Antiganglioside antibodies in patients with neuropsychiatric systemic lupus erythematosus.

Antiganglioside antibodies (AGA) were determined in sera and cerebrospinal fluids (CSF) from 50 systemic lupus erythematosus (SLE) patients, and age-matched normal controls. The SLE patients were subdivided according to the type of clinical manifestation into two groups: neuropsychiatric SLE and active SLE without neuropsychiatric manifestation. The presence of these antibodies showed a significant correlation between IgG AGA in the CSF and IgM AGA in the serum and neuropsychiatric SLE. Fifteen patients had this antibody in the CSF without detectable levels in the serum. No correlation was seen between anticardiolipin antibodies in the serum of CSF and neuropsychiatric SLE. The present work suggests that antibodies against gangliosides may be a marker for neuropsychiatric SLE and that intrathecal antibody production can result in the development of this manifestation.


The sequelae of chronic cutaneous lupus erythematosus.

Eighty-six patients with chronic cutaneous lupus erythematosus were examined. Twelve of these also suffered from systemic lupus erythematosus. The mean duration of the disease was 15.1 years. Fifty-seven percent of patients (49/86) had scarring of some kind producing destruction and deformity; 47% (41/86) had scarring of glabrous surfaces and 35% (30/86) had scarring alopecia; 35% (30/86) were also suffering from pigmentary disturbance. The details and treatment of these and other non-scarring sequelae are discussed.


Management of antiphospholipid antibody positivity and elective orthopedic procedures.

Current management of primary or secondary antiphospholipid antibody (aPL) syndromes with known embolic phenomena requiring anticoagulation is empiric in the setting of elective orthopedic procedures. Short-term withdrawal of warfarin with continuance of aspirin and glucocorticoid therapy was undertaken for sequential bilateral knee replacements in a lupus patient with aPL. Her course was successfully managed without thrombo-embolic complications.


Plasma sex hormones and aromatase activity in tissues of patients with systemic lupus erythematosus.

Clinical observations and experimental data suggest that sex hormones influence the development of systemic lupus erythematosus (SLE). An imbalance between androgen and estrogen plasma levels may suggest an abnormality in the aromatase activity involved in estradiol synthesis. Aromatase activity in skin and subcutaneous tissue and plasma sex-hormone levels (testosterone, androstenedione, estrone, estradiol, dehydrosterone sulfate, cortisol) were measured in 15 SLE patients (nine female, six male) who had never received corticosteroid treatment and in eight (four female, four male) healthy control subjects. There was a tendency toward an increase in aromatase activity in SLE patients when compared to control subjects. Among SLE patients the aromatase activity varied inversely with disease activity. Patients with SLE had decreased androgen and increased estrogen levels. Aromatase activity in SLE patients had significant direct correlation with estrogen levels. These data suggest that abnormal regulation of aromatase activity may partially explain the abnormalities of estrogen synthesis in SLE.


Multiorgan thrombotic disorders in systemic lupus erythematosus: a common link?

Multiorgan thrombotic disorders have been described in a variety of conditions including systemic lupus erythematosus. Among these are the 'catastrophic' antiphospholipid syndrome, disseminated intravascular coagulation and the thrombotic thrombocytopenic purpura-haemolytic uraemic syndrome. In this review we briefly analyse some specific clinical and haematological characteristics of these conditions and attempt to uncover common links between them.


Can experienced clinicians predict the outcome of lupus nephritis?

The ability of four experienced clinicians to predict short-term outcome (serum creatinine level at 1 year) and long-term outcome (renal insufficiency) was evaluated in 87 patients with lupus nephritis. The correlational agreement and the accuracy of their predictions were contrasted with the actual outcomes observed and with statistically generated prognostic regression models. In contrast to previously published data, all four clinicians predicted both short-term outcomes (P < 0.001) and long-term outcomes (P < 0.02) well. The clinicians' predictions approximated that of a statistically generated computer model for both agreement and accuracy for renal function at 1 year. The four clinicians identified nearly identical clinical variables as important in determining prognosis. Provision of biopsy data to the clinicians improved short-term and long-term prediction slightly. The value of the statistical models was 'validated' by demonstrating that three of the four clinical variables identified by the models, but not by the clinicians, could enhance clinical prediction (P < 0.05). In addition, the extent of tubulo-interstitial involvement on biopsy, a predictor that has recently received increased attention, could improve the long-term clinical predictions of all four clinicians (P < 0.05).


I-123 iofetamine SPECT scan in systemic lupus erythematosus patients with cognitive and other minor neuropsychiatric symptoms: a pilot study.

Accurate diagnosis of central nervous system (CNS) lupus remains difficult, especially when the manifestations are of subtle cognitive and affective changes. This pilot study reports on the use of I-123 iofetamine single photon emission computerized tomography (SPECT) scans in 18 such patients with documented systemic lupus erythematosus. Eight of the 18 scans were abnormal (44%), four in a diffuse bi-temporo-parietal pattern previously noted only in Alzheimer's disease, and four with large focal deficits. Neither the existence of the abnormal scan nor the particular pattern of abnormality correlated with the results of other diagnostic tests. These preliminary results raise the possibility that SPECT scans may offer an additional valuable diagnostic instrument in CNS lupus, although further studies are necessary to delineate their precise role.


