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A F Ofulue

Publications and source records attributed to A F Ofulue.

18 recordsLinked to original sources
PubMed1999

Effects of depletion of neutrophils or macrophages on development of cigarette smoke-induced emphysema.

The aim of this study was to ascertain the putative roles of neutrophils or macrophages in the pathogenesis of cigarette smoking-induced emphysema on the basis of effects of anti-neutrophil (anti-PMN) antibody or anti-monocyte/macrophage (anti-MoMac) antibody on the development of emphysema in cigarette smoke-exposed rats. Rats were treated with rabbit anti-PMN or anti-MoMac antibody and exposed 7 days/wk for 2 mo to cigarette smoke inhalation; rats treated with nonimmunized rabbit IgG (control antibody) and exposed to cigarette smoke or normal room air served as controls. Antibody treatments began 24 h before the start of smoke or air exposure and was continued with 1 treatment/wk. Total and differential cell counts in bronchoalveolar lavage fluid and collagenase-dissociated lung and determinations of the elastinolytic activity of lung neutrophils or macrophages in [(3)H]elastin-coated wells indicated specific suppression of neutrophil accumulation and neutrophil-related elastinolytic burden in the lungs of the anti-PMN antibody-treated smoke-exposed rats, in contrast to specific suppression of macrophage accumulation and macrophage-related elastinolytic burden in the lungs of the anti-MoMac antibody-treated smoke-exposed rats. Cigarette smoke exposure-induced lung elastin breakdown (quantitated by immunologic assay of levels of elastin-derived peptides and desmosine in lavage fluid) and emphysema in the lungs (based on morphometric analysis of alveolar mean linear intercepts and alveolar tissue density in fixed lungs) were not prevented in the lungs of anti-PMN antibody-treated smoke-exposed rats but was clearly prevented in lungs of the anti-MoMac antibody-treated smoke-exposed rats. These findings implicate macrophages rather than neutrophils as the critical pathogenic factor in cigarette smoke-induced emphysema.

Animals↗
PubMed1998

Time course of neutrophil and macrophage elastinolytic activities in cigarette smoke-induced emphysema.

The aim of this study was to compare the time course of neutrophil and macrophage elastinolytic potentials in the lungs of rats exposed daily to cigarette smoke inhalation for 1-6 mo in relation to the onset and progression of cigarette smoke-induced emphysema. Normal room air-exposed rats served as controls. Morphometric data of lung histological sections showed evidence of emphysema lesions in the lungs of smoke-exposed rats at month 2 and continuing to month 6. Data of total and differential cell counts in bronchoalveolar lavage fluid and collagenase-dissociated lung showed an increased number of lung neutrophils at month 1 of smoke exposure, but this was reduced to control levels at months 2-6. In contrast, an increased number of lung macrophages was evident in the smoke-exposed rats at month 2 of exposure and continued to month 6. Data of the elastinolytic activities of the neutrophils and macrophages, determined in [3H]elastin-coated culture wells, showed that the elastinolytic activity of lung neutrophils in the smoke-exposed rats was similar to that of the control air-exposed rats at months 1-6 of exposure. In contrast, the elastinolytic activity of lung macrophages in the smoke-exposed rats was increased at month 2 of exposure and remained increased at month 6. Excessive in vivo lung elastin breakdown (judged by increased levels of elastin-derived peptides and desmosine in lavage fluid, determined immunologically) was observed in the smoke-exposed rats at months 2-6 of exposure. These data indicate that the time course of increased macrophage-directed elastinolytic activity in the lung, not that of neutrophils, is more closely associated with the evolution of cigarette smoke-induced emphysema.

Animals↗
PubMed1998

Relationship of alveolar macrophage plasminogen activator and elastase activities to lung function and CT evidence of emphysema.

