THE CHANGES OF WOUND MACROPHAGES IN PATIENTS WITH DIABETES

Purulent-necrotic lesions of the extremities require amputation in 30-50% of cases. Among all cases of lower extremity amputations 50-70% are due to diabetes. Moreover, 5 out of 6 amputations, not related to the traumatic injury of the limb, are performed in patients with diabetes. The mortality rate among patients with diabetes, who undergo amputation varies from 28 to 40%, and 5-year surveillance is only 10-25%. The study of ultrastructural changes of macrophages on the 3rd day of treatment revealed masses of chaotically located fibrillar structures in the cytoplasm of macrophages that occasionally had an increased electron density. This phenomenon was observed in the purulent-necrotic areas of soft tissues of patients from the main group, compared to the control group. In all cases, mitochondria were enlarged in size, swollen, with a light matrix and contained a reduced amount of cristae. The cristae were deformed and shortened. Swollen matrix in mitochondria led to the formation of vacuoles on their place containing fine-grained contents. The nucleus had a usual form and size with the presence of single invaginations. Chromatin was predominantly concentrated in the form of solid electron-dense masses or evenly distributed throughout the nucleus. There were nuclei with partial chromatin dispersion. The contents of the nuclei included granular, fibrillar, and fine vacuolar material. The nuclear membrane folding did not fluctuate significantly. The folds did not cover the whole surface of the nucleus. In some areas invaginates were represented by the continuation of perinuclear space only. The nuclear envelop pores, which connect the contents of the cytoplasm and nucleoplasm, have been observed. The cytoplasm between the zones of the plate complex was occupied by small mitochondria, single polysomal rosettes, and cisternae of granular endoplasmic reticulum, which was represented by extended intracellular channels and vacuolar formations. The smooth endoplasmic reticulum was predominantly located in the central part. The surface of macrophages in the process of their differentiation from monocytes was relatively plane. Occasionally there occurred small processes or pseudopodia. The number of pinocytic vesicles surrounded by a border was reduced in poorly differentiated cells. Ozone therapy stimulates the functional activity of wound macrophages, as it causes destructive changes in these cells without necrotic lesions. Intravenous introduction of ozonized saline con-tributes to the elimination of wound macrophag-es, mainly through genetically programmed cell death (apoptosis), which plays a significant role in the regulatory mechanisms of the inflammato-ry process.

Ultrastructural and Molecular Changes in the Developing Small Intestine of the ToadBufo regularis

The ontogenetic development of the small intestine of the toadBufo regulariswas investigated using twofold approaches, namely, ultrastructural and molecular. The former has been done using transmission electron microscope and utilizing the developmental stages 42, 50, 55, 60, 63, and 66. The most prominent ultrastructural changes were recorded at stage 60 and were more evident at stage 63. These included the appearance of apoptotic bodies/nuclei within the larval epithelium, the presence of macrophages, swollen mitochondria, distorted rough endoplasmic reticulum, chromatin condensation, and irregular nuclear envelop, and the presence of large vacuoles and lysosomes. The molecular investigation involved examining DNA content and fragmentation. The results showed that the DNA content decreased significantly during the metamorphic stages 60 and 63 compared with both larval (50 and 55) and postmetamorphic (66) stages. The metamorphic stages (60 and 63) displayed extensive DNA laddering compared with stages 50, 55, and 66. The percentage of DNA damage was 0.00%, 12.91%, 57.26%, 45.48%, and 4.43% for the developmental stages 50, 55, 60, 63, and 66, respectively. In conclusion, the recorded remodeling of the small intestine represents a model for clarifying the mechanism whereby cell death and proliferation are controlled.

Nucleocytoplasmic movement of fluorescent tracers microinjected into living salivary gland cells.

The permeability of the nuclear envelop of a somatic cell, the C. thummi larval salivary gland cell, was studied by intracellular microinjection of fluorescent molecular tracers. As shown previously in oocytes (4,5,15,16), the envelop is permeable to a wide variety of materials, including molecules which are large enough to possess condiderable biological specificities and to play important roles in regulation of cellular activities. The envelop exhibits transport selectivity on the basis of size in the range of naturally occurring intracellular materials and it may thus perform important controlling functions in nucleocytoplasmic exchange. The nucleus to cytoplasm movement of in vivo ribonucleoprotein particulates in these synthetically active cells probably requires conformational changes in the particulates and/or the envelope pore complexes; morphological evidence exists for such processess in these cells (20).