Electron microscopic and radioautographic study of the bronchi in chronic inflammation as affected by helium-neon laser treatment

Structural-metabolic changes were studied in 98 biopsies of large bronchial mucosa from 39 male patients with chronic suppurative lung diseases. It was shown (by the level of DNA and RNA synthesis) that the proliferative and metabolic processes were induced by endobronchial treatment of the damaged epithelium with helium-neon laser which passing through a number of transitional stage recovers its structure and differentiation to ciliary and goblet cells with normal ultrastructure. In lamina propria hyperemia, intensive leukodiapedesis, leukocyte infiltrates and granulations develop and proliferative and metabolic activity of endotheliocytes and interstitial cells increases, which results in the formation of delicate fibrous connective tissue. Simultaneous reorganization of the epithelium and underlying connective tissue is interpreted in terms of parenchymatous-stromal interactions.     

An electron microscopic and radioautographic study of hypostomal regeneration inHydra viridis

Summary The gastrodermal secretory cells inHydra viridis are limited to specific regions in the body column. There are two types of mucous cells present, and they are limited to the hypostome. The zymogen cells are absent from the hypostome, but they extend along the body column from the tentacles to the peduncle. Transection beneath the tentacles produces a proximal portion of the hydra devoid of mucous cells. This piece regenerates new tentacles and a normal hypostome, filled with mucous cells, within four days.The following events were observed during regeneration. The zymogen cells formed an aggregate within twenty-four hours in the region of the presumptive hypostome. These cells organized and formed lobes of zymogen cells that were positioned similarly to the arrangement of mucous cells in the normal animal. Sparsely distributed small basophilic cells were also present in the reforming hypostome. Using corresponding thick and thin sections we identified the cells incorporating radiosulfate: 1) The zymogen cells in the distal aggregate. 2) Small basophilic cells, some filled with free ribosomes, and others with a well-developed E. R. 3) Secretory cells containing both mucous and serous granules. 4) Secretory cells with granules similar to the granules in mouse Paneth cells.The fate of the secretory granules in the zymogen cells in the distal aggregate is unknown. Some are autolysed within the cell, and others are extruded. However, some observations suggest that there may be a direct transformation of some of the serous granules to mucous granules. The E. M. observations, the radiosulfate incorporation data, and the migrations of cells to the wound site, suggest that both the zymogen cells and basophilic cells transform to mucous cells. Identification of the early stages of mucous synthesis in these basophilic cells enabled us to study the sequence of mucous granule maturation of both the hypostomal mucous cells.The two most significant questions which we feel remain unaswered are: 1) What are the ultrastructural events during the zymogen cell transformation to a mucous cell ? 2) What is the origin of the small gastrodermal basophilic cells ?

---

Zusammenfassung Die gastrodermalen sekretorischen Zellen von Hydra viridis kommen nur in spezifischen Regionen der Körpersäule vor. Es gibt zwei Typen muköser Zellen, und diese findet man ausschließlich im Hypostom. Zymogene Zellen gibt es nicht im Hypostom, aber erstrecken sich längs der Körperachse von den Tentakeln zum Pedunkulus. Sektion unterhalb der Tentakel produziert eine proximale Region von Hydra ohne muköse Zellen. Dieses Stück regeneriert neue Tentakel und ein normales Hypostom mit mukösen Zellen innerhalb vier Tagen.Die folgenden Vorgänge wurden beobachtet während der Regeneration. Die Zymogenzellen bildeten ein Aggregat in der Gegend des präsumptiven Hypostoms innerhalb 24 Std. Diese Zellen bildeten Lappen von Zymogenzellen in ähnlicher Anordnung wie die mukösen Zellen im Normaltier. Ebenfalls vorhanden im neu sich bildenden Hypostom waren locker verteilte, kleine basophile Zellen. Durch Verwendung alternierender dicker und dünner Schnitte identifizierten wir die Zellen, die radioaktives Sulfat einbauten: 1. Zymogenzellen im distalen Aggregat. 2. Kleine basophile Zellen, einige mit freien Ribosomen angefüllt, andere mit gut entwickelten endoplasmatischen Retikulum. 3. Sekretorische Zellen mit mukösen und serösen Granula. 4. Sekretorische Zellen mit Granula, die ähnlich aussehen wie die Granula von Maus Paneth-Zellen. Das Schicksal der sekretorischen Granula in den Zymogenzellen des distalen Aggregates ist unbekannt. Einige werden innerhalb der Zellen autolysiert, andere ausgestoßen. Es scheint aber, daß einige der seriösen Körner sich direkt in muköse umwandeln. Die elektronenoptischen Bilder, die Ergebnisse des Sulfat-Einbaus, und die Wanderung von Zellen zur Wunde weisen darauf hin, daß sowohl Zymogenzellen, als auch basophile Zellen sich in muköse Zellen verwandeln. Identifikationen der Frühstadien von Mukus-Synthese in diesen basophilen Zellen erlaubte uns, die Sequenz der Reifung der Mukus-Granula beider hypostomalen Mukuszellen zu studieren.Die zwei wichtigsten, noch unbeantworteten Fragen verbleiben: 1. Was für feinstrukturelle Veränderungen finden statt während der Transformation von Zymogenzellen in muköse Zellen ? 2. Woher stammen die kleinen gastrodermalen, basophilen Zellen ?

