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Biomedical subjects

B Agostini

Publications and source records attributed to B Agostini.

At least 19 recordsLinked to original sources

Platelet-neutrophil interactions in uremic patients: effects on neutrophil superoxide anion production and chemiluminescence.

Isolated resting platelets are able to limit neutrophil activation and then can control the tissue-damaging potential of activated neutrophils. In the present study, platelet-neutrophil interactions have been evaluated in 10 uremic patients; the blood samples have been collected before the hemodialysis session. Twelve normal subjects served as controls. Platelets and neutrophils have been isolated and recombined in an autologous ex vivo system. Anion superoxide production and chemiluminescence (which is related to hypochlorous acid production) have been evaluated after stimulation with N-formyl-methionyl-leucyl-phenylalanine. Coincubation of platelets from normal subjects with autologous neutrophils led to a dose-dependent inhibition of both superoxide anion generation induced by N-formyl-methionyl-leucyl-phenylalanine and chemiluminescence. Instead, platelets from uremic patients have not affected superoxide anion production by autologous neutrophils. The chemiluminescence was reduced by coincubation with autologous platelets only at the highest platelet-neutrophil ratio (100:1). In conclusion, the modulation exerted by platelets towards neutrophil activation can be impaired in chronic uremia. Therefore, the tissue-damaging potential of circulating neutrophils, due to toxicity by superoxide anion and hypochlorous acid, may be increased.

Adult

Sculpting present and future: a systemic intervention model applied to psychosomatic families.

We present a therapeutic intervention model for use with psychosomatic families. This method, the result of our extensive research on various psychosomatic disorders, uses family sculptures of the "present" and "future": each family member is requested to represent the family as it now "is," and how it "will be" in the future. We discuss the theoretical reasons for our choice of this method: (a) the opportunity to use a therapeutic language that is similar to the nonverbal language of the psychosomatic symptom, and (b) the usefulness of reinserting temporal dimensions into family systems that seem to have lost their evolutionary potential and to be in a sort of "time lock." Two clinical cases are discussed (a child with chronic asthma and an anorexic adolescent); the use of sculptures in both cases revealed the underlying problems and made positive therapeutic interventions possible. Finally, we point out how the use of sculpture as a therapeutic technique enables therapists to deal with multiple systemic levels.

Adolescent

Calcium uptake.

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Animals

Dipsogenic factors operating in chronic uremics on maintenance hemodialysis.

Thirst and hyperdipsia of anuric chronic uremics on maintenance hemodialysis and the possible dipsogenic factors were studied. Exaggerated thirst was present in 213 (86%) of the 247 studied patients. It usually started 4-6 h after the end of the dialysis session, persisted during the whole interdialytic period and often disappeared during the subsequent dialysis. Hyperdipsia, as indicated by the high body weight gain (> 4%) in the interdialytic periods, was present in 33.6% of patients. The highest rate of increase of body weight occurred in the first hours following the end of dialysis sessions. Hypernatremia, potassium depletion, increasing plasma urea levels and elevated plasma angiotensin II levels were considered as the possible dipsogenic factors of a nonpsychic nature. Sodium is certainly of paramount importance for its obliged extracellular position, and when sodium intake is elevated, hypernatremia is very likely the cause of exaggerated thirst and weight gain in patients on hemodialysis. Potassium depletion may cause thirst in animals, but this condition is extremely rare in patients on maintenance hemodialysis, who often accumulate it. In these patients it is, therefore, unlikely that potassium depletion is a dipsogenic factor. Increasing serum urea levels exert an evident dipsogenic effect in anephric rats and urea, when infused into normal volunteers, stimulates thirst. The extracellular urea levels in the interdialytic period are certainly higher than the intracellular ones, as a consequence of its continuous accumulation, and this creates an osmotic gradient with a dipsogenic effect. When this gradient is reversed, following hemodialysis (which removes first the extracellular urea), the dipsogenic effect disappears.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Expression of myosin heavy and light chains and phosphorylation of the phosphorylatable myosin light chain in the heart ventricle of the European hamster during hibernation and in summer.

We investigated the expression of myosin subunits (myosin heavy chains) as well as light chains and the in vivo phosphorylation of the phosphorylatable myosin light chain in the heart ventricle of the adult male European hamster (Cricetus cricetus L.). Two myosin heavy chain isoenzymes could be detected under native and denaturing electrophoretic conditions having high (alpha-myosin heavy chain) and low (beta-myosin heavy chain) enzymatic activity. Enzymatic activity of alpha- and beta-myosin heavy chain revealed a different temperature dependency. When temperature increased ATPase activity of the alpha-myosin heavy chain isoenzyme increased relatively more than ATPase activity of the beta-myosin heavy chain isoenzyme. Summer animals expressed predominantly the beta-myosin heavy chain (79% of total myosin) while during hibernation the alpha-myosin heavy chain expression increased to 53% of total myosin. Winter-active hamsters kept at 22 degrees C and 12 h day/night rhythm showed the same myosin heavy chain isoenzyme pattern as summer-active animals. Two myosin light chain forms were expressed in the ventricle of all animal groups. The in vivo phosphorylation level of the phosphorylatable myosin light chain decreased from 45% in summer-active hamster to 23% during hibernation.

Animals

On the problem of season and cold dependence of calcium transport by skeletal muscle sarcoplasmic reticulum.