Prevalence of acne rosacea in a rheumatic skin disease subspecialty clinic.

There are many causes of malar erythema besides the classic butterfly rash of acute cutaneous lupus erythematosus (LE). Twenty-one patients (6.7% of new patient visits) referred to a dermatology department-based rheumatic skin disease subspecialty clinic over a 5-year period in whom a diagnosis of cutaneous LE had been entertained were found to have diagnoses other than autoimmune connective tissue diseases. Sixteen of the patients in this cohort (76%) had acne rosacea (rosacea), while the remaining five had other dermatologic disorders. Review of their records revealed that upon referral nine of these 21 patients (43%) had positive antinuclear antibody (ANA) assays, most with insignificant or marginal titers by our laboratory standards. On repeat ANA testing in our laboratory, all of these patients had insignificant ANA titers. Physicians may be giving too much weight to low-titer ANAs in assessing patients with isolated malar erythema. These issues are discussed in the overall context of the differential diagnosis of malar erythema. A simple punch skin biopsy can be very helpful in distinguishing cutaneous LE from other causes of malar erythema.


Monoclonal autoantibodies to cardiolipin derived from SLE mice.

The objective of this study was to clarify the specificity of anticardiolipin antibodies (aCL). Eighteen monoclonal hybridoma aCL from systemic lupus erythematosus (SLE)-prone MRL/Mp-lpr/lpr mice were established, and the reactivity of monoclonal aCL to phospholipids, DNA, nuclei of human epithelial cells, platelets, vascular endothelial cells, heparin, protein C and thrombomodulin was examined. All the 18 monoclonal aCL reacted with phosphatidylserine and some showed reactivity to phosphatidylinositol and phosphatidylcholine. Six of 16 monoclonal aCL were demonstrated to have the property of lupus anticoagulant. Monoclonal aCL were classified into three categories, in terms of DNA-binding specificity. Ten of 18 aCL had characteristics of antinuclear antibodies. Six of 11 aCL reacted with platelets. Three of 18 aCL were bound to vascular endothelial cells and to heparin. No monoclonal aCL reacted with protein C or thrombomodulin. Therefore, the conclusion was made that monoclonal aCL from SLE mice showed a polyspecific nature.


Two sisters producing anti-U1RNP exhibit serological concordance and clinical discordance.

Two sisters had autoimmune responses to the U1RNP particle that were quantitatively similar and/or identical in molecular characteristics. No other autoantibodies were demonstrable. Both sisters immunoprecipitated only U1RNA, had a reaction of identity in gel diffusion, bound the 68-kDa band in HeLa cell extract in Western blot, and reacted almost equally to a rabbit anti-idiotypic reagent made against either sister's isolated anti-U1RNP Fab fragments. They both carried a DR4 allele, which has been associated with anti-U1RNP production in several studies. While the sisters both had Raynaud's phenomenon, their clinical pictures were otherwise dissimilar. One had a seizure disorder (Ju); the other had polymyositis and features of scleroderma (Je). In sister Ju, Raynaud's phenomenon was manifest for the first time in association with the appearance of precipitating anti-U1RNP.


Bacterial lipopolysaccharide causes variable deposits of diverse immunoglobulin isotypes in kidneys of lupus-prone mice.

We investigated the role of immunoglobulin isotypes in the exacerbation of lupus nephritis associated with exposure to bacterial lipopolysaccharide. The data indicate that enhanced polyclonal B-cell activation and exacerbated autoimmune disease evoked by lipopolysaccharide are associated with an increase in the concentration of isotypes in plasma and in renal eluate, that this isotype response is polyclonal and preferential but not restrictive, that all B cells are responsive but all are not equally sensitive to the effects of lipopolysaccharide, and that some expanded isotypes may be more nephritogenic in certain strains of lupus-prone mice.


Monoclonal gammopathy in systemic lupus erythematosus.

In order to ascertain the incidence of monoclonal gammopathy (MG) in systemic lupus erythematosus (SLE), 120 SLE patients were evaluated. Four patients (3.3%), two of whom were under 50 years of age, were found to have a monoclonal immunoglobulin in their serum. Our data agree with the previous literature in demonstrating a higher incidence of MG in SLE patients in comparison with the general population. Neither the clinical importance nor the pathogenetic significance of this association is known.


Prevalence of anticardiolipin antibodies in subacute cutaneous lupus erythematosus.

We examined the prevalence of the antibodies to cardiolipin measured by solid-phase enzyme immunoassay during a prospective study of patients with subacute cutaneous lupus erythematosus (SCLE). Seven of 44 (16%) consecutive patients with SCLE had positive anticardiolipin antibodies; of these only three satisfied the American Rheumatism Association's revised criteria for the classification of systemic lupus erythematosus. Clinical findings probably associated with the positive anticardiolipin antibodies were found in four cases, including clotting abnormalities, thrombocytopenia, hemolytic anemia, livedo reticularis, chilblain lupus erythematosus lesions, migraine, leg venous thrombosis and pulmonary embolism after surgery, and spontaneous abortion. Our data suggest that it is reasonable to screen SCLE patients for these antibodies to confirm the presence of the antiphospholipid syndrome.