OBJECTIVE: To evaluate the relationship between alveolar macrophage (AM) elastase and plasminogen activator (PA) activities (considered to be potential pathogenetic factors in emphysema) and the development of emphysema in smokers. PARTICIPANTS: Thirty-four healthy smokers >35 years of age (mean+/-SD, 46+/-7 years), with a mean+/-SD of 33+/-10 pack-years of smoking, who were recruited as volunteers. METHODS: Subjects had lung function testing and BAL to obtain AMs; limited high-resolution CT scans of the chest were obtained in 32 subjects to assess the presence of emphysema. Macrophage PA and elastase were determined using AM cultured on (131)I-fibrin-coated plates and 3H-elastin-coated plates, respectively. RESULTS: The number of AMs recovered per milliliter of BAL was significantly greater in the 16 subjects with CT evidence of mild emphysema than the 16 subjects without evidence of emphysema (669+/-301 x 10(3)/mL vs 414+/-268x 10(3)/mL; p=0.01). There was no significant difference between AM elastase or PA activities in the 16 subjects with CT evidence of mild emphysema, when compared with the 16 subjects who had no CT evidence of emphysema (elastase, 2.72+/-1.35 microg vs 2.49+/-0.91 microg elastin per 10(6) AMs per first 24 h; PA, 0.375+/-0.126 vs 0.344+/-0.096 urokinase units/10(6) AMs). There was no significant correlation between levels of PA or elastase activities and FEV1, FEV1/FVC, forced expiratory flow rate between 25% and 75% of the FVC; PA activity but not elastase activity had a significant negative correlation (r=-0.47, p<0.01) with diffusion of carbon monoxide (DCO). The macrophage count in BAL had a significant negative correlation with DCO percent predicted (r=-0.61, p<0.001). CONCLUSIONS: The findings suggest that the number of AMs recovered per milliliter of BAL (presumably indicating the number in the alveolar spaces) is related to the development of emphysema in smokers as indicated by CT scan of the chest and DCO. The results also suggest that the level of PA enzyme activity in AMs may be a pathogenetic factor in the decrease in DCO in smokers.

Adult↗
PubMed1995

Effects of streptozotocin-induced diabetes on postpneumonectomy lung growth and connective tissue levels.

Diabetes is generally accompanied by abnormal levels of growth hormone and adrenal steroids, hormones known to modulate postpneumonectomy (post-PNX) compensatory lung growth. Thus, we examined the possibility that diabetes may affect post-PNX lung growth processes. Left PNX was performed in young diabetic rats (streptozotocin-induced; 75 mg/kg body weight) (DM-PNX) and in control rats (C-PNX), for comparison with sham-operated control rats (C-SHAM). The rats were permitted free access to food and water. Examination at day 7 after surgery showed that right lung absolute dry weight and absolute DNA, collagen and elastin contents were increased in C-PNX and DM-PNX rats (but only C-PNX values reached those of both lungs in C-SHAM rats). Body weights (BW) of DM-PNX rats were lower than those of C-PNX and C-SHAM rats. Lung DNA/BW in C-PNX and DM-PNX rats were comparable, and matched values for both lungs in C-SHAM rats. Lung dry weight/BW, collagen/BW, and elastin/BW in DM-PNX rats exceeded values in C-PNX rats and even more for values of both lungs in C-SHAM rats. Data of another experiment, comparing DM-PNX rats with body weight-matched (by food limitation) PNX and control rats (WMC-PNX and WMC-SHAM rats, respectively) indicated comparable lung absolute DNA contents in DM-PNX and WMC-PNX rats (which matched values for both lungs in WMC-SHAM rats), however, lung absolute collagen and elastic contents were greater in the DM-PNX rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Assessment of the antielastase activities in bronchoalveolar lavage fluid: effect of assay buffer ionic strength.