---

---

This paper was prepared from a thesis submitted in partial fulfillment for the degree of Master of Arts.

---

This work was supported by the National Institutes of Health grant No GM-11218 to Dr. A. L.Burnett.     

Electron microscopic and radioautographic study of the wall of the large bronchi in chronic inflammatory processes in the lungs

Synthesis of DNA, RNA and protein in the structures of large bronchi in patients with chronic inflammatory processes was studied. It was shown, that nuclei of all cell types of the tegmental bronchial epithelium, capillary endotheliocytes and stromal cells actively included 3H-uridine. By the intensification of bronchial wall sclerosis index and hardness of label with 3H-uridine were reduced in epitheliocytes, endotheliocytes, pericytes, labrocytes, macrophages, but were increased in fibroblasts which also had a high level of 3H-proline inclusion. High index of label with 3H-thymidine was found in basal cells of epithelium and capillary endotheliocytes. In conditions of chronic inflammation of bronchial wall the greatest metabolic and proliferative activity was found in the capillary endotheliocytes and cells of a pericapillary zone. In sclerosis the proliferative activity of basal cells was changed, that predetermined the character of tegmental epithelium reorganization.     

Radioautographic study of DNA synthesis in bronchoalveolar lavage macrophages during chronic inflammatory processes in the lungs

The cells obtained by broncho-alveolar lavage from patients with chronic pulmonary inflammations were investigated using light and electron microscopy and radioautography. Proliferative activity of lavage alveolar macrophages was studied by 3H-thymidine in vitro incorporation. The labelling index of alveolar macrophages was up to 0.2% in chronic bronchitis and 1.25-3.33% in chronic destructive inflammation. The enhancement of human alveolar macrophage proliferation in chronic pulmonary destructive inflammation is of great importance for the maintenance of the number of mononuclear phagocytes responsible for cellular protection of respiratory lung structures.     

Electron microscopic and electron radioautographic study of cells in glandular hyperplasia and cancer of the endometrium

Peculiarities of the ultrastructure and RNA synthesis in the epithelial endometrial cells of nine patients with hyperplasia and the uterine cavity cancer have been studied by the electron-radioautographic method. Quantitative differences are revealed in the ultrastructures of the cells. Amplification of the nucleoli labelling and asynchronism of the RNA synthesis in the nucleoli with glandular hyperplasia and endometrial cancer are observed.     

An improved method of bronchoalveolar lavage of lungs of small laboratory animals: short report

This report describes a semi-automatic method for standardized bronchoalveolar lavage of small laboratory animals. In essence the method is constructed from a dispenser and a pressure chamber.     

An electron microscopic radioautographic study of the uptake of tritiated serotonin by nerve fibres in the posterior salivary duct and gland of cephalopods

In the posterior salivary duct and gland of Octopus vulgaris and of Eledone cirrhosa, the duct secretory nerve trunks and their ramifications in the gland tubules include many fibres that incorporate labelled serotonin. However, there are also unlabelled secretory fibres, which cannot be discriminated from incorporating fibres on morphological grounds. Neuroglandular junctions are not apparently established by incorporating fibres. In the duct, the motor nerve trunks contain a small number of labelled fibres, and nerve bundles supplying the duct muscle contain, in variable proportions, serotonin incorporating fibres. Both labelled and unlabelled nerve fibres reach the duct muscle fibres, but neuromuscular junctions involve only unlabelled presynaptic fibres. The nerve fibres which join the gland muscle are usually unlabelled, and the small quota of incorporating fibres in the motor trunks apparently supply only duct tissues. Both secretory and motor trunks, originating from different ganglia, can be considered to contain heterogeneous fibres, releasing different neurotransmitters at the terminals. Certain of these fibres could be serotoninergic.     

