Role of radiotherapy in the management of rare gynaecological cancers.

There are a large number of gynaecological cancers with rare histologies, for which the available data are limited and usually retrospective. Because of their rarity and poor prognosis, the management of these cancers must be centralized in expert centres, for both histological diagnosis and treatment. With the exception of sarcomas, most endometrial or cervical cancers with rare histologies respond to the same radiation treatment modalities than cancers with more common histologies, although there are some specificities regarding treatments such as neuroendocrine carcinomas (chemotherapy with platinum and etoposide, major role of surgery). For localized or locally advanced ovarian cancer, external beam radiotherapy has a role in the management of hypercalcaemic small cell carcinoma of the ovary. This article summarizes the current role of external beam radiotherapy and brachytherapy in the management of cancers of the uterine cervix, uterine corpus and ovaries, with rare or very rare histologies, and with localized or locally advanced stages.

[MRI evaluation of residual breast carcinoma after neoadjuvant chemotherapy]. / Cancer du sein traité par chimiothérapie néoadjuvante: évaluation du reliquat tumoral par l'IRM mammaire.

PURPOSE: This study aims to evaluate the sensibility and specificity of MRI in the detection and size measuring of residual breast cancer in patients treated with neoadjuvant chemotherapy before surgery. PATIENTS AND METHODS: This is a retrospective study of 32 women, who underwent breast MRI before and after neoadjuvant treatment. MRI has been confronted to surgical pathology results. RESULTS: The sensibility of MRI to assess pathologic Complete Response (no invasive residual tumor) was excellent (100%) but the specificity was low (55,5%). There was no false negative case and four false positive cases (Two ductal carcinomas in situ and two scars-like fibrosis). When MRI outcomes were compared with the presence or absence of invasive or in situ residual carcinoma, only one false negative case was noticed (one "in situ" residual tumor). The correlation between tumor size measured by MRI and histopathology was low (r=0,32). Underestimations of tumor size were due to non-continuous tumor regression or invasive lobular carcinoma or association of invasive carcinoma and intra ductal breast cancer. Over estimations of tumor size were due to chemotherapy-induced changes. CONCLUSION: MRI is a sensitive but poorly specific method to assess the pathological complete response after neoadjuvant chemotherapy. Estimation of tumor size and detection of isolated residual in situ carcinoma are fare. Therefore, surgical intervention remains necessary whatever the MRI outcomes.

Inhibition of the histamine-induced Ca2+ influx in primary human endothelial cells (HUVEC) by volatile anaesthetics.

BACKGROUND AND OBJECTIVE: Vasoactive substances such as histamine, acetylcholine or ATP increase the [Ca2+]i of endothelial cells, which leads to the activation of nitric oxide synthase (eNOS). The NO produced by this enzyme relaxes the underlying smooth muscle. Evidence suggests that eNOS activation is dependent on agonist-induced Ca2+ entry. Recently we have shown that in human endothelial cells (HUVEC), this Ca2+ entry is sensitive to isoflurane. The objective here was to study the mechanism by which volatile anaesthetics can depress the histamine-induced Ca2+ entry into HUVEC cells. METHODS: HUVECs on coverslips were loaded with the Ca2+ indicator Fluo-3 and inserted in a gastight, temperature-controlled perfusion chamber. Excitation was at 488 nm and fluorescence signals were monitored with a confocal laser scanning microscope (MRC1024, Biorad). Direct measurement of the Ca2+ influx was with Mn2+ as surrogate for calcium at 360 nm in cells loaded with Fura-2. RESULTS: Addition of histamine induces a biphasic [Ca2+]i increase consisting of Ca2+ release from internal stores and a Ca2+ influx from the external medium (plateau phase). The plateau phase was dose-dependently inhibited by enflurane and sevoflurane (13.7 resp. 21.9% inhibition by 1 MAC anaesthetic). Direct measurement of the Ca2+ influx using the Mn2+ quench of the Fura-2 fluorescence gave similar results. The inhibition of the anaesthetics was not reduced by inhibition of the cGMP pathway, inactivation of protein kinase C, depolarization of the cells or the presence of specific Ca2+-dependent K+ channel inhibitors. Interestingly, unsaturated fatty acids inhibit the histamine-induced Ca2+ influx in a similar way as the volatile anaesthetics. CONCLUSIONS: Volatile anaesthetics dose-dependently inhibit the histamine-induced Ca2+ influx in HUVECs by a mechanism that may involve unspecific perturbation of the lipid bilayer.

Volatile anesthetics stimulate the phorbol ester evoked neurotransmitter release from PC12 cells through an increase of the cytoplasmic Ca2+ ion concentration.

