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Biomedical subjects

J C Belton

Publications and source records attributed to J C Belton.

11 recordsLinked to original sources

Regulation of lung fibroblast proliferation and protein synthesis by bronchiolar lavage in experimental silicosis.

Male Sprague-Dawley rats were subjected to a single, intratracheal instillation of 30 mg Min-U-Sil silica in sterile saline and were sacrificed 3, 7, or 14 days following instillation. Control animals were instilled with sterile saline only. Silica instillation produced an inflammatory reaction followed by histological changes characteristic of lung fibrosis. Thickened alveolar septa associated with inflammatory cells transforming into large multifocal fibrotic nodules were detected in silica-exposed animals. Increased numbers of bronchoalveolar cells (principally macrophages), elevated levels of protein (principally serum albumin), and lysozyme, proteolytic (trypsin-like), and myeloperoxidase activities were detected in lavage fluids obtained from animals instilled with silica. These factors (except for lysozyme activity) were elevated above control levels from 3 to 7 days postinstillation and declined to near control levels by Day 14. The rate of DNA, collagen, and noncollagen protein synthesis was significantly elevated in lung tissue minces from silica-treated rats 3 and 7 days after instillation. Elevated levels of total protein, and lung collagen in particular, were observed 9 weeks after insult. Lavage fluid from silica-instilled rats stimulates DNA synthesis in cultures of proliferating and quiescent rat lung fibroblasts. Lavage fluid from silica-instilled rats also stimulates lung fibroblasts to increase collagen and noncollagen protein synthesis.

Animals↗

The effects of dipalmitoyl phosphatidyl choline on the precipitation of native fibrils and segment-long-spacing aggregates from collagen solution.

The effect of dipalmitoyl phosphatidyl choline (DPPC), the major phospholipid component of pulmonary surfactant, on the precipitation of collagen in the form of native fibrils and segment-long-spacing (SLS) aggregates was studied in vitro. The effects of DPPC on both phases of collagen fibrillogenesis were analyzed spectrophotometrically, and alterations in the morphology of precipitated fibrils and SLS aggregates were ascertained by transmission electron microscopy (TEM). Low concentrations of DPPC inhibited the growth phase of fibrillogenesis, while higher concentrations were required to inhibit nucleation. Both the meshwork density and mean width of precipitated fibrils were altered by DPPC, as was the size of SLS aggregates. Segment-long-spacing aggregates prepared from pepsin-treated collagen were inhibited to a greater degree than SLS aggregates prepared from untreated collagen, indicating that the pepsin-susceptible residues of the telopeptide extensions of tropocollagen molecules stabilize SLS aggregates against the effects of DPPC. Based on these results and the inhibition of the growth phase at lower concentrations than those which inhibited the nucleation phase of fibrillogenesis, it was concluded that the primary mechanism of DPPC inhibition is electrostatic interference between the positively charged phospholipid molecules and the net positive charge of collagen. It is proposed that pathological conditions involving the pulmonary epithelium may allow interaction between surfactant and collagen, which could further weaken the interstitial connective tissue.

Animals↗

Macromolecular synthesis in organ cultures of neonatal rat lung.

Organ cultures of newborn rat lungs synthesize and accumulate DNA, RNA, collagen and noncollagenous proteins almost at a linear rate for at least 5 days. During this period the synthesis of collagen consistently exceeds the synthesis of noncollagenous proteins in a pattern similar to neonatal lung growth in vivo. Although some morphological characteristics of lung architecture are distorted after culture, fundamental structural similarities to lungs growing in intact animals are retained. When these cultures are maintained in atmospheres rich in oxygen, increased collagen synthesis is observed, a response similar to that of lungs in intact animals exposed to high oxygen concentrations in vivo. Our studies suggest that lung organ cultures may be a suitable system for investigating the biochemical aspects of lung tissue-environmental interaction.

Animals↗

Ultrastructural alterations in collagen associated with microscopic foci of human emphysema.

Postmortem and biopsy lung samples were examined with a dissecting microscope and classified into three categories: no overdistended air spaces visible, mild destruction and some fenestration of the septa with minimal confluence of air spaces, and more advanced breakdown of alveolar walls with at least a sixfold increase in air space to a 1300 mu diameter. Prominent ultrastructural tissue alterations were found within the focal regions where fenestration of the alveolar wall and some confluence of air spaces were identified. Edema and rearrangement of the supportive tissues were noted in the alveolar septa adjacent to foci having the greatest alveolar enlargement. Randomized electron dense collagen fibrils were found only in these areas, whereas the normal alveolar septa contained uniformly parallel collagen fibrils. Many of the randomized fibrils had sufficient swelling to display prominent internal spiraling. Other changes in structure and cell numbers were consistently found in association with the altered collagen. These included: separation of the type I epithelial cells from the basal lamina and some autolysis of the cytoplasmic organelles, numerous flattened type II epithelial cells, and aggregations of pulmonary macrophages.

Adult↗

Idiopathic spontaneous pneumothorax. Electron microscopic study.

A lung biopsy obtained from a patient following a second occurrence of idiopathic spontaneous pneumothorax was studied by electron microscopy. A remarkable increase in septal mass was a notable finding. Proliferation of the interstitium was due to an abundance of collagen and elastic fibers. In additon, a proliferation of type 1 and type 2 epithelial cells was seen.

Adult↗

Ultrastructural changes in chloroplasts resulting from fluctuations in NaCl concentration: freeze-fracture of thylakoid membranes in Dunaliella salina.

The structure of chloroplasts isolated from Dunaliella salina has been studied with respect to changing concentrations of sodium chloride in the culture medium. Freeze-fracture replicas and thin sections of intact chloroplasts do not exhibit any noticeable changes in structure at concentrations ranging between 3.5 and 25% NaCl. Chloroplasts isolated from algal cells that have been acclimatized to the higher salt concentration show a change in the thylakoid membranes. The thylakoid membranes appear compressed over a major portion of the membrane surface, with only the end of the thylakoid membranes unappressed. The number of particles per unit area on the B face is also altered by the salt concentration. The chloroplasts acclimatized to 25% NaCl have about 3 times the number of particles per unit area on a B face of end-membranes as on a comparable face of thylakoid membranes acclimatized to low (3.5% NaCl) salt concentration. These morphological changes can be reversed if the chloroplasts acclimatized to high or low salt concentrations are returned to a medium of different salt concentration prior to freeze-fracturing.

Adaptation, Biological↗

Freeze-etch study of collagen. I. Native collagen from tendon and lung of rats.

The freeze-fracture technique was used to study the fine structure of native collagen from rat tendon and lung. The collagen filaments were organized into a spiraled lamellar substructure within each fibril. Small particles, representing the broken ends of the collagen filaments, were found in nearly straight rows when the lamellae were broken tranversely. Filamentous connections were shown which span the interfibrillar matrix and unite all of the fibrils into a reticular network. The fibrils were coated with material that was greatly hydrated in vivo. Sublimation occurred more rapidly from some portions of the coating substance than from others, resulting in a banding pattern with a 610 A repeat distance.

Animals↗

Regulation of lung fibroblast proliferation and collagen synthesis by alveolar macrophages in experimental silicosis. I: Effect of macrophage conditioned medium from silica instilled rats.

Alveolar macrophages were lavaged from silica or saline instilled rats 0, 3, 7 and 14 days after exposure. Macrophages were cultured for twenty-four hours and the conditioned media assayed for the ability to stimulate rat lung fibroblast proliferation and collagen synthesis. Macrophages remained viable throughout the culture period. DNA synthesis was significantly elevated by macrophage conditioned media (MCM) from silica instilled rats (S-MCM) compared to untreated fibroblasts or fibroblasts exposed to MCM from saline instilled control animals (C-MCM). Stimulation of DNA synthesis was not observed when S-MCM was exposed to non-proliferating fibroblasts. Collagen synthesis quantitated as 3H-hydroxyproline accumulation or percent collagen synthesis was also increased by day 3 and day 7 S-MCM. Specific activity measurements of intracellular 3H-proline minimized the possibility that the increase in 3H-proline incorporation into collagen was a reflection of increased proline transport. Non-collagen protein synthesis was also increased in fibroblasts exposed to day 14 S-MCM. These results suggest that alveolar macrophages elaborate factors following silica exposure capable of altering the DNA and protein synthetic activity of lung fibroblasts. These changes in fibroblast DNA and protein metabolism are similar to those observed for lung tissue in vivo and lend further support to the hypothesis of macrophage mediation of the pulmonary response following silica exposure.

Animals↗