Mutant WD-repeat protein in triple-A syndrome.

Triple-A syndrome (MIM 231550; also known as Allgrove syndrome) is an autosomal recessive disorder characterized by adrenocorticotropin hormone (ACTH)-resistant adrenal insufficiency, achalasia of the oesophageal cardia and alacrima. Whereas several lines of evidence indicate that triple-A syndrome results from the abnormal development of the autonomic nervous system, late-onset progressive neurological symptoms (including cerebellar ataxia, peripheral neuropathy and mild dementia) suggest that the central nervous system may be involved in the disease as well. Using fine-mapping based on linkage disequilibrium in North African inbred families, we identified a short ancestral haplotype on chromosome 12q13 (<1 cM), sequenced a BAC contig encompassing the triple-A minimal region and identified a novel gene (AAAS) encoding a protein of 547 amino acids that is mutant in affected individuals. We found five homozygous truncating mutations in unrelated patients and ascribed the founder effect in North African families to a single splice-donor site mutation that occurred more than 2,400 years ago. The predicted product of AAAS, ALADIN (for alacrima-achalasia-adrenal insufficiency neurologic disorder), belongs to the WD-repeat family of regulatory proteins, indicating a new disease mechanism involved in triple-A syndrome. The expression of the gene in both neuroendocrine and cerebral structures points to a role in the normal development of the peripheral and central nervous systems.

Identification of human cancer-related genes by naturally occurring Hepatitis B Virus DNA tagging.

Proviral tagging has been used in animals as a powerful tool for cancer genetics. We show that a similar approach is possible in patients with hepatocellular carcinoma (HCC) infected by Hepatitis B Virus (HBV), a human pararetrovirus which may act by insertional mutagenesis. In this work, the HBV genome is used as a probe to identify cancer-related genes. By using HBV-Alu-PCR, we obtained 21 HBV/cellular DNA junctions from 18 different patients. In six of 21, we found the HBV DNA integrated into a cellular gene: (1) Sarco/Endoplasmic Reticulum Calcium ATPase1 Gene; (2) Thyroid Hormone Receptor Associated Protein 150 alpha Gene; (3) Human Telomerase Reverse Transcriptase Gene; (4) Minichromosome Maintenance Protein (MCM)-Related Gene; (5) FR7, a new gene expressed in human liver and cancer tissues; and (6) Nuclear Matrix Protein p84 Gene. Seven junctions contained unique cellular sequences. In the remaining eight, the HBV DNA was next to repetitive sequences, five of them of LINE1 type. The cellular genes targeted by HBV are key regulators of cell proliferation and viability. Our results show that studies on HBV-related HCCs allow to identify cellular genes involved in cancer. We therefore propose this approach as a valuable tool for functional cancer genomic studies in humans.

Recurrent de novo mitochondrial DNA mutations in respiratory chain deficiency.

Starting from a cohort of 50 NADH-oxidoreductase (complex I) deficient patients, we carried out the systematic sequence analysis of all mitochondrially encoded complex I subunits (ND1 to ND6 and ND4L) in affected tissues. This approach yielded the unexpectedly high rate of 20% mutation identification in our series. Recurrent heteroplasmic mutations included two hitherto unreported (T10158C and T14487C) and three previously reported mutations (T10191C, T12706C and A13514G) in children with Leigh or Leigh-like encephalopathy. The recurrent mutations consistently involved T-->C transitions (p<10(-4)). This study supports the view that an efficient molecular screening should be based on an accurate identification of respiratory chain enzyme deficiency.

[Laparoscopic promontofixation technique]. / Technique de promontofixation laparoscopique Laparoscopic promontofixation technique.

Laparoscopic promontofixation often remains possible whatever the previous history of pelvic surgery, including the placing of prosthetic material. Preoperative care is standardized and is accompanied by antibiotic prophylaxis, preventive antithrombotic treatment and in the event of a history of pelvic surgery, a digestive preparation. Positioning of the patient must plan a 30 degrees Trendelenbourg position. After the introduction of trocars, initial surgery comprises interrectovaginal dissection to free the whole posterior surface of the vagina. This is followed by the installation of a posterior mesh pre-cut in an arch. The anterior face of the promontory is then freed after incision of the posterior peritoneum with the patient placed beforehand in a Trendelenbourg position. After intervesical vaginal dissection, the anterior prosthesis comprising a precut polyester mesh is fixed avoiding excess traction. The end of the surgery involves careful reperitonization of all the prosthetic parts. Possible specific surgical complications are vascular and visceral wounds. Postoperative secondary haemorrhage and gastrointestinal occlusion may occur. Occurrence of an inflammatory syndrome and low back pain suggests spondylodicitis and MRI should be performed. Vaginal erosion on the prosthesis (1.6 to 10% depending on the series) may occur after several months and seems relatively independent of the prosthetic material used.

Follow-up study of the use of antihypertensive drug treatment and of the population treated.

We carried out a study describing the extent and patterns of use of antihypertensive drugs and identifying common diseases in a cohort of workers. The population was monitored on a yearly basis by a questionnaire to assess the use of antihypertensive drugs. The extent and causes of absenteeism for medical reasons and diagnoses of cancer were taken from systematic records from the company. Four cross-sectional analyses covering a period of 5 years are presented. For the 17,244 respondents in 1990--12,731 men aged 41 to 51 years and 4,513 women aged 36 to 51 years--the prevalence of antihypertensive drug use was 6.5%, and 5 years later, antihypertensive drug use had doubled. This increase involved calcium channel inhibitors in particular. In all analyses, excess absenteeism for all health-related disorders was observed among those treated with antihypertensive drugs compared with the remainder of the population, who were almost all nonhypertensive: 44% v 34% (P < .000) in 1990, 42% v 34% (P < .000) in 1991, 41% v 34% (P < .000) in 1993, and 37% v 29% (P < .000) in 1994. Psychiatric disorders and fatigue were among the most frequent causes of absenteeism and were more frequent in individuals treated with antihypertensive drugs.

[Rehabilitation of the knee movement after ligamentoplasty using Mac Intosh's procedure augmented by Kennedy-Lad: a comparison between recent and old rupture of the anterior cruciate ligament]. / La récupération de la mobilité du genou après ligamentoplastie type Mac Intosh renforcé par Kennedy-Lad: étude comparative entre rupture récente et rupture ancienne du ligament croisé antérieur.

About 60 ligamentoplasties of the A.C.L. using the Mac Intosh procedure with augmentation by the Kennedy-Lad, the authors compare the rehabilitation of knee motion between fresh tears (30 cases) and old tears (30 cases) of the A.C.L. operated on by the same procedure. All the operations were performed by the same surgeon. The rehabilitation program was the same for everybody; no plaster cast, total weight-bearing after the 15th day, no more crutches-stick after the 21th day and beginning of flexion on the 12th day, 88 per cent of the knees were rehabilitated by the same physiotherapists. The plaster cast is usually incriminated to be the main reason of post-operative knee stiffness. But no plaster cast for fresh A.C.L. tear also give such a stiffness (16.5 per cent). The authors think that the initial injury increased by the surgical trauma, for a non conditioned patient are the main factors of post-operative stiffnesses. This study justifies the late reconstruction of "isolated" A.C.L. tears (between the 2nd and 3rd month), after "cooling down" of the lesions.

[Laparoscopic promontofixation]. / Promontofijación laparoscópica. Nuestra técnica.

OBJECTIVES: The pathology of the pelvic floor, including the urinary incontinence, the anal incontinence and the genital prolapse, is very dominant, concerning approximately a third of the adult women. It is fundamental that this musculature supports a good function, because of the weakness of the pelvic floor produces urinary incontinence, cysto and rectocele, genital prolapses and sexual dysfunctions. The above mentioned pathology can be corrected by laparoscopic promontofixation, whatever the previous history of pelvic surgery, including the placing of prosthetic material. In this article we describe the above mentioned intervention. MATERIAL AND METHODS: Preoperative care is standardized and is accompanied by antibiotic prophylaxis, preventive antithrombotic treatment and in the event of a history of pelvic surgery, a digestive preparation. Positioning of the patient must plan a 30° Trendelenbourg position. After the introduction the trocars, initial surgery comprises anterior dissection of promontory after incision of the posterior peritoneum with the patient placed beforehand in a Trendelembourg position. After that, we make interrectovaginal dissection to free the whole posterior surface of the vagina. This is followed by the installation of a posterior mesh pre-cut in an arc. After intervesical vaginal dissection, the anterior prosthesis comprising a precut polyester mesh is fixed avoiding excess traction. The end of the surgery involves careful reperitonization of all the prosthetic parts. Possible specific surgical complications are vascular and visceral wounds. RESULTS Y CONCLUSIONS: The technique allows the correction of the dysfunction of the pelvic floor and incontinence with good anatomical and functional results. Postoperative secondary haemorrhage and gastrointestinal occlusion may occur. Occurrence of an inflammatory syndrome and low back pain suggests spondylodicitis and MRI should be performed. Vaginal erosion on the prosthesis may occur after several months and seems relatively independent of the prosthetic material used.

Nerve sparing laparoscopic radical prostatectomy: our technique.

PURPOSE: To describe our technique of nerve sparing laparoscopic radical prostatectomy (LRP). We present the oncological and functional results (potency and urinary continence). MATERIAL AND METHODS: LRP has become standard at our institution based on experience with more than 2800 consecutive cases operated on between 1997 and 2005. From May 2003 to March 2005 a total of 677 LRP were performed, 425 consecutive patients candidates for a nerve sparing technique have been operated using the intrafascial approach. The challenge of our technique is to remove the prostate without any thermic and mechanic traumatism, avoiding dissection of outer layer. Oncological data were assessed by pathological examination and post-operative PSA level. Functional results were assessed with a self questionnaire. RESULTS: By pathological stage, 2 pT2a specimens (7.4%), 7 pT2b specimens (21%), 44 pT2c specimens (24%), 63 pT3a specimens (43%), 11 pT3b specimens (46%) were found to have positive surgical margins (SMs). In 86 specimen (59%) positive SMs were focal inframillimetric. Median follow-up was 11 months (range 1-22). The continence rate (no leakage/no pad) was 95% at 6 months, confirmed at 12 months among 202 patients. For 137 patients, potency rate was 58.5% at 12 months. CONCLUSION: Intrafascial LRP provides satisfactory results in regard to recovery of continence and sexual function. Long-term progression and survival outcome are necessary before this procedure should be offered as a replacement for interfascial nerve sparing technique.

Restriction map construction using a 'complete sentences compatibility' algorithm.

We have developed a new algorithm 'Complete sentences compatibility' (CSC) which uses single and double digestion fragments to rapidly determine restriction maps of circular DNA. From possible combinations of fragments of each simple digestion, which we call 'sentences of decomposition', we construct a restriction map which combines the sentences while taking into account compatibility rules. The algorithm can also deal with experimental errors of fragment weight and can suggest solutions that account for non-readable bands (fragments of zero length or multiple bands) on the gel. Because experiments using pairs of restrictive enzymes often result in multiple solutions, a complementary algorithm tries to reduce the number of proposed solutions by establishing consensus maps. The restriction map construction algorithm was tested on real cases, some containing more than fifteen fragments. Execution times range from 1-10 s on an IBM PC compatible microcomputer.

[Subcutaneous rupture of long finger flexor pulleys in rock climbers. 12 case reports]. / Les ruptures sous-cutanées des poulies des fléchisseurs des doigts longs chez les grimpeurs de haut niveau. A propos de douze cas.

Rock climbing generates a new type of microtraumatic, stress pathology involving all of the osteoarticular chain of the upper limb. The hand is the site of specific lesions: pulley ruptures (A2) are the most frequent. 12 top-level climbers (7a and above) were reviewed out of a series of 23 cases. In 7 cases, the ring finger was affected and in 5 cases the middle finger was affected. All cases were treated by rest and protection of the A22 pulley with an external rigid ring. Progressive return of climbing was allowed after the 45th day. The effects on the level of performance, residual pain and its consequences, and on associated lesions of the upper limb were investigated. 8/12 patients regained at least their previous level of performance and 6/12 no longer experienced any pain. 5/12 patients experienced minor pain which did not affect their level of performance. One patient had to stop top-level climbing because of persistent pain. Protection of the pulley, possibly combined with syndactylisation and complete rest, not only from sports activities, for at least 45 days are the best way of ensuring a satisfactory functional result in these particularly demanding subjects. Physiotherapy and non-steroidal anti-inflammatory treatment can also be combined.

A computer program for the design of optimal synthetic oligonucleotide probes for protein coding genes.

A computer program has been written in FORTRAN 77 to locate on a protein sequence a region with optimum length and limited degeneracy in order to design artificial oligonucleotide probes for use in molecular cloning. In addition the program checks for regions of homology between this probe and any other base sequence found in nucleotide sequence data banks. There are options in the program to eliminate rare codons or to make preferential choices of bases in order to minimize the degeneracy of probes.

In vitro synthesis of 17,20 beta,21-trihydroxy-4-pregnen-3-one by ovaries of turbot (Scophthalmus maximus L.) during oocyte maturation.

Ovaries from female turbot (Scophthalmus maximus L.), a serial spawner, were incubated in vitro with 17-hydroxy[1,2,6,7-3H]progesterone or [7-3H(N)]pregnenolone (P5-3H) during the spawning season. Several metabolites comigrated on TLC and HPLC with known reference steroids and were identified after chemical reaction and crystallization. Incubation with P5-3H generated many 4-ene steroids, accounting for 55% of total radioactivity, indicating strong 3 beta-HSD activity. The major steroids produced by ovaries were testosterone, androstenedione, and 17,21-dihydroxy-4-pregnene-3,20-dione. In addition, 17,20 alpha-dihydroxy-4-pregnen-3-one was also identified, while small quantities of 17,20 beta,21-trihydroxy-4-pregnen-3-one (20 beta-S) (maximum 1.7% of the total radioactivity) were also synthesized. The identity of 20 beta-S was confirmed in incubates with nonlabeled 17-hydroxyprogesterone by mass spectrometry. Production of 17,20 beta-dihydroxy-4-pregnen-3-one was not apparent.

[The evaluation of the Artem grip in its function assessment]. / Evaluation de la pince Artem dans sa fonction bilan.

The aim of our study is to assess the Artem pincers efficiency, which is both a new grip strength instrument and a hand reeducation instrument. It is original through its pleasant good looking and it offers many different programs of use. This study was carried out involving 60 healthy subjects. We compared the Artem system with two other measurement instruments of reference (Jamar for grip strength and Pinch-Gauge for pinch strength). With this Artem system we also studied two other originals tests: 1) Evolution of hand and finger strength within 15 seconds. 2) Tiredness test. Our measurements demonstrated good high correlation and high reliability between reference instruments and Artem system dealing with strength measurements. We can't conclude about the two original program's statistics, but it would probably be interesting to compare these with a group of patients in course of reeducation.

Metabolism of quinine in children with global malnutrition.

Malnutrition and malaria are two important public health problems in Africa. Quinine is one of the major treatments of chloroquine-resistant malaria. Although some authors have shown that quinine clearance is decreased in kwashiorkor, this type of malnutrition is caused by protein deficiency that differs from global protein-energy malnutrition. In rats, hepatic metabolism of many drugs is decreased in protein deficiency and increased in global food restriction. Several studies have found that human hepatic metabolism of many drugs is decreased in kwashiorkor, but, as yet, no study has focused on human global energy-protein malnutrition. Thus, as quinine is a drug with a narrow therapeutic index, we compared the pharmacokinetics of quinine in two groups. One group included children with global malnutrition and the other was a control group of children with normal nutrition. Volume of distribution and plasma concentrations of unbound quinine did not differ between children with global malnutrition and children with normal nutritional status. Clearance was significantly faster, half-life shorter, and concentrations, 12 h after the beginning of treatment, lower in malnourished children compared with control subjects. The ratio between area under the curve of hydroxyquinine (metabolite of quinine in man) and area under the curve of quinine was significantly increased in malnourished children and correlated with mid-arm/ head circumference ratio (marker of malnutrition in children). Thus, as metabolism of quinine is increased in children with global malnutrition, we suggest that the administration interval should be reduced in these children to obtain the same plasma concentrations of quinine found in normally nourished children. A safe and effective dosing strategy is postulated.

[Diffuse diverticulosis of the small intestine and malabsorption syndrome. Apropos of a case]. / Diverticulose diffuse du grêle et syndrome de malabsorption. A propos d'une observation.

A case of acquired jejuno-ileal malabsorption complicated by vitamin B12 malabsorption with macrocytic anemia and posterior column dysfunction is reported. Few such observations have been published in the medical literature. In the light of published studies and case-reports the authors review the pathogenic hypotheses concerning the formation of diverticula, the part played by bacterial infection in the mechanisms of malabsorption and the value of antibiotic therapy in the initial treatment of this condition.

Screening human EST database for identification of candidate genes in respiratory chain deficiency.

Disorders of mitochondrial oxidative phosphorylation (OXPHOS) are now recognized as major causes of human metabolic diseases and several mutations of mitochondrial and nuclear genes encoding respiratory chain components have been reported. Interestingly, mutations of nuclear genes involved in mitochondrial respiratory chain assembly, protein trafficking, and iron metabolism are also known to alter oxidative phosphorylation. While several hundred of these genes have been described in yeast, only a few nuclear genes have been hitherto identified in humans. Yeast gene databases present therefore an invaluable tool for identification of human homologues that should be regarded as candidate genes in OXPHOS diseases. In an attempt to identify the human counterparts of yeast genes, we developed a systematic comparison of yeast protein sequences to the GenBank dbEST database. Starting from 340 yeast protein sequences as templates, we searched the human dbEST counterparts using the BLAST similarity searching program and identified 102 groups of human EST likely to represent orthologues of yeast genes because of significant homology. This collection of human genes possibly related to mitochondrial OXPHOS may help identify nuclear genes responsible of mitochondrial disorders.

Cloning and expression analysis of a novel gene, RP42, mapping to an autism susceptibility locus on 6q16.

We isolated a novel mouse gene, RP42, in a systematic search for genes expressed in proliferating neuroblasts whose human orthologs map to susceptibility loci for autism. This gene is intronless and encodes a putative 259-amino-acid protein that exhibits 30-36% overall sequence identity to a fission yeast and a nematode protein (GenPept Accession Nos. CAA17006 and CAB54261). Nevertheless, no homology to any known gene was found. RP42 has developmentally regulated expression, particularly in proliferating neuroblasts from which neocortical neurons originate. Its human ortholog is located in a cluster of embryonic neuronally expressed genes on the 6q16 chromosome, making it a positional candidate susceptibility gene for autism.