose the capability of normal development after fertilization. Ethanol activation of mature oocytes led to the disappearance of p39mos. beta-Tubulin, but not p34cdc2, was co-precipitated with Mos when extracts of metaphase II-stage bovine oocytes were incubated with Mos antiserum. These results demonstrated that Mos is present and actively synthesized in mature bovine oocytes and that oocytes aged beyond the optimal time for fertilization seem to lose the ability to synthesize the Mos protein. beta-Tubulin was found to be associated with Mos, which suggests a possible role for the cytoskeletal protein in maintaining the meiotic arrest in mature bovine oocytes.

Amino Acid Sequence↗

PubMed1997

Age and hormonal dependence of acquisition of oocyte competence for embryogenesis in prepubertal calves.

This research was designed to improve our understanding of oocyte maturation and acquisition of developmental competence of oocytes using prepubertal heifers as a model. Oocytes were collected by ultrasound-guided transvaginal oocyte retrieval from 20 age-matched calves at 5, 7, 9, and 11 mo of age that had or had not received gonadotropin stimulation. Numbers of oocytes recovered from unstimulated heifers decreased with age, being 15 +/- 2, 12 +/- 1, 7 +/- 1, and 7 +/- 1 (for 5-, 7-, 9-, and 11-mo-old calves, respectively). Corresponding numbers for the gonadotropin-stimulated heifers were 18 +/- 2, 16 +/- 2, 13 +/- 1, and 15 +/- 2. Cumulus-oocyte complexes were graded from A to D on the basis of their morphology, and the better-grade A and B oocytes were used for in vitro maturation and fertilization. A higher proportion of grade A and B oocytes was found for the stimulated vs. unstimulated prepubertal calves at 5 mo of age (92% vs. 49%, p < 0.05) and 7 mo of age (96% vs. 63%, p < 0.05), but the improvement of this parameter by stimulation was not significant for peri- and postpubertal calves at 9 (53% vs. 38%, p > 0.05) and 11 (53% vs. 38%, p > 0.05) mo of age. Embryo development to morula and blastocyst stages was poorer (0-11%, p < 0.05) for oocytes collected from unstimulated calves at 5-9 mo of age than for those from the age-matched but gonadotropin-stimulated groups (10%, 39%, and 31%, at 5, 7, and 9 mo of age, respectively). In calves 11 mo of age, embryo development to morula and blastocyst stages was similar with and without gonadotropin stimulation (48% vs. 40%, p > 0.05) and was comparable to that of adult cow oocytes (45%, p > 0.05). The data suggest that the acquisition of oocyte competence for normal embryo development in prepubertal calves is influenced by animal age and hormonal treatment.

Aging↗

PubMed1997

Glomerular sequence of events concerning monocyte/macrophage accumulation and ICAM-1 expression during experimental serum sickness nephritis in the rat.

Serum sickness nephritis was induced in Fischer rats by preimmunization and repeated immunization with chicken egg albumin. This experimental model is characterized by marked accumulation of monocytes/macrophages (MO) and deposition of immune complexes (IC) in glomeruli during the inflammatory stage and, thereafter, the advancement to glomerulosclerosis. The correlations between glomerular tissue damage, MO participation, intercellular adhesion molecule-1 (ICAM-1) expression and IC deposition were analyzed during the long-term disease process. The grade of ICAM-1 expression was well correlated with MO accumulation and IC deposition, and its distribution was observed on the glomerular endothelial layer, mesangium, and along the parietal epithelial layer of the Bowman's capsule. It is suggested that glomerular MO accumulation is largely affected by the ICAM-1 expression on glomeruli and, underneath such adhesion molecules, MO may play a role in subendothelial or mesangial migration, mesangial cell activation, inducing sclerosis and monocytic-epithelial crescent formation.

Animals↗

PubMed1997

Cross-spectral and temporal factors in the precedence effect: discrimination suppression of the lag sound in free-field.

