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R Bianchi

Publications and source records attributed to R Bianchi.

At least 91 records · Page 5Linked to original sources
PubMed1998

Expression of Fos immunoreactivity in the rat supraspinal regions following noxious visceral stimulation.

We used immunohistochemical detection of the Fos protein to study the neuronal activation in the brain of methoxyfluorane-anesthetized rats after noxious deep somatic or visceral stimulation. The anesthesia was effective in triggering gene induction in many brain regions. Nevertheless, Fos appeared de novo in several brain nuclei following noxious stimulation in anesthetized animals. This could be of clinical relevance, as it suggests that the gas anesthetic does not suppress noxious stimulus-evoked reactivity in brain neurons. Two types of visceronociceptive stimuli were used to compare the effects of a diffuse visceral inflammation (peritoneal inflammation) with those of a more restricted inflammation (urinary bladder inflammation). In the same supraspinal areas, there were very few immunostained neurons in unstimulated controls, whereas Fos-positive cells were slightly more numerous in anesthetized controls and significantly more numerous after noxious stimulation. The peritoneal inflammation induced more Fos-labeled neurons than the restricted visceral stimulation. Labeled cells were found in these cases mainly in the ventrolateral medulla, parabrachial complex, dorsal raphe nucleus, periaqueductal gray, several hypothalamic and thalamic nuclei, amygdaloid complex, and cortex. Altogether these findings indicated that somatic and visceral inputs generally activate the same neuronal groups. However, a separation between the activation of somatic and visceral pathways was found in some brain nuclei, such as the parabrachial complex, hypothalamic, and thalamic nuclei.

Animals↗
PubMed1998

Mitochondrial membrane potential and DNA stainability in human sperm cells: a flow cytometry analysis with implications for male infertility.

Sperm cells from control donors of proven fertility and men from barren couples were studied by conventional procedures, i.e., light microscopy as well as flow cytometry. Light microscopy analysis of semen included the measurement of spermatozoa concentration, morphology, and motility. All the men from barren couples were asthenozoospermic at the conventional analysis of semen samples. Flow cytometry was applied to study two important parameters of sperm cells: mitochondrial membrane potential (MMP) assessed by the cationic dye JC-1 and DNA stainability with propidium iodide (PI). JC-1 staining was more reliable than the classical procedure used for this purpose, i.e., rhodamine 123 (Rh123) staining, and allowed us to show a positive correlation between MMP and spermatozoa motility. Regarding DNA analysis, a higher relative percentage of immature spermatozoa, showing a high accessibility of DNA to the intercalating PI fluorochrome, was found in men from barren couples compared to donors of proven fertility. The relative percentage of immature spermatozoa was significantly higher in semen from oligoasthenozoospermic subjects. Moreover, a positive correlation was found between immature spermatozoa, as evaluated by PI staining, and cells with depolarized mitochondria, as evaluated by JC-1 staining, suggesting that spermatozoa defective for nuclear maturity could be functionally defective cells. No correlation between immature spermatozoa determined by FCM and immature spermatozoa determined by light microscopy was found, suggesting that these two techniques assess sperm cell maturity at different levels.

Adult↗
PubMed1998

Induction of NADPH-diaphorase activity in the rat periaqueductal gray matter after nociceptive visceral stimulation.

Nitric oxide (NO) is a neuronal messenger that it is thought to be involved in the nociceptive transmission modulation. The activity of nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) was shown to be identical to NOS activity in the brain. Since the periaqueductal gray matter (PAG) plays an important role in pain perception and antinociception this study was carried out to monitor the expression of NADPH-d in PAG after nociceptive visceral stimulation. Our data showed that the noxious visceral stimulation significantly increased NADPH-d positive neurons and that these neurons were localized in the ventrolateral areas of the PAG. These findings suggest that NO in the PAG may play a role in pain modulation and antinociception.

Acetic Acid↗
PubMed1998

Dendritic arborization and spines of the neurons of the cat and human periaqueductal gray: a light, confocal laser scanning, and electron microscope study.

