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Biomedical subjects

F Franconi

Publications and source records attributed to F Franconi.

At least 19 recordsLinked to original sources

Further insights into the anti-aggregating activity of NMDA in human platelets.

1. In the present study the effect of N-methyl-D-aspartate (NMDA) on thromboxane B2 synthesis and on [Ca2+]i was studied in human platelets. 2. NMDA (10(-7) M) completely inhibited the synthesis of thromboxane B2 from exogenous arachidonic acid (AA), while it did not interfere with the aggregating effect of the thromboxane A2 receptor agonist U-46619. 3. NMDA (0.1 microM - 10 microM) dose-dependently increased intracellular calcium in washed platelets preloaded with fura 2 AM, and this effect was not additive with that of AA. 4. NMDA shifted the dose-response curve of AA to the right. At the highest AA concentrations platelet aggregation was not inhibited. 5. The antiaggregating effect of NMDA was not antagonized by N(G)-monomethyl-L-arginine (L-NMMA), a nitric oxide synthase (NOS) inhibitor. 6. Finally, NMDA (0.01 nM - 100 nM) associated with either aspirin or indomethacin significantly potentiated the antiaggregating activity of both cyclo-oxygenase inhibitors. 7. It was concluded that NMDA is a potent inhibitor of platelet aggregation and thromboxane B2 synthesis in human platelet rich plasma (PRP).

Aspirin

Recording of EEG during fMRI experiments: patient safety.

The acquisition of electroencephalograms (EEG) during functional magnetic resonance imaging (fMRI) experiments raises important practical issues of patient safety. The presence of electrical wires connected to the patient in rapidly changing magnetic fields results in currents flowing through the patient due to induced electromotive forces (EMF), by three possible mechanisms: fixed loop in rapidly changing gradient fields; fixed loop in a RF electromagnetic field; moving loop in the static magnetic field. RF-induced EMFs were identified as the most important potential hazard. We calculated the minimum value of current-limiting resistance to be fitted in each EEG electrode lead for a representative worst case loop, and measured RF magnetic field intensity and heating in a specific type of current-limiting resistors. The results show that electrode resistance should be > or = 13 k(omega) for our setup. The methodology presented is general and can be useful for other centers.

Electroencephalography

Acanthamoeba castellanii metabolites increase the intracellular calcium level and cause cytotoxicity in wish cells.

Previous studies have shown that trophozoites of the pathogenic free-living amoeba Acanthamoeba castellanii rapidly lyse a variety of cells in vitro. However, the role played by cytolitic molecules that may participate in Acanthamoebal cytopathogenicity has yet to be completely elucidated. The aim of this work was to study whether soluble molecules released by A. castellanii trophozoites could induce cytopathic effect in human epithelial cells in vitro. The results obtained indicate that A. castellanii trophozoites constitutively elaborate and release soluble factors that immediately elicit a cytosolic free-calcium increase in target cells. This phenomenon is induced by low molecular weight amoebic metabolites and depends on a transmembrane influx of extracellular calcium. Morphological changes, cytoskeletal damage, cell death and cytolysis followed the elevation of cytosolic free-calcium levels. Calcium ions are very important for cell homeostasis, in fact, they control the functions of a variety of cellular responses, including secretion, cell proliferation and apoptosis. Our results suggest that the substained elevation of the cytosolic free-calcium in response to A. castellanii metabolites might play a fundamental role in target cell damage during Acanthamoeba infections.

Acanthamoeba

Three-point Dixon method with a MISSTEC sequence.

We propose an adaptation of the MISSTEC sequence (simultaneous acquisition of a spin echo and several stimulated echoes) for performing fat suppression with a three-point Dixon method. In vivo measurements were performed on healthy volunteers using a sequence of three echoes (one spin echo and two stimulated echoes) within a third of the acquisition time taken by the regular three-point method, with the same spatial resolution and with half the signal-to-noise ratio (SNR). Homogeneity of fat suppression was greatly increased in comparison with the two-point method.

Algorithms

Raised erythrocyte polyamine levels in non-insulin-dependent diabetes mellitus with great vessel disease and albuminuria.