Maternally transmitted diabetes and deafness associated with a 10.4 kb mitochondrial DNA deletion.

Diabetes mellitus (DM) is one of the most common chronic disorders of children and adults. Several reports have suggested an increased incidence of maternal transmission in some forms of DM. Therefore, we tested a pedigree with maternally transmitted DM and deafness for mitochondrial DNA mutations and discovered a 10.4 kilobase (kb) mtDNA deletion. This deletion is unique because it is maternally inherited, removes the light strand origin (OL) of mtDNA replication, inhibits mitochondrial protein synthesis, and is not associated with the hallmarks of mtDNA deletion syndromes. This discovery demonstrates that DM can be caused by mtDNA mutations and suggests that some of the heterogeneity of this disease results from the novel features of mtDNA genetics.


Mutations in the 70K peroxisomal membrane protein gene in Zellweger syndrome.

The peroxisomal membrane protein, with a relative molecular mass of 70,000 (M(r) 70K) (PMP70), is an important component of peroxisomal membranes and an ATP-binding cassette protein. To investigate its possible involvement in Zellweger syndrome (ZS), an inborn error of peroxisome assembly, we cloned and sequenced cDNAs for human PMP70 and mapped the gene to chromosome 1. Amongst 32 probands with ZS or related disorders, we found two mutant PMP70 alleles in single ZS probands from the same complementation group. One allele has a donor splice site mutation and the second a missense mutation. Our results suggest that PMP70 plays an important role in peroxisome biogenesis and that mutations in PMP70 may be responsible for a subset of ZS patients.


Rapid generation of region specific probes by chromosome microdissection and their application.

The strategy presented here to identify unequivocally cryptic chromosomal rearrangements has relevance to both prenatal and postnatal cytogenetic analysis as well as the analysis of tumour-associated chromosome rearrangements. Microdissection and in vitro amplification of specific chromosomal regions are performed, followed by labelling for fluorescent in situ hybridization (FISH) to normal metaphase chromosomes (Micro-FISH). Micro-FISH probes have been used successfully to determine the derivation of chromosome segments unidentifiable by standard chromosome banding analysis. Micro-FISH probes (created in less than 24 hours) now make it possible to identify explicitly the chromosome constitution of virtually all cytologically visible chromosome rearrangements.


Gene dosage is a mechanism for Charcot-Marie-Tooth disease type 1A.

Charcot-Marie-Tooth disease type 1A (CMT1A) is the most common inherited peripheral neuropathy in humans, characterized electrophysiologically by decreased nerve conduction velocities (NCVs). CMT1A is associated with a large submicroscopic DNA duplication in proximal 17p. In this report we demonstrate that a patient with a cytogenetically visible duplication, dup(17)(p11.2p12), has decreased NCV. Molecular analysis demonstrated this patient was duplicated for all the DNA markers duplicated in CMT1A as well as markers both proximal and distal to the CMT1A duplication. These data support the hypothesis that the CMT1A phenotype can result from a gene dosage effect.


Automated DNA sequencing and analysis of 106 kilobases from human chromosome 19q13.3.

A total of 116,118 basepairs (bp) derived from three cosmids spanning the ERCC1 locus of human chromosome 19q13.3 have been sequenced with automated fluorescence-based sequencers and analysed by polymerase chain reaction amplification and computer methods. The assembled sequence forms two contigs totalling 105,831 bp, which contain a human fosB proto-oncogene, a gene encoding a protein phosphatase, two genes of unknown function and the previously-characterized ERCC1 DNA repair gene. This light band region has a high average density of 1.4 Alu repeats per kilobase. Human chromosome light bands could therefore contain up to 75,000 genes and 1.5 million Alu repeats.


Definition of a consensus binding site for p53.

Recent experiments have suggested that p53 action may be mediated through its interaction with DNA. We have now identified 18 human genomic clones that bind to p53 in vitro. Precise mapping of the binding sequences within these clones revealed a consensus binding site with a striking internal symmetry, consisting of two copies of the 10 base pair motif 5'-PuPuPuC(A/T)(T/A)GPyPyPy-3' separated by 0-13 base pairs. One copy of the motif was insufficient for binding, and subtle alterations of the motif, even when present in multiple copies, resulted in loss of affinity for p53. Mutants of p53, representing each of the four "hot spots" frequently altered in human cancers, failed to bind to the consensus dimer. These results define the DNA sequence elements with which p53 interacts in vitro and which may be important for p53 action in vivo.


Human CCAAT displacement protein is homologous to the Drosophila homeoprotein, cut.

Human CCAAT displacement protein (CDP), a putative repressor of developmentally regulated gene expression, was purified from HeLa cells by DNA binding-site affinity chromatography. cDNA encoding CDP was obtained by immunoscreening a lambda gt11 library with antibody raised against purified protein. The deduced primary amino acid sequence of CDP reveals remarkable homology to Drosophila cut with respect to the presence of a unique homeodomain and "cut repeats". As cut participates in determination of cell fate in several tissues in Drosophila, the similarity predicts a broad role for CDP in mammalian development.


Cretinism with combined hormone deficiency caused by a mutation in the PIT1 gene.

