Twenty sperm samples from five dogs were frozen in liquid nitrogen at -196 degrees C in 16 different media, two control media containing 20% egg yolk and 6% low-density lipoproteins (LDL); 10 test media containing 6% LDL (the active cryoprotective ingredient of chicken egg yolk) combined with 10, 20, 30, 40, 50, 60, 70, 80, 90, and 100 mmol of glutamine respectively at 4%, 5%, 7%, and 8% LDL. Following thawing, sperm mobility was assessed using an image analyser, HAMILTON THORN CERROS 12. The percentage of mobile spermatozoa was 62.05% in the 6% LDL + 20 mmol glutamine medium compared with 48.90% in the egg yolk-based medium (p < 0.05) or 57.55% for the 6% LDL medium (p < 0.05). Furthermore, in most cases, the motility parameters (average path velocity, curvilinear velocity, straight line velocity) in the 6% LDL + 20 mmol glutamine medium, were superior, to a statistically significant extent, to those in the control media. Finally, the 6% LDL + 20 mmol glutamine combination provides spermatozoa with better protection during freezing than egg yolk or the 6% LDL medium alone in terms of acrosome integrity (fluorescein isothiocyanate--Pisum sativum agglutinin test: p < 0.05), the flagellar plasma membrane (hypo-osmotic test: p < 0.05 for 6% LDL), the DNA (acridine orange test; no significant difference) and the integrity of the acrosome (Spermac test: no significant difference).

Background— Serotonin (5-hydroxytryptamine; 5-HT) overproduction is responsible for cardiac valvular disease in patients with carcinoid tumors. Reduced 5-HT inactivation is one proposed mechanism of the valvulopathy observed in individuals treated with the appetite suppressants fenfluramine and phentermine. One key protein limiting systemic availability of 5-HT is the 5-HT transporter (5-HTT) expressed by platelets and pulmonary vascular cells; 5-HTT is responsible for 5-HT uptake and subsequent inactivation of the amine passing through the lung. Here we investigated whether 5-HTT–deficient (5-HTT-KO) mice developed structural and/or functional cardiac abnormalities and valvulopathy. Methods and Results— Cardiac endothelial cells expressed large amounts of 5-HTT in wild-type mice. 5-HTT deficiency appeared to be associated with marked interstitial, perivascular, and valvular fibrosis as evidenced by staining of cardiac collagen in 5-HTT-KO mice. Histological analysis provided evidence for valvulopathy characterized by valvular hyperplasia and prominent fibrosis at the attachment site and base of the leaflets. Echocardiography revealed an increase in left ventricular lumen diameter and a decrease in left ventricular diameter fractional shortening. Although 5-HT 1B receptors mediated the 5-HT–induced collagen secretion by human cardiac myofibroblasts, the contribution of this receptor type to valvulopathy was ruled out because double-KO mice deficient in both 5-HTT and 5-HT 1B receptors showed the same cardiac alterations as 5-HTT-KO mice. Conclusions— The present results establish a link between 5-HTT and the development of cardiac fibrosis and valvulopathy in vivo. 5-HTT-KO mice represent an especially relevant model for studying the mechanisms by which 5-HT induces valvulopathy.

An efficient electromagnetic method for the optimization of shaped reflectors is presented. The repeated computation of scattering from a reflector surface at each iteration, using the conventional numerical techniques of either physical optics (PO) or aperture integration (AI), usually makes the iterative procedures very inefficient for the synthesis of large reflectors. An asymptotic Gaussian beam (GB) technique has been developed and applied successfully to the fast analysis of reflector antennas of various shapes (Cabbage, H.-T. et al., Proc. IEEE Ant. and Propag. Society URSI Symp., vol. 4, p.2336-9, 1999). This GB technique completely avoids numerical integration and thus makes the analysis very efficient. Our method uses the GB technique, coupled with a local description of the primary source and the reflector. The primary source radiation is expanded using a modified Gaussian beams basis. An antenna pattern calculation is demonstrated on a reflector that is described by local parameters in a novel way. By virtue of these local properties and the use of a steep step descent algorithm, a basic display of antenna pattern optimization is presented to illustrate the effectiveness of our method.

We present two distinct truncated variants of ankyrin associated with mild to moderate hereditary spherocytosis. Ankyrin Saint-Etienne 1 was manifested by an additional band located between bands 2.1 and 2.2. It was associated with a nonsense mutation in exon 39: TGG→TGA; W1721X. Ankyrin Saint-Etienne 2 appeared as two faint bands underlining bands 2.1 and 2.2. It was associated with a nonsense mutation in exon 41: CGA→TGA; R1833X. Overall ankyrin was diminished in splenectomized patients. Messenger RNAs Saint-Etienne 1 and 2 amounted to 20 and 37% of the total ankyrin mRNA, respectively. Ankyrin molecules truncated in their C-terminal region retain some ability to bind to the membrane whereas the bulk of nonsense mutations, located in more upstream regions, result in the mere disappearance of one haploid set of ankyrin. In the present cases, it was not possible to apportion the roles of ankyrin reduction and truncation in the pathogenesis of hereditary spherocytosis. Am. J. Hematol. 58:36–41, 1998. © 1998 Wiley-Liss, Inc.