In an anechoic chamber, subjects were required to discriminate a 20 degrees azimuthal change in a lag sound's position in the presence of a lead sound coming from a different direction. Delay between lead and lag sounds was adaptively varied in several conditions to track discrimination suppression thresholds. In experiment 1, lead and lag stimuli were 5-ms, 1-octave, A-weighted noise bursts (65 dB), with lead and lag parametrically set to center frequencies of 0.5, 2.0, or 3.0 kHz. Discrimination suppression thresholds were higher when lead and lag center frequencies coincided (mean: 11.3 ms) than when they did not coincide (mean: 2.9 ms). These results support the "spectral overlap hypothesis" of Blauert and Divenyi [Acustica 66, 267-274 (1988)], but not the "localization strength hypothesis" later proposed by Divenyi [J. Acoust. Soc. Am. 91, 1078-1084 (1992)]. Spectral overlap and localization strength appear to be two relatively independent factors governing discrimination suppression. It is proposed here that localization strength is weighted more when stimuli are presented via headphones and the only cue to lateral position is the interaural temporal difference, while spectral overlap is weighted more for free-field presented stimuli. In experiment 2, lead and lag stimuli were 8-ms, 1.5-kHz A-weighted tone bursts (65 dB), with lead and lag rise times parametrically set to 0, 2, or 4 ms. In this case the amount of discrimination suppression increased as lead rise time became more abrupt or as lag rise time became more gradual. These results support the localization strength hypothesis: The greater the localization strength of the lead stimulus (independently assessed by measuring its minimum audible angle in isolation), the greater suppression it exerted on discriminability of the lag sound's position. It appears that for stimuli presented in the free-field, spectral overlap is the primary factor affecting discrimination suppression, but when overlap is held constant, abruptness of stimulus onsets governs the amount of suppression.

Adult↗

PubMed1997

Autoimmunity to heat shock protein 60 and antigen-specific production of interleukin-10.

The immunopathologic sequelae of chlamydial infection are correlated with immune responses to the Chlamydia trachomatis heat shock protein 60 (hsp60). One pathogenic mechanism that may explain this association is the induction of autoimmune responses to self hsp60, since these two proteins share a high degree of amino acid sequence identity. To investigate the conditions under which autoimmune responses can be generated against self hsp60, groups of CBA mice were immunized with recombinant mouse hsp60, recombinant chlamydial hsp60, or both proteins. The data show that autoimmune responses characterized by strong T-cell proliferation and high titers of antibody to self hsp60 are induced only by concurrent immunization with mouse and chlamydial hsp60. Immunization with mouse hsp60 alone induced lymphocytes that secreted high levels of interleukin-10 (IL-10) but did not proliferate in response to in vitro stimulation with mouse hsp60; coimmunization with mouse and chlamydial hsp60s induced lymphocytes that proliferated strongly in response to mouse hsp60, secreted 6-fold less IL-10, and exhibited a 12-fold increase in the ratio of gamma interferon/IL-10 production. Switches in cytokine production patterns may mediate the pathogenesis of hsp60-associated diseases such as C. trachomatis immunopathology.

Animals↗

PubMed1997

The Drosophila bifocal gene encodes a novel protein which colocalizes with actin and is necessary for photoreceptor morphogenesis.

Photoreceptor cells of the Drosophila compound eye begin to develop specialized membrane foldings at the apical surface in midpupation. The microvillar structure ultimately forms the rhabdomere, an actin-rich light-gathering organelle with a characteristic shape and morphology. In a P-element transposition screen, we isolated mutations in a gene, bifocal (bif), which is required for the development of normal rhabdomeres. The morphological defects seen in bif mutant animals, in which the distinct contact domains established by the newly formed rhabdomeres are abnormal, first become apparent during midpupal development. The later defects seen in the mutant adult R cells are more dramatic, with the rhabdomeres enlarged, elongated, and frequently split. bif encodes a novel putative protein of 1063 amino acids which is expressed in the embryo and the larval eye imaginal disc in a pattern identical to that of F actin. During pupal development, Bif localizes to the base of the filamentous actin associated with the forming rhabdomeres along one side of the differentiating R cells. On the basis of its subcellular localization and loss-of-function phenotype, we discuss possible roles of Bif in photoreceptor morphogenesis.

Actins↗

PubMed1997

Altered metabolism and superoxide generation in neural tissue of rat embryos exposed to high glucose.