OBJECTIVE: To evaluate the effect of assay buffer ionic strength on assessment of the antielastase activities in bronchoalveolar lavage fluid. METHOD: An improved assay protocol in which elastase (in Tris-HCI buffer) is added to increasing volumes of test samples (made up to equal volume with phosphate-buffered saline) was used. RESULTS: The percent NE activity inhibited by BALF decreased with increasing NaCl concentration of the buffer. Inhibition of pancreatic elastase (PE) was not affected. One hundred percent inhibition of NE by pure AAT and SLPI standards occurred at molar ratios of 0.91 +/- 0.03 for AAT-to-NE and 0.83 +/- 0.02 for SLPI-to-NE when assayed in buffer with < or = 0.15 mol NaCl/L, compared to ratios of 0.99 +/- 0.02 and 1.06 +/- 0.02, respectively, for assays in buffers with 0.50-1.00 mol NaCl/L (p < 0.05 for AAT-to-NE; p < 0.02 for SLPI-to-NE). The AAT-to-PE molar ratio at 100% inhibition of PE was not affected. Assays in buffer with < or = 0.15 mol NaCl/L indicated that 86.9 +/- 4.1% of AAT and 100.9 +/- 4.9% of SLPI in BALF were active against NE, while assays in buffer with 0.50-1.00 mol NaCl/L showed that 84.4 +/- 3.5% of AAT and 81.6 +/- 5.9% of SLPI present were active. AAT inhibited NE and PE equally only in buffer with 0.50-1.00 mol NaCl/L. CONCLUSIONS: The results of assays of BALF antielastase activities depend on the assay buffer NaCl concentration, which may account for the conflicting reports in the literature. The buffer with 0.50-1.00 mol NaCl/L appear to be optimal for valid quantitation of anti-NE activities in BALF.

Adsorption↗
PubMed1994

Effect of assay conditions on measurement of elastolytic activity of alveolar macrophages in culture and characterization with proteinase inhibitors.

We have investigated whether varied assay conditions account for the conflicting reports on measured elastolytic activity of alveolar macrophages (AM) cultured in direct contact with the 3H-elastin substrate coated onto 16-mm wells in serum-containing media. The data indicate that measured elastolytic activity in this assay system was dependent on the amount of 3H-elastin/culture well. 3H-elastin > 350 micrograms/well, in contrast to the < or = 200 micrograms/well commonly used in this assay system, resulted in optimal measurement of elastolytic activity that was linear with respect to culture time (up to 72 h examined) and was directly proportional to number of AM/well (up to 1.0 x 10(6) examined). The sensitivity of measured elastolytic activity to tissue inhibitor of metalloproteinases (TIMP) and to Z-phe-phe (a specific cysteine proteinase inhibitor) was not affected by amount of 3H-elastin/well, but appears to be dependent on the time period of AM culture. TIMP (at 5 microM, maximal dose examined) inhibited the measured elastolytic activity by 25% in 24-h cultures compared to 69% in 72-h cultures; Z-phe-phe (at 10 microM, dose at which maximal effect was obtained) inhibited the elastolytic activity by 45% in the 24-h cultures compared to 34% in the 72-h cultures. These findings indicate that differences in substrate levels and in culture time have a significant effect on the results obtained in measurement of AM-mediated elastolytic activity in culture, which may account for the conflicting reports in the literature. Thus standard optimal assay condition are required for valid interpretation of results of AM-mediated elastolytic activity measurements.

Biological Assay↗
PubMed1993

Role of calmodulin as an endogenous initiatory factor in compensatory lung growth after pneumonectomy.

Compensatory lung growth after pneumonectomy is a well established phenomenon in young humans and experimental animals. To date, the cellular initiating and/or regulatory factor(s) responsible for this growth response have yet to be established. We have studied changes in lung content and activity of calmodulin, a calcium-dependent regulatory protein in relation with lung mass and DNA content during postpneumonectomy compensatory lung growth in 4-week-old rats. We observed that after left pneumonectomy, right lung calmodulin content (measured by radioimmunoassay) and calmodulin activity (measured by cyclic nucleotide phosphodiesterase activation) were increased at days 1 and 2 after surgery in the pneumonectomized rats, preceding the increase of lung mass and DNA content which started at day 3. Treatment of the pneumonectomized rats with a highly specific calmodulin antagonist, trifluoperazine, immediately after the surgery, resulted in diminished lung calmodulin activity, sparing the calmodulin content, and in concomitant reduction of lung mass and DNA content to values intermediate between those of controls and the pneumonectomized animals. Based on these findings, we conclude that calmodulin may be an important intracellular (possibly, autocrine) initiatory or facilitatory factor in compensatory hyperplastic lung growth after pneumonectomy.

Animals↗
PubMed1991

Morphometric and biochemical changes in lungs of growing rats treated with a calmodulin antagonist.