An electron microscopic study of the human infundibulum

Summary For a limited period during the oogenesis of Protopterus, blebs of the perinuclear cistern contain, in addition to other inclusions, a special kind of microtubular elements. Most of these blebs face parts of multiple nucleolar bodies that extend toward and make contact with the inner nuclear membrane. The microtubular lumen contains a finely dispersed material of moderate electron density which seems to be in contact with this nucleolar material.Aside from these intracisternal structures there are, within both the perinuclear cytoplasm and the nucleoplasm, similar microtubular arrays without apparent connection with the nuclear envelope. These are either enclosed by membranes derived from those of the envelope or unconfined, having escaped through breaks in their respective bounding membranes. Extracisternal tubules are presumed to have passed their period of putative functional activity and to be undergoing a process of regression and subsequent disintegration.Among possible roles attributable to the intracisternal microtubular apparatus are the following: (1) It may serve for the transport of special nucleolar components to the cytoplasm, possibly to be incorporated in the matrix of developing perinuclear mitochondria; (2) it may provide openings in the nuclear membranes for the direct passage of particulate elements between nucleus and cytoplasm; (3) it may be instrumental in the breakdown of parts of the nuclear envelope prior to its restitution during the subsequent phase of oogenesis.Supported by grants NB-00840 and NB-05219 from the U.S.P.H.S.     

An electron microscopic radioautographic study of the viability of human cells after death. The postmortem activation of RNA synthesis in neutrophils]

RNA synthesis in human neutrophils isolated from blood 2.5-67 hours after death of patients was studied by means of light and electron microscopy radioautography. The rate of RNA synthesis in neutrophils isolated from blood of cadavers was higher in comparison with the same parameter in neutrophils isolated from survivor's blood. The significance of this observation for resuscitation and transplantology is discussed.     

An electron microscopic study of the human epidermal keratinocyte

Summary 1. The epidermis of the flexor surface of the upper arm of human subjects was studied with the electron microscope. 2. The cytoplasm of the keratinocytes in the basal layer contained many tonofilaments, ribosomes and other cell organelles. The tonofilaments were arranged singly or in loose bundles and many were attached to the inner membrane of the desmosomes. Along the basal border of the cells pinocytotic vesicles could be seen at different stages of development. 3. The keratinocytes in the stratum spinosum differed from those in the basal layer in two main ways: (a) The tonofilaments were grouped together into large compact bundles known as tonofibrils and it was possible to determine a definite beading or cross banding along the length of some of the filaments. (b) The cells were assuming a flattened shape. 4. The keratinocytes in the stratum granulosum possessed large numbers of irregularly shaped keratohyaline granules. The granules were strongly osmiophilic and were always situated on a meshwork of tonofibrils. The keratohyaline granules had no internal structure. The nuclei and mitochondria showed evidence of degeneration. 5. The keratinocytes in the stratum corneum were long and flattened. The cell walls showed increased electron density and were considerably thickened. The cytoplasm was filled with closely packed fibres separated by a small amount of lucent matrix. The fibres were grouped together in bundles running in different directions within the flattened squames. The fibres had along their entire length alternating areas of high and low electron density. The keratohyalin granules had disappeared and nothing remained of the nuclei or the organelles. In the deepest cells of this region the fibres were sometimes loosely packed leaving large irregular open spaces. This area corresponded to the stratum lucidum. In the most superficial layers of the stratum corneum the fibres appeared to be breaking down so that little remained within the keratinocyte except large lucent spaces. The desmosomes showed distinct structural changes. 6. An attempt was made to correlate the structural changes in the different epidermal layers with the process of keratinization. The possible part that keratohyalin may play in the process of thickening of the cell walls was discussed. The relationship between the desmosome and its dynamic environment was considered.I wish to express my sincere thanks to Dr. David Hilding of the Department of Otolaryngology for the use of an R.C.A. electron microscope and other facilities in his laboratory. This research was supported by the United States Public Health Service and American Cancer Society grants. USPHS CA 04679-07, NB 03995.     

Abnormal collagen fibril structure of skin in chronic haemodialysis patients: An electron microscopic study

The structure of collagen fibrils of skin in chronic haemodialysis patients was studied by electron microscopy. Although, in all patients, the parallel packing of the fibrils remained, there were areas with disorganization. The periodicity D as well as the banding pattern were normal. The most conspicuous finding of the work presented here is that in all patients the fibril diameters were significantly smaller than those from normal age-matched control subjects. Also, the former showed a much higher degree of variability in width of collagen fibrils than the latter.     

An electron microscopic study of surface polysaccharides in Bacteroides

The electron microscopic appearance of the cell surface of Bacteroides strains and Klebsiella pneumoniae stained with ruthenium red or colloidal iron is described. The effect of polymyxin B (PMB) was also registered. It was found that all Bacteroides strains have a polysaccharide lined 'micro-capsule' external to the outer membrane which could aggregate and form blebs. The blebs so formed were distinct from other types of bleb formed in Klebsiella involving the outer membrane and induced by PMB. Such types of PMB alterations were not induced in Bacteroides.