[Comparison between the terminal motor pudendal nerve terminal motor latency, the localization of the perineal neuralgia and the result of infiltrations. Analysis of 53 patients]. / Comparaison entre la latence distale motrice du nerf pudendal, la topographie de la douleur périnéale et le résultat d'infiltrations. Analyse pour 53 patients.

AIM: The aim is to study the value of the pudendal nerve terminal motor latency (PNTML) testing, in respect to the painful side in patients with pudenda neuralgia, and to determine whether a possible increased latency in the painful side is predictive of a good result with the infiltration. METHOD: This retrospective study included 53 outpatients (42 women, mean age 62) with suffering from pudendal neuralgia, who were seen between 2000 and 2004. The mean duration of the pain was 30+/-47 months. The PNTMPNL was measured by the Saint-Mark hospital technique, by the same operator. The following criteria have been were defined: significant increased latency greater than above 6 ms, significant difference of 2 ms in latency between 2 sides from 2 ms, and side of the infiltration corresponding to the side of the neuralgia. The infiltrations were performed either by perineal (30 cases) or transgluteal (8 cases) way. The results on pain were have been considered as good when a substantial reduction of the pain was observed for 6 months or more. Statistical analysis involved was done by the exact Fischer's test to seek for a possible relation between variables. RESULTS: Of 53 patients (42 women, 11 men, mean age 62) suffered from a with perineal neuralgia. The duration of the neuralgia was 30+/-47 months. It was bilateral in 10 cases and unilateral in 43 cases. In 43 patients with When the pain was unilateral pain, PNTML we find that the MDLPN was increased in both sides in 39.5% of the population, in the painful side in 14% and in the side opposite side of the to pain in 11%. In 10 patients with the neuralgia was bilateral pain, in 10 patients. Among then, 4 had a bilateral increase of the latency, one patient had an increase only on the right side, and another one an increase only on the left side. We did not find any correlation between the increased of the PNTML TMPNL and, either neither the duration of the neuralgia nor the result of the infiltrations, whatever the method way of the infiltration. CONCLUSION: The PNTML can be increased whether it corresponds or not to an entrapment of the pudendal nerve. Thus, the management of perineal pain is based mainly, from us, on clinical findings.

Value of peritoneal cytology after hysteroscopy in surgical stage I adenocarcinoma of the endometrium.

Nineteen clinical stage I adenocarcinoma of the uterus with favourable histological prognosis factors (low grade, no myometrial extension, and no pelvic node involvement) were diagnosed using a pre-operative hysteroscopy. During the laparotomy, peritoneal cytology was performed systematically. The frequency of positive peritoneal washings was abnormally high (7 cases) with cytologic findings showing grouped cells in large clusters. However, these patients have not experienced peritoneal recurrences. The endoscopic procedures may have facilitated the transtubal malignant cell dissemination and are questionable in endometrial carcinoma.

[Value of peritoneal cytology after hysteroscopy in endometrial adenocarcinoma stage I]. / Valeur de la cytologie péritonéale après hystéroscopie dans les adénocarcinomes endométriaux de stade I.

Nineteen clinical stage I adenocarcinoma of the uterus with favourable histological prognosis factors (low grade, no myometrial extension, and no pelvic node involvement) were diagnosed using a preoperative hysteroscopy. During the laparotomy, a peritoneal cytology was performed systematically. The frequency of positive peritoneal washings was abnormally high (7 cases) with cytologic findings showing grouped cells in large clusters. However, these patients have not experienced peritoneal recurrences. The endoscopic procedures may have facilitated the transtubal malignant cell dissemination and are questionable in front of endometrial carcinoma.

[Gynecologic allergy to spermatic fluid. A case report]. / Allergie gynécologique au liquide spermatique. A propos d'une observation.

Seminal fluid allergy is a possible diagnosis in front vulvovaginal inflammations occurring rapidly after coitus. The type I reaction (revealed by the immediate hypersensitivity Prick-test) is most frequently encountered. In case of systemic symptoms, the circulating Ig E specific antibodies can be increased. Treatment is palliative (condoms, antihistamines cream). In case of systemic disease, desensibilisation can be proposed. This infrequent but uncomfortable syndrome is also in question in some cases of immunologic infertility.

Lateral diffusion and conductance properties of a fluorescein-labelled alamethicin in planar lipid bilayers.

In order to follow alamethicin diffusion within membranes under conditions of pore-formation, a fluorescein isothiocyanate (FITC) analogue was synthesized. To test the influence of the fluorescent probe addition on the pore-forming activity of the new analogue, macroscopic and single-channel experiments into planar lipid bilayers were performed. Although the apparent mean number of monomers per conducting aggregate was equivalent, the voltage-dependence of the new analogue was slightly reduced and hysteresses were broader, in agreement with the much longer duration of the open single-channels. Thus, the conducting aggregates seem to be stabilized by the introduction of the probe, presumably through the interaction of the conjugated cycles with the lipid headgroups, while the added steric hindrance may account for the slightly higher conductances of the open substates. Lateral diffusion of the labelled peptide associated with the bilayer was then investigated by the fluorescence recovery after photobleaching technique. Under applied voltage, associated with high conductance, D, the lateral diffusion coefficient, was reduced by 50% when compared to peptide at rest. These results provide new independent experimental evidence for a voltage-driven insertion of the highly mobile surface-associated peptide into the bilayer as a prominent step in pore formation.

Ion channel stabilization of synthetic alamethicin analogs by rings of inter-helix H-bonds.

Rings of inter-helix H-bonds due to Gln at position 7, a highly conserved residue in all pore-forming peptaibols, have been suggested to play an important role in the stabilization of alamethicin channels. In an attempt to test this hypothesis, experimental studies have been undertaken on four synthetic alamethicin non-Aib analogs (Alm-dUL) in which the Gln at position 7 (Q7) is substituted by Ala, Asn, or Ser (Q7A, Q7N, or Q7S). Voltage-dependent pore formation by these analogs in planar lipid bilayers is compared at the macroscopic and single-channel conductance levels. As anticipated, the Q7A substitution abolished all channel-forming activity. The voltage dependence of macroscopic current-voltage curves was conserved with the Q7N substitution but reduced in the Q7S analog. Normalized single-channel conductance ratios between substates follow the same pattern, with the Q7S analog yielding the highest unit conductances. Channel lifetimes were the most significantly modulated parameter with markedly faster kinetics when Gln or Asn was replaced by Ser. The effect of the Q7S substitution on channel lifetimes may be explained through a reduced stabilization of bundles by inter-helix H-bonds.

Sequencing of hydrophobic peptides as lithiated adducts using liquid secondary-ion mass spectrometry without tandem mass spectrometry.

Lithium cationized hydrophobic peptides in the molecular weight range 1900-2650 Da have been studied by liquid secondary-ion mass spectrometry. The mass spectra exhibit characteristic lithiated sequence ions. N-terminal [a(n)+Li-H]+ and [dn+Li-H]+ ions together with limited C-terminal [ypro+Li+H]+ ions lead to an easy sequencing of the peptides studied, without the need for tandem mass spectrometry.

[Breast cancer after treatment of Hodgkin's disease]. / Cancer du sein après traitement de maladie de Hodgkin.

OBJECTIVES: Development of diagnostic, therapeutic and preventive measures for breast cancer after cure of Hodgkin disease. STUDY TYPE: Presentation of 4 patients treated conjointly by the Radiotherapy and Gynaecology Surgery Departments of the Rennes University Hospital. RESULTS: Illustrations of difficult management of breast cancer at different stages of diagnosis and therapy. CONCLUSION: A past history of treated Hodgkin disease is a factor of risk for breast cancer and suggests the need for annual mammography screening 10 years after the end of treatment. Though more difficult, mastectomy is recommended over conservative radiosurgical treatment. The choice of drugs for adjuvant chemotherapy should rely on Hodgkin protocols and take into account heart function. Long-term carcinogenic effects of Hodgkin disease treatments requires modulation of the different treatment protocols as a function of stage, clinical and histological factors of prognosis and patient age.

Implication of segment S45 in the permeation pathway of voltage-dependent sodium channels.

A 34-mer peptide, encompassing the S4 and S45 segments of domain IV of the electric eel voltage-dependent sodium channel, was synthesized in order to test the potential implication of S45 in the gating or permeation pathway. The secondary structure of peptide S4-S45 assessed by circular dichroism was found mainly helical, both in organic solvents and in lipid vesicles, especially negatively-charged ones. The macroscopic conductance properties of neutral and negatively-charged Montal-Mueller planar lipid bilayers doped with S4-S45 were studied and compared with those of S4. With regard to voltage-dependence, the most efficient system was S4-S45 in neutral bilayers. Voltage thresholds for exponential conductance development were found to correlate with the background or "leak" conductance. Assuming that the latter reflects interfacial peptide concentration, the mean apparent number of monomers per conducting aggregate could be estimated to be 3-5. In single-channel experiments, the most probable events had amplitudes of 8 pS and 5 pS in neutral and negatively-charged bilayers respectively. Ionic selectivity under salt gradients conditions, both at macroscopic and single-channel levels, was in favour of sodium ions (PNa/PK = 3). These properties compare favourably to previous reports dealing with peptide modelling transmembrane segments of voltage-dependent ionic channels. Specifically, when compared to S4 alone, the reduced unit conductance and the increased selectivity for sodium support the implication of the S45 region in the inner lining of the open configuration of sodium channels.

Prolines are not essential residues in the "barrel-stave" model for ion channels induced by alamethicin analogues.

In the "barrel-stave" model for voltage-gated alamethicin channels in planar lipid bilayers, proline residues, especially Pro14, are assumed to play a significant role. Taking advantage of a previous synthetic alamethicin analogue in which all eight alpha-aminoisobutyric acids were replaced by leucines, two new analogues were prepared in order to test the effects of Pro14 and Pro2 substitutions by alanines. The alpha-helical content of the three analogues in methanol solution remains predominant (between 63 and 80%). Macroscopic conductance experiments show that a high voltage dependence is conserved, although the apparent mean number of monomers forming the channels is significantly reduced when the substitution occurs at position 14. This is confirmed in single-channel experiments which further reveal faster fluctuations for the modified analogues. These results demonstrate that, although prolines, especially Pro14, are favorable residues for alamethicin-like events, they are not absolute prerequisites for the development of highly voltage-dependent multistate conductances.

Gram-scale enzymatic synthesis of a peptide bond.

The synthesis reaction of the peptide, N-Cbz-L-tryptophanyl-glycineamide, catalyzed by alpha-chymotrypsin was performed in a 20% water/80%, 1,4-butanediol mixture. The synthesis yield reached 90.9% at the end of the reaction and 72.3% after purification. The effects on the yield of both pH and the ratio between total initial concentrations of glycineamide and N-Cbz-L-tryptophan are examined. The high yield, specificity, simplicity and reproducibility of this method make it complementary of the chemical methods.

Design and conformation of non-Aib synthetic peptides enjoying alamethicin-like ionophore activity.

Analogues of alamethicin, a 20-mer amphipathic helical peptide with ionophore activity, in the sequence of which all Aib residues were substituted by Ala (A1) or Leu (L1), were synthesized by the solid phase method, purified by high performance liquid chromatography and characterized by fast atomic bombardment mass spectrometry. Infrared and CD studies showed that A1 easily underwent a transconformation to beta-structure whereas L1 displayed a predominant alpha-helical character, thus being a potential ionophore model. Its voltage-dependent multistate activity in model membranes showed that Aib is not a requisite residue to observe an alamethicin-like behavior. However, as the lifetime of the single channels was much shorter than for alamethicin, the peptide chain was lengthened by a Leu (LL1) or a Ser (SL1) residue. The last peptide gave an increased channel lifetime, but the design of other non-Aib peptides, taking into account the hydroxyl C-terminus and side-chain interactions between helices in a barrel-stave bundle, is desirable to approach more closely the alamethicin activity.

Ionophore properties of a synthetic alpha-helical transmembrane fragment of the mitochondrial H+ ATP synthetase of Saccharomyces cerevisiae. Comparison with alamethicin.

A 22-amino acid polypeptide was synthesized to model the central transmembrane segment of subunit 8 of the H+ ATP synthetase of Saccharomyces cerevisiae and to test ionophore properties. Solid-phase synthesis was conducted on benzhydrilamino resin, and purification followed by high pressure liquid chromatography allowed the isolation of the pure product whose NH2 terminal was acetylated and whose molecular weight determined by Fast Atomic Bombardment was the expected 2,666. The infrared spectrum of this peptide in the solid state reveals a fully alpha-helical conformation, whereas in low dielectric constant solvents the alpha-helical content is 60%, as determined by circular dichroism studies. Macroscopic current-voltage curves displayed by different planar lipid bilayers (monomyristoleoyl-glycerol and phosphatidylethanolamine) doped with this peptide suggest a weakly voltage-dependent conductance. Only one conductance level is observed in any given single-channel conductance experiment. However, a series of experiments shows a distribution of conductance states, most often 440 or 3,000 pS, and occasionally 80, 1,200, or 6,500 pS. This behavior contrasts with the usual behavior of alamethicin, chosen as a model of "aggregating-helices" ionophore and whose conductance fluctuates continually between substates, through uptake and release of monomers. Nevertheless, alamethicin too can display, under certain conditions, long-lived and mono-level conductance states similar to those reported here for the newly synthesized peptide. These properties could possibly be explained by the formation of large domains of helical rods with a set of allowed and independent ionic pathways.