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Biomedical subjects

F Robert

Publications and source records attributed to F Robert.

At least 73 records · Page 4Linked to original sources
PubMed1996

Cytokine production by human thymic epithelial cells: control by the immune recognition of the neurohypophysial self-antigen.

Oxytocin (OT) has been shown to be the dominant peptide of the neurohypophysial family expressed by thymic epithelial and nurse cells (TEC/TNC) in various species. Thymic OT is not secreted but, after translocation of a hybrid neurophysin/MHC class I protein, is integrated within the plasma membrane of TEC, thus allowing its presentation to pre-T cells. In order to further demonstrate that thymic OT behaves like a membrane antigen, we assessed the effect of mAbs to OT on cytokine productions by cultures enriched in human TEC. 75-85% pure TEC cultures were prepared from human thymic fragments. Using immunofluorescence and confocal microscopy, ir-OT, ir-interleukin-1 beta (IL-1 beta), ir-interleukin-6 (IL-6) and ir-leukemia inhibitory factor (LIF) could be detected in these TEC cultures. ir-OT was restricted to TEC, while some ir-IL-6 and ir-LIF were also seen in occasional fibroblasts. In basal conditions, ir-IL-6 and ir-LIF (but not ir-OT and ir-IL-1 beta) were detected in the supernatants of human TEC cultures. MAbs to OT induced a marked increase of ir-IL-6 and ir-LIF secretion in TEC cultures. No significant effect was observed using mAbs against vasopressin, mouse immunoglobulins, or control ascitic fluid controls. These data show that OT is fully processed and recognized by specific mAbs at the outer surface of TEC plasma membrane. They further support that thymic OT behaves as the self-antigen of the neurohypophysial family.

Antibodies, Monoclonal↗
PubMed1996

Microdialysis monitoring of variations in extracellular levels of serotonin, GABA and excitatory amino acids in the frontal cortex of awake rats in response to a single peripheral or central administration of dexfenfluramine.

The effects of a single dexfenfluramine (D-fen) administration on the release of endogenous serotonin (5-hydroxytryptamine, 5-HT), excitatory (glutamate, Glu, aspartate, Asp) and inhibitory (gamma-aminobutyric acid, GABA) amino acids from the frontal cortex were studied by using in vivo microdialysis in freely-moving rats. Extracellular levels of these neurotransmitters were measured with HPLC coupled to electrochemical detection or with capillary electrophoresis coupled to laser-induced fluoresence detection (CE-LIFD). In a first study, single intraperitoneal administration of D-fen (0.5, 1.3, 5 and 10 mg/kg) increased extracellular 5-HT levels in a dose-dependent manner (maximal increase by 982% over baseline for the highest dose) while changes in Glu, Asp or GABA never reached statistical significance. In a second study, 73 nM of D-fen applied locally through the frontocortical dialysis probe, at a flow rate of 1.5 microliters/min in 30 microliters of perfusion fluid for 20 min, increased extracellular 5-HT and Asp levels [the maximal increases were to 1804% and 280% of the respective basal values (100%)] without altering extracellular levels of Glu and GABA. Thus, the order of magnitude of the changes induced by systemic administration or local infusion of D-fen on frontocortical extracellular levels of several neurotransmitters (5-HT > > Asp > GABA = Glu) demonstrate that D-fen, an indirect serotoninergic agonist, mainly increases 5-HT release while producing slight (Asp) or no (Glu, GABA) short-term in vivo variations in amino acid extracellular levels in the rat frontal cortex.

Animals↗
PubMed1996

Four IgG anti-islet human monoclonal antibodies isolated from a type 1 diabetes patient recognize distinct epitopes of glutamic acid decarboxylase 65 and are somatically mutated.

