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Biomedical subjects

M A Russo

Publications and source records attributed to M A Russo.

69 records · Page 4Linked to original sources

Ouabain-resistant mechanism of volume control and the ultrastructural organization of liver slices recovering from swelling in vitro.

We have studied the net extrusion of water by liver slices recovering from swelling at 1 degree C and have attempted to relate this to ultrastructural alterations. Special attention was paid to the ouabain-resistant extrusion of water. The restoration of many details of intracellular architecture was dissociated from the net loss of water, since an osmotic stimulus (produced by 5% inulin) caused a passive withdrawal of water with little recovery of structure. Also, a similar recovery of structure was produced during active extrusion of water in the absence and presence of ouabain, even though ouabain reduced the water extrusion by 50%. The time-course of water extrusion in the presence of ouabain was correlated with the formation of cytoplasmic vesicles. Incubation without K+ in the medium had similar effects to those caused by ouabain. Colchicine had little effect on water extrusion in presence or absence of ouabain except at concentrations which reduced tissue ATP levels and caused much necrosis. Cytochalasin B alone had little effect on water extrusion, but led to the accumulation of many vesicles in the cytoplasm and appeared to abolish the access of such vesicles to the canaliculi. In the presence of ouabain, cytochalasin B had a similar effect on ultrastructure, and totally prevented the ouabain-resistant water extrusion. Ni2+ had rather similar effects to cytochalasin B both in the presence and absence of ouabain, although to a smaller degree. The results support our previous suggestion that the ouabain-resistant water extrusion proceeds by secretion of water into cytoplasmic vesicles, followed by the exocytotic expulsion of the vesicular contents into the bile canaliculi. Microfilaments appear to play an important role in the process.

Animals↗

Surface interaction in vitro between Sertoli cells and germ cells at different stages of spermatogenesis.

The presence of surface-recognition mechanisms between somatic and germ cells of the seminiferous epithelium has been studied in the rat by an assay in vitro based on the ability of homogeneous populations of spermatogenic cells, at specific differentiative stages, to adhere to monolayers of cultured Sertoli cells. The results show that germ cells adhere specifically to Sertoli cells and that the adhesion is dependent on the differentiative stage of the germ cells. Pachytene spermatocytes show the highest ability to adhere and form typical junctional specializations with the underlying Sertoli cells, while round spermatids adhere much less to the substrate. The possible regulative role of a somatic cell-germ interaction is discussed.

Animals↗

Immunochemical and ultrastructural study of multiple myeloma with a heavy chain protein in the serum.

A patient with multiple myeloma had antigenically related monoclonal Fc-gamma fragments and complete IgG-kappa molecules in the serum. The urine contained only Fc-gamma fragments in the absence of Bence-Jones protein. The two distinct M-components in the serum showed electrophoretic identity but could be separated by chromatography. The simultaneous presence of complete monoclonal IgG molecules and Fc-gamma fragments, though difficult to detect, could be a frequent occurrence in multiple myeloma, and it could be defined as 'double paraproteinaemia'. A detailed ultrastructural study was performed in this case and showed fibril bundles being released from the malignant plasma cells; such fibrils could be the supramolecular organisation of the neosynthesised heavy chain fragments.

Aged↗

Observations on the regulation of cell volume and metabolic control in vitro; changes in the composition and ultrastructure of liver slices under conditions of varying metabolic and transporting activity.

Liver slices incubated at 1 degree C underwent swelling of both cellular and intercellular compartments, as judged by electronmicroscopy. The ultrastructure showed marked changes, including disorganization of the cytocavitary network and plasma membrane and alterations of mitochondria. Restoration of metabolically favorable conditions (oxygenated medium at 38 degrees C) caused a nearly complete recovery of ultrastructure closely associated with extrusion of water; measurements of inulin space and electronmicroscopy both indicate a recovery of cell volume, with intercellular spaces remaining somewhat expended. The fluid lost was a roughly isotonic solution of Na+ and Cl-, while K+ was reaccumulated in exchange for Na+. Cyanide prevented recovery. Ouabain and oligomycin each partially prevented fluid extrusion, but had little effect on ultrastructural recovery except to induce intracellular vesicles containing particles of thorium dioxide derived from sinusoidal spaces. The vesicles were, however, markedly different in form with each inhibitor. There are, thus ouabain-sensitive and insensitive components of volume regulation; the former appears to depend on the coupled transport of Na+ and K+ and the latter, we suggest, on a secretion of Na+ and Cl- into vesicles which release their contents into the bile canaliculi by an oligomycin-sensitive mechanism. Mitochondria showed conformational changes between orthodox and condensed forms, but these could not be directly related to tissue energy states; the numbers of mitochondrial dense granules bore a closer relation to tissue ATP.

Animals↗

Subcellular reactions to injury. I. Ultrastructural and biochemical investigations on the hepatic cellular damage produced by haemorrhagic shock in dogs.

The ultrastructural changes produced by haemorrhagic shock in hepatocytes have been studied in dogs. The associated biochemical and biophysical system alterations allow us to propose a unitary pathogenetic mechanism of the cellular injury based on oxygen deprivation and its effect on the function and structure of the cellular compartments.

Animals↗

The metabolism-dependent maintenance of cell volume and ultrastructure in slices of Morris hepatoma 3924A.