Cretinism is marked by irreversible mental and growth retardation. We describe here an entirely new case of cretinism showing combined pituitary hormone deficiencies of thyrotropin, growth hormone and prolactin that appears to be caused by homozygosity for a nonsense mutation in the gene for the pituitary specific transcription activator, Pit-1/GHF-1 (designated PIT1 in humans for pituitary specific factor 1). This is the first report in humans of a defect in a transcription activator causing deficiency of multiple target genes.


A mutation in adenylosuccinate lyase associated with mental retardation and autistic features.

We have examined the molecular basis of three cases of severe mental retardation with autistic features in one family. A point mutation in a purine nucleotide biosynthetic enzyme, adenylosuccinate lyase (ASL), segregates with the disorder. The affected children are homozygous for the point mutation while the parents and all four unaffected children are heterozygous. The point mutation is absent in control subjects. The point mutation results in a Ser413Pro substitution which leads to structural instability of the recombinant mutant enzyme, and this instability lowers ASL levels in lymphocytes. These observations suggest that the instability of ASL underlies the severe developmental disorder in the affected children, and that mutations in the ASL gene may result in other cases of mental retardation and autistic features.


Aberrant splicing of the CHM gene is a significant cause of choroideremia.

Choroideremia (CHM) is an X-linked progressive degeneration of the choroid and retina. 12% of unrelated male patients carry deletions of the partially cloned CHM gene. In Finland, there are more than 120 living CHM patients belonging to eight apparently unrelated pedigrees. Molecular deletions involving the CHM gene have been detected in three families. We have screened the remaining five families for point mutations. In one large family a single nucleotide (T) insertion into the donor splice site of exon C leads to two aberrantly spliced mRNAs both producing a premature stop codon. The mutation can be assayed easily by amplification and digestion with Msel. Our findings provide additional evidence for the pathogenetic role of CHM mutations and provide a diagnostic tool for one fifth of the world's known CHM patients.


A survey of expressed genes in Caenorhabditis elegans.

As an adjunct to the genomic sequencing of Caenorhabditis elegans, we have investigated a representative cDNA library of 1,517 clones. A single sequence read has been obtained from the 5' end of each clone, allowing its characterization with respect to the public databases, and the clones are being localized on the genome map. The result is the identification of about 1,200 of the estimated 15,000 genes of C. elegans. More than 30% of the inferred protein sequences have significant similarity to existing sequences in the databases, providing a route towards in vivo analysis of known genes in the nematode. These clones also provide material for assessing the accuracy of predicted exons and splicing patterns and will lead to a more accurate estimate of the total number of genes in the organism than has hitherto been available.


Caenorhabditis elegans expressed sequence tags identify gene families and potential disease gene homologues.

A database containing mapped partial cDNA sequences from Caenorhabditis elegans will provide a ready starting point for identifying nematode homologues of important human genes and determining their functions in C. elegans. A total of 720 expressed sequence tags (ESTs) have been generated from 585 clones randomly selected from a mixed-stage C. elegans cDNA library. Comparison of these ESTs with sequence databases identified 422 new C. elegans genes, of which 317 are not similar to any sequences in the database. Twenty-six new genes have been mapped by YAC clone hybridization. Members of several gene families, including cuticle collagens, GTP-binding proteins, and RNA helicases were discovered. Many of the new genes are similar to known or potential human disease genes, including CFTR and the LDL receptor.


Construction of a mouse yeast artificial chromosome library in a recombination-deficient strain of yeast.

We have constructed a new generation yeast artificial chromosome (YAC) library from female C57BL/10 mice in a recombination-deficient strain of Saccharomyces cerevisiae carrying a mutation in the RAD52 gene. The YAC library contains 41,568 clones with an average insert size of 240 kilobases, representing a greater than threefold coverage of the mouse genome. Currently, the library can be screened by polymerase chain reaction and we have isolated positive clones at a number of loci in the mouse genome. This rad52 library should enable a long-term assessment of the effect of one of the yeast recombination pathway genes on both, genome-wide YAC clone stability and the frequency of chimaeric clones.


A novel GC-rich human macrosatellite VNTR in Xq24 is differentially methylated on active and inactive X chromosomes.

A new X chromosome-specific repetitive sequence, a 3 kilobase HindIII clone with a base composition of 63% C+G, has been isolated. The sequence is organized as a hypervariable tandem repeat cluster ranging in size from 150-350 kilobases, with outlying single copies. This locus, designated DXZ4 and mapped to chromosome band Xq24, may consist of as many as 50 variable-length alleles. It represents a class of variable number of tandem repeat polymorphism which may be termed 'macrosatellite'. The cluster is highly methylated on the active X chromosome and hypomethylated on the inactive X.


Germline intronic and exonic mutations in the Wilms' tumour gene (WT1) affecting urogenital development.

Denys-Drash syndrome is a rare human developmental disorder affecting the urogenital system and leading to renal failure, intersex disorders and Wilms' tumour. In this report, four individuals with this syndrome are described carrying germline point mutations in the Wilms' tumour suppressor gene, WT1. Three of these mutations were in the zinc finger domains of WT1. The fourth occurred within intron 9, preventing splicing at one of the alternatively chosen splice donor sites of exon 9 when assayed in vitro. These results provide genetic evidence for distinct functional roles of the WT1 isoforms in urogenital development.


A donor splice site mutation in the parathyroid hormone gene is associated with autosomal recessive hypoparathyroidism.

Investigation of one kindred with autosomal recessive isolated hypoparathyroidism, which had resulted from a consanguineous marriage, has identified a g to c substitution in the first nucleotide of intron 2 of the parathyroid hormone (PTH) gene. This donor splice mutation could be detected by restriction enzyme cleavage with Ddel, and this revealed that the patients were homozygous for the mutant alleles, the unaffected relatives were heterozygous, and unrelated normals were homozygous for the wild type alleles. Defects in messenger RNA splicing were investigated by the detection of illegitimate transcription of the PTH gene in lymphoblastoid cells. The mutation resulted in exon skipping with a loss of exon 2, which encodes the initiation codon and the signal peptide, thereby causing parathyroid hormone deficiency.


Evolution of the mammalian G protein alpha subunit multigene family.

Heterotrimeric guanine nucleotide binding proteins (G proteins) transduce extracellular signals received by transmembrane receptors to effector proteins. The multigene family of G protein alpha subunits, which interact with receptors and effectors, exhibit a high level of sequence diversity. In mammals, 15 G alpha subunit genes can be grouped by sequence and functional similarities into four classes. We have determined the murine chromosomal locations of all 15 G alpha subunit genes using an interspecific backcross derived from crosses of C57BL/6J and Mus spretus mice. These data, in combination with mapping studies in humans, have provided insight into the events responsible for generating the genetic diversity found in the mammalian alpha subunit genes and a framework for elucidating the role of the G alpha subunits in disease.


Rescue of erythroid development in gene targeted GATA-1- mouse embryonic stem cells.

Development of definitive (fetal liver-derived) red cells is blocked by a targeted mutation in the gene encoding the transcription factor GATA-1. We used in vitro differentiation of GATA-1- mouse embryonic stem (ES) cells to reveal a requirement for GATA-1 during primitive (yolk sac-derived) erythropoiesis and to establish a rescue assay. We show that the block to development includes primitive, as well as definitive, erythroid cells and is complete at the level of globin RNA expression; that the introduction of a normal GATA-1 gene restores developmental potential both in vivo and in vitro; and that efficient rescue is dependent on a putative autoregulatory GATA-motif in the distal promoter. Use of in vitro differentiated ES cells bridges a gap between conventional approaches to gene function in cell lines and analysis of loss of function mutations in the whole animal.


The Huntington's disease candidate region exhibits many different haplotypes.

Analysis of 78 Huntington's disease (HD) chromosomes with multi-allele markers revealed 26 different haplotypes, suggesting a variety of independent HD mutations. The most frequent haplotype, accounting for about one third of disease chromosomes, suggests that the disease gene is between D4S182 and D4S180. However, the paucity of an expected class of chromosomes that can be related to this major haplotype by assuming single crossovers may reflect the operation of other mechanisms in creating haplotype diversity. Some of these mechanisms sustain alternative scenarios that do not require a multiple mutational origin for HD and/or its positioning between D4S182 and D4S180.


Genetic linkage of vitelliform macular degeneration (Best's disease) to chromosome 11q13.

Macular degeneration is the most common cause of legal blindness in older patients in developed countries. Best's vitelliform dystrophy is an early-onset, autosomal dominant form of macular degeneration characterized by an egg-yolk-like collection of lipofuscin beneath the pigment epithelium of the retinal macula. Fifty-seven members of a five-generation family affected with this disease were studied. A combination of ophthalmoscopy and electro-oculography was used for diagnosis; 29 patients were found to be affected and 16 unaffected. Linkage analysis mapped the disease-causing gene to chromosome 11q13. Three markers in this region were found to be significantly linked (Zmax > 3.0) to the disease. Multipoint analysis yielded a maximum Lod score of 9.3 in the interval between markers INT2 and D11S871.


Defective colour vision associated with a missense mutation in the human green visual pigment gene.

All red/green colour vision defects described so far have been associated with gross rearrangements within the red/green opsin gene array (Xq28). We now describe a male with severe deuteranomaly without such a rearrangement. A substitution of a highly conserved cysteine by arginine at position 203 in the green opsins presumably accounted for his colour vision defect. Surprisingly, this mutation was fairly common (2%) in the population but apparently was not always expressed. In analogy with nonexpression of some 5'green-red hybrid genes in persons with normal colour vision, we suggest that failure of manifestation occurs when the mutant gene is located at a distal (3') position among several green opsin genes. This mutation might also predispose to certain X-linked retinal dystrophies.


Evidence of founder chromosomes in fragile X syndrome.

The mutation responsible for fragile X syndrome and myotonic dystrophy involves the amplification of a simple trinucleotide repeat sequence, which increases in successive generations of affected pedigrees accounting for increasing penetrance of both disorders. This common molecular basis suggests that the two diseases may share other genetic features, but whereas myotonic dystrophy exhibits a significant founder chromosome effect, fragile X syndrome apparently has a very high mutation frequency. By haplotype analysis of microsatellite markers which flank the fragile X unstable element, we have uncovered evidence of founder chromosomes of the fragile X 'mutation'. Disorders caused by heritable unstable elements may therefore exhibit common genetic properties including anticipation and founder chromosomes.