PC12 cells preloaded with [3H]norepinephrine release this neurotransmitter at a slow rate (basal release). This rate is increased by the addition of phorbol myristate acetate (PMA), but not by a biologically inactive phorbol ester. This effect most likely is mediated by protein kinase C, since desensitization of this kinase abolished the stimulation of the neurotransmitter release by PMA. Unexpectedly, clinical concentrations of the volatile anesthetics halothane, enflurane, isoflurane and methoxyflurane stimulated the PMA evoked neurotransmitter release in good correlation with their anesthetic potency. Since the volatile anesthetics increased the cytoplasmic Ca2+ concentration of the PC12 cells in a dose dependent manner it seems very likely that the effect of the anesthetics on the PMA-evoked neurotransmitter release is mediated by this rise in Ca2+ concentration.

Effects of addition of derived 40 S subunits on translation rate and polysome profile of the reticulocyte lysate.

We have investigated the way in which the addition of exogenous 40 S subunits to a reinitiating cell-free translation system, prepared from reticulocytes, may affect translational parameters of the system. The disturbance of the system's subunit stoichiometry resulted in the following changes in the ribosome profile: (1) rapid exhaustion of the pool of native 60 S subunits; (2) appearance of humps on the peaks of the polysome profile, which probably represent unusually long-lived [40 S. polysomal] complexes; (3) at higher doses of exogenous particles, the amount of polysomes decreased. This latter effect reflected a corresponding decrease in the overall translation (i.e. initiation) rate. The phenomena are interpreted as follows: exogenous 40 S subunits combine with 60 S subunits, forming idle 80 S ribosomes. The shortage of 60 S subunits delays the utilization of [40 S. polysomal] complexes, which is compensated for by a pool increase of these complexes. At high 40 S subunit doses this compensatory mechanism fails, and the 60 S shortage begins to determine the overall translation rate. The observations underline that the various translational parameters of the lysate function in an optimally concerted manner, so that only small amounts of derived 40 S subunits are tolerated by the system for analysis.

Volatile anesthetics inhibit the ion flux through Ca2+-activated K+ channels of rat glioma C6 cells.

Ca2+-activated K+ channels in rat glioma C6 cells were investigated using monolayers of these cells in petri dishes. The ion flux through the channels was studied with 86Rb+ after addition of a Ca2+-ionophore to the incubation medium. Both the influx and efflux of 86Rb+ through these Ca2+-activated K+ channels were inhibited by the general anesthetic halothane (at clinical concentrations). Other volatile anesthetics such as isoflurane, enflurane and methoxyflurane also inhibited the Ca2+-activated K+ channels at clinical concentrations. Inhibition of these channels by general anesthetics could have profound effects on signal transmission in the brain.

Lack of selectivity between anesthetic stereoisomers for an inhibitory site which is located on a membrane protein.

Lack of selectivity towards anesthetic stereoisomers is one of the few criteria available for the identification of an anesthetic target site. Until now this criterion has not been tested for anesthetics that directly interact with sensitive membrane proteins which are considered the targets of anesthetic action. In this communication we show that stereoisomers of 2-butanol and 2,4-pentanediol did not differ in their inhibitory potency for a site located on the Na+/K+/Cl- co-transporter protein. This suggests that an inhibition site on a membrane protein can fulfill the criterion of lack of stereoisomer selectivity.

Lipid solubility is not the sole criterion for the inhibition of a Ca2(+)-activated K+ channel by alcohols.

The effect of a homologous series of n-alkanols (C2-C8) on the 86Rb+ influx through charybdotoxin sensitive Ca2(+)-activated K+ channels of rat glioma C6 cells was investigated. The lipid solubility of the n-alkanols was not the sole determinant of the inhibitory potency of these substances for ion flux inhibition. 1-Hexanol for example was about 8-times less potent than one would expect on the basis of its lipid solubility. The introduction of a second OH-group in this molecule (giving 1,6-hexanediol) or a structural shift in the OH-group of 1-hexanol from position 1 to 3 strongly increased the potency of the alcohol. The above data cannot be explained by a pure lipid site of action of the alcohols. Therefore it seems likely that direct effects on protein are involved in the inhibitory action of some of the alcohols.

Characterization of an Na+/K+/Cl- co-transport in primary cultures of rat astrocytes.

The furosemide- and bumetanide-sensitive component of the 86Rb+ uptake into primary cultures of rat astrocytes was fully dependent on the simultaneous presence of Na+ and Cl- in the incubation mixture and is therefore most likely an Na+/K+/Cl- co-transporter. As expected for such a co-transporter, its activity is insensitive to 0.1 mM amiloride and to 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid, and of the tested anions, only Br- could partly replace Cl-. The K0.5 values for K+, Na+ and Cl- activation were 2.7, 35 and 40 mM, respectively. The activity of the co-transporter was stimulated 1.5-times in hyperosmolar (500 mosM) medium.

Inhibition of the elongation step of protein synthesis by vaccinia virus.

Vaccinia cores inhibit translation in cell-free protein synthesis systems at two stages: initiation; and, as shown here, elongation. The former effect tends to obscure the latter. Elongation control could, however, be revealed as follows: when, in a reticulocyte of L-cell lysate, initiation was blocked by a drug (edein), the residual [35S]methionine incorporation was severely reduced by the subsequent addition of vaccinia cores. The elongation block could also be demonstrated by analysis of ribosome profiles: treatment with edein alone permitted ribosomal run-off; treatment with either the elongation inhibition anisomycin or with cores preserved the polyribosomes.

Halothane inhibits the neurotoxin stimulated [14C]guanidinium influx through 'silent' sodium channels in rat glioma C6 cells.

We have investigated the effect of pharmacological agents on [14C]guanidinium ion influx through sodium channels in C6 rat glioma and N18 mouse neuroblastoma cells. The sodium channels of the N18 cells can be activated by aconitine alone, indicating that they are voltage-dependent channels. In contrast, sodium channels in the C6 cells require the synergistic action of aconitine and scorpion toxin for activation and are therefore characterized as so-called silent channels. The general anesthetic halothane used at clinical concentrations, specifically inhibited the ion flux through the silent sodium channel of C6 rat glioma cells. The voltage-dependent channels of the N18 cells were insensitive to halothane at the concentrations tested.

Volatile anesthetics depress the depolarization-induced cytoplasmic calcium rise in PC 12 cells.

In the rat pheochromocytoma cell line PC 12, the effects of four volatile anesthetics (halothane, isoflurane, enflurane, methoxyflurane) on the K+-evoked intracellular calcium [( Ca2+]i) rise were investigated using the Ca2+-sensitive fluorescence dye fura-2. The [Ca2+]i rise was depressed, at clinical concentrations, by all anesthetics with almost identical aqueous IC50 values. The study extends to neuronal cells the observation made previously in cardiac tissue that volatile anesthetics may interfere with Ca2+ fluxes through voltage-gated channels.

Preliminary characterization of an Na+,K+,Cl- co-transport activity in cultured human astrocytes.

We have studied the potassium uptake using 86Rb+ into monolayers of secondary cultures of human astrocytes prepared from cerebral hemispheres of a 4-month-old fetus. With the use of inhibitors we could attribute 30-40% of the 86Rb+ uptake to an Na+,K+-ATPase, 50-60% to an anion-cation co-transporter and 10% to potassium leak channels. The anion-cation co-transporter was dependent on the simultaneous presence of both sodium and chloride in the incubation medium and is therefore most likely an Na+,K+,Cl- co-transporter. This is the first evidence of such an Na+,K+,Cl- co-transport in human astrocytes.

Presence of a charybdotoxin sensitive Ca2+-activated K+ channel in rat glioma C6 cells.

A study was made of the 86Rb+ influx and efflux through Ca2+-activated K+ channels of intact rat glioma C6 cells after addition of a Ca2+ ionophore to the incubation medium. Half-maximal activation of the channels was obtained at a cytoplasmic Ca2+ concentration of approximately 400 nM. The 86Rb+ ion flux through the Ca2+-activated K+ channels was insensitive to apamin, but was inhibited by low concentrations of charybdotoxin (IC50 = 1.6 nM). This is the first evidence for the presence of charybdotoxin-sensitive Ca2+-activated K+ channels in glial cells.

The volatile anesthetic isoflurane suppresses spontaneous calcium oscillations in vitro in rat hippocampal neurons by activation of adenosine A1 receptors.

Primary cultures of rat hippocampal neurons were loaded with the Ca(2+)-indicator fluo-3 and studied with a confocal laser microscope. In Mg(2+)-free medium the cultures showed spontaneous synchronized calcium oscillations. These oscillations derived from excitatory signal transmission by N-methyl-D-aspartate and (+/-)-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid/kainate receptors and were modulated by gamma-aminobutyric acid(A) receptors. The oscillations were dose-dependently depressed by adenosine (IC50=2 microM) or by 2-chloro-N6-cyclopentyladenosine a specific adenosine A1 receptor agonist (IC50=40 nM). These effects were reverted by 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), a specific adenosine A1 receptor antagonist. The volatile anesthetic isoflurane also depressed these spontaneous calcium oscillations in a dose dependent manner (IC50=0.25 MAC, Minimum Alveolar Concentration). The isoflurane-induced inhibition was partly reversed in 29-38% of the neurons by DPCPX, indicating that the anesthetic activates this receptor possibly by increasing the extracellular concentration of adenosine.

The volatile anesthetic enflurane activates capacitative Ca2+ channels in rat glioma C6 cells.

(1) The volatile anaesthetics halothane and enflurane released Ca2+ from thapsigargin sensitive stores in rat glioma C6 cells, but only enflurane induced a significant Ca2+ influx. (2) The reason for this can be explained by a different inhibitory effect of the anesthetics on the capacitative Ca2+ influx. (3) Halothane inhibited the capacitative Ca2+ influx with an IC50 of 1.9 vol.%, whereas enflurane only marginally affected the ion flux through the channel.

Influence of cadmium ions on endothelin-1 binding and calcium signaling in rat glioma C6 cells.

Cadmium ions did not influence the binding of endothelin-1 (ET-1) to its receptor on the surface of rat glioma C6 and rat aorta A10 cells. This was studied (a) by the binding of 1251-ET-1 to intact cells in the absence or presence of cadmium (Cd2+) and (b) by analysis of the receptor/ET-1 complex after crosslinking with disuccinimidyl suberate (DSS) or ethylene glycol-bis-(succinimidyl succinate) (EGS) on SDS PAGE. Using Fura-2 and Quin-2 loaded C6 rat glioma cells, it was shown that Cd2+ ions strongly interfered with the ET-1 induced Ca(2+)-influx in C6 glioma cells (IC50 approximately 10 microM).

Influence of the carcinogenic oestrogen diethylstilboestrol on the intracellular calcium level in C6 rat glioma cells.

The synthetic oestrogen diethylstilboestrol (DES) causes a dose-dependent elevation of the cytoplasmic Ca(2+) concentration in C6 rat glioma cells. This Ca(2+) rise is caused neither by Ca(2+) influx nor by release from the Ca(2+) stores of the endoplasmic reticulum. Therefore it seems likely that DES mobilizes Ca(2+) from a mitochondrial source. The DES-induced Ca(2+) signal is remarkably similar to the one induced by the tumour promotor thapsigargin. As this compound causes leakage of calcium from the endoplasmic reticulum it seems possible that DES induces a similar leakage from mitochondrial Ca(2+) stores. It remains to be established whether the DES-mediated rise in intracellular calcium is causally related to the tumour-promoting properties of this compound.

Role of extracellular calcium and calcium stores in the intracellular calcium rise induced by diethylstilboestrol in C6 rat glioma cells.

The aneuploidy-inducing carcinogenic oestrogen diethylstilboestrol (DES) causes an elevation of the cytoplasmic Ca(2+) concentration of C6 rat glioma cells. It is known that the intracellular calcium concentration is a critical factor in mitosis. DES induces mitotic disturbances resulting in aneuploidy; the mechanisms of these events are still largely unknown. It is therefore important to identify the calcium source for the DES-mediated calcium rise. Experiments with isolated rat liver mitochondria showed no release of calcium after DES treatment. However, DES releases calcium from the same stores as the tumour promotor thapsigargin, which are the inositol 1,4,5-triphosphate- and GTP-sensitive Ca(2+) stores. In contrast to DES, thapsigargin also induces a calcium influx into the cell. Administration of DES following thapsigargin treatment strongly reduced the thapsigargin-mediated calcium entry. Furthermore, DES inhibits the decay of the ionomycin-induced calcium signal, suggesting an interference of DES with the extrusion of calcium from the cell. In conclusion, thapsigargin and DES both cause release of calcium from the endoplasmic reticulum. However, for each compound the prolongation of the calcium signal is based on different mechanisms.

Malignant lymphoma of mucosa associated lymphoid tissue: a new etiology of amyloidosis.

Non Hodgkin Malignant lymphomas (NHML) of mucosa associated lymphoid tissue (MALT) are known to have multiple involvement of the digestive tract. We report one case, presenting with an infiltrative process of the jejuno-ileum, associating lymphoplasmacytoid proliferating cells and amyloidosis. The plasmacytoid cells expressed Alpha and scarce Mu heavy chains, and lambda light chain. Lympho-epithelial lesions were more obvious at the second site of involvement, in the gastric mucosa. The amyloid substance was negative with the Amyloid A component antibody and gave a background noise with Alpha, Mu and lambda chains. No similar report of amyloidosis associated with MALT NHML has been found in the literature.