The changes in water and electrolyte content of slices of Morris hepatoma 3924A induced by various conditions of incubation have been compared with the ultrastructural appearance of the tissue. Incubation at 1 degrees led to an increase of water, Na+, and Cl- content and to a loss of K+. There was simultaneous increase in size of the cells and intercellular spaces, loss of junctional complexes, increase in the number of microvilli, and fragmentation and dilation of the cytocavitary network. Subsequent incubation at 38 degrees in oxygenated medium led to a substantial reversal of all of these changes of composition and structure, which was well advanced within 10 min and largely complete by 60 min. The presence of 20 mM glucose in the medium somewhat enhanced the degree of recovery. A reduction of cell volume and intercellular spaces was evident both from the electron microscopic observations and measurements of the volume of inulin distribution. The presence of ouabain inhibited the net accumulation of K+ and much of the Na+ extrusion, but permitted about 50% of the net extrusion of water (accompanied by Na+ and Cl-) and had little effect on the ultrastructural recovery. The presence of glucose increased the resistance of volume and structural recovery of ouabain without releasing the inhibition of K+ accumulation. A marked feature of the recovering tissues was the Golgi apparatus, which assumed an appearance suggestive of increased activity when water extrusion was active. In slices using only endogenous substrate, cyanide and (to a lesser extent) oligomycin greatly inhibited the recovery of volume and structure. The presence of glucose permitted some recovery in the presence of cyanide. The control of cell volume in hepatoma 3924A appears to involve two separate components of water transport, one of which is sensitive, and one insensitive to ouabain. The ouabain-insensitive component appears to be especially related to the recovery of cell ultrastructure after incubation at 1 degrees, to be more sensitive to paucity of adenosine 5'-triphosphate, and to proceed by secretion of water, Na+, and Cl- into vesicles that fuse with the Golgi apparatus. This mechanism may be related to that for bile secretion in normal liver. The ouabain-sensitive component of water transport is a function of the mechanism for the coupled transport of Na+ and K+.

Anaerobiosis↗

Interaction of Na+ and K+ transport with aerobic energy metabolism in slices of Morris hepatoma 3924A.

Addition of increasing concentrations of glucose to slices of Morris hepatoma 3924A greatly stimulated aerobic lactate production and reduced respiration by 20%. Neither the adenine nucleotide content of the slices nor the calculated rate of adenosine 5'-triphosphate synthesis was altered. Ouabain reduced the rate of O2 uptake (by 20 to 25%) and of aerobic lactate production (by 25 to 50%) without affecting adenine nucleotide contents. The reduction by ouabain of the calculated rate of adenosine 5'-triphosphate synthesis was similar whether the slices were utilizing only endogenous substrate or exogenous glucose also. Raising the medium K+ concentration (and correspondingly reducing Na+) partially overcame the inhibition of ion transport by ouabain and partially restored the rates of respiration and aerobic lactate production toward control levels. Electron microscopic observations of mitochondria within the slices incubated under different conditions showed variations in configuration between "orthodox," "condensed" and degenerating forms. Slices preincubated at 1 degrees showed mitochondria in the condensed form: they were restored to the orthodox configuration during incubation at 38 degrees in oxygenated medium. Oligomycin and glucose enhanced the transition, but ouabain reduced the number of mitochondria undergoing the change. The results suggest that in hepatoma 3924A utilization of adenosine 5'-triphosphate by ion transport exerts a simultaneous control of both respiration and aerobic glycolysis, which is presumably mediated by alterations in the availability of adenosine 5-diphosphate. The mitochondria undergo conformational transitions under conditions likely to affect local availability of adenosine 5'-diphosphate within cell compartments, but the transitions are not all externally added adenosine diphosphate on isolated mitochondria.

Adenine Nucleotides↗

Hormonal regulation of cytokine release by human fetal membranes at term gestation: effects of oxytocin, hydrocortisone and progesterone on tumour necrosis factor-alpha and transforming growth factor-beta 1 output.

Inflammatory cytokines can play an important role in the biomolecular processes leading to labour by regulating prostaglandin production in intrauterine tissues. In the setting of intrauterine infection, an increased production of these cytokines by placenta, decidua and fetal membranes occurs and is responsible for the onset and maintenance of preterm labour. However, the factors involved in the control of cytokine release by these tissues in normal pregnancy at term are still largely unknown. We investigated the possibility that the synthesis and release of tumour necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) by human fetal membranes at term gestation is regulated by several hormones potentially involved either in the maintenance of pregnancy or in the parturitional process. In the present study, the effects of hydrocortisone, progesterone and oxytocin on TNF-alpha and TGF-beta1 release by explants of fetal membranes at term gestation were evaluated. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to assess the effect of the above hormones on mRNA expression; TNF-alpha and TGF-beta1 release in culture medium was quantitifed by ELISA assays. Results show that both tissue mRNA expression for TNF-alpha and TNF-alpha release in culture medium were significantly increased by oxytocin, but not by hydrocortisone and progesterone. On the contrary, all the hormones tested increased both tissue TGF-beta1 mRNA expression and release in culture medium. These findings suggest that TNF-alpha and TGF-beta1 production by human fetal membranes in uncomplicated pregnancy at term is selectively modulated by oxytocin, hydrocortisone and progesterone.

Extraembryonic Membranes↗

Acute myopathy associated with chronic licorice ingestion: reversible loss of myoadenylate deaminase activity.

A patient with acute rhabdomyolysis and absence of myoadenylate deaminase (MADA) associated with chronic licorice intoxication is presented. Clinical and laboratory examination of the patient and morphologic study over skeletal muscle were performed. The major effect of licorice intoxication is hypokalemia, which may explain most of the observed clinical symptoms and morphological changes. The absence of MADA may be a consequence of the direct toxic effect of licorice glycosides. To our knowledge, this is the first report in which a lack of MADA and chronic licorice intoxication has been shown to be associated with clinical, histochemical, biochemical, and morphological changes, which were completely reversed with potassium supplementation and licorice withdrawal.

AMP Deaminase↗