Neurons of the periaqueductal gray (PAG) have an extensive dendritic tree which plays an important role in the neuronal circuits supporting the functional activities of this region. The complexity of the local circuits is increased by the occurrence of dendritic spines. We have compared the dendritic and spine organization in the cat with that of man in order to verify whether an inverse relationship exists between dendritic tree extension and spine density and complexity. Sections of cat and human PAG prepared according to the Golgi-Cox method were studied with the conventional light microscope (LM) and the confocal laser scanning microscope (CLSM). The cat PAG was also studied at the electron microscopic level. The light microscopic study provided the morphoquantitative characteristics of the dendritic arborization and spines of the multipolar and fusiform neurons of the human and cat PAG. The CLSM methodology, thanks to the three-dimensional reconstruction of the neurons and the rotation of the reconstructed images, brought into view dendritic branches and spines that could not have been observed at the LM, thereby showing a wider dendritic tree and more numerous spines. The data combined from LM and CLSM demonstrate that in both species most spiny neurons are multipolar and probably projection neurons. In man, the multipolar neurons show a more extensive dendritic tree due to a wider secondary ramification, which would seem to be balanced by more numerous spines in cat. At the electron microscopic level, axo-dendritic synapses are numerous and show symmetrical and asymmetrical junctions in equal proportions; furthermore, the great majority of the spines are in contact with synaptic boutons which contain round vesicles and make predominantly asymmetrical contacts features which indicate excitatory activity. The combined use of different techniques gave a complete picture of the dendritic tree and spines of the neurons of human and cat PAG and showed a wider dendritic surface available for the receipt of the synaptic contacts than had been reported previously. Furthermore, our findings demonstrate that the PAG dendritic spines are important and specific structures in the synaptic complex of the neuropil, suggesting that they might create a local device to modulate and integrate the afferent inputs, probably in an excitatory way. The differences observed in the two species suggest that afferent information might be handled in different ways in human and cat PAG.

Adult↗
PubMed1998

Serum macrophage colony-stimulating factor (M-CSF) levels in patients with epithelial ovarian cancer.

The preoperative macrophage colony-stimulating factor (M-CSF) levels were measured in serum samples from 56 patients with epithelial ovarian cancer and 68 patients with benign ovarian disease who had undergone laparotomy. M-CSF values were significantly higher in the former (median, range: 2.18, 0.70-10.00 ng/ml versus 1.19, 0.17-5.54 ng/ml, P < 0.0001), and were not significantly related to stage, histology, grade of differentiation, age, and residual disease after first surgery. M-CSF concentrations were also measured in 163 serum samples drawn from patients with stage III-IV epithelial ovarian cancer at different times since the first surgery. M-CSF values were higher in the 81 samples from patients with clinically evident disease than in the 82 samples from patients with no clinical evidence of disease (median, range: 2.13, 0.60-10.00 ng/ml versus 1.05, 0.40-10.00 ng/ml, P < 0.0001). M-CSF levels before second-look laparotomy were similar in the 18 patients who showed persistent disease at surgical reevaluation and in the 11 patients who achieved pathological complete response (median, range: 1.26, 0.70-3.27 ng/ml versus 0.94, 0.46-4.23 ng/ml, P = NS). M-CSF concentrations were raised (> or = 1.70 ng/ml) only in 1 (14.3%) of the 7 samples from patients with clinically evident disease and serum CA125 < 35 U/ml, and only in 5 (38.5%) of the 13 samples from patients with positive second-look findings and serum CA125 < 35 U/ml. In conclusion, serum M-CSF levels correlated with the clinical status of disease in patients with epithelial ovarian cancer. However, the concomitant determination of serum M-CSF seems to add little to the CA125 assay alone in the monitoring of patients with this malignancy.

Adult↗
PubMed1998

Changes in diastolic function and collagen content in normotensive and hypertensive rats with long-term streptozotocin-induced diabetes.

We investigated the relationship between left ventricular diastolic function and interstitial collagen content in the endocardium, mesocardium and epicardium of transverse sections of the heart, using an image analysis system in normotensive and hypertensive long-term streptozotocin (STZ) diabetic rats. STZ-induced diabetes was characterised by elevated blood glucose, polyuria, polydypsia and loss of body weight. In vivo systolic blood pressure was 165 +/- 4, 136 +/- 3 and 129 +/- 7 mmHg in hypertensive and normotensive diabetic rats and age-matched controls, respectively. Heart rate was significantly lower (P < 0.01) in diabetic rats (283 +/- 8 and 280 +/- 10 beats min-1 in normotensive and hypertensive rats, respectively) than controls (393 +/- 18 beats min-1). Pressure-volume (P-V) curves were studied in isolated Langendorff perfused hearts at rest and after 20 min global ischaemia and 30 min reperfusion 6 months after induction of diabetes. Left ventricular volumes were significantly smaller in diabetic rats than age-matched controls, but volumes normalised for heart weight were higher in normotensive (by 28%) and hypertensive (by 10%) diabetic rats. Slopes of end-diastolic P-V curves were similar between groups in basal conditions, but left ventricular systolic P-V curves were steeper in normotensive and flatter in hypertensive diabetic hearts. Post-ischaemic left ventricular end-diastolic pressure was significantly higher than the pre-ischaemic value at comparable increments of volume in each group. Collagen content significantly increased in the heart of rats with STZ-diabetes both in the free left ventricular wall and septum, and suggested this may play a role in the cardiac defects in contractility and relaxation in our experimental conditions. These results indicate that diabetes, irrespective of associated hypertension, can cause major changes in cardiac performance and susceptibility to ischaemia and reperfusion.

Animals↗
PubMed1998

Trocar site tumor recurrences. May pneumoperitoneum be responsible?

BACKGROUND: Port site metastasis following laparoscopy for cancer is reported with increasing frequency and represents one of the most important limitations of the technique. METHODS: A scintigraphic model was utilized to evaluate a possible role of pneumoperitoneum in tumor cell dissemination. Labeled red blood cells (RBC) were injected at the level of the gallbladder bed during laparoscopic cholecystectomy (LC) performed for symptomatic cholecystolithiasis. LC was performed in two groups with standard CO2 pneumoperitoneum: in one group an endobag for retrieval of the specimen was utilized. In one group a gasless LC with endobag was performed. RESULTS: Radioactivity in the area of the trocar introduction was observed in almost all the patients who underwent standard (CO2) LC but represented a rare event in patients treated with the gasless method. The utilization of a protective bag for the extraction of the surgical specimen did not modify significantly the results. Moreover all patients treated with pneumoperitoneum demonstrated a wide intraperitoneal diffusion of the tracer not observed in gasless patients. CONCLUSIONS: The results of this study confirm that pneumoperitoneum may play an important role in the evolution of port site metastasis after laparoscopy for gastrointestinal cancer.

Cholecystectomy, Laparoscopic↗
PubMed1998

IL-12 acts directly on DC to promote nuclear localization of NF-kappaB and primes DC for IL-12 production.

We analyzed the expression of an IL-12 receptor by fresh dendritic cells (DC) and a DC line. Using RT-PCR, RNAse protection, and electrophoretic mobility shift assay analysis, we found that DC possess an IL-12 receptor with beta1 subunit (downstream box 1)-related differences from that on T cells. IL-12 signaling through this receptor involved members of the NF-KB but not STAT family. The unique properties of the IL-12 receptor on DC, characterized by a single class of binding sites with a Kd of about 325 pM, may underlie rather unique effects, such as IFNgamma-independent augmentation of class II antigen expression and priming for LPS-induced production of IL-12.

Animals↗
PubMed1998

Supraspinal connections and termination patterns of the parabrachial complex determined by the biocytin anterograde tract-tracing technique in the rat.