To determine the role of calmodulin in postnatal lung growth and development, 4-week-old rats were injected intraperitoneally on consecutive days with trifluoperazine (TFP), a potent and specific calmodulin antagonist, for a period fo 3 weeks and studied in comparison with normal controls and undernourished weight-matched animals. TFP treatment resulted in stunting of lung growth such that observed normal increments in morphometrically determined total number of alveoli and alveolar surface area and in biochemically determined DNA, elastin, and collagen contents of the lungs were diminished in comparison with age-matched normal controls. However, the TFP treatment also resulted in reduced daily food intake and body weight gain. In the TFP group, lung weight and lung volume were also reduced compared with the weight-matched control group. This resulted in reduced alveolar surface area, total number of alveoli, DNA, collagen, and elastin in the TFP group compared with values in the weight-matched controls. Thus the TFP-induced lung changes were not due to inanition and/or reduced somatic growth. The TFP treatment resulted in reduced activities of calmodulin and cyclic adenosine monophosphate (cAMP)-phosphodiesterase in the lungs of the animals, independent of their nutritional status. Based on these findings, we suggest that calmodulin may be an important regulatory component of postnatal lung growth and development.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1988

Experimental diabetes and the lung. I. Changes in growth, morphometry, and biochemistry.

Diabetes induced by streptozotocin at 3 wk of age in rats resulted in diminished somatic growth by 7 wk of age. Specific lung volume and weight (volume or weight per 100 gram body weight) were increased. The amount of lung DNA was decreased, and collagen and elastin were increased. The volume proportion of alveolar walls was increased at the expense of alveolar air. Air spaces were diminished in size, and alveoli increased in number. Total phospholipids and disaturated phosphatidylcholine (DSPC) were decreased, but they were normal relative to alveolar surface area. These changes reverted towards normal as a result of insulin treatment. Rats matched in weight (undernourished animals) for the diabetic animals showed relative preservation of lung weight, increased DNA, and reduced nonconnective tissue protein, RNA, collagen, and elastin. Air spaces were enlarged, alveolar surface area was decreased, and alveoli were decreased in number. Total phospholipids and DSPC were decreased, but normal when expressed per alveolar surface area. We conclude that diabetes and undernourishment have different effects on connective tissue synthesis in the lung that affect lung growth and structure, providing further evidence for the "fishnet" hypothesis of alveolar growth.

Animals↗
PubMed1988

Experimental diabetes and the lung. II. In vivo connective tissue metabolism.

In vivo lung connective tissue and DNA synthesis, following intraperitoneal injection of [14C]proline and [3H]thymidine, were studied in streptozotocin-induced diabetic rats fed ad libitum. Insulin-treated diabetic and normal rats similarly fed, and undernourished rats weight-matched to the untreated diabetics, served as comparison groups. The ratio of unbound [14C]hydroxyproline to total [14C]hydroxyproline in the lung was used to assess connective tissue degradation. Compared to the normal group, the untreated diabetic animals showed similar synthesis of collagen and elastin, reduced synthesis of total protein and DNA, and decreased degradation of connective tissue. When compared with the normal group, the undernourished animals showed diminished synthesis of total protein, collagen, elastin, and DNA, and increased degradation of connective tissue. In comparison to the undernourished group, the untreated diabetic animals indicated increased synthesis of collagen and elastin, and diminished degradation of connective tissue; total protein and DNA syntheses were similar. The insulin-treated diabetic animals showed increased collagen and DNA synthesis. We conclude that experimental diabetes has a profound effect on lung connective tissue metabolism, supporting previous observations of connective tissue abnormalities and morphometric changes. The increase in lung collagen and elastin in diabetes is in part due to reduced breakdown of the connective tissue proteins.

Animals↗
PubMed1987

Does the PI polymorphism alone control alpha-1-antitrypsin expression?

Whether genetic factors other than the protease-inhibitor (PI) polymorphism itself contribute to variation in alpha-1-antitrypsin is of both theoretical and practical interest. We have measured the quantity of alpha-1-antitrypsin (by an immunoturbidometric assay) and its activity (by assaying elastase inhibitory capacity [EIC]) in 583 individuals from 114 twin kinships who were also typed for PI by isoelectric focusing. Models of variation were fitted directly to the raw observations by a maximum-likelihood method. Specification of phenotypic means led to highly significant improvements in fit over models including only individual environment variance and additive genetic variance. The 29 phenotype means could also be described as the appropriate additive combinations of the 12 allelic effects. Only small improvements in fit could then be obtained by addition of polygenic components of variance. We conclude that nearly all genetic variation in alpha-1-antitrypsin quantity and activity can be explained by detectable variation at the PI locus and that this variance is largely additive. Bivariate analysis of alpha-1-antitrypsin and EIC revealed marginal evidence for differences in specific activities of molecules coded by different PI alleles. The correlation between environmental deviations for the two measures was only .63, which may reflect, in part, the rather low reliability of the assays and account for the modest heritabilities (less than .5) of the two measures. An intriguing finding was the presence of significant differences in E1 variance for different PI types, suggesting that different phenotypes have differing capacities to react to environmental challenges.

Adolescent↗
PubMed1984

Lung changes resulting from intraperitoneal injections of porcine pancreatic elastase in suckling rats.

We administered 1 mg of purified porcine elastase per 100 g body weight intraperitoneally twice weekly to male rats during the first 4 wk of life. This altered the structure of the lungs so that the proportion of alveolar duct air (the "core" of air internal to the mouths of alveoli in alveolar ducts) increased and the proportion of alveolar air decreased. The alveolar surface-to-volume ratio was decreased, and the lungs had too few alveoli. The lungs were also hyperexpandable, containing more air and saline per gram of lung tissue. No biochemical alterations were noted in elastin or collagen content in elastase-treated animals. In a separate experiment, we showed that elastaselike activity appears in the blood 1 h after intraperitoneal injection of elastase and, although low, was still significantly raised 24 h after injection. We feel that elastase has interfered with lung growth and alveolar development by altering mainly the elastin in the collagen-elastin network. Intraperitoneal elastase injection at the time of active elastin synthesis and alveolar multiplication may be a suitable model to test the hypothesis that lungs injured in infancy may be more susceptible to damage in later life.

Animal Population Groups↗
PubMed1981

Calmodulin activation of rat lung adenylate cyclase is independent of the cytoplasmic factors modulating the enzyme.

Adenylate cyclase activity in the rat lung membranes washed with 150 microM-EGTA was stimulated by calmodulin in the presence of 100 microM-Ca2+. The calmodulin activation of the enzyme was concentration-dependent; however, at high concentrations the activation was diminished. Activation of adenylate cyclase by calmodulin was immediate, reversible and due to an increase in the Vmax. without apparent effect on the affinity of the enzyme for ATP. The rat lung supernatant produced additive activation of the adenylate cyclase that was already maximally stimulated by calmodulin, indicating that either calmodulin and cytoplasmic factors act at different sites on adenylate cyclase or different adenylate cyclases may be involved. The data further support our previous conclusion that calmodulin is not involved in the activation of adenylate cyclase by cytoplasmic factors in rat lungs.

Adenylyl Cyclases↗
PubMed1981

Influence of streptozotocin-induced diabetes on the cytoplasmic factors modulating adenylate cyclase activity in rat lungs.

Basal adenylate cyclase activity in rat lung alveolar tissue particulate fraction was depressed during streptozotocin-induced diabetes. However, the activation of the particulate adenylate cyclase by the cytoplasmic factor(s) was markedly increased in lungs from the diabetic rats. The increased activation of basal adenylate cyclase in the diabetic tissue appeared to be due to an increase in the activity of the cytoplasmic factor(s) and not due to an increase in the sensitivity of the particulate enzyme to the cytoplasmic factor(s). Insulin treatment of the diabetic animals restored the activation of adenylate cyclase by the supernatant activator to the control values. The cytoplasmic factor(s) did not appear to be related to the ubiquitous calcium-dependent regulator protein, calmodulin.

Adenylyl Cyclases↗
PubMedDate not supplied

Lung growth in rats subsequent to administration of intraperitoneal elastase during the first 4 weeks of life.

Male rats received intraperitoneal injections of porcine pancreatic elastase twice weekly during the first 4 weeks of life. Saline-injected male rats served as controls. After a 4-week recovery period, the rats were sacrificed, and the excised lungs were studied using saline- and air-filled volume-pressure curves, morphology, and biochemistry. Both air- and saline-filled volume-pressure curves showed loss of elastic recoil in the elastase-treated animals. In elastase-treated animals the mean linear intercept was increased, the numbers of alveoli per unit area, per unit volume, and per lung decreased, and alveolar surface area decreased. These animals also showed a disproportionate increase of alveolar duct air at the expense of the proportion of alveolar air. Morphometric differences between the elastase-treated animals and the controls were somewhat smaller in this experiment than in a previous one in which the animals were sacrificed immediately after 4 weeks of intraperitoneal elastase. We attribute this difference to alveolar multiplication between 4 and 8 weeks of age. However, evidence is presented that these alveoli were abnormal. Loss of recoil properties was more evident after the 4-week recovery period than at the termination of elastase administration, suggesting that the newly formed alveoli were functionally abnormal. Collagen content was increased in the elastase group, but elastin was significantly decreased. This may be a good elastase model for assessing the hypothesis that lungs minimally damaged in infancy may have an increased susceptibility to environmental damage in later life.

Animals↗
PubMedDate not supplied

Changes of growth hormone, somatomedin C, and bombesin following pneumonectomy.

Left pneumonectomy (PX) was performed on 14-day-old pregnant rats. Serum growth hormone (GH), lung somatomedin-C-like immunoreactivity (SmC), and lung bombesin-like immunoreactivity (BLI), using optimized radioimmunoassays and lung protein concentration (P), were measured 3 h, and 1, 2, 3, 5, and 7 days following pneumonectomy. These levels were compared to two groups of similar animals: sham operated animals and animals not subjected to surgery. Serum GH, lung SmC, and BLI levels were similar in the last two groups of animals, suggesting that surgery had no effect on GH, SmC, and BLI levels. These two control groups were combined and compared to the post-pneumonectomy animals. The post-pneumonectomy animals had significantly higher levels of serum GH at postoperative day 3 and significantly higher levels of SmC at days 2 and 5 without any significant difference in total BLI level and body weight. These results suggest that, first, GH and SmC may play a part in post-pneumonectomy compensatory lung growth and these two may also be interrelated in this response and, second, BLI material(s) perhaps do not play a role in post-pneumonectomy lung growth.

Animals↗
PubMedDate not supplied

Diminished levels of insulin-like growth factor-I in lungs in streptozotocin-induced diabetes: relation to nutritional status and growth.

It is yet unknown whether the impaired nutritional status of streptozotocin-induced diabetic rats influences changes in levels of insulin-like growth factor-I (IGF-I) in this experimental model of diabetes. To explore this possibility, simultaneous studies were undertaken of rats made diabetic by streptozotocin (75 mg/kg body wt, intraperitoneally) and undernourished control rats with similar somatic growth rate (determined by body weight gain), in comparison with normal controls. Serum IGF-I levels were diminished in the untreated diabetic and undernourished control animals, but more so in the diabetic group. Lung IGF-I levels (per lung and per lung DNA) and DNA contents were diminished to similar degrees in the untreated diabetic animals and the undernourished control group. Lung dry weights of the diabetic rats were greater than those of the undernourished control group, such that lung IGF-I/100 mg tissue dry wt in the former was significantly lower than in the latter group. Insulin treatment of the diabetic rats restored their body weights, serum and lung IGF-I levels, and DNA contents to normal control values. Lung IGF-I levels in the diabetic rats correlated strongly with serum glucose (r = .75) and body weight (r = .79), and moderately with lung weight (r = .43) and lung DNA (r = .58). These findings suggest that the diminished lung IGF-I levels in streptozotocin-induced diabetes may be related to the impaired nutritional status and/or somatic growth of the experimental animals, and that this relationship may be responsible, at least in part, for the diminished lung cellular proliferation observed in experimental diabetic animals.

Animals↗