Oxygen uptake and glucose utilization of embryonic and fetal neural tissue of normal and diabetic rat pregnancy were studied. Exposure to 50 mM glucose inhibited oxygen uptake of embryonic neural tissue of normal rats by 28% at gestational day 9 (P < 0.001) and 20% at days 10-12 and 15 (P < 0.001) and stimulated glucose utilization by 132% at day 9 (P < 0.001), 50% at days 10 and 11 (P < 0.01), 168% at day 12 (P < 0.001), and 338% at day 15 (P < 0.001), indicating a Crabtree effect. The glucose-altered metabolism led to production of superoxide by the tissue, which was 1.8 to 2.4 nmol.h-1.microgram DNA-1 at days 9-12 and 1.2 nmol.h-1.microgram DNA-1 at day 15. The embryonic neural tissue of diabetic rats showed a diminished metabolic sensitivity to high glucose exposure, suggesting an impaired mitochondrial function. Consequently, the glucose-induced superoxide production was not detected in the tissue of embryos of diabetic rats. The data suggest that high concentration of glucose alters embryonic and fetal metabolism and causes generation of superoxide. Prolonged duration of the glucose-induced metabolic changes may impair cellular function and lead to embryonic dysmorphogenesis.

Animals↗

PubMed1997

The new economic policy and permanent migration in Zhejiang Province, China.

"This article examines the impact of economic reforms on the volume and characteristics of permanent migration in Zhejiang Province, China. The data suggest that the new economic policy induced a surge in permanent migration during the post-reform years. Such positive impact of the reform on permanent migration has started to fade away in 1985, as government relaxed its control over residence. Moreover, market mechanisms started playing a more important role in employment, exchange, and consumption. The data also suggest that the new economic policy has particularly favored the better educated, and thereby increased educational differentials between permanent migrants and nonmigrants. For all other characteristics examined, the results show consistently that post-reform migrants are less differentiated from nonmigrants than their pre-reform counterparts." An earlier version of this paper was presented at the 1994 Annual Meeting of the Population Association of America.

Asia↗

PubMed1997

Increased expression of Gs(alpha) enhances activation of the adenylyl cyclase signal transduction cascade.

Expression of the stimulatory G protein, G(S)alpha, can vary over a 3-fold range in human tissues and in rodent central nervous system. In fact, the offspring of alcoholics have higher levels of G(S)alpha expression in certain tissues compared with the offspring of nonalcoholics. The aim of this research was to test the hypothesis that a causal relationship exists between the level of expression of G(S)alpha and induction of the adenylyl cyclase (AC) cascade. The methodology employed transient transfection of HEK 293 cells with a cDNA for the 52-kDa form of G(S)alpha under regulation by inducible metallothionein promoters. Transfectants were exposed to varying concentrations (0-125 microM) of zinc sulfate that produced a 3-fold range of membrane G(S)alpha expression. The range of G(S)alpha expression produced was found to mimic a physiologically relevant spectrum of G(S)alpha expression in membranes derived from human tissues and rat brain. It was observed that induction of G(S)alpha expression increased constitutive as well as stimulated cAMP accumulation. Moreover, induction of G(S)alpha expression increased events distal to the accumulation of cAMP including the phosphorylation of the transcription factor, cAMP response element binding protein and transcriptional activation of cAMP-dependent reporter genes. In summary, these studies show that the amount of G(S)alpha expression has a marked impact on the level of activity of the AC cascade from the membrane through to the nucleus. It is hypothesized that individuals who differ in G(S)alpha expression may also differ in the expression of certain cAMP-dependent genes.

1-Methyl-3-isobutylxanthine↗

PubMed1997

Postmitotic cells fated to become rod photoreceptors can be respecified by CNTF treatment of the retina.

Lineage analyses of vertebrate retinae have led to the suggestions that cell fate decisions are made during or after the terminal cell division and that extrinsic factors can influence fate choices. The evidence for a role of extrinsic factors is strongest for development of rodent rod photoreceptors ('rods'). In an effort to identify molecules that may regulate rod development, a number of known factors were assayed in vitro. Ciliary neurotrophic factor (CNTF) was found to have a range of effects on retinal cells. Addition of CNTF to postnatal rat retinal explants resulted in a dramatic reduction in the number of differentiating rods. Conversly, the number of cells expressing markers of bipolar cell differentiation was increased to a level not normally seen in vivo or in vitro. In addition, a small increase in the percentage of cells expressing either a marker of amacrine cells or a marker of Muller glia was noted. It was determined that many of the cells that would normally differentiate into rods were the cells that differentiated as bipolar cells in the presence of CNTF. Prospective rod photoreceptors could make this change even when they were postmitotic, indicating that at least a subset of cells fated to be rods were not committed to this fate at the time they were born. These findings highlight the distinction between cell fate and commitment. Resistance to the effect of CNTF on rod differentiation occurred at about the time that a cell began to express opsin. The time of commitment to terminal rod differentiation may thus coincide with the initiation of opsin expression. In agreement with the hypothesis that CNTF plays a role in rod differentiation in vivo, a greater percentage of cells were observed differentiating as rod photoreceptors in mouse retinal explants lacking a functional CNTF receptor, relative to wild-type littermates.