Characterization of the myotonic dystrophy region predicts multiple protein isoform-encoding mRNAs.

The mutation underlying myotonic dystrophy (DM) has been identified as an expansion of a polymorphic CTG-repeat in a gene encoding protein kinase activity. Brain and heart transcripts of the DM-kinase (DMR-B15) gene are subject to alternative RNA splicing in both human and mouse. The unstable [CTG]5-30 motif is found uniquely in humans, although the flanking nucleotides are also present in mouse. Characterization of the DM region of both species reveals another active gene (DMR-N9) in close proximity to the kinase gene. DMR-N9 transcripts, mainly expressed in brain and testis, possess a single, large open reading frame, but the function of its protein product is unknown. Clinical manifestation of DM may be caused by the expanded CTG-repeat compromising the (alternative) expression of DM-kinase or DMR-N9 proteins.


Complete coverage of the Schizosaccharomyces pombe genome in yeast artificial chromosomes.

The genome of the fission yeast, Schizosaccharomyces pombe, consists of some 14 million base pairs of DNA contained in three chromosomes. On account of its excellent genetics we used it as a test system for a strategy designed to map mammalian chromosomes and genomes. Data obtained from hybridization fingerprinting established an ordered library of 1,248 yeast artificial chromosome clones with an average size of 535 kilobases. The clones fall into three contigs completely representing the three chromosomes of the organism. This work provides a high resolution physical and clone map of the genome, which has been related to available genetic and physical map information.


Generation of region- and species-specific expressed gene probes from somatic cell hybrids.

Human genes expressed in interspecific somatic cell hybrids can be cloned specifically by subtractive cDNA hybridization. This approach is based on the observation that cDNA fragments from noncoding segments of mature human transcripts do not form stable heteroduplexes with their rodent homologues under high-stringency hybridization conditions. Thus, small, oligo-dT primed cDNAs from a rat/human hybrid retaining a fragment of human chromosome 17 were enriched for human sequences by hybridization with RNA from a sister clone containing a smaller human chromosome fragment. The enriched probe was used to screen a human cDNA library, and nine expressed genes from within the non-overlap region were obtained. This method should be useful for cloning active human genes from defined chromosome segments.


Selection of a human chromosome 21 enriched YAC sub-library using a chromosome-specific composite probe.

The subdivision of total genomic human yeast artificial chromosome (YAC) libraries into specific chromosome clone collections will greatly facilitate the construction of an integrated genetic, physical and transcriptional map of the genome. We report the isolation of 388 YAC clones from a human library with an average insert size of 620 kilobases (kb) by the hybridization of a composite chromosome 21 probe to a high-density array of YAC clones. Roughly half of these clones hybridize to chromosome 21 by fluorescence in situ hybridization. These clones represent a twofold coverage of the chromosome. The technique offers the potential of sub-dividing whole genomic YAC libraries into their chromosomal elements to produce high-resolution tools for genome mapping.


The gene responsible for familial hypocalciuric hypercalcemia maps to chromosome 3q in four unrelated families.

Familial hypocalciuric hypercalcemia (FHH) is an autosomal dominant syndrome of unknown aetiology characterized by lifelong elevation in serum calcium concentration and low urinary calcium excretion. These features suggest that the causal gene is important for maintenance of extracellular calcium homeostasis by the parathyroid gland and kidney. To identify the chromosomal location of FHH gene(s), we clinically evaluated 114 individuals in four unrelated affected families and performed linkage analyses. The disease gene mapped to the long arm of chromosome 3 in each family (combined maximum multipoint lod score = 20.67). We suggest that this is the predominant FHH locus and anticipate that identification of the FHH gene will improve our understanding of the molecular basis for physiologic and pathologic regulation of calcium.


The human PAX6 gene is mutated in two patients with aniridia.

Aniridia is an inherited ocular disorder of variable expressivity characterized by iris hypoplasia. A candidate aniridia gene, AN, which is the human homologue of the mouse Pax-6 gene, has recently been isolated by positional cloning from the WAGR region of 11p13. Here we describe mutations in this gene in two cases of sporadic aniridia, one detected at the DNA level and one at the RNA level, both of which are predicted to affect protein function. Mutations in Pax-6 have been described previously in Small eye, the proposed mouse model for aniridia. We present new phenotypic evidence for the validity of this mouse model.


Kallmann syndrome due to a translocation resulting in an X/Y fusion gene.

The X-linked Kallmann syndrome gene was recently cloned and homologous sequences of unknown functional significance identified on the Y chromosome. We now describe a patient with Kallmann syndrome carrying an X;Y translocation resulting from abnormal pairing and precise recombination between the X-linked Kallmann syndrome gene and its homologue on the Y. The translocation created a recombinant, non-functional Kallmann syndrome gene identical to the normal X-linked gene with the exception of the 3' end which is derived from the Y. Our findings indicate that the 3' portion of the Kallmann syndrome gene is essential for its function and cannot be substituted by the Y-derived homologous region, although a 'position' effect remains a formal possibility.


Fragile X syndrome without CCG amplification has an FMR1 deletion.

