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C Gardi

Publications and source records attributed to C Gardi.

32 records · Page 2Linked to original sources
PubMed1989

A biochemical and morphological investigation of the early development of genetic emphysema in tight-skin mice.

The tight-skin (Tsk) mouse has recently been proposed as a genetic model of emphysema. In the present study, the development of emphysema was investigated in these mice with histological, biochemical, and ultrastructural methods at 4 days and at 1 and 2 months of life. At 4 days after birth, histological examination of the lungs revealed only a mild enlargement of the primary sacculi. Neither biochemical nor ultrastructural changes were seen however at this time. At 1 month of age, the histological examination showed marked emphysema-like changes, characterized by enlargment of air spaces accompanied by destruction of alveolar walls. Biochemical analysis showed a marked decrease in insoluble elastin content and a significant increase in salt-extractable collagen. Ultrastructural investigation revealed edema fluid in the interstitium and broken and disorganized elastic fibers. All these findings strikingly resemble the changes which occur in the lungs early after an instillation of elastase. In the 2-month-old Tsk mice the histological lesion progressed in severity. The ultrastructural findings were similar to those observed at 1 month, and the biochemical changes showed no signs of recovery. Thus, in these mice, the emphysematous lesion develops very rapidly between 4 days and 1 month of life and shows the characteristics of an elastolytic process which is still ongoing at 2 months of age.

Animals↗
PubMed1989

A 16-month study of the development of genetic emphysema in tight-skin mice.

The tight-skin (tsk) mouse has been recently proposed as a model of genetically determined emphysema. In the present study, the development of this lung lesion was investigated morphometrically in both male and female tsk mice from 4 days to 16 months of age. "Pallid" (pa) mice with a closely related genetic background served as controls. Despite small differences in the mean linear intercept seen at 1 and at 16 months, both male and female tsk mice were found to be similarly susceptible to the development of the emphysematous lesion. Three main phases were observed in the evolution of the emphysema. The first phase, from 4 days to 2 months after birth, included enlargement of the peripheral lung units, already evident at 4 and at 15 days, and destruction of the lung parenchyma, which occurred between 15 days and 1 month and rapidly progressed for as long as 2 months. A second phase of stabilization or of mild progression of the emphysematous lesion occurred between 2 and 8 months of age. A third phase, in the advanced age between 8 and 16 months, included a further exacerbation of the parenchymal destruction. At 16 months of age, the lungs of the pa mice also showed some patchy areas of air-space enlargement with destruction of the alveolar septa.

Animals↗
PubMed1988

The saccharin method for testing mucociliary function in patients suspected of having primary ciliary dyskinesia.

In order to evaluate the clinical value of the saccharin test as a practical and simple measure of mucociliary clearance, nasal mucociliary clearance (NMCC) and ciliary ultrastructure were studied in 22 patients suspected of having primary ciliary dyskinesia (PCD) based on the saccharin test. Ten patients fulfilling the diagnostic criteria of PCD had a pathological response to the saccharin test (transport time greater than 60 minutes), and this was consistently associated with ultrastructural defects, specific for PCD. These results validate the suitability of the clinical use of the saccharin test as a screening procedure for NMCC. The false-negative results obtained in three cases of PCD, all with borderline values, cannot be ascribed to ineffectiveness of the test, but rather to the persistence of some motility by certain defective cilia, detectable by microphoto-oscillographic investigation of specimens obtained by nasal biopsy or brushing.

Adolescent↗
PubMed1988

Cilia-lacking respiratory cells in ciliary aplasia.

This report describes the ultrastructural alterations observed in the nasal and bronchial mucosa of an 11-yr-old male suffering from immotile cilia syndrome (ICS). The morphological features observed in this patient are consistent with a ciliary aplasia. In fact, ciliated cells appeared to be replaced by columnar cells lacking cilia and basal bodies, and bearing on their surface cilium-like projections without any internal axonemal structure. In spite of the absence of basal bodies, centrioles, and kinocilia, these cells unexpectedly showed mature striated roots and centriolar precursor material scattered throughout the apical cytoplasm. These data suggest that control over basal body assembly is distinct from control over striated root formation. The presence of the above-reported structures in cells otherwise presenting many morphological features of normal ciliated cells is discussed on the basis of current knowledge of respiratory cilia biogenesis.

Bronchi↗
PubMed1988

Isolation and partial characterization of a proteinase with elastolytic activity from mouse blood leukocytes.