Calcium transport of skeletal muscle sarcoplasmic reticulum from golden hamsters was studied in January and in June on animals kept at 22 degrees C under natural photoperiod and in January after cold-acclimation at +/- 2 degrees C in the dark for 55 days. Crude homogenates from psoas and soleus muscles and from mixed skeletal muscles were used. No differences were observed in the calcium storing capacity of sarcoplasmic reticulum among the three groups of animals. Kinetic studies on the dependence of the calcium uptake rate on the concentration of free calcium revealed a significant increase of the uptake rates and a decrease of the calcium affinity in the control animals sacrificed in winter as compared to those killed in June. Cold-acclimation in winter leads to a further small reduction of the calcium affinity. This shift of calcium uptake rate and affinity in the sense of that of a fast-twitch muscle may be related to the functional demands of the cold season and cold-acclimation respectively.

Acclimatization

The formation of intravesicular calcium phosphate deposits in microsomes of smooth muscle. A comparison with sarcoplasmic reticulum of skeletal muscle.

The calcium uptake in the microsomial fraction isolated from the smooth muscle of the antrum of the pig stomach is stimulated by phosphate. The microsomial vesicles which are loaded with calcium phosphate can be purified by differential centrifugation. A purification of 36 times in terms of calcium content was reached. Electron microscopy of the freshly prepared material revealed calcium phosphate deposits in the form of needles of crystalline calcium phosphate. This structure differs from that of the deposits which appear in the fragmented sarcoplasmic reticulum of skeletal muscle. Their morphology is that of non-crystalline calcium phosphate. However, on standing these deposits convert slowly into crystalline calcium phosphate. This difference reflects different kinetics of crystallization of the precipitates in the two preparations. After negative staining of the calcium phosphate loaded microsomes of skeletal and of smooth muscle, only few deposits are preserved because a release of calcium occurs as a consequence of the action of the stain and also of the dilution and warming up of the suspension. Smooth muscle microsomes partially purified by loading with calcium phosphate were studied by freeze etching and rotary replication. Membrane fragments displaying subunit intramembrane particles similar to those observed in sarcoplasmic reticulum of skeletal muscle could be identified. However, in the smooth muscle microsomes the intramembrane particles were much less densely packed. Part of these particles could correspond to calcium transport sites.

Animals

Electron cytochemistry of oxalate-stimulated calcium uptake in microsomes from the smooth muscle of the pig stomach.

A microsomal fraction was isolated from the smooth muscle of the antrum of the pig stomach by differential centrifugation. Electron microscopy of the negatively stained material showed that this fraction is heterogeneous in composition. The microsomes accumulated calcium in the presence of ATP, magnesium and oxalate. The amount of calcium taken up per mg protein was in the same range as observed for other smooth muscle microsomal preparations. Although this amount is much smaller than that in the microsomal fraction of skeletal muscle, calcium oxalate crystals were formed in some vesicles, as occurs in the skeletal muscle fragmented sarcoplasmic reticulum. Through the presence of the calcium oxalate crystals, many of these vesicles acquired sufficient mass and density to allow them to be isolated by centrifugation. A purification of about 40 fold in terms of calcium content was reached.

Adenosine Triphosphate

Evaluation by electron microscopy of the integrity of iodinated plasmalipoproteins.

Iodination of proteins and lipoproteins is a widely used "in vitro" labelling procedure in metabolic, autoradiographic and various other studies. However, all available iodination techniques have involved the possible damage to the proteins by self-irradiation, oxidizing agents, the alkaline milieu or by the introduction of iodine into the molecular structure itself. To evaluate the integrity of iodinated lipoprotiens, we observed the electron microscopic appearance of normal and iodinated rabbit very low density lipoproteins (VLDL) by negative staining with phosphotungstic acid. Iodination up to a molar iodine/protein ratio of 2.89 did not results in any change of shape, size or aggregating tendency of the particles. No stacks or disk-like particles like those of various hyperlipoproteinemic states were found. We conclude that electron microscopy is a valuable tool in assessing the morphological appearance of lipoprotein iodination, but it should be complemented by other techniques.

Iodoproteins

Effect of 1,25-dihydroxycholecalciferol on impaired calcium transport by the sarcoplasmic reticulum in experimental uremia.

In the fragmented sarcoplasmic reticulum from skeletal muscle of rabbits with experimental uremia, defective calcium ion transport is found. An impairment of all parameters is observed (initial rate of uptake, storing capacity with and without oxalate, and concentrating ability). In vivo administration of 1,25-dihydroxycholecalciferol (1,25-(OH)2-vitamin D3)(2 X 27 ng X kg of body wt-1 X day-1 and 6 X 27 ng X kg-1 X day-1, respectively) improved the kinetic parameters. The low dose improved storing capacity, and the higher dose, in addition to the storing capacity, also corrected concentrating ability and the initial rate of uptake. It is concluded that active calcium transport in the sarcoplasmic reticulum is impaired by uremia and that this defect is responsive to the administration of 1,25-(OH)2-vitamin D3.

Animals

Cytochemical studies on sarcoplasmic reticulum of heart and skeletal muscle.

Ultrastructural aspects and cytochemical localization of Ca uptake and of -SH groups involved in the calcium transport of vesicular fragments of sarcoplasmic reticulum (SR) from rabbit skeletal muscle and from dog myocardium were studied by positive and various negatively-staining techniques. Size, shape, and membrane structure as well as proportion of calcium-containing vesicles vary a great deal depending on the method applied. On negatively stained material sarcoplasmic reticulum derivatives are identifiable in both skeletal muscle as well as in heart preparations by the presence on the surface of the vesicular membrane of 40-A particles and the capability of accumulating calcium. The amount of these vesicles is lower in cardiac SR than in the skeletal SR; however, no qualitative differences can be observed. Vesicles of cardiac SR labeled with the electron dense -SH group reagent, Hg-phenyl azoferritin, also bind asymmetrically the marker only at the outer surface of the membrane-like microsomes of skeletal muscle.

Animals