Animals↗

PubMed1997

Murine 86-kDa heat shock protein gene and promoter.

The class of 90 kDa heat shock proteins (Hsp90) is among the most abundant heat shock proteins (Hsps) in eukaryotic cells. In vertebrates, Hsp90 is encoded by two distinct gene families giving rise to products of 84 and 86 kDa. In mice the expression of these two genes, hsp84 and hsp86, vary with respect to each other in responses to stress, and also in response to signals for growth and development. Therefore, as a step towards understanding the molecular basis for the differential regulation of these two genes, we have isolated and characterized genomic clones of the murine hsp86 gene and its 5' flanking region. The gene is composed of eleven exons interrupted by 10 introns. The 5' region contains consensus TATA, several stimulatory protein-1 binding site (SP1) elements as well as six consensus heat shock elements (HSE) 5' of the transcription start site. An 806 bp fragment of the 5' promoter region conferred constitutive expression upon a reporter gene and this expression was increased upon heat shock.

Animals↗

PubMed1997

Echo suppression and discrimination suppression aspects of the precedence effect.

The precedence effect is a phenomenon that may occur when a sound from one direction (the lead) is followed within a few milliseconds by the same or a similar sound from another direction (the lag, or the echo). Typically, the lag sound is not heard as a separate event, and changes in the lag sound's direction cannot be discriminated. The hypothesis is proposed in this study that these two aspects of precedence (echo suppression and discrimination suppression) are at least partially independent phenomena. Two experiments were conducted in which pairs of noise bursts were presented to subjects from two loudspeakers in the horizontal plane to simulate a lead sound and a lag sound (the echo). Echo suppression threshold was measured as the minimum echo delay at which subjects reported hearing two sounds rather than one sound; discrimination suppression threshold was measured as the minimum echo delay at which subjects could reliably discriminate between two positions of the echo. In Experiment 1, it was found that echo suppression threshold was the same as discrimination suppression threshold when measured with a single burst pair (average 5.4 msec). However, when measured after presentation of a train of burst pairs (a condition that may produce "buildup of suppression"), discrimination suppression threshold increased to 10.4 msec, while echo suppression threshold increased to 26.4 msec. The greater buildup of echo suppression than of discrimination suppression indicates that the two phenomena are distinct under buildup conditions and may be the reflection of different underlying mechanisms. Experiment 2 investigated the effect of the directional properties of the lead and lag sounds on discrimination suppression and echo suppression. There was no consistent effect of the spatial separation between lead and lag sources on discrimination suppression or echo suppression, nor was there any consistent difference between the two types of thresholds (overall average threshold was 5.9 msec). The negative result in Experiment 2 may have been due to the measurements being obtained only for single-stimulus conditions and not for buildup conditions that may involve more central processing by the auditory system.

Adult↗

PubMed1997

[Culture and characterization of rat lung microvascular endothelial cells].

Cultures of rat lung microvascular endothelial cells (RLMECs) were obtained from perepheral lung tissue. The lung tissue was cut into small pieces and cultured with RPMI-1640 containing 20% bovine calf serum, 90 micrograms/ml heparin, 4 mmol/L L-glutamine, 100 mu/ml penicillin and 100 micrograms/ml streptomycin. Erythrocytes and leukocytes left the tissue first, followed by RLMECs. Fibroblasts and other cells grew after 72 hours of culture. After 60 hours of culture, the lung tissue was discarded. RLMECs in flask showed regular cobblestone morphology and positive for binding of the lectin Bandeiraea simplicifolia I and indirect immunofluoresence staining with factor VIII antiserum.

Animals↗