Leukocyte elastase has been implicated in the etiology of pulmonary emphysema. Recently, two genetic models of emphysema have been described, in mouse, which may enhance our understanding of the pathogenesis of emphysema. We therefore sought to purify mouse leukocyte elastase in order to characterize its biochemical properties. Leukocyte enzyme has been purified by a two-step procedure involving salt extraction of granular fraction, followed by preparative isoelectric focusing on Sephadex G-75 Superfine. The enzyme hydrolyses elastin and synthetic substrates for elastase, even if to a different extent. Inhibition studies indicates that the enzyme is a serine proteinase. Mouse elastase has a single isoelectric point of 8.65 and it behaves on sodium dodecyl sulphate polyacrylamide gel electrophoresis as a major band (molecular weight 29,000) and two minor bands (molecular weight 27,000 and 25,800, respectively.

Animals↗
PubMed1987

Purification and partial characterization of elastase activity from rat alveolar and peritoneal macrophages.

Macrophage elastase was purified from conditioned media from alveolar and thioglycollate-elicited peritoneal macrophages. The enzyme was purified to apparent electrophoretic homogeneity by preparative isoelectric focusing after a purification step consisting of low ionic strength dialysis and sequential batch fractionation on DEAE-Sephadex A-50. The proteinase activities isolated from alveolar and peritoneal macrophages showed the same physical and biochemical properties. This fact suggests that the same enzyme activity is present in rat macrophages of two different anatomical sites. The molecular weight and isoelectric point of the enzyme were estimated to be 22,500 and 8.3, respectively. The enzyme, characterized as a metallo proteinase, had elastolytic activity, as well as activity toward Suc-(Ala)3-NA. It is inhibited by o-phenanthroline, chicken ovoinhibitor, and EDTA, but not by phenylmethylsulfonyl fluoride or soybean trypsin inhibitor. The macrophage enzyme possesses biochemical and biophysical properties different from the rat pancreatic and granulocyte elastases (which are serine proteinases), and from the metallo proteinase with elastolytic activity isolated from rat platelets.

Animals↗
PubMed1987

Bromotrichloromethane-induced damage to bronchiolar Clara cells.

Administration of bromotrichloromethane (BrCCl3) to rats results in selective damage to bronchiolar non-ciliated Clara cells; ciliated bronchiolar cells and pneumocytes were unaffected. The cellular alterations begin very early (10 min) after poisoning. Lipid peroxidation, as measured by the malonic dialdehyde (MDA) content in the lung, is greatly increased 10 min after BrCCl3 administration. A histochemical technique to detect, in vitro, lipid peroxidation in frozen sections was used to demonstrate whether sufficient activation of BrCCl3 occurs in lung tissue. The positivity for the histochemical reaction was observed in bronchiolar epithelium in which cytochrome P450-dependent monooxygenase activity is predominantly located. The data obtained strongly support that BrCCl3 is highly metabolized in bronchiolar Clara cells.

Animals↗
PubMed1986

Isolation and partial characterization of rat elastolytic enzymes from various cells and tissues.

Different elastolytic enzymes were isolated from rat aorta and platelets, as well as from granulocyte and pancreatic extracts. The active fractions were purified to electrophoretic apparent homogeneity by precipitation with ammonium sulfate, sequential batch fractionation on DEAE-Sephadex A-50, and finally by isoelectric focusing (IF) on Sephadex G-75 Superfine. The molecular weight and the isoelectric point of the isolated enzymes were estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and by analytical IF, respectively. All the enzymes have elastolytic activity as well as activity toward Suc-(Ala)3-NA. The inhibition profile of the different isolated enzymes toward various inhibitors indicates that aortic, pancreatic, and granulocyte enzymes all belong to the group of serine proteinases, unlike the platelet elastase which is a metalloproteinase.

Animals↗
PubMed1986

Effect of the novel synthetic protease inhibitor furoyl saccharin on elastase-induced emphysema in rabbits and hamsters.

Furoyl saccharin, a novel heterocyclic acylating agent which has been previously found to possess a potent inhibitory capacity in vitro for elastase and other serine proteases, has been investigated in vivo in two acute animal models of emphysema. In hamsters, intratracheal (i.tr.) administration of 0.1 mg porcine pancreatic elastase resulted seven days later, in a 42% increase of the mean linear intercept (Lm). Addition of 0.3 mg to 0.3 mg furoyl saccharin to elastase exhibited a partial, not dose-related, but statistically significant inhibition of the increase of LM. Addition of 1 mg furoyl saccharin (equivalent to a dose of 12.5 mg/kg) completely abolished the increase in Lm. In the rabbit i.tr. instillation of 3.7 mg porcine pancreatic elastase induced within seven days, a 48% increase of the Lm, a 27% decrease of the internal surface area (ISA) of the lungs and a 33% decrease of the ISA corrected to an arbitrary total lung volume of 70 ml (ISA70). Furoyl saccharin given i.tr. 15 min prior to elastase at the doses 3, 10 and 20 mg prevented significantly in a dose-related manner, the changes in Lm, ISA and ISA70. The highest furoyl saccharin dose (equivalent to a dose of 10.8 mg/kg) completely protected against the emphysematous lesion. Additionally furoyl saccharin (20 mg i.tr.) prevented in the rabbit model the depletion in lung insoluble elastin and the increase in salt soluble collagen induced by the elastase administration. These results show that furoyl saccharin also in vivo has a marked antielastase activity.

Animals↗
PubMed1985

Pulmonary vascular injury in pancreatitis: evidence for a major role played by pancreatic elastase.

Using an experimental model of pancreatitis in the rat, the role of trypsin and elastase in mediating lung vascular injury in this condition was examined. The induction of pancreatitis by injection of sodium cholate in the pancreas resulted in a significant decrease in serum trypsin inhibitory capacity, and in a complete saturation of serum elastase inhibitory capacity matched by the appearance of endothelial injury of pulmonary capillaries and edema formation. The complete lack of serum elastase inhibitory capacity was associated with the presence of elastase activity in serum and bronchoalveolar lavage (BAL) fluids. The pretreatment of animals with N-furoyl saccharin (a potent inhibitor of many serine proteinases) prevented lung capillary injury and the imbalance of serum proteinase-anti-proteinase activities as well as the appearance of any elastolytic activity in serum and BAL fluids. These findings which clearly demonstrate the protease dependence of the pulmonary vascular injury in our experimental model, strongly suggested a major role for elastase(s). The suppression, in the experimental model, of the serum elastase inhibitory capacity by using chloramine-T resulted in an earlier onset of lung vascular damage, a marked worsening of pulmonary lesions, and an increase of elastolytic levels in serum and BAL fluids. Furthermore the physical properties of the protein molecule with enzyme activity detected in BAL fluids were consistent with those of rat pancreatic elastase. The reported data strongly support the hypothesis that pancreatic elastase plays a major role in the development of pulmonary vascular injury after acute pancreatitis.

Animals↗
PubMed1985

Severe teratospermia in an infertile man with bronchiectasis.

Morphologic examination of sperm samples from an infertile man with bronchiectasis revealed a severe teratospermia characterized by rare abnormalities that were present in most of his spermatozoa. These abnormalities were represented by round-headed spermatozoa, changes in chromatin condensation, acrosomal alterations, multiple tails, and complete subcellular derangement. The similarity between the above-reported abnormalities and those observed in various species of laboratory animals strongly suggests that the observed defects might be the result of a defective function of the manchette during spermiogenesis. The absence of specific defects of the axonemal morphologic features in sperm tails and in respiratory cilia, together with some laboratory data, suggests that association between infertility and bronchiectasis cannot be related, in our patient, to a generalized syndrome.

Adult↗
PubMed1984

Detection of elastase activity with a zymogram method after isoelectric focusing in polyacrylamide gel.

A zymogram method for detecting elastase activity following isoelectric focusing in polyacrylamide gel is described. After enzyme activity has been visualized, the gel itself is available for protein staining and for analysis in sodium dodecyl sulfate-polyacrylamide gel electrophoresis in second dimension. The zymogram method is suitable for detecting microgram amounts of elastase and has one step only. It can be used with the purified enzyme as well as with crude extracts of tissue containing elastases showing activity toward succinyl-(Ala)3-p-nitroanilide. By this method a major component of elastase in both porcine and rat pancreas was detected. In addition, two forms of elastase with isoelectric points of 8.2 and 8.8, respectively, were identified in rat leukocyte extracts.

Agar↗
PubMedDate not supplied

Skin biopsies for cell cultures from Mediterranean free-ranging cetaceans.

The aim of this study was to develop a useful method for obtaining viable tissue samples for establishing cell cultures from skin biopsies of free-ranging cetaceans. The skin biopsies were performed by two methods: dart from an air gun and dart from a crossbow. The dart tip was modified to collect tissue. The tissue was kept in tissue culture medium at ambient temperature, then processed within 24 h. Many modifications in culture technique, with respect to conventional culture methods for human fibroblasts, were made. The cultures thus obtained can be used for many purposes, including genetic and toxicological studies. In toxicology they are an alternative in vitro system for studying threatened animals such as marine mammals. In particular, fibroblasts can be used to test the vulnerability of cetaceans and pinnipeds to different environmental contaminants such as organochlorine compounds, heavy metals and polycyclic aromatic hydrocarbons.

Animals↗