The selective destruction by an autoimmune process of the beta cells in the pancreas is the hallmark of the type 1 insulin-dependent diabetes mellitus. What triggers islet cell-specific autoreactive T and B cells, however, remains unclear. Identification of the targets of the anti-islet cell autoantibodies frequently found in insulin-dependent diabetes mellitus patients and analysis of their sequences should provide some insights into the nature of this disease. We have combined EBV transformation with CD40 activation of peripheral B cells from one patient with insulin-dependent diabetes mellitus to isolate four B cell clones that secrete islet cell-specific autoantibodies, These four human monoclonal autoantibodies are of the IgG1 isotype, and they each recognize a different epitope of the glutamic acid decarboxylase enzyme. Analysis of their variable gene sequences shows that, while clonally unrelated, three of the four human monoclonal autoantibodies use a member of the VH4 family, and two have rearranged the same delta light chain variable gene. The IgG1 isotype of the four autoantibodies as well as the presence of somatic mutations in both heavy and light chain genes provide concrete evidence for their derivation by a T cell-dependent immune response.

Adult↗
PubMed1996

Localization of subunits of transcription factors IIE and IIF immediately upstream of the transcriptional initiation site of the adenovirus major late promoter.

The assembly of a preinitiation complex containing RNA polymerase II on promoter DNA is a complex process that involves several general transcription factors. Using 5-[N-(p-azidobenzoyl)-3-aminoallyl] photocross-linking, we previously determined the locations of the two large subunits of transcription factor (TF) IIA (A35 and A21), TATA box-binding protein (TBP), RNA polymerase II-associated protein (RAP) 30, and TFIIB along the Ad2 ML promoter. We have now localized TFIIE34 and RAP74 just upstream of the transcription start site. The two subunits of TFIIF, RAP74 and RAP30, cross-linked to nucleotides that probed adjacent spaces on the same face of the DNA helix beginning just downstream of TBP at -19 and extending to -5. Specific photocross-linking of TFIIE34 required the presence TFIIE56. In addition, TFIIE and RAP74 strongly stimulated cross-linking of RAP30 and the large subunits of RNA polymerase II to position -19. Our topological data support the idea that RAP74 and TFIIE34 may be involved in melting of the promoter DNA upstream of the initiation site.

Adenoviruses, Human↗
PubMed1996

[Retrospective evaluation of the detection of Toxoplasma gondii by polymerase chain reaction in AIDS patients].

OBJECTIVES: We retrospectively evaluated routine detection of Toxoplasma gondii by polymerase chain reaction (PCR) amplification of the B1 gene and the TGR 1E sequence in blood and CSF samples from patients with acquired immune deficiency syndrome (AIDS) and suspected toxoplasmosis. METHODS: From January 1993 to February 1994, 93 blood, 33 cerebrospinal fluid (CSF) and 1 aqueous humor samples were obtained from 83 HIV-positive patients with CD4 counts under 200/mm3 and suspected toxoplasmosis. RESULTS: Authentic cerebral toxoplasmosis was confirmed by response to specific treatment in 18/29 patients with typical focal brain lesions. Blood samples were available in 17/18 and CSF in 6. PCR was positive for both B1 and TGR (23.5% sensitivity, 100% specificity) in 4/17 blood samples and for either B1 or TGF (58.9% sensitivity, 72.7% specificity) in 10/18. PCR of CSF was positive in only 1/6 patients with cerebral toxoplasmosis. PCR (TGR 1E only) was positive in 3/11 blood samples and in one CSF sample from patients without cerebral toxoplasmosis. Five out of 21 patients with diffuse neurologic symptoms and presumed HIV encephalitis had positive Toxoplasma detection in blood or CSF. However, no clinical improvement was obtained after specific antitoxoplasmic therapy. Two out of 38 patients with unexplained fever with or without pneumonia had a proven disseminated toxoplasmosis. These two patients had PCR-positive blood samples. One of them was cured by a specific anti-toxoplasmic treatment and the other died two days later. Seven other patients had a positive PCR result in blood or CSF. Three of them improved with specific treatment and 2 died before treatment could be initiated. The one patient with toxoplasmic retinitis was PCR-positive both in blood and aqueous humor. CONCLUSION: PCR detection of Toxoplasma gondii appears to add little to the diagnosis of cerebral toxoplasmosis but could be helpful in the diagnosis of cerebral toxoplasmosis associated with extracerebral reactivation and in disseminated toxoplasmosis.

AIDS-Related Opportunistic Infections↗
PubMed1996

High-speed separation of subnanomolar concentrations of noradrenaline and dopamine using capillary zone electrophoresis with laser-induced fluorescence detection.

Capillary zone electrophoresis with laser-induced fluorescence detection has been shown to give rapid separations with high resolution and sensitivity. These advantages are documented for catecholamines analysis in the present work, which shows that separation of subnanomolar concentrations of dopamine and noradrenaline (detection limits of 8.6 x 10(-11) M, corresponding to a detected amount of 143 zeptomoles of each catecholamine) can be performed in 82 s with an efficiency of several million theoretical plates.

Dopamine↗
PubMed1996

Phase I and pharmacologic study of 7- and 21-day continuous etoposide infusion in patients with advanced cancer.

PURPOSE: This phase I study was undertaken to evaluate the safety and tolerability of prolonged infusional etoposide, and to evaluate its pharmacokinetic/pharmacodynamic profile in patients with advanced cancer. METHODS: A group of 17 patients received a 7-day infusion of etoposide (schedule A) every 21 days at doses from 30 to 75 mg/m2 per day, and a second group of 37 patients a 21-day infusion (schedule B) every 28 days at doses from 18 to 40 mg/m2 per day. Patients had a median Karnofsky performance status (PS) of 80%, and 34 patients had no prior chemotherapy. Etoposide concentrations at steady state (Css) and other pharmacokinetic parameters (plasma clearance, CLp; area under the curve, AUC) were determined during the first treatment cycle. Correlation coefficients were calculated to measure the relationship between variables. RESULTS: Myelosuppression was the major toxicity, and was associated with three deaths. The maximum tolerated dose due to neutropenia was 75 mg/m2 per day for schedule A and 40 mg/m2 per day for schedule B. There was significant interpatient pharmacokinetic variability in both infusional schedules. Even though etoposide dose levels did not significantly correlate with plasma levels, the Css was > or = 1 microgram/ml in the majority of the patients. A significant correlation between AUC and neutrophil absolute decrease was noted only in schedule B (r = 0.56, P = 0.003). There were several marginal relationships in schedule B: PS versus Css (r = 0.31, P = 0.058), PS versus AUC (r = -0.38; P = 0.058) and age versus CLp (r = -0.31, P = 0.057). CONCLUSION: Overall, significant correlations were found for several hematologic variables and etoposide dose levels, but not with the Css values. One major problem with the application of pharmacodynamic models to predict hematologic toxicity in clinical practice is the presence of significant interpatient variability.

Adult↗
PubMed1996

Prolonged infusion of etoposide in patients with advanced non-small cell lung cancer.

We assessed the efficacy and toxicity of two etoposide infusional schedules in patients with advanced non-small cell lung cancer (NSCLC). Twenty-six patients were treated with a 21-day infusion every 28 days at a dose of 25-40 mg/m2/d, and six patients with a 7-day infusion every 21 days at a dose of 45-75 mg/m2/d. Sixty-three percent of patients had a Karnofsky status of 80% or better, and only five (15%) patients had prior chemotherapy. Plasma etoposide concentrations were determined in 26 patients. Sixty-nine treatment cycles were administered. Two patients (6.3%; 90% confidence interval, 1.1-18.4%) had partial responses; with response durations of 2 and 7 months, respectively. The median survival was 4 months. Grade 3 or 4 neutropenia occurred in 13 of 69 cycles (19%) and was associated with three toxic deaths. Ten patients required RBC transfusions. Nausea was common, but was associated with vomiting in only 7% of all cycles. The interpatient variability of etoposide concentrations at steady state was significant. We conclude that the antitumor activity of prolonged infusion of etoposide is not superior to standard dose and schedule in advanced NSCLC.

Adult↗
PubMed1996

Investigation of immunofluorescence cross-reactions against Trichinella spiralis by western blot (immunoblot) analysis.

Immunofluorescence cross-reactions in Trichinella spiralis serodiagnosis are sometimes difficult to identify. We compared the results of an indirect immunofluorescence assay and the profiles obtained by Western blot (immunoblot) analysis for three groups of patients: 10 T. spiralis-infected patients, 10 patients with autoimmune diseases, and 7 patients with parasitic diseases other than trichinellosis. The degree of immunofluorescence cross-reaction was variable. Western blotting allowed us to differentiate Trichinella infection from other parasitic diseases. In 3 of 10 serum samples from patients with autoimmune diseases, bands which had the same sizes as Trichinella bands were observed, and they could correspond to shared epitopes such as heat shock proteins.

Animals↗
PubMed1996

[Catheter ablation and modulation of the atrioventricular junction; current aspects].

Ablation of the atrioventricular junction consists in creating a therapeutic AV block to facilitate the treatment of symptoms caused by atrial arrhythmias refractory to drug therapy. The technical performance of ablation has been improved by restricting the indication to atrial fibrillation, by using radiofrequency currents, by choosing a nodal rather than His bundle ablation site, and by improving the function of cardiac pacemakers (rate adapting, back-up). The functional results are excellent but the outcome is punctuated by rare cases of sudden death, the cause of which is not fully understood (dependance, ventricular arrhythmias, ...). To avoid permanent pacing, it has been suggested that atrioventricular conduction should be modulated rather than completely interrupted. Modulation of the fast pathway has been shown to be ineffective; that of the more complex, slow pathway, seems to be more promising. Although this obviates the need for a pacemaker, it does not suppress irregularity of the ventricular rhythm, the main cause of symptoms in paroxysmal atrial fibrillation and of the haemodynamic changes associated with permanent atrial fibrillation.

Actuarial Analysis↗
PubMed1996

[Voltametric detection of cerebral NO in rats. Variations of the signal throughout the sleep-wakefulness cycle].

Nitric oxide (NO) is synthesized in the neurons by constitutive NO synthase (NOS). Within given neuronal sets, this enzyme is colocalized with different other neurotransmitters such as, for example, GABA, acethylcholine or serotonin. Our attention has been focused on the fact that serotoninergic neurons, well known for their involvement in sleep triggering and maintenance, synthesize also NO. In order to evaluate the modalities of release of this compound throughout the rat sleep-waking cycle, we prepared a sensor allowing its specific detection in freely moving animals. The active part of this sensor is a carbon fiber (phi = 30 microns) successively coated with porphyrin nickel and nafion. In vitro, together with differential normal pulse voltammetric measurements, it allows the detection of a 650 mV signal varying linearly in NO solutions ranging from 5.10(-7) to 10(-4) M. At physiological concentrations, L-arginine, L-citrulline, nitrites and nitrates do not yield a signal at 650 mV. Similarly, the compounds administered to the animals, hydroxylamine, L-arginine p-nitroanilide (L-ANA) and L-N omega-nitro arginine methyl ester (L-NAME) are not electroactive at 650 mV. L-ANA and L-NAME, also appear to be trapping agents for NO while leaving the electrochemical properties of the sensor untouched. In vivo, in the frontal cortex of the anesthetized rat, a signal is measured at 650 mV. The administration of hydroxylamine (40 mg/kg, i.p.) induces a 100% increase in its height. The administration of L-ANA (100 mg/kg, i.p.) produces its complete disappearance within 50 min. Finally, the administration of L-NAME (100 mg/kg, i.p.) is without effect. This last aspect might be dependent upon the inability of L-NAME to cross the blood brain barrier. On the contrary, the increase in the signal height obtained with hydroxylamine and its disappearance with L-ANA support that it might depend upon NO. In vivo, and in animals also equipped with polygraphic electrodes, the signal measured in the same area of the cortex exhibits the highest height during the waking state and decreases during either slow-wave sleep (-6%) or paradoxical sleep (-9%). These mild variations might represent the mean of several NO sources (cortical GABAergic interneurons, cholinergic and serotoninergic axonal nerve endings), each of them varying differently throughout the sleep-waking cycle.

Animals↗
PubMed1996

Spinal metastases as a first presentation of malignant astrocytoma.

A 46-year-old man presented with low dorsal pain and paresthesia. Computed tomography showed an osteolytic lesion involving most of the vertebral body and the left pedicle of the 12th thoracic vertebra (T12). Contrast-enhanced magnetic resonance imaging (MRI) of the spine showed an enhancing soft-tissue mass that involved the T11 and T12 vertebral bodies, as well as that of the first lumbar vertebra; the mass caused cord compression. Another lesion was identified at T9. The findings of percutaneous needle aspiration biopsy of the lesion were consistent with metastatic astrocytoma, a diagnosis confirmed at surgery. MRI of the brain showed an asymptomatic lesion of the left temporal lobe; histologic confirmation of malignant astrocytoma was obtained by stereotactic biopsy. This report shows that metastatic bone disease secondary to malignant astrocytoma may manifest itself before the primary lesion becomes symptomatic. This presentation of astrocytoma was unusual because there were no symptoms of the intracranial tumour and because metastatic disease to the bones is less common than to the chest and the lymph nodes.

Astrocytoma↗
PubMed1995

Capillary zone electrophoresis with laser-induced fluorescence detection for the determination of nanomolar concentrations of noradrenaline and dopamine: application to brain microdialysate analysis.

Determination of catecholamines by capillary zone electrophoresis with laser-induced fluorescence detection was performed on low-concentration samples, which were derivatized with naphthalene-2,3-dicarboxaldehyde to give highly fluorescent compounds. When the borate concentration in the derivatization medium was decreased from 130 to 13 mM, sensitivity for noradrenaline (NA) and dopamine (DA) was greatly enhanced while resolution between these two compounds decreased. A 50 mM borate concentration in derivatization medium was chosen since it provided maximal resolution between NA and DA, together with a high separation efficiency (3.1 million theoretical plates per meter for DA). The injection of 2.4 nL of a NA and DA solution derivatized at 10(-9) M produced peaks with signal-to-noise ratio of 8:1 and 3:1, respectively, corresponding to 1.8 amol of each catecholamine. The calibration curves were linear when NA and DA solutions were derivatized at concentrations ranging from 10(-6) to 10(-9) M. This method was used to determine NA in brain extracellular fluid: a peak corresponding to a basal level of 5 x 10(-9) M endogeneous NA was observed in microdialysates from the medial frontal cortex of the rat, and its nature was confirmed by both electrophoretic and pharmacological validations.

Animals↗
PubMed1995

Interstellar water in meteorites?

D/H ratios of two meteorites (Renazzo CR and Semarkona LL3), which are known to exhibit the largest departures from the terrestrial hydrogen isotopic ratios, have been determined with the CRPG Nancy ion microprobe. Correlations between the D/H ratios and the chemical compositions (H2O, K, Si, C/H) of plausible hydrogen carriers were observed. From these correlations, it is possible to show that, contrary to previous interpretations, phyllosilicates are the carriers of the deuterium-rich hydrogen in Semarkona and Renazzo: 870 x 10(-6) > or = D/H > or = 670 x 10(-6) (+4600 > or = deltaD > or = 3300%) and > or = 320 x 10(-6) (deltaD > or = 1050%), respectively. Hydrogen is also present in the chondrules of these two deuterium-rich meteorites. The large differences in D/H ratios between matrix (up to 700 x 10(-6), deltaD up to +3500%) and chondrules (from 120 x 10(-6) (deltaD = -230%) to 230 x 10(-6) (deltaD = +475%)) show that hydrogen in chondrules cannot originate from the matrix by simple contamination or diffusion processes. The high D/H ratios measured in water-bearing minerals could not have been produced thermally within a dense solar nebula. Chemical reactions (i.e., involving ions or radicals), taking place in interstellar space or in the outer regions of the nebula at 110-140K are presently the only conceivable mechanisms capable of yielding such isotopic enrichments. Water in these meteorites should no longer be considered as a simple product of nebular condensation under equilibrium thermodynamic conditions at T > or = 160K.

Carbon↗
PubMed1995

Development of a schwannoma within a facial nerve neurofibroma: a case report and literature review.

We report the presence of a schwannoma within a neurofibroma of the intratemporal facial nerve. This neurofibroma recurred 39 years after its first excision in the parotid gland. Although some believe that schwannomas and neurofibromas represent the same entity, these tumors present distinctive histopathologic and clinical characteristics, which are discussed. The extreme rarity of a schwannoma developing within a neurofibroma is underlined. This is the first report of such an association occurring within a cranial nerve.

Cranial Nerve Neoplasms↗