NADP+ catabolic enzymes in differentiating rabbit erythroid cells

NADP-glycohydrolase and NADP-pyrophosphates activities were examined during the rabbit erythroid cell differentiation. The former is high in erythroblast lysates, especially in the erythroblast nuclei. As erythroid cell maturation proceeds, the activity of NADP-glycohydrolase decreases. At the first step (erythroblast-reticulocyte transformation), this activity falls down more than by 20 times, whereas at the second step (reticulocyte-erythrocyte transformation) it decreases no more than twice. NADP-glycohydrolase is associated with the stroma of erythroid cells throughout their maturation, being bound with the nucleus in erythroblasts. NADP-pyrophosphatase activity has been detected in reticulocytes and mature cells only. The role of NADP- and NAD-glycohydrolases for characterization of the intracellular metabolic pools is discussed.     

Changes in the enzymes related to 2,3-bisphosphoglycerate metabolism in developing erythroid cells

Measurements of glycerate-2,3-P2 and hemoglobin in the developing erythroid cells indicated that the glycerate-2,3-P2 level rose during erythroid differentiation in a linear relationship to the hemoglobin level, suggesting the presence of regulation to accumulate both substances synchronously. The accumulation of glycerate-2,3-P2 was found to be primarily attributable to the increase in glycerate-2,3-P2 synthase activity. The activities of phosphofructokinase and pyruvate kinase changed so as to be favourable for glycerate-2,3-P2 accumulation. The increase in glycerate-2,3-P2 synthase activity was shown to be caused by an increase in the enzyme protein. Synthesis of glycerate-2,3-P2 synthase protein was proved in bone marrow erythroid cells and in reticulocytes.     

Microglial cells of the rat brain in postnatal period (comparative immunocytochemical analysis)

In the course of the brain’s development, distribution of microglial cells was studied in rats using immunocytochemical detection. To identify the microglial cells, antibodies to lipocortin 1 (LC1) and phosphotyrosine (PT) were used. On postnatal day 1, LC1-positive microglial cells of an ameboid shape were distributed mainly in the subventricular zone; their mean density was 31±8 cells/mm² (counted across the total area of frontal sections). On postnatal day 7, microglial cells of an intermediate type were located throughout the whole brain; their density was 54±15 cells/mm². On the 15th day, LC1-positive cells were of a ramified shape, and their density reached 104±20 cells/mm² (the microglial cell density in the mature normal brain was 103±3 cells/mm²). On postnatal day 7, PT-positive cells were similar in their morphology to LC1-positive cells of an intermediate type, while their mean density was 32 cells/mm². In the mature brain, the density of PT-positive microglia was 53±5 cells/mm²; the shape of the cells in the white and gray matter of the brain was, on the whole, similar to that of LC1-positive microglia. Therefore, LC1 is a specific marker for different types of microglial cells in the developing brain. Our data about 3D distribution and morphological peculiarities of microglial cells at different stages of postnatal development are consistent with the hypothesis on the neuroectodermal origin of microglia.     

Selenium as a sulfhydryl redox catalyst and survey of potential selenium-dependent enzymes

The ability of selenium (Se) to act as a redox catalyst is an important factor in understanding the biological function of selenoproteins in addition to that of GSH peroxidase. Selenocystine at micromolar levels exhibited pseudothiotransferase activity by enhancing the reduction of 5,5-dithiobis-(2-nitrobenzoic acid) (DTNB) by thiols. In contrast, selenite inhibited the reduction of DTNB by thiols. Selenite was more catalytic than selenocystine in the reduction of cytochrome c by GSH, whereas GSH peroxidase was a weak catalyst. Tissues from Se-deficient and Se-supplemented rats were assayed for activities of GSH-thiotransferase, NADPH cytochrome c reductase, formaldehyde dehydrogenase, and a hypothesized GSH cytochrome c reductase. GSH-thiotransferase activity was significantly increased in the liver of Se-deficient rats. No appreciable activity of this enzyme was found in the kidney of rats from either dietary group. No enzymatic activity for cytochrome c reduction by GSH was detected in cytosols, mitochondria, or microsomes from liver and kidney of Se-deficient or Se-supplemented rats. Formaldehyde dehydrogenase was significantly higher in liver cytosols from Se-supplemented rats than from Se-deficient rats. The higher activity was not attributed to Se-containing proteins, but to an unknown small molecular-weight factor. This study did not support the hypothesis that physiological levels of Se may be involved in sulfhydryl-disulfide exchange reactions in vivo, or that selenium may enhance cytochrome c reduction by GSH in vivo.     

Intestinal enzymes activities in isolated villus and crypt cells during postnatal development of the rat

Summary A modification of Weiser's (1973) cell isolation method was used in order to study the developmental pattern of various intestinal enzyme activities in villus and crypt cells of normal rats from 5 days after birth until 8 weeks. Alkaline phosphatase and enterokinase activities were always located in the upper villus zone during postnatal development. Enterokinase activity was higher in the upper villus cells during the third week of life than after this period. Aminopeptidase activity was located in the crypt cells during the first week, its maximum activity remained in this area until the third week. At this time, sucrase activity appeared in the crypt cells, then aminopeptidase and sucrase activities rose to the villus zone during the fourth week. Amylase activity was detected along the entire crypt-villus axis 5 days after birth, reaching maximum activity in crypt cells at the end of the first week and in the upper villus cells after the fourth week. In contrast with the other enzymes studied almost all amylase activity was soluble in the youngest animals whereas at weaning most of the activity appeared in a particulate form in the villus cells. But in the crypt cells the ratio between particulate and soluble form remained unchanged until the adult stage. Various hypotheses are advanced to explain the patterns of evolution of the different enzymes.     

Activity of some mitochondrial enzymes in the progeny of rabbits treated with dichlorvos in the gestation period.

The activities of cytochrome oxidase (CYO) and succinate dehydrogenase (SDH) in brains of progeny of rabbits treated with dichlorvos (DDVP) 6 mg/kg/day during last ten days of pregnancy were investigated. Biochemical estimation of mitochondrial fraction of the brain tissue and the histochemical studies of fresh cryostat slices were carried out in 1, 8 and 16 day old rabbits. In comparison with control animals the decrease in CYO and SDH activity and the disturbances in "enzymatic maturation" rythm were observed in progeny of rabbits submitted to treatment with DDVP. In neuropil the changes were more pronounced than in pericarium of the nerve cells. In the brain structures which are sensitive to the metabolic disturbances (Ammon horn) SDH activities decreased.     

Combined reaction of the lymphoid organs and the adrenals to stress in mature animals subjected to cold in the early period of postnatal ontogeny

Structural-functional organization of the lymphoid organs and functional state of the adrenals have been studied in animals, subjected to cold in early postnatal period, as well as changes of the parameters mentioned to a short and prolonged cooling in mature rats. For the animals increase in the thymus mass and in reproduction rate of lymphocytes in the thymus, spleen and lymph nodes is specific against the background of high corticosteroid secretion. When the control animals are kept in cold for a long time, after the phase of an acute stress, accompanied with hypercorticism and a pronounced lymphatic effect, during the period of an increased cold stability, the high secretion of glucocorticoid hormones is accompanied with a certain activation of thymus-dependent zones in the peripheral lymphoid organs. In the mature rats, subjected to cold at early ontogenesis both stress-reaction to cooling and rearrangement in the regulatory systems studied does not develop at adaptation to cold.     

Activity of pentose phosphate pathway enzymes in alkane-oxidizing yeast cells]

The activity of the key enzymes of the pentose phosphate pathway (glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, transketolase) was determined in cell-free homogenates of Candida lipolytica 695 and Candida tropicalis 303 growing on different carbon sources. The activity of these enzymes remained almost the same in the course of growth of both cultures. The activity of the enzymes differed only slightly in the cells metabolizing hexadecane and glucose. The activity of glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in the cell-free homogenates of C. tropicalis 303 was twice as high as in the cells of C. lipolytica 695. The activity of transketolase was the same in both cultures. The main role of the pentose phosphate pathway is presumed to consist not in catabolism of the carbon source, but in biosynthesis of pentoses and other important intermediates.