The molecular characterization of the major polar tube protein gene from Encephalitozoon hellem, a microsporidian parasite of humans.

The microsporidia are obligate intracellular protozoan parasites of increasing importance as human pathogens, which are characterized by a small resistant spore with a single polar filament that coils around the sporoplasm. When stimulated, the polar filament rapidly everts out of the spore to form a hollow polar tube through which the sporoplasm passes, thus serving as a unique mechanism of transmission. A genomic library of the human microsporidium Encephalitozoon hellem was screened using a polyclonal rabbit antibody (anti-PTP Eh55) produced to the major HPLC purified polar tube protein (PTP) of E. hellem. This antibody localized to intrasporal polar filaments and extrasporal polar tubes of E. hellem by immunogold electron microscopy confirming the polar tube specificity of the antibody. A total of 14 anti-PTP Eh55 reactive genomic clones were identified and purified. A PTP gene was identified consisting of 1362 bp coding for 453 amino acids. The N-terminus of the translated protein consists of aputative N-terminal signal sequence of 22 amino acids, which when cleaved results in a mature protein of 431 amino acids with a predicted molecular mass of 43 kDa. The protein has a high proline content (14.6%) and contains a central domain of six alternating tandem repeats of 20 amino acids. After ligation of the gene into a glutathione S-transferase (GST) expression vector, a fusion protein was produced that reacted by immunoblotting with the polar tube specific anti-PTP Eh55. The gene was present as a single copy in the genome and there was no homology with other known genes. As the polar tube is a critical structure for the transmission of this organism to a new host cell, further study of PTPs may lead to the development of new therapeutic strategies and diagnostic tests.

220 cases of distal hypospadias: results of MAGPI and Duplay procedures. Respective place of glanduloplasty and urethroplasty.

220 distal hypospadias have been operated over a period of 10 years: 53 glandular, 144 coronal and 23 anterior penile subcoronal hypospadias with no chordee. Although with 131 MAGPI procedures we had only 3 urethral complications (one fistula, one total and one partial disunion), it does not seem to be a well adapted procedure for the correction of coronal and anterior penile hypospadias: the new meatus is not exactly apical, the glans remains short on its inferior side, and we often noticed a distal curvature at the end of the intervention. The urethroplasty technique that we performed 89 times avoids these disadvantages: the urethra is reconstructed in 2 layers as far as the summit of the glans according to Duplay's procedure, the urethrocutaneous adhesions are freed down the penis, the distal thin urethra is covered by an intermediate layer and the shortage of ventral skin is compensated for by the formation of a circular esthetic mucous annulus. There have been no complications with glandular hypospadias (7 cases), 4 with coronal hypospadias (64 cases) and 4 with anterior penile hypospadias (18 cases). While the MAGPI procedure remains an excellent technique for the correction of glandular hypospadias, the Duplay's procedure is better adapted for the correction of coronal hypospadias, in spite of the occurrence of more frequent (easily treated) fistula (6.25%). The juxta coronal position of some anterior penile hypospadias is still better repaired by the Mathieu's or the Snyder's procedures.

Single-stage correction of posterior hypospadias (178 cases). Comparison of three techniques: free skin graft, free bladder mucosal graft, transverse pedicle preputial graft.

Three techniques have been developed for single-stage repair of posterior hypospadias: free skin grafts, free bladder mucosal grafts, and pedicle preputial grafts. This multicenter retrospective study of 178 children who underwent surgery for posterior hypospadias was designed to compare the results achieved with these procedures. Free skin grafts (15 cases) resulted in the most frequent complications, and in particular the most severe strictures; in our opinion this technique should be abandoned. Pedicle preputial grafts (133 cases) gave the greatest number of successes from the outset, and should be preferred whenever the dimensions of the prepuce are sufficient for urethroplasty. Results with bladder mucosal grafts (30 cases) were not as good as with preputial grafts, but this technique remains the only solution when the meatus is in a very posterior position, and for children who have already undergone multiple operations.

[A triple association: rectocolonic adenomatosis, gastroduodenal polyposis and secreting adrenocortical adenoma]. / Triple association: adénomatose rectocolique, polypose gastro-duodénale et adénome cortico-surrénalien sécrétant.

A case of adenomatosis coli, gastroduodenal polyps, and functional adrenocortical adenoma in a 28 year-old woman is reported. The association of cystic fundic gland polyposis and duodenal adenomatosis was demonstrated endoscopically. The particular point of interest in this case report is the functional nature of the adrenocortical adenoma. Endocrine tumors associated with adenomatosis coli are generally discovered at autopsy. No similar case has been previously published.

[Endoscopic treatment of vesicorenal reflux in children. Review of three years experience and prospects for the future]. / Traitement endoscopique du reflux vésico-rénal chez l'enfant. Le point sur trois années d'expérience et perspectives d'avenir.

Endoscopic injection of teflon paste, used since 1984, represents an alternative to the surgical treatment of vesico-renal reflux. The success rate varies from 75 to 89% after a single injection and from 85 to 95% after two injections, according to the teams. The results are less satisfactory for the high grades and for refluxes on duplicity (25 to 45% failure rate). This technique is particularly attractive in the case of a difficult surgical intervention, notably when a reimplantation has failed, in the case of reflux on neurological bladder, urethral valves or exstrophy. Various causes of failure or relapse are considered. The advantages of the technique are obvious (low morbidity, very short hospital stay) and surgery is always possible in the case of failure. After one or two years, the reflux reappears in 2 to 9% cases. Long term results cannot be assessed as the follow-up period is too brief at the present time. For this reason, and although the endoscopic and surgical findings are reassuring, tolerance to teflon both locally and at a distance requires confirmation and other products such as collagen are now being used.

[Management and treatment of uropathies detected in utero. Evaluation of 8 years' experience]. / Prise en charge et traitement des uropathies dépistées in utero. Bilan de 8 années d'expérience.

Over an 8 year period, the antenatal diagnosis of uropathy was made in 125 fetuses, but confirmed in only 22%; in the other cases, it was inadequate (50%) or wrong (28%). It was only in a few cases that surgery was any use in the first days of life (sectioning urethral valves, and temporary nephrostomy); in most cases, surgery was carried out between 1 and 3 months of age: total or partial nephrectomy (mainly for multicystic dysplasia and ureteral duplication with a ureterocele or ectopia), and usually conservative surgery (resection of the pyelo-ureteral junction, uretero-vesical implantation because of reflux or mega-ureter). The long-term results depend upon the degree of renal dysplasia: the prognosis was good in all cases of hydronephrosis that were operated on, but it is often poor when uropathies are characterized by early obstructive phenomena in intra-uterine life as occurs with urethral valves. In those cases, renal insufficiency cannot always be prevented by neonatal surgery.

[Improved prognosis and treatment of hydronephrosis due to ureteropelvic junction syndrome in children]. / Amélioration du pronostic et du traitement de l'hydronéphrose par maladie de jonction pyélo-urétérale chez l'enfant.

Ureteropelvic junction obstruction in children: evolution of diagnosis and treatment. 180 hydronephrosis due to ureteropelvic junction obstruction in 162 patients have been referred to our hospital. The patients were all infants and children with an age ranging between 1 day to 15 years. Out of 9 abstinences (grade I or 11) and 7 nephrectomies for destroyed kidneys (3 of them after temporary nephrostomy), we performed 164 ureteropelvic junction resections and anastomosis, 51 of them with associated pyeloplasty. The use of a stenting catheter passing through the anastomosis and coming out as a nephrostomy in 81 cases gave us 5 mechanical complications while the complete absence of drainage gave only 3 fistulaes. 2 of these fistulaes had a spontaneous rapid healing after endoscopic ureteral catheterism. Out of 160 follow-up cases, we reoperated 6 failures with success: 5 ureteropelvic reanastomoses and one ureterocalycostomy. This study shows us that the early treatment due to antenatal ultrasound diagnosis improves the prognosis. Renal scanning with furosemide test permits to specify the surgical indication in limited cases and the presence of a residual obstruction in case of persistance of renal pelvis dilatation in the postoperative period. The posterior approach and the absence of internal drainage simplify the treatment, improve the patient comfort and limit the hospitalisation period to 5 days.

[Analysis of the failure of endoscopic treatment of vesico-renal reflux in children using injections of teflon and collagen and the preliminary results of injections of Macroplastic]. / Analyse des échecs du traitement endoscopique du reflux vésico-rénal chez l'enfant par injections de téflon ou de collagène et résultats préliminaires des injections de Macroplastique.

This study of 785 cases of vesicorenal reflux in 494 children treated endoscopically over a 7-year period was designed to evaluate the results obtained with three products used successively: Teflon, collagen and Macroplastic. Following Teflon injection, despite a 90% short-term success rate, recurrent reflux was subsequently observed in 16.71% of the ureters reviewed. The failure rate was 52.63% after collagen injection and 11.77% after Macroplastic. After one or two injections, complete resolution of reflux was obtained in 48% of children treated with collagen, versus 85.72% with Teflon and 93.33% with Macroplastic. In one half of cases, failure was related to the quality of the product and its modifications after injection. The marked resorption of collagen accounts for the poor results despite the large doses injected. Apart from one case of partial resorption of Teflon paste, the failures with this product were due to lateralisation or secondary elimination of the product from the injection site due to its fluidity. Macroplastic, due to its higher viscosity and absence of retraction, currently provides the best results with doses of less than 0.20 ml in children.

[Malformations of Wolffian duct derived male genital organs (epididymis, vas deferens, seminal vesicules, ejaculatory ducts)]. / Malformations des organes génitaux masculins issus du canal de Wolff (épididyme, déférent, vésicule séminale, canal éjaculateur).

OBJECTIVES: To analyse embryological phenomena, in order to define a practical approach to management. METHODS: The authors reviewed the files of eight boys with an abnormality of Wolffian duct derived genital organs, consisting of three cases of vas deferens agenesis, two of which were accompanied by homolateral renal agenesis, one case of ectopic vas deferens with contralateral renal agenesis, three cases of duplicated vas deferens and one congenital seminal vesicle cyst with renal agenesis. DISCUSSION: Congenital malformations of the epididymis consist of cysts and agenesis or partial atresia. Cysts essentially raise a problem of differential diagnosis. Treatment is only required in the case of severe discomfort. Vas deferens agenesis is the commonest lesion, an incidental finding in children. An experimental treatment is proposed in adults. Duplications, interdeferential communications and deferentomegaly are much rarer lesions. Seminal vesicle cysts are well visualized by transrectal ultrasonography and should be treated surgically. Lastly, ejaculatory ducts may present urethroseminal reflux or may be ectopic. CONCLUSION: The possibility of absent or ectopic vas deferens should be considered in the case of unilateral renal agenesis. Vas deferens agenesis justifies examination of the contralateral side and investigation to exclude renal agenesis. Congenital cysts of the epididymis only require surgery when they are symptomatic. The presence of ipsilateral renal agenesis should be investigated in the case of cystic dilatation of the seminal vesicle.

[High frequency ventilation in neonatal surgery]. / Ventilation par oscillation à haute fréquence en chirurgie néonatale.

High frequency oscillatory ventilation (HFOV) is one of the artificial ventilation techniques used for the treatment of severe respiratory distress in the neonatal period. We studied (between 1987 and 1992) 57 newborn infants ventilated by HFOV during surgery. The aim was to facilitate the surgical procedure without having any deleterious effects on the gas exchange or the hemodynamic state of the infant. Three groups were studied. Group I (thoracic procedures: oesophageal atresia and pulmonary malformations, n = 25), group II (diaphragmatic hernia, n = 22), group III (abdominal procedures, n = 10). In the thoracic surgery group, no deleterious effect related to the HFOV was reported. The hyperventilation noticed after anaesthetic induction was rapidly corrected by placing the patient in a lateral position. In the three groups, no intraoperative complications related to the HFOV were observed. The diaphragmatic hernia group was divided into two subgroups according to the timing of surgery (emergency surgery, surgery after stabilization). In the latter subgroup, surgery was performed when the ventilation and gas exchange conditions had significantly improved. This subgroup showed the highest survival rate. This technique of peroperative ventilation is specifically used in the thoracic and diaphragmatic procedures of term and preterm newborn infants. It achieves a more stable operative field with less pulmonary expansions and diaphragmatic movements. In spite of the fact that HFOV is seldom used during anaesthesia, this study confirms its safety as well as the simplicity of its monitoring.

[Tumors of the liver in children]. / Les tumeurs du foie de l'enfant.

This paper is a concise review of tumors of the liver, which represent 2 to 5% of all tumors in children. Both benign and malignant forms are reviewed. The relationship between hepatitis virus B and carcinoma, cirrhosis, toxic agents and cancer is discussed.

Treatment of vesicoureteral reflux in children by endoscopic injection of Teflon. Review of 2 years of experience.

Endoscopic subureteral injection of Teflon was performed in 113 children (217 ureters) to correct all types of vesicoureteral reflux. In all cases immediate disappearance of the reflux was noted on an operative cystogram. Two-hundred and thirteen ureters were controlled at 1 month. Reflux had disappeared in 85.11% of the cases after a single injection and after 2 injections in 89.3% of the cases. The results remained stable in 93.22% of the cases at 1 year. Most relapses were explained by an insufficient quantity of Teflon and difficult anatomical conditions. Indications for this technique and the safety of the paste used are discussed.

The ultrastructure of spores (Protozoa: Microsporida) from Lophius americanus, the angler fish.

Spinal and cranial ganglia of American angler fish, Lophius americanus, are often infected with microsporidia. This protozoon elicits the formation of large, spore-filled, hypertrophied host cells, cysts. Previous reports of microsporidia in European lophiids identify the parasite as Spraguea lophii, a genus which has recently been shown to be dimorphic. The spores from L. americanus are monomorphic (2.8 X 1.5 micron) and uninucleate. Each spore contains a polar tube that forms six to nine coils. Spraguea lophii differs from the microsporidium described in L. americanus in several ways. Spraguea lophii has two spore types: a large spore (4.0 X 1.25 micron) containing a diplokaryon and three to four polar tube coils and a smaller uninucleate spore (3.5 X 1.5 micron) with five to six polar tube coils. Because of these major differences, the microsporidium from L. americanus is removed from the genus Spraguea and returned to its original genus, Glugea, as a new species, G. americanus n. sp. Other ultrastructural characteristics of G. americanus are included: the posterior vacuole encloses two distinct membranous structures; one is tubular and resembles a "glomerular tuft" and the second is lamellar and composed of concentric membrane whorls, additionally, the straight or manubroid portion of the polar tube proceeds beyond the posterior vacuole before it turns anteriorly and begins to coil.

[Correction of posterior hypospadias by a transverse vascular preputial graft, evolution of technics and results. Apropos of 42 cases]. / Correction des hypospadias postérieurs par greffon préputial transverse vascularisé, évolution des techniques et résultats. A propos de 42 observations.

During more than seven years 42 hypospadias were treated in on stage with different technics of urethroplasty by transverse preputial gauntlet grafts. With original Duckett's technic only 21,5% of patients had a good result. The authors reduced the risk of devascularisation first time by a personal modification then by using the double face flap described by Duckett himself; then the percentage of success after one operation overtake 75%. The different complications and their treatment are described and some operative details are pointed out to prevent complications.

The effects of elevated temperatures and various time-temperature combinations on the development of Brachiola (Nosema) algerae N. Comb. in mammalian cell culture.

Nosema algerae Vávra and Undeen 1970, a microsporidian known to cause infection in mosquitoes, develops in mammalian cell cultures at 24-35 degrees C and in the tails and footpads of athymic mice. More recently it has been reported to grow at 38 degrees C in human cell culture. The present study is a two-part temperature/development examination. The first part examines the development of N. algerae in rabbit kidney cell culture at 29 degrees C, which permits the formation of functional spores within 72 h, and compares the effect of elevated temperatures (36.0, 36.5, 37 degrees C) on parasite development. At these elevated temperatures, N. algerae infects but undergoes only one or two proliferative divisions, with no evidence of sporogony by 72 h post-inoculation. During this time, however, the host cells continue to divide resulting in fewer infected cells over time and giving the appearance of a diminished parasitemia. Additionally, at 37 degrees C some organisms degenerate/hibernate by 72 h while others remain viable/active. It is not until 96 h that the parasites appear in large clusters of proliferative stages in the few host cells that are infected. By 120 h post-inoculation, proliferative cells, sporoblasts, and early spores are observed. These results suggest that elevated temperatures impede proliferation rates and the onset of sporogony. The second part of this study evaluates developmental changes in N. algerae when incubation temperatures and times are varied during parasite growth, resulting in abnormal parasite morphology. These abnormalities were not present when parasites were grown at constant temperature (29-37 degrees C). This report demonstrates that N. algerae can successfully develop at high temperatures (37 degrees C), justifying its taxonomic relocation to the genus Brachiola.