We describe a case of spherocytosis in a French child splenectomized at age 10 years. The parents were devoid of any clinical, hematological, or biochemical abnormalities. Following splenectomy, the proposita exhibited a reduction of red cell membrane ankyrin. The variable number of dinucleotide repeats associated with the erythroid ankyrin gene ( ANK1 ) were studied at the genomic level. The father, the mother, and the proposita had the AC 14 /AC 11 , AC 14 /AC 14 , and AC 14 /AC 11 genotypes, respectively, although the proposita exhibited a pattern consistent with an AC 14 ,-combination at the cDNA level. We thought there could be a de novo mutation in the ANK1 allele of paternal origin (AC 11 ). A false paternity seemed most unlikely. Based on PCR-amplification of exons, SSCP analysis, and, when appropriate, nucleotide sequencing, we found a one-nucleotide deletion in codon 146 (exon 6): 521delC, A???→AG. This placed in phase a TAG triplet normally overlapping codons 150 and 151. Early interruption of translation presumably accounted for the premature degradation of mutant mRNA. Restriction analysis confirmed the presence of the mutation in the proposita and its absence in the parents. The variant was designated ankyrin Bugey. Am. J. Hematol. 54:242–248, 1997 © 1997 Wiley-Liss, Inc.

Severe intrauterine growth retardation and suggestive dysmorphic and malformation syndrome in 1 twin with a normal karyotype during a monozygotic, monochorionic pregnancy led to the diagnosis of discordant Russell-Silver syndrome (RSS). Placental anomalies (velamentous cord insertion, single umbilical artery, placental hypotrophy) for the small-for-date twins and a twin-twin transfusion syndrome confirmed at 21 weeks of amenorrhea suggest that early hemodynamic disorders may reveal or trigger RSS which has been considered to be of genetic origin. The prognosis for RSS is favorable, except for diminished adult size (less than -2 standard deviations).

Journal Article VNTR at the DXYS14 locus Get access M.-G.Le Roux, M.-G.Le Roux 1Laboratoire de Biologie MoléculaireCHU Hotel-Dieu, BP 1005, 44035 Nantes Cédex 01, France Search for other works by this author on: Oxford Academic PubMed Google Scholar O. Pascal, O. Pascal 1Laboratoire de Biologie MoléculaireCHU Hotel-Dieu, BP 1005, 44035 Nantes Cédex 01, France2Laboratoire de Génétrque des Espéces, CHU Hotel-DieuBP 1005, 44035 Nantes Cédex 01, France Search for other works by this author on: Oxford Academic PubMed Google Scholar A. Lostanlen, A. Lostanlen 1Laboratoire de Biologie MoléculaireCHU Hotel-Dieu, BP 1005, 44035 Nantes Cédex 01, France Search for other works by this author on: Oxford Academic PubMed Google Scholar I. Bérard, I. Bérard 1Laboratoire de Biologie MoléculaireCHU Hotel-Dieu, BP 1005, 44035 Nantes Cédex 01, France Search for other works by this author on: Oxford Academic PubMed Google Scholar O. Vergnaud, O. Vergnaud 2Laboratoire de Génétrque des Espéces, CHU Hotel-DieuBP 1005, 44035 Nantes Cédex 01, France Search for other works by this author on: Oxford Academic PubMed Google Scholar J.-P. Moisan J.-P. Moisan * 1Laboratoire de Biologie MoléculaireCHU Hotel-Dieu, BP 1005, 44035 Nantes Cédex 01, France2Laboratoire de Génétrque des Espéces, CHU Hotel-DieuBP 1005, 44035 Nantes Cédex 01, France *To whom correspondence should be addressed Search for other works by this author on: Oxford Academic PubMed Google Scholar Human Molecular Genetics, Volume 3, Issue 2, February 1994, Page 389, https://doi.org/10.1093/hmg/3.2.389-a Published: 01 February 1994

Four men and one woman of the same family with Kennedy-type-bulbo-spinal amyotrophy have been followed up for 7 to 20 years. The genetic marker: insertion of repeated sequences of trinucleotide Cytosine-Adénine-Guanine described by Fischbeck and La Spada in Nature (1991), in the coding region of the androgen receptor gene, on the long arm of X chromosome, has been demonstrated here by DNA extraction and PCR amplification.

Muckle-Wells syndrome is a rare autosomal dominant disorder characterized by chronic recurrent urticaria, periodic arthritis, sensorineural deafness, general signs of inflammation, and secondary amyloidosis (AA type). We report on a 4-generation family with 7 persons sharing various signs of this syndrome associated with bipolar aphthosis in 5 cases and cystinuria in one. Two other relatives in the family had ichthyosis.

Journal Article Missense mutation in the choroideremia gene Get access P. Donnelly, P. Donnelly Search for other works by this author on: Oxford Academic PubMed Google Scholar H. Menet, H. Menet Search for other works by this author on: Oxford Academic PubMed Google Scholar C. Fouanon, C. Fouanon 112 Rue Guepin44000 Nantes, France Search for other works by this author on: Oxford Academic PubMed Google Scholar O. Herbert, O. Herbert Search for other works by this author on: Oxford Academic PubMed Google Scholar J.P. Moisan, J.P. Moisan Search for other works by this author on: Oxford Academic PubMed Google Scholar M.G.Le Roux, M.G.Le Roux Search for other works by this author on: Oxford Academic PubMed Google Scholar O. Pascal O. Pascal * * To whom correspondence should be addressed Search for other works by this author on: Oxford Academic PubMed Google Scholar Human Molecular Genetics, Volume 3, Issue 6, June 1994, Page 1017, https://doi.org/10.1093/hmg/3.6.1017 Published: 01 June 1994 Article history Received: 04 March 1994 Revision received: 07 April 1994 Accepted: 07 April 1994 Published: 01 June 1994

Four men and one woman of the same family with Kennedy-type-bulbo-spinal amyotrophy have been followed up for 7 ro 20 years. The generic marker: insertion of repeated sequences of trinucleotide Cytosine-adenine-Guanine described by Fischbeck and La Spada in Nature (1991), in the coding region of the androgen receptor gene, on the long arm of X chromosome, has been demonstrated here by DNA extraction and PCR amplification.

Journal Article A new (old) deletion in the choroideremia gene Get access O. Pascal, O. Pascal * * To whom correspondence should be addressed Search for other works by this author on: Oxford Academic PubMed Google Scholar P. Donnelly, P. Donnelly Search for other works by this author on: Oxford Academic PubMed Google Scholar C. Fouanon, C. Fouanon 1Genetic Laboratory, University HospitalBP 1005, 44035 Nantes and 12 rue Guepin, 44000 Nantes, France Search for other works by this author on: Oxford Academic PubMed Google Scholar O. Herbert, O. Herbert Search for other works by this author on: Oxford Academic PubMed Google Scholar M.G.Le Roux, M.G.Le Roux Search for other works by this author on: Oxford Academic PubMed Google Scholar J.P. Molsan J.P. Molsan Search for other works by this author on: Oxford Academic PubMed Google Scholar Human Molecular Genetics, Volume 2, Issue 9, September 1993, Page 1489, https://doi.org/10.1093/hmg/2.9.1489 Published: 01 September 1993 Article history Received: 27 May 1993 Revision received: 18 June 1993 Accepted: 18 June 1993 Published: 01 September 1993

We have determined the frequency of the cystic fibrosis (CF) ΔF508 mutation in a large sample of CF patients originating from different areas of France, including the greater Paris, Brittany, Alsace, Lorraine and Rhône-Alpes regions. A total of 422 CF chromosomes were studied, and the defect was found to account for 75% of the mutant alleles. In the course of the survey, a rare nucleotide sequence polymorphism leading to an isoleucine to valine substitution at position 506 of the CF transmembrane conductance regulator protein has been characterized in an unaffected individual. Our data enable the evaluation of the probabilities that a chromosome negative for the ΔF508 mutation carriers another CF defect.

Poly(A+)-RNAs were extracted from human hyperplasic adrenocortical tissue and translated in a wheat germ cell-free system in the presence of [35S]-methionine. Labeled immuno-reactive adrenodoxin (ADX)-like material was immunoisolated and examined following mono and bi-dimensional electrophoretic analysis. Bovine mRNA translation products were analysed under similar conditions. While it was confirmed that bovine ADX was synthesized as a precursor of Mr 21 kDa, human pro-ADX was characterized for the first time as a somewhat larger moiety (24 kDa). On the other hand, both human and bovine mature mitochondrial ADX showed a Mr of 12 kDa. Electrophoretic study disclosed that the human, as well as the bovine pro-ADX could be resolved into several components differing by their pHi (6.5 and 6.9 for h-proADX and 5.9, 6.1 and 6.2 for b-proADX, respectively). This molecular heterogeneity might be explained by discrete disparity in the pro-adrenodoxin amino acid contents.

We have shown that the precursors of porcine LH alpha and beta subunits have similar apparent molecular weights (Biochem. Biophys. Res. Comm., 1984, 118, 254). The aim of the present study was to characterize physiochemical features which permit to distinguish these precursors from each other. We report here that these precursors can be clearly distinguished by both their tryptic digests analyzed by one-dimension electrophoresis and their behavior in two-dimension electrophoresis whose first dimension was made in a non equilibrium pH gradient (2D-NEPHGE). The 2D-NEPHGE method developed in this study appears also useful to estimate simultaneously the translation activities of pituitary hormone mRNAs without a previous immunoprecipitation.