We have re-evaluated, using the anterograde tracer biocytin, supraspinal efferent projections from the parabrachial complex (PBN) to gain new information about the nature of its connections and nerve terminal patterns. We selectively injected biocytin into the 3 main regions of the nucleus (lateral PBN, medial PBN and Kölliker-Fuse nucleus). We observed distinct groups of ascending and descending fibres of different calibre from the PBN running throughout the brain and reaching many brain areas involved in the regulation of autonomic function. Here we detected labelled bouton-like terminals and fibres with en-passage varicosities. The ascending efferents from the lateral PBN mainly reached the reticular, raphe and thalamic nuclei, the zona incerta (ZI), central nucleus of the amygdala (CeA) and lateral area of the periaqueductal grey (PAG). Thin descending efferents reached the ventral region of the solitary tract nucleus (STN). The ascending efferents from the medial PBN were seen in the raphe nuclei, reticular nuclei, ventral and lateral areas of the PAG, thalamic nuclei, and in the medial and lateral nuclei of the amygdala. Descending efferents were seen in the STN and in some reticular nuclei. The ascending projections from the Kölliker-Fuse targeted the ventral area of PAG, CeA, ZI, lateral hypothalamic area, ventromedial thalamic nucleus and, with only a few terminals, the ipsi and contralateral reticular area. A large number of descending efferents reached STN, caudal and paragigantocellular reticular nuclei. The higher sensitivity of biocytin compared with other types of markers allowed us to determine more effectively the distribution, nature and extent of the supraspinal PBN connections. This suggested that in several nerve circuits the PBN probably plays a more important role than previously thought.

Animals↗
PubMed1998

Acute exogenous elevation of plasma free fatty acids does not influence the plasma magnesium concentration.

OBJECTIVE: Plasma non-esterified (free) fatty acid concentrations rise as a consequence of stimulated endogenous lipolysis and are inversely related to the plasma magnesium concentration when plasma adrenaline concentration is increased. The aim of the study was to test whether high plasma non-esterified fatty acid concentration after infusion of non-esterified fatty acids decreases plasma magnesium concentration. METHODS: Twelve healthy subjects received 500 ml Intralipid or saline in a randomised, cross-over, double-blind design. Infusion of Intralipid results in an isolated elevation of plasma non-esterified fatty acid concentration. Plasma magnesium concentration was determined at baseline and every 30 minutes; plasma non-esterified fatty acid and triglyceride concentrations at baseline and after 120 minutes. RESULTS: Initial plasma magnesium, non-esterified fatty acid, and triglyceride concentrations were similar in both groups. A significant increase in plasma non-esterified fatty acids (2.42 +/- 0.96 mmol/l vs 0.58 +/- 0.23 mmol/l, p = 0.00013) and triglyceride (median and 95th percentile 5.36 (7.35) mmol/l vs 1.18 (1.92) mmol/l, p = 0.003) concentrations was seen with Intralipid. Plasma magnesium concentration increased significantly after Intralipid (0.89 +/- 0.09 mmol/l vs 0.81 +/- 0.06 mmol/l, p = 0.007). No significant changes were seen with saline. A positive association was found between the change in plasma magnesium and triglyceride concentrations (r = 0.85, p = 0.001). CONCLUSION: Acute infusion of non-esterified fatty acids from an exogenous source does not result in a fall in plasma magnesium concentration, indicating that the circulating non-esterified fatty acids play no part in a decrease in plasma magnesium concentration. The high circulating non-esterified fatty acid levels and the fall in plasma magnesium concentration are both a consequence of intracellular lipolysis.

Adult↗
PubMed1998

Dendritic cells, interleukin 12, and CD4+ lymphocytes in the initiation of class I-restricted reactivity to a tumor/self peptide.

Cell-mediated immunity involving CD8+ lymphocytes is effective in mediating rejection of murine mastocytoma cells bearing P815AB, a tumor-associated and self antigen showing similarity to tumor-specific shared antigens in humans. Although this antigen may act as an efficient target for class I-restricted responses in immunized mice, neither P815AB expressed on tumor cells nor a related synthetic nonapeptide will activate unprimed CD8+ cells for in vivo reactivity, measured by skin test assay. We review evidence showing that the failure of P815AB to initiate CD8+ cell reactivity may be due to defective recruitment of accessory and Th1-like cells to the afferent phase of the response initiated by transfer of mice with dendritic cells pulsed in vitro with the P815AB peptide. Although the copresence of a T helper peptide in dendritic cell priming in vitro with P815AB may compensate for the poor generation of accessory and Th1 cells in the adoptively transferred mice, recombinant IL-12 can replace the helper peptide in both effects. Effective priming to P815AB in vivo is achieved by either exposing dendritic cells to IL-12 prior to P815AB priming or administering the recombinant cytokine in vivo. Different approaches suggest that IL-12 may act both on accessory cells to improve presentation of previously undescribed class II-restricted epitopes of P815AB and on CD4+ cells to improve recognition of such epitopes. In particular, at the CD4+ cell level, IL-12 apparently acts as an adjuvant and an inhibitor of anergy induction. These data offer useful information for developing vaccination strategies using dendritic cells and class I-restricted tumor peptides in humans.

Adjuvants, Immunologic↗
PubMed1998

Serum levels of tumor associated trypsin inhibitor (TATI) and glomerular filtration rate.

TATI (tumor associated trypsin inhibitor) is a low molecular weight protein employed as a tumor marker. To evaluate the role of the kidney in the clearance of TATI, we studied the rat kidney uptake of 125I-TATI. Total body scan demonstrated a high radioactivity in the kidneys of the rats and none in other organs. The relationship between serum TATI and glomerular filtration rate (GFR) was studied in man. For comparison serum beta 2-microglobulin (beta 2M) arid plasma creatinine were also determined. The decrease in GFR was accompanied by an increase in the other parameters. Serum TATI increased in patients with renal failure (GFR < 20 mL/min) 12.4 times with respect to subjects with normal renal function (p < 0.001, non-parametric Mann-Whitney test), beta 2M increased 7.6 times (p < 0.001) and creatinine 4.7 times (p < 0.001). The increase in TATI is statistically significant already in patients with GFR 60-40 mL/min (p < 0.005). These results suggest that TATI is handled by the kidney. It is a sensitive marker of reduction in renal function.

Adolescent↗
PubMed1998

[Sinusoscopy in stomatology. Review of a 5-year clinical experience].

BACKGROUND: Investigative sinus endoscopy for purely diagnostic purposes has become an important therapeutic tool for disorders of the paranasal system thanks to the enormous technological developments of the last twenty years. METHODS: In a retrospective study, sinus endoscopies carried out between 1990 and 1994 on 57 patients, mean age 43 years, were subjected to critical re-evaluation, with the aim of evaluating the efficacy of endoscopic treatment. In the first phase, the documentation available for each patient was examined; it included clinical data, preoperative traditional and CT radiographs, rhinoscopic and sinus endoscopic findings. Once the analysis of the available documentation had been completed the patients were recalled and subjected to clinical and radiological evaluation. Radiographic examination was carried out by traditional radiography. RESULTS: CT scanning was only performed for those patients in whom the clinical findings indicated a possible relapse. Immediate and delayed complications occurring after the endoscopic sinus surgery were taken into consideration. CONCLUSIONS: In conclusion, sinus endoscopy is a fundamental instrumental investigation in the diagnosis of rhinosinusopathies. It can no longer be considered as only a diagnostic investigation able to provide clear visual images, it also allows specimens to be taken for bacteriological, micological, cytological and histological analysis. Furthermore, endoscopic surgery can treat pathologies for which, until a few years ago, operations were necessary which respected the physiology of the paranasal system to a much lesser extent (Denker-Caldwell-Luc).

Endoscopy↗
PubMed1997

Circulating levels of IL-10 are critically related to growth and rejection patterns of murine mastocytoma cells.

Previously tumorigenic P815 tumor cells are rejected by histocompatible mice after transfection with a mutated retroviral gene, and the host is made resistant to subsequent challenge with tumorigenic (control) cells transfected with the nonmutated sequence. To functionally characterize the class I-restricted response to the tumor cell vaccine, we have assessed the in vitro (by CD8+ cells) and in vivo production of type 1 or type 2 cytokines in mice injected with either type of transfected P815 derivative. IL-12 and IL-10 were selectively or preferentially expressed by the regressor mice, and this correlated with the detection of functional type 1 reactivity in vivo (i.e., delayed-type hypersensitivity). Other cytokines were produced by the regressor mice only in vitro (IFN-gamma) or were not detected at all with either type of tumor recipient (IL-4). By means of monoclonal antibody-mediated neutralization or enhancement of endogenous cytokine levels, IL-10 was found to serve an important role in the growth and rejection patterns of the transfected P815 derivatives. In addition to previous evidence for an IL-12 requirement in promoting anti-P815 reactivity, these data establish IL-10 as an important cytokine in permitting optimal expression of this reactivity, which apparently develops in the absence of a strong bias toward a type 1 or type 2 cytokine response.

Animals↗
PubMed1997

A tumor-associated and self antigen peptide presented by dendritic cells may induce T cell anergy in vivo, but IL-12 can prevent or revert the anergic state.

Ag-specific CD8+ cell responses, including delayed-type hypersensitivity in vivo and IFN-gamma production in vitro, are initiated by host immunization with P815AB, a self peptide bearing CTL epitopes and expressed by murine mastocytoma cells. Using P815AB-pulsed dendritic cells (DC) and monitoring class I-restricted skin test reactivity in DC-primed mice, we have previously shown that the development of a Th1-like response to P815AB requires T helper effects, such as those mediated by coimmunization with class II-restricted (helper) peptides or by the use of rIL-12. The adjuvanticity of IL-12 was suggested to involve improved recognition of class II-restricted epitopes of P815AB. In the present study, we provide evidence for the occurrence of I-A(d)-restricted epitopes in the tumor peptide. We also show that in the absence of helper peptide or rIL-12, P81 5AB not only failed to initiate CD8+ cell responses in vivo and in vitro, but resulted in a transient state of functional unresponsiveness, characterized by a profound inability of CD4+ cells to produce IL-2 in vitro. Ag-specific T cell anergy was also observed after neutralization of endogenous IL-12 at the time of priming with P815AB plus helper peptide. All of these effects were reversed by rIL-12, which was added to DC cultures and administered to the DC-recipient mice. Anergy induction may thus contribute to P81 5AB unresponsiveness in vivo. IL-12 may act to prevent or revert anergy to this tumor-associated and self peptide.

Animals↗
PubMed1997

Cyclosporine and pregnancy in the rat.

Female Wistar rats mated with male Wistar rats were given 15 mg/kg cyclosporine (CsA) per day during pregnancy (to 21 days after coitus). Other rats were mated and were used as controls. The pups born from treated mothers were fed by control mothers and those born from control mothers were fed by treated mothers. The newborn rats showed temporary defects in the development of some hemopoietic organs (the thymus, spleen, liver, and bone marrow). There was no difference in the structure of the organs studied between the litters fed by control mothers and those fed by treated mothers. The structure of the hemopoietic organs resembled those of the controls 30 days after birth. Our findings show that the delay in the development of the organs studied is due to the transplacental passage of CsA and suggest that it could be due to CsA interference in the maternal immune response to specific embryonic antigens.

Animals↗
PubMed1997

p75 neurotrophin receptor induction and macrophage infiltration in peripheral nerve during experimental diabetic neuropathy: possible relevance on regeneration.

In this study we examined the expression of the neurotrophin receptor p75 (p75NTR) and the activation of macrophages in the sciatic nerve of rats at different time points after the induction of diabetes with streptozotocin (STZ). Northern blot and immunocytochemical analysis showed that p75NTR was not detectable in the sciatic nerve by Week 2 after STZ treatment. At this time, single nerve fiber immunostaining using ED1 monoclonal antibody revealed that active macrophages were infiltrating the endoneurium, which had a normal morphological aspect. By Weeks 5 and 15 p75NTR mRNA and protein were induced in the endoneurium of diabetic animals. Immunocytochemical analysis of teased single nerve fibers showed that p75NTR protein was distributed uniformly along isolated fibers with no pathological evidence of axonal degeneration or myelin disruption. At this time, cells of the phagocyte lineage had already disappeared from the nerve. These data show that during experimental diabetic neuropathy, the endoneurial induction of p75NTR is localized along isolated nerve fibers showing no morphological alterations, and in time, follows the recruitment of active macrophages in the nerve, suggesting that these cells, directly or through their products, can influence p75NTR induction. This process might play an important role in STZ diabetic neuropathy, as a response to decreased levels of neurotrophins such as NGF and promoting nerve regeneration in the early phases of the disease.

Animals↗