We describe a patient with typical clinical features of the fragile X syndrome, but without cytogenetic expression of the fragile X or an amplified CCG trinucleotide repeat fragment. The patient has a previously uncharacterized submicroscopic deletion encompassing the CCG repeat, the entire FMR1 gene and about 2.5 megabases of flanking sequences. This finding confirms that the fragile X phenotype can exist, without amplification of the CCG repeat or cytogenetic expression of the fragile X, and that fragile X syndrome is a genetically homogeneous disorder involving FMR1. We also found random X-inactivation in the mother of the patient who was shown to be a carrier of this deletion.


A pathogenic mutation for probable Alzheimer's disease in the APP gene at the N-terminus of beta-amyloid.

Mutations at codon 717 in exon 17 of the beta-amyloid precursor protein (APP) gene have previously been shown to segregate with early onset Alzheimer's disease in some families. We have identified a double mutation at codons 670 and 671 (APP 770 transcript) in exon 16 which co-segregates with the disease in two large (probably related) early-onset Alzheimer's disease families from Sweden. Two base pair transversions (G to T, A to C) from the normal sequence predict Lys to Asn and Met to Leu amino acid substitutions at codons 670 and 671 of the APP transcript. This mutation occurs at the amino terminal of beta-amyloid and may be pathogenic because it occurs at or close to the endosomal/lysosomal cleavage site of the molecule. Thus, pathogenic mutations in APP frame the beta-amyloid sequence.


Adenovirus-mediated in vivo gene transfer and expression in normal rat liver.

Replication deficient, recombinant adenovirus (Ad) vectors do not require target cell replication for transfer and expression of exogenous genes and thus may be useful for in vivo gene therapy in hepatocytes. In vitro, primary cultures of rat hepatocytes infected with a recombinant Ad containing a human alpha 1-antitrypsin cDNA (Ad-alpha 1AT) synthesized and secreted human alpha 1AT for 4 weeks. In rats, in vivo intraportal administration of a recombinant Ad containing the E. coli lacZ gene, was followed by expression of beta-galactosidase in hepatocytes 3 days after infection. Intraportal infusion of Ad-alpha 1AT produced detectable serum levels of human alpha 1AT for 4 weeks. Thus, targeted gene expression has been achieved in the liver, albeit at low levels, suggesting that adenovirus vectors may be a useful means for in vivo gene therapy in liver disorders.


A methodological review of "method skeptic" reports.

The literature purporting to demonstrate that clinical neuropsychology is of limited validity in the forensic setting is reviewed critically and alternative interpretations are discussed. The methodological, procedural, conceptual, data analytical and survey/research design limitations are evaluated.


Effects of ethanol, dexamethasone and RU 486 on expression of cytochromes P450 2B, 2E, 3A and glutathione transferase pi in a rat hepatoma cell line (Fao).

The aim of our study was to investigate the suitability of Fao cells, derived from the Reuber H35 rat hepatoma as a tool for studying regulation of drug-metabolizing enzymes and drug metabolism. Fao cells express P450 2B, 2E, 3A and GST pi and were used to study the effects different inducers on these enzymes. Ethanol considerably increased the amounts of P450 2E and, to a lesser extent, P450 2B and GST pi mRNA and protein. Dexamethasone decreased the amounts of P450 2B, 3A and GST pi mRNAs, but had no appreciable effect per se upon the protein concentration of these enzymes. However, it antagonized the induction of P450 2E, 2B and GST pi by ethanol, even at the protein level. RU 486 decreased P450 2B protein and P450 2E mRNA and protein levels without effecting P450 3A and GST pi expression. RU 486 did not antagonize the dexamethasone effects, suggesting that at least some of these effects are not mediated by the glucocorticoid receptor. These data indicate that these cells constitute a suitable tool for studying the regulation of drug-metabolizing enzyme expression and drug metabolism.


Deficient nifedipine oxidation: a rare inherited trait associated with cystic fibrosis kindreds.

Previous studies have indicated that there is weak genetic linkage between the defective gene in cystic fibrosis (CFTR) and the gene encoding the nifedipine metabolizing enzyme P4503A4 which are both located on chromosome 7. To examine further this possible association, nifedipine metabolism was investigated in a group of 59 volunteers, and 17 adult cystic fibrosis patients and 37 of their relatives. In agreement with the majority of previous studies, the volunteer group showed a unimodal distribution of recoveries for the major metabolite M-II ranging from 33 to 78% excretion in 8 h. In the case of both the cystic fibrosis patients and their parents, the distribution of recoveries was shifted to the left with five out of 20 parents and three out of 11 unrelated cystic fibrosis patients showing recoveries below the range observed in the volunteer group. This poor metabolism appeared to be both reproducible and heritable and did not appear to be a consequence of mutations in the CFTR gene.


Clinical implications of variable antiarrhythmic drug metabolism.

Due to their narrow therapeutic indices, antiarrhythmic drugs have a great potential for adverse outcome. This is amplified by extreme inter-individual variability in their disposition and their pharmacological actions. Genetically determined inter-individual differences in metabolism account for a great deal of this variability. However, because of active metabolites, chirally-specific actions and chirally-specific metabolism, it is not possible to generalize about the outcome of phenotypic differences in the metabolism of a given drug. Careful study of these factors can enable physicians to understand the spectrum of potential responses to a drug. Newly developed molecular biology techniques now make it possible to determine the genotype for the CYP2D6 gene that controls metabolism of many antiarrhythmic drugs. This information, combined with a full understanding of the drugs' clinical pharmacology now makes it possible to predict the clinical outcome for drugs such as encainide, flecainide, mexiletine, propafenone and combinations of these drugs with quinidine.


Alcohol, head injury, and neuropsychological function.

Evidence is reviewed indicating that the extent of alcohol abuse alone cannot account for the neuropsychological deficits observed in alcoholics, and that alcohol abuse and head injury may interact in some patients to influence neuropsychological status. Alcohol abuse both increases the risk for head trauma and potentiates the resulting brain injury, which can lead to negative neuropsychological consequences. Clinicians involved in the treatment of addiction should assess patients for history of head injury, and neuropsychological deficits consequent to both head injury and ethanol. These deficits may limit patient ability to comply with addiction rehabilitation programs. Conversely, clinicians in acute care and rehabilitation of the sequelae of head trauma should routinely assess their patients for substance abuse, because such abuse can have a significant impact on recovery from brain injury.


Polymorphic hydroxylation of S-mephenytoin and omeprazole metabolism in Caucasian and Chinese subjects.

Twelve Caucasian healthy men and women, of whom six were poor metabolizers (PMs) and six were extensive metabolizers (EMs) of S-mephenytoin, together with 13 Chinese healthy men and women (five PMs and eight EMs), received a single oral 20 mg dose of omeprazole. Plasma levels of omeprazole and its two main metabolites, omeprazole sulphone and hydroxyomeprazole, were determined by HPLC. The mean (+/- SD) area under the plasma concentration-time curve (AUC) for omeprazole was 11.1 +/- 2.6 and 0.9 +/- 0.4 microM h in the Caucasian PMs and EMs, respectively. Corresponding values for elimination half-life were 2.3 +/- 0.4 and 0.7 +/- 0.4 h. In the Chinese PMs and EMs the AUC of omeprazole as 13.3 +/- 5.6 and 2.6 +/- 1.8 microM h. The AUCs of omeprazole were significantly higher in the Chinese EMs than in the Caucasian EMs possibly due to the higher proportion of heterozygotes in the former than in the latter group. The elimination half-life in the Chinese PMs and EMs was 2.4 +/- 0.2 and 0.8 +/- 0.2 h--similar to the observations in the Caucasian subjects. The maximum plasma concentration of hydroxyomeprazole was five-fold and four-fold higher in EMs compared to PMs among Caucasians and Chinese, respectively. The elimination half-life of the hydroxy metabolite was also longer in PMs than in EMs in both populations. The ratio between AUC for omeprazole and AUC for hydroxyomeprazole was 11.9 +/- 2.1 and 0.6 +/- 0.1 in Caucasian PMs and EMs, respectively. Corresponding values in the Chinese were 13.1 +/- 2.9 and 1.6 +/- 0.5.(ABSTRACT TRUNCATED AT 250 WORDS)


Molecular characterization of the murine Coh locus: an amino acid difference at position 117 confers high and low coumarin 7-hydroxylase activity in P450coh.

Coumarin 7-hydroxylase (P450coh) and steroid 15 alpha-hydroxylase (P450(15 alpha) are encoded by members within the mouse 2A subfamily. Since P450coh activity is regulated by the Coh locus, we characterized P450coh cDNAs in strains having high coumarin 7-hydroxylase activity (CohH homozygote) including 129/J and DBA/2J, and compared them with P450coh cDNAs in low activity strains (CohL homozygote) C57BL/6J, C3H/HeJ and AKR/J. The nucleotide sequences of these two cDNAs differ by a single base, which results in an amino acid difference at position 117 (Val in P450cohH and Ala in P450cohL). The CohH phenotype exhibits approximately 10-fold greater Vmax and four-fold lower Km values than those in the CohL. Male 129 AKF1/J expresses approximately equal amounts of P450cohH and P450cohL mRNAs, associated with two Coh alleles. The levels of P450coh and P450(15 alpha) mRNAs in the F1 offspring suggested that a trans-acting factor(s) appeared to regulate the expressions of the P450 genes. A recent duplication in the ancestral mouse established the line of descent to P450(15 alpha) from the ancestral P450coh gene. During evolution, amino acid substitutions have selectively occurred at positions which alter the enzyme's substrate specificity and increase in the specific activity. Consistent with an important role of natural selection in the evolution of these genes is the relatively high nonsynonomous substitution rates on the P450(15 alpha) and the P450coh branches. As a result of these evolution events, the gene family consists of members which exhibit an extremely high degree of structural similarity, but very divergent hydroxylase activities and modes of regulation.


Variability of acetaminophen metabolism in Caucasians and Orientals.

Acetaminophen (paracetamol) is extensively conjugated with glucuronic acid and sulfate prior to renal excretion. A minor metabolic route involves microsomal oxidation of acetaminophen to a hepatotoxic reactive intermediate, which subsequently undergoes glutathione (GSH) conjugation, yielding cysteine and mercapturate conjugates, both of which are excreted in the urine (Slattery et al., 1987). Data collected by de Morais et al. (1989) indