[Exact determination of the lecithin/sphingomyelin ratio in amniotic fluid. Suggestion of a reference methods (author's transl)]. / Determination precise du rapport lecithine/sphingomyeline dans le liquide amniotique: Proposition d'une methode de reference.

Pulmonary maturity of the fetus can be evaluated by the lecithin/sphingomyelin (L/S) ratio in amniotic fluid. To existing methods of lipid extraction, precipitation with acetone and chromatography, we add a simple and accurate estimation of sphingomyelins (S) and precipitated lecithins (Lp) without acid digestion. The method is reproducible (C.V. less than 9%) for the measurement of Lp/S ratio and gives with accuracy the concentrations of Lp, avoiding possible errors in interpretation of Lp/S. Our results show that at 35 weeks of normal gestation, Lp/S ratio is about 2 and Lp concentration, 10 mg/1.

[Assay of proteins in the urine and cerebrospinal fluid. Choice of a method in centrifugal analysis]. / Dosage de la protéinurie et de la protéinorachie. Choix d'une méthode en analyse centrifuge.

The study consists in adapting two methods allowing estimation of proteins in cerebrospinal fluid and urine on a centrifugal analyzer using Coomassie Blue-SDS and benzethonium chloride. Results obtained with the two reagents are compared with turbidimetric technic using trichloroacetic acid, by statistic methods.

[Peroperative biopsy of the sub-apical urethra during prostatectomy for cancer]. / Etude par biopsie extemporanée de l'urètre sous-apexien au cours de la prostatectomie pour cancer.

INTRODUCTION AND OBJECTIVES: Between 35 to 45% of radical prostatectomies (RP) will show positive margins and approximately 50% of patients with positive margins will progress following RP. Peroperative diagnosis of positive margin possibly can improve the currability of RP. We report here the incidence and management of positive urethral margins diagnosed by frozen sections during radical prostatectomy. METHODS: Frozen sections were performed systematically to analyse the surgical margins during 130 RP performed for localized prostate cancer from january 1989 to december 1993. Peroperative analysis of distal (urethral) and proximal (bladder neck) margins, as well as posterior (Denonvilliers fascia) margins, and every surrounding tissue that was macroscopically abnormal were performed. RESULTS: Analysis of the distal margins showed no prostatic gland in 68 cases (52%), normal prostatic glands in 56 cases (43%) and neoplastic prostate glands in 6 cases (5%). Further urethral sections with frozen section analysis were performed in 62 cases until the distal margin was free of prostatic glands. Three out of six patients with positive urethral margins had another positive margin that was excised as well. Five out of six patients with positive urethral margins were pT3, NO, MO and one pT2, NO, MO. In the group of patients, with positive urethral margins, the average preoperative PSA serum level was 37 ng/ml and the average tumor weight was 14 g. Postoperative pelvic radiation therapy (45 grays) was performed in 5 of these patients. The mean follow-up is 36 months (12-68). PSA serum level is undetectable in 4 cases, and 0.4 ng/ml in 1 case. Urinary PSA level is undetectable in 5 out of 6 cases. CONCLUSION: Peroperative analysis of surgical margins, and particularly urethral margins, during RP allows to a better staging and to perform any tumor excision complement if possible. Surgical margins analysis when performed during surgery should help for the choice of the most adapted surgical procedure. In our experience, nerve-sparing RP are performed only when surgical margins are negative on frozen sections.

[Primary percutaneous approach in staghorn kidney calculus]. / Abord per-cutané premier dans la lithiase rénale coralliforme.

36 staghorn calculi were treated percutaneously under ultrasound guidance between 1983 and 1992. Each stone had a renal pelvic element and at least two caliceal branches. The area of each stone was measured on the plain abdominal x-ray (mean: 1,020 mm2) and the total length of the various caliceal branches was measured from the pelvic element (mean: 50.2 mm). These 36 procedures represented 8.2% of the 438 percutaneous nephrolithotomies performed over the same period. The stone was able to be entirely removed by nephrolithotomy in 12 patients. Of the 24 residual stones after percutaneous nephrolithotomy, 16 were treated by extracorporeal shock-wave lithotripsy, which eliminated all stones in 12 of these patients. Eight of the remaining 12 patients were lost to follow-up and treatment was not completed, and 4 present a residual stone (11% of failures). These results are compared with those of other series and are comparable to those of surgery which gives a similar residual stone rate of 16% in the AFU 1982 report [6]. The primary percutaneous approach to staghorn calculi therefore represents an effective therapeutic modality, whose use and results must be weighed up with those of surgery.

[Surveillance of renal function by scintigraphy with 99m Tc-DMSA after a trans-ileal cutaneous ureterostomy]. / Surveillance de la fonction rénale par scintigraphie au 99m Tc-DMSA après urétérostomie cutanée trans-iléale.

The renal function of sixty one adult patients was monitored by 99mTc-DMSA renal scan after cystectomy for a malignant tumour followed by ileal conduit urinary diversion. The postoperative follow-up period was 10 years. The stenosis rate of the uretero-ileal anastomosis was 12.8% and the incidence of pyelonephritis was 8% The mean value for overall renal function and for the function of each kidney did not deteriorate significantly (p = 0.1 and p = 0.7, respectively) over time. However, 26% of kidneys evaluated at 1 year and at 5 years showed a markedly decreased uptake on the renogram. In 70% of cases, this decreased uptake was related to the development of stenosis of the uretero-ileal anastomosis or pyelonephritis. Renal function remained stable at 5 years in group A, corresponding to patients with good initial renal function (n = 22, p = 0.07), and in group B, corresponding to patients with poor initial renal function (n = 7, p = 0.9). Similarly, the function of solitary kidneys did not deteriorate over the 5-year postoperative follow-up period (n = 7, p = 0.5). The functional value of the kidneys was therefore not globally altered after ileal conduit urinary diversion. The existence of a mechanical or infectious complication should be systematically investigated in the presence of a deterioration in renal function. DMSA isotope renal scan is a complete, qualitative and quantitative, follow-up examination after this type of urinary diversion.(ABSTRACT TRUNCATED AT 250 WORDS)

[Trans-ureteral pyelostomy: 5 cases]. / Trans-urétéro-pyélostomie: 5 observations.

The authors report 5 cases of extensive loss of substance of the distal ureter due to radiation fibrosis, operative trauma or tumour invasion; these lesions occurred during the course of a genital tumour in 4 cases and rectal prolapse in one case. The lesion was bilateral and radiation-induced in 2 cases and unilateral in 3 cases. The patients presented with renal colic in 2 cases, renal failure in 1 case and infection in 2 cases (pyelonephritis, septic shock). All patients presented with secondary hydronephrosis. The extent of the lesions made vesical anastomosis impossible, especially as the bladder was irradiated and/or absent in 3 cases. Anastomosis of the proximal ureteric stump onto the contralateral ureter was impossible in the presence of bilateral lesions requiring diversion of the contralateral ureter or due to the short donor ureter, or because of the discordant diameter of the two ureters. Transureteropyelostomy was therefore performed in these 5 cases. The postoperative course was uneventful 4 cases, with a transient anastomotic fistula in only one case. Four of the 5 patients achieved long-term survival with an excellent clinical and radiological result.

Results of radical cystectomy and pelvic lymphadenectomy for bladder cancer with pelvic node metastases.

OBJECTIVES: To study the results on survival of radical cystectomy and pelvic lymphadenectomy in bladder cancer with pelvic node metastases in a retrospective study. PATIENTS AND METHODS: Of 248 patients with transitional cell carcinoma of the bladder treated by cystectomy with pelvic lymphadenectomy from 1970 to 1990, 40 (16%; 37 males, 3 females) had pelvic nodal metastases. Fifteen patients had one positive lymph node (stage pN1) and 25 patients had more than one positive node (stage pN2). Preoperative pelvic radiotherapy was realized in 14 patients. RESULTS: The perioperative mortality rate was 5% and the perioperative morbidity rate 15%. The overall 1- and 5-year survival rates for patients with nodal metastases were, respectively, 42 and 14%. Overall median survival was 9 months (range: 1-288 months). Three patients are still alive with follow-ups of 7, 12 and over 20 years. Kaplan-Meier survival curves revealed no survival advantage for stage pN1 over pN2 disease (p = 0.10). The 5-year survival rate for patients with stage pN1 was 22% with a median of survival of 17 months (range: 1-288 months). The 5-year survival rate for patients with pN2 disease was 8% with a median of survival of 7 months (range: 1-144 months). The cause of the death was tumoral progression in 26 patients (75%) and local recurrence in 3 patients (10%). A preoperative radiotherapy didn't influence survival. CONCLUSION: In our experience, radical cystectomy with pelvic lymphadenectomy provided long-term progression-free survival for a few patients with nodal involvement. Efficacious adjuvant therapy has to be found to improve the results.

[Left varicocoele and iliac venous stasis (author's transl)]. / Varicocèle gauche et stase veineuse iliaque

Idiopathic left varicocele is not always due to renospermatic reflux, as is shown by left renal phlebography. Furthermore, radiological exploration of the left iliac venous axis has shown the frequency of main iliac obstruction, compressed by the right iliac artery. The lumen of the iliac vein frequently bears traces of this compression (May and Thurner). The multiplicity of the colateral pathways draining the iliac vein in stasis, is evidence of the reigning increase in pressure. The authors propose the hypothesis that the varicocoele may be due to iliospermatic reflux. This occurs through anastomoses which are well known to anastomists and found on 2 occasions in the series presented. If this hypothesis were verified, it would incite an approach to the cord via the inguinal canal in order to ligature the spermatic veins above their anastomoses with the epigastric.

[Total prostatectomy for stage D1 cancers]. / La prostatectomie totale dans les stades D1.

Nineteen patients with stage D1 (TXN/MO) prostatic adenocarcinomas were treated with radical prostatectomy and adjuvant radiation therapy. The latter was aortoiliac for stage B tumors and aortoiliac plus pelvic for stage C tumors. No hormonal therapy was associated to this. Lymph node invasion was multifocal in 10 of 19 cases, bilateral in 3 cases, and massive with rupture of the capsule and invasion of the fat surrounding the lymph nodes in 3. The maximum follow-up was 17 years, with a minimum follow-up of 1 year. The dosage of the prostatic specific antigen (PSA) has been an essential element of surveillance since 1987. Eight patients died of their cancer, 11 are alive, including 9 alive and healthy (47%). Among the latter, 5 (26%) are alive without any treatment, with a prostatic antigen level around zero at a follow-up of 6 to 12 years. The review of the literature shows that nonsurgical treatments have a palliative effect for stage D1 tumors, and that surgery alone is not always sufficient. Surgery must be followed by radiation therapy, the only modality likely to complete the effect of radical surgery with the same curative purpose.

[Congenital segmental duplication of the lumbar ureter (author's transl)]. / Dédoublement segmentaire congénital de l'uretère lombaire

In a 23-year-old man attacks of nephritic colic led to the discovery of an obstruction on the left lumbar ureter. Segmental resection of the ureter was performed, removing 10 mm of malformed, obstructed ureter. This was an incomplete duplication, the two ureteral segments lying side-by-side, each with its own musculature, for a distance of 7mm. Above and below the anomaly, the ureter was normal. This exceptional malformation is compared with other internal obstructions of the ureter.

[Extemporaneous histological control of margins in prostatectomy for cancer]. / Le contrôle histologique extemporané des marges dans la prostatectomie pour cancer.

A histological study of the margins of radical prostatectomy specimens has been made intraoperatively by frozen section in a series of 66 patients. In 8 of them (12% of all) a positive margin was discovered 10 times at the urethral [3], vesical [2], deferential [1] level, at the level of the Santorini [1] and of the rectoprostatic wall [3]. Further resection was immediately performed until histologically normal tissue was reached in 6 of these patients, for whom positive margins were thus made negative; and the prostatectomy was performed. The prostate was not removed in two patients. Only one positive margin not screened intraoperatively was discovered on definitive specimen examination. The rate of postoperative PSA was lowered in the 6 patients whose margins had been made negative, which is compatible with complete resection. However, 7 of these 8 patients received radiation (45 gray). We made a comparison between the groups with negative (57 patients) and positive margins (9 patients) for preoperative PSA levels, Gleason's scores, and invasion of the capsule, of the seminal vesicles and of the lymph nodes. There was no significant difference between both groups for the PSA levels, Gleason's score and the involvement of the seminal vesicles. Conversely, there was a significant difference between both groups (corrected chi 2 test) for invasion of the capsule (p = 0.02) and of lymph nodes (p = 0.02). Intraoperative histological control allows immediately recognizing and treating some positive margins, in order to choose during the operation whether resection should be widened or abandoned.

Histone H4 from cuttlefish testis is sequentially acetylated. Comparison with acetylation of calf thymus histone H4.

The differently acetylated subfractions of histone H4 isolated from cuttlefish testis and from calf thymus were separated by ion exchange chromatography on sulfopropyl-Sephadex, using a shallow linear gradient of guanidine hydrochloride in the presence of 6 M urea at pH 3.0. The tetra-, tri-, di-, mono-, and nonacetylated forms of cuttlefish H4 represent 2, 6.4, 18, 32.2, and 41.4% of the whole histone, respectively. The di-, mono-, and nonacetylated forms of calf H4 represent 11.7, 41.3, and 44% of the whole histone, respectively. The acetylation sites were determined in each subfraction by identification of the acetylated peptides. In each acetylated H4 subfraction, the acetylated tryptic peptides were identified by peptide mapping and amino acid analysis with reference to the peptide map of nonacetylated H4. In cuttlefish testis H4, lysine 12 is the main site of acetylation in the monoacetylated subfraction; lysines 5 and 12 are found acetylated in diacetylated H4; lysines 5, 12, and 16 are found acetylated in triacetylated H4. From these results and the stoichiometry of the different H4 subfractions, it can be concluded that lysine 5 is acetylated after lysine 12. In calf thymus, lysine 16 is the only site of acetylation in the monoacetylated subfraction. All the diacetylated forms are acetylated in lysine 16, the second site of acetylation being, in decreasing order, lysine 12, lysine 5, or lysine 8. These observations suggest that acetylation occurs in a sequential manner. Moreover, the sites of acetylation depend upon the biological event in which acetylation is involved.

The amino-acid sequence of histone H2A from cuttlefish Sepia officinalis.

The amino acid sequence of cuttlefish testis histone H2A (124 residues) was established from structural data obtained by automated sequencing of large peptides generated by the cleavage of the protein with V8 staphylococcal protease or by limited chymotryptic hydrolysis. Compared to the calf thymus homologous histone, cuttlefish H2A shows 14 substitutions (most of them conservative) and 5 deletions. Extensive evolutionary changes were mainly observed in the basic amino-terminal and carboxy-terminal regions of the molecule, which are the primary DNA-binding sites. Few punctual changes are observed in the central region (residues 18-118), which interacts strongly with histone H2B to form the dimer H2A-H2B.

Isolation and characterization of a small putative zinc finger protein from cuttlefish epididymal sperm cells.

At the end of spermiogenesis, sperm chromatin stabilization is ensured by protamine dephosphorylation and, in mammals, by the formation during epididymal transit, of intra- and inter-molecular disulfide bridges between protamines. In cuttlefish, the nuclear protein transition histones-->spermatid-specific protein T-->protamine Sp is very similar to that occurring in mammals during spermiogenesis. However, in cuttlefish, the protamine Sp is devoid of cysteine residues. The protein complement of cuttlefish epididymal sperm nuclei has been investigated. A minor basic protein, called protein E, has been isolated. Its primary structure was established from sequence analysis and mass spectrometry data of the protein and its fragments. Protein E contains a motif -Cys-Xaa2-Cys-Xaa23-His-Cys-Xaa2-Cys- which is likely to adopt a zinc finger conformation. Reduced protein E does fix zinc whereas alkylation of cysteine residues abolishes this ability. The sequence of protein E does not correspond to that of any known protein, but presents some similarities with a part of ZFY protein, a putative human transcription factor specifically expressed in germinal cells and which could be involved in spermatogenesis.

Cuttlefish sperm protamines. 1. Amino acid sequences of two distinct variants.

The amino acid sequences of two cuttlefish protamine variants Sp1 and Sp2 have been established from automated sequence analysis and mass spectrometry data. Sp1 (57 residues) and Sp2 (56 residues) have molecular masses of 8410 and 8253 Da, respectively. They are almost identical proteins which differ only by one residue of arginine and the position of two of the serine residues (14 and 37 in Sp1; 13 and 35 in Sp2). With an arginine content of about 77%, cuttlefish protamine is one of the most basic proteins which have ever been characterized and the first typical protamine sequenced in invertebrates. It is closely similar to sperm basic proteins identified in squids but strongly differs from the protamine-like components isolated from the sperm of bivalve molluscs.

Cuttlefish spermatid-specific protein T. Molecular characterization of two variants T1 and T2, putative precursors of sperm protamine variants Sp1 and Sp2.

In cuttlefish, as in selachians and mammals, spermiogenesis is characterized by the double nuclear protein transition histones----intermediate protein (protein T)----protamine (protein Sp). The cuttlefish protein T, which consists of two structural variants phosphorylated at different degrees, is the first invertebrate spermatid-specific protein to be fully characterized and sequenced. The primary structures of these two variants were established from sequence analysis and mass spectrometric data of the proteins and their fragments. T1 and T2 are two highly related proteins of 78 and 77 residues, respectively, which differ only by four conservative substitutions, two inversions Ser in equilibrium with Arg, and the deletion of 1 residue of arginine in variant T2. The asymmetrical distribution of the hydrophobic and basic residues determines two well defined domains: an amino-terminal domain (residues 1-21) devoid of arginine and aromatic residues and containing all the aliphatic hydrophobic residues and a highly basic carboxyl-terminal domain (residues 22-77 or 78) that contains 77% of arginine, all the tyrosine residues, and most of the phosphorylated serine residues present in the protein. The complete structural identity of the basic carboxyl-terminal domain of spermatidal proteins T1 and T2 with the protamine variants Sp1 and Sp2 isolated from cuttlefish spermatozoa strongly suggests that T1 and T2 could be precursors of Sp1 and Sp2, respectively.