Influence of X-ray on the autophagic-lysosomal system in rat pancreatic acini

Lysosomes have an important role in radiation injury of cells and tissues. Activation of autophagy is frequently observed in different types of pathological tissue degeneration. In radiation response it increases in some cases, and lysosomes are responsible for regulated degradation of the autophagic vacuoles. Lysosomes are also involved in ionizing radiation induced cell death. In apoptosis lysosomes degrade content of the phagocytotic vacuoles derived from engulfed apoptotic blebs. On the other hand lysosomal enzymes discharged from disintegrated cells have a key role in induction of necrotic changes. In this work we investigate autophagy and lysosomal protein degradation in the relatively radiation insensitive exocrine pancreatic acini in vivo and in vitro. Type of cell death induced by X-ray was also examined in relation to the changes of the lysosomal processes. In 5h after 16 Gy in vivo whole body irradiation we observed significant increase in the cytoplasmic volume fraction of autophagic vacuoles and in the number of apoptotic cells in vivo. But in the acini isolated from irradiated rats we could not detect a change in the lysosomal degradation of intracellular proteins. Therefore irradiation probably influences the autophagy in an earlier step than lysosomal degradation. Extended necrotic lesions were not observed in vivo as long as 48 h. Isolated pancreatic acini usually contain more autophagic vacuoles than in vivo, but we could not observe additional increase in autophagy after 8 Gy, in vitro irradiation. Lysosomal degradation of intracellular proteins was also unaltered after 8 Gy, in vitro irradiation. Other biochemical functional parameters of the isolated pancreatic acini, like protein synthesis and amylase secretion were not changed either after 8 Gy, in vitro X-ray treatment. These results indicate that pancreatic acinar cells in vitro have a high tolerance to irradiation. The observed in vivo radiation induced changes of the exocrine pancreas are possibly indirectly induced by injuries of more sensitive mechanisms.     

Morphological aspects of ionizing radiation response of small intestine

Knowledge of the acute and late ionizing radiation exposure damage to the gastrointestinal tract, particularly injury of the small intestine, is of great significance in radiotherapy, as is management of accidental radiation exposure. Irradiation (X-ray, neutron, cobalt gamma) induces a series of events in this rapidly renewing tissue resulting in the well-known symptoms of the gastrointestinal (GI) radiation syndrome, such as GI haemorrhage, endotoxemia, bacterial infection, anorexia, nausea, vomiting, diarrhoea, and loss of electrolytes and fluid. In spite of the significant advances that have occurred in research on underlying mechanisms over the last two decades, the overall etiology and pathogenesis of the GI-syndrome still remains unclear. Currently, to our knowledge, these symptoms are probably due to a rapid modification of the intestinal motility and to the structural alteration of the intestinal mucosa (cell loss and altered crypt integrity). Several evidences suggest that radiation-induced dysfunctions and structural changes of this organ (either changes in subcellular, cellular, and histological structure) are mediated by concerted and interrelated changes of a plethora of various extracellular mediators and their intracellular messengers. The aim of this review is to summarize our current knowledge about the pathomorphology and cell biology of the ionizing radiation response of the GI tract with a focus on the small intestine.     

Radiation response of cell organelles

The cellular responses to various form of radiation, including ionizing- and UV-irradiation or exposure to electromagnetic fields is manifested as irreversible and reversible structural and functional changes to cells and cell organelles. Moreover, beside the morphological signs related to cell death, there are several reversible alterations in the structure of different cell organelles. The radiation-induced changes in the supramolecular organization of the membranes, including plasma membrane, and different cell organelle membranes, play a significant role in the development of acute radiation injury. These signs of radiation-induced reversible perturbation biological membranes reflect changes in the organization and/or composition of the glycocalix, modified activity and/or distribution of different membrane domains, including enzymes and binding sites. The observed changes of the cell surface micromorphology and the alteration of intercellular connections are closely related to the reorganization of the cytoskeletal elements in the irradiated cells. The mitochondria, endoplasmic reticulum, Golgi-complex, the lysosomal system have long been considered to be direct intracellular targets of irradiation. The listed morphological alterations of nuclear chromatin (e.g. changes of fine structure, altered number of nucleolar organizing regions and micronuclei, development of chromosome aberrations) may originate from the radiation-induced damage to the supramolecular organization of DNA and/or nucleus specific proteins. These endpoints of radiation effects resulted as direct consequence(s) of absorbed radiation energy, and indirectly altered intra-, intercellular communication or modified signal transduction. Some complementary data suggest that all these effects are not strictly specific to radiation and may be best considered as general stress responses, similar to those observed after application of various injurious agents and treatments to cells. Moreover, they may be equally responsible for direct degradation of supramolecular component of cells, altered signal transduction, or changes in the amount or ratio of any extracellular mediators upon irradiation. Nevertheless, qualitative and/or quantitative evaluation of any changes of chromosomes by different techniques (morphological analysis of metaphase chromosomes, fluorescent in situ hybridization, development of micronuclei etc.) are useful biological indicators as well as "biological dosimeters" of radiation injury. It is suggested, that some modern methods such as immunohistochemical detection of different proteins, specific markers of cell organelles and cytoskeleton, inspection of distribution of cell surface charged sites and different membrane domains and application of tracer substances may all be included into protocols for evaluation of cell alterations induced by different types and intensities of radiation.     

INVESTIGATIONS ON BIOLOGICAL EFFECT OF POLARIZED LIGHT

Abstract— The biological effects of single and 4-time irradiation of primary human embryo fibroblasts with 4 J/cm² polarized light emitted by a halogen light source were investigated. The functional state of the plasma membrane was examined by means of lectin-binding and polycationized ferritin-binding techniques. It was established that the Con A binding of the cells did not change, whereas the number of negatively charged binding sites increased to a significant degree in relation to the untreated (control) samples and cell cultures exposed to diffuse (non-polarized) light. The micromorphological examinations showed no ultrastructural deviations. The quantitative increase of negative surface charges may be regarded as an indication of the biological effect of polarized light exerted on the cell membrane. The modifying effect of polarized light on the survival of E. coli exposed to the ionizing radiation was manifested in decreased anoxic radiation response.