Erythrocyte content of polyamines has been previously found increased in insulin-dependent diabetes mellitus with microalbuminuria. Since increased urinary albumin excretion (AER) is associated with the presence of vascular diseases in non-insulin-dependent diabetes mellitus (NIDDM) the aim of this study was to verify the hypothesis that the presence of increased urinary albumin excretion (AER), and of macroangiopathy in NIDDM would be related to a significant modification in polyamine erythrocyte levels. The erythrocyte content of spermine and spermidine was measured by a HPLC method in 39 patients affected with NIDDM and in 24 age- and sex-matched healthy control subjects, evaluating the relationship between erythrocyte polyamines of NIDDM patients with the presence of macroangiopathy as well as with retinopathy or increased AER (> or = 20 micrograms/ml). Both spermidine and spermine were not modified in the group of NIDDM patients while the presence of raised urinary AER was characterised by an increase in erythrocyte spermine (11 +/- 1.7 vs. 7.7 +/- 1.7 nmol/ml packed erythrocytes; P = 0.04) and spermidine (18.9 +/- 1.7 vs. 12.6 +/- 1.5 nmol/ml packed erythrocytes; P = 0.02), being both polyamines significantly related to AER and to metabolic control. Erythrocyte spermidine and spermine were moreover significantly higher in the group of patients with macroangiopathy (22.8 +/- 1.5 vs. 12.3 +/- 1.5 nmol/ml; P = 0.0001 and 11.5 +/- 1.7 vs. 7.8 +/- 1.7 nmol/l packed erythrocytes; P = 0.04) and being, moreover, erythrocyte spermidine augmented in patients with retinopathy (24.2 +/- 1.5 vs. 12.2 +/- 1.5 nmol/ml packed erythrocytes; P = 0.009). In conclusion the levels of erythrocyte spermine and spermidine are both associated with the presence of albuminuria and macroangiopathy in NIDDM, while spermidine is on the average increased in the group of diabetic patients with retinopathy.

Albuminuria

Carbon centered radicals reduce the density of L-type calcium channels in rat cardiac membranes.

The carbon centered radicals generated by preincubation of cumene hydroperoxide with rat cardiac membranes dose dependently reduced the Bmax of [3H]nitrendipine binding, while KD was unchanged. The reduction induced by cumene hydroperoxide was prevented by 2,6-di-t-butlyl-4-methyl-phenol. The generation of OH did not influence the [3H]nitrendipine binding; also other oxidative agents (H2O2 and HClO4) did not modify this binding. Therefore the reduction in [3H]nitrendipine binding sites is not attributable to generic oxidative stress but to the formation of carbon centered radicals.

Animals

Frequency offset corrected inversion (FOCI) pulses for use in localized spectroscopy.

Gradient localized spectroscopy techniques suffer from a well documented spatial localization error caused by the difference in chemical shifts between resonances. This results in the acquisition of spectra from partially overlapping spatial regions of the sample, with each resonance representing a different region. The image-selected in vivo spectroscopy technique uses hyperbolic secant inversion pulses, where the main limitation in reducing this error is in the RF power available for application of the selective RF pulse. This spatial localization error may be dramatically reduced by increasing, and temporally shaping, the gradient pulse during slice-selective spin inversion. The performance of these RF pulses have been experimentally verified.

Magnetic Resonance Spectroscopy

Lactate, N-acetylaspartate, choline and creatine concentrations, and spin-spin relaxation in thalamic and occipito-parietal regions of developing human brain.

Previous studies of the brains of normal infants demonstrated lower lactate (Lac)/choline (Cho), Lac/creatine (Cr), and Lac/ N-acetylaspartate (Naa) peak-area ratios in the thalamic region (predominantly gray matter) compared with occipitoparietal (mainly unmyelinated white matter) values. In the present study, thalamic Cho, Cr, and Naa concentrations between 32-42 weeks' gestational plus postnatal age were greater than occipito-parietal: 4.6 +/- 0.8 (mean +/- SE), 10.5 +/- 2.0, and 9.0 +/- 0.7 versus 1.8 +/- 0.6, 5.8 +/- 1.5, and 3.4 +/- 1.1 mmol/kg wet weight, respectively: Lac concentrations were similar, 2.7 +/- 0.6 and 3.3 +/- 1.3 mmol/kg wet weight, respectively. In the thalamic region, Cho and Naa T2s increased, and Cho and Lac concentrations decreased, during development. Lower thalamic Lac peak-area ratios are principally due to higher thalamic concentrations of Cho, Cr, and Naa rather than less Lac. The high thalamic Cho concentration may relate to active myelination; the high thalamic Naa concentration may be due to advanced gray-matter development including active myelination. Lac concentration is higher in neonatal than in adult brain.

Aspartic Acid

NMDA receptors play an anti-aggregating role in human platelets.

Receptors for different monoamines, peptides and other neurohormones are present in the plasma membrane of platelets, and the sophisticated process of haemostasis is regulated by the interplay of their physiologic agonists (1). The recent report of a platelet binding site for phencyclidine (2) suggested a possible role of N-methyl-D-aspartate (NMDA) receptors in platelet function. Isotherms of [3H]-glutamate (GLU), [3H]-CGP-39653, [3H]-glycine (GLY) and [3H]-MK-801 carried out in platelet membranes yielded Bmax and Kd values for these ligands similar to those present in neurons, and NMDA only partially displaced [3H]-GLU. In neurons [3H]-MK-801 binding is potentiated by GLU and/or GLY and, being specific for the open NMDA receptor channel species, it has a functional meaning. In platelet membranes neither GLU and/or GLY increased [3H]-MK-801 binding; thus suggesting that NMDA receptors in platelets are different from those present in neurons. GLU or NMDA alone did not induce platelet aggregation. However, both amino acids were antagonistic on the aggregating activity of arachidonic acid (AA), NMDA being 3 orders of magnitude more active than GLU, and NMDA also antagonized adenosine diphosphate (ADP) and platelet aggregating factor (PAF) induced platelet aggregation. Finally, NMDA increased cAMP levels in intact platelets, and such an effect did not occur in a Ca(2+)-free medium; yet, cAMP increase was not antagonized by the calmodulin inhibitor trifluoperazine (TFP). It was concluded that platelet membranes carry an NMDA receptor, functionally distinct from the neuronal one, which seems to play an anti-aggregating role.

Cyclic AMP

[Use of a stimulated echo sequence in the MRI study of the brain and spine].

We describe in this paper how the STEAM sequence can be an efficient tool to obtain images free of flow artifacts in anatomical situation where the spin echo failed. The simplest way to eliminate flow artifacts is to exploit the dephasing induced by motion in magnetic field gradients and to reduce to zero the signal from moving tissues. This can be achieve by increasing the time elapsed between the spin excitation and the signal observed. Because of T2 relaxation, such an increase results in a signal decrease when the spin echo sequence is used. The STEAM sequence has the unique property that the time elapsed between observation and excitation can be increased without change in T2 value and so allows a good suppression of signals from the moving spins with short TE. Our results demonstrate that, although the stimulated echo intensity is only half that of a spin echo taken at the same read out time, the advantages of STEAM imaging can compensate for this partial loss in signal to noise in some particular clinical situations. The influence of mixing time on contrast has been evaluated using thoracic spine imaging and it has been shown that contrast between spine and CSF can be significantly improved (+ 60%) when TM is increased (from 17 ms to 50 ms). In the same time, the contrast between spine and fat issue decreases (40%). This last effect facilitates the adjustment of contrast window. Suppression of motion artifacts has first been evaluated with thoracic spine imaging, using a whole body coil. Suppression of artifacts was better than that obtained with a flow compensated spin echo sequence, especially in the case of kyphotic patients when a presaturation band was inefficient. In a second step suppression of motion artifacts has been evaluated from posterior fossa examination after injection of a paramagnetic contrast agent. The images obtained with the stimulated echo sequence show a dramatic reduction of signal from blood in the lateral sinus, and therefore an increase of quality by elimination of motion artifacts.

Adipose Tissue

Kinetic changes of alpha B crystallin expression in neoplastic cells and syngeneic rat fibroblasts at various subculture stages.

Alpha B crystallin, a structural at variable levels, in many extraocular tissues where it plays a protective role in stress conditions. In fact, heat or toxic shocks, as well as pathological states, increase alpha B crystallin levels in many cell types. Here we show that alpha B crystallin expression is also modulated in subcultures of rat fibroblasts and Galliera sarcoma cells. Western blots analysis with anti alpha B crystallin antibodies reveals the presence of the protein in both cell populations, although the kinetic pattern of expression is different. Galliera fibroblasts constitutively express the protein up to the 70th subculture and afterwards the synthesis ceases. On the other hand, Galliera sarcoma cells do not contain alpha B crystallin in the early stages of the culture, but there is a progressive increases between the 20th and 40th cell subculture. Differences also exist concerning the intracellular distribution: alpha B crystallin is diffusely localized in the cytoplasm of fibroblasts while in sarcoma cells it localizes mainly to the perinuclear region. Alpha B crystallin is totally recovered as soluble protein in the supernatants obtained after low speed centrifugation of fibroblast homogenates, while in sarcoma cells a portion of the protein is also recovered in the insoluble pellet. Intracellular pH measurements show an alkaline cytosol in sarcoma cells compared to fibroblasts. Heat shock treatment of fibroblast subcultures constitutively expressing alpha B crystallin induces an over-expression of the protein, while in fibroblasts whose biosynthetic capacity is lost, heat shock is unable to activate the crystallin gene. Correlation between alpha B crystallin expression and proliferative rate shows that highly proliferating fibroblasts do not express alpha B crystallin, while neoplastic cells do.

Animals

Effects of taurine and hypotaurine on lipid peroxidation.

It has been suggested that hypotaurine might inhibit lipid peroxidation in vivo by scavenging the initiator OH. The results presented demonstrate that hypotaurine affects other reactions relevant to the initiation, propagation and termination phases of lipid peroxidation. Hypotaurine a) decreases Fe2+ autoxidation, either spontaneous or catalyzed by Fe3+, that may generate perferryl iron; b) decreases Fe2+ oxidation, by cumene hydroperoxide, that forms the alkoxy radical; c) inhibits the lipid hydroperoxide dependent lipid peroxidation, favoring the onset of the termination phase. Hypotaurine does not affect the autoxidation of Fe2+ bound to phosphatidic acid containing liposomes. Taurine is ineffective in all the experimental systems tested.

Dimyristoylphosphatidylcholine

Alpha B crystallin is constitutively expressed in cultures of bovine articular chondrocytes.

alpha B crystallin is a small heat shock protein constitutively expressed in the mammalian lens and in a variety of extraocular tissues. We report here the presence of alpha B crystallin also in bovine articular chondrocytes by means of an immunoblot and immunofluorescence analysis carried out with anti-alpha B crystallin polyclonal antibodies. The expression level of alpha B crystallin can be further induced by a short heat shock treatment of chondrocytes as well as cell treatment with cadmium bromide or calcium ionophore A 23187. The level of alpha B crystallin expression is not modified by treating chondrocytes with interleukin-1 and phorbol 12-myristate 13-acetate. In some preparations the antibodies recognise two bands of alpha B crystallin, probably corresponding to different degrees of protein phosphorylation, but in cells treated with phorbol ester a single band is constantly observed, indicating a complete phosphorylation of alpha B crystallin.

Animals

Chemical shift imaging from simultaneous acquisition of a primary and a stimulated echo.

An imaging method is presented for obtaining chemical shift images from only one acquisition. Images are acquired with the same spatial resolution as in regular spin-echo imaging. The sequence is based on the simultaneous acquisition of a spin echo and a stimulated echo in a single pass. The use of 90 degrees radiofrequency pulse flip angles results in the same proton density, T2 and T1 weighting for the two echoes. Application of this sequence to chemical shift imaging is discussed for three fat suppression techniques (chopper, CHESS, and hybrid). Imaging was performed on phatoms and volunteers. The image quality was the same as those obtained by the chopper and the hybrid methods and the acquisition time was reduced by a factor of two.

Adipose Tissue

Plasma and platelet taurine are reduced in subjects with insulin-dependent diabetes mellitus: effects of taurine supplementation.

Plasma and platelet taurine concentrations were assayed in 39 patients with insulin-dependent diabetes mellitus (IDDM) and in 34 control subjects matched for age, sex, and both total and protein-derived daily energy intake. Platelet aggregation induced by arachidonic acid in vitro at baseline and after oral taurine supplementation (1.5 g/d) for 90 d was also studied. Plasma and platelet taurine concentrations (mean +/- SEM) were lower in diabetic patients (65.6 +/- 3.1 mumol/L, or 0.66 +/- 0.07 mol/g protein) than in control subjects (93.3 +/- 6.3 mumol/L, or 0.99 +/- 0.16 mol/g protein, P < 0.01). After oral supplementation, both plasma and platelet taurine concentrations increased significantly in the diabetic patients, reaching the mean values of healthy control subjects. The effective dose (mean +/- SEM) of arachidonic acid required for platelets to aggregate was significantly lower in diabetic patients than in control subjects (0.44 +/- 0.07 mmol compared with 0.77 +/- 0.02 mmol, P < 0.001, whereas after taurine supplementation it equaled the mean value for healthy control subjects (0.72 +/- 0.04 mmol). In in vitro experiments, taurine reduced platelet aggregation in diabetic patients in a dose-dependent manner, whereas 10 mmol taurine/L did not modify aggregation in healthy subjects.

Adult

Measurement of epidermal moisture content by magnetic resonance imaging: assessment of a hydration cream.

The moisture content of the epidermis was measured by magnetic resonance imaging (MRI), using transverse relaxation time. The spatial resolution was 86 microns, allowing a quantitative, accurate and localized determination of variations in epidermal hydration. The wrists of 15 volunteers were studied before and after application of a hydration cream. Results showed an increase of 15% of epidermal T2 after application of the cream. Moisture content curves varied according to different degrees of skin dryness. This study demonstrates that MRI is a useful tool in evaluation of epidermal hydration.

Administration, Cutaneous

Glutathione, glutathione utilizing enzymes and thioltransferase in platelets of insulin-dependent diabetic patients: relation with platelet aggregation and with microangiopatic complications.

Reduced glutathione (GSH) and activity of GSH related enzymes play a key role in defence against oxygen free radicals, whose production is, as known, raised in patients affected by diabetes mellitus, and at the same time they may contribute to the process of platelet aggregation. The purpose of this study was to evaluate GSH levels and activity of glutathione peroxidase (GSH-Px), glutathione reductase (GSSG-Red), glutathione transferase (GSH-Tr), glucose-6-phosphate-dehydrogenase (G6PDH), and thioltransferase (TT) in platelets of insulin-dependent diabetic patients in fair metabolic control (mean glycated haemoglobin: 6.5%), as related to presence of retinopathy, neuropathy or nephropathy and to platelet aggregation by arachidonic acid (AA) in vitro. Mean effective dose (ED50) of AA was on average significantly lower in the group of insulin-dependent diabetic patients (0.41 +/- 0.02 mM (SEM), n = 46) as compared with that of control subjects strictly matched for age, sex and weight (0.77 +/- 0.02, n = 51; P = 0.0001). Mean platelet GSH as well as the activity of GSH related enzymes expressed as geometric mean (95% confidence intervals) were similar in diabetic patients and in controls, except for GSSG-Red whose activity was significantly higher in diabetic subjects (28.5 (14.4-57.5) mU 10(-9) platelets vs. 20.3 (8.7-56) mU 10(-9) platelets; P = 0.01). In the diabetic group TT was reduced when compared with healthy controls (3.8 (0.9-12.2) mU 10(-9) platelets vs. 6 (1.6-26.1) mU 10(-9) platelets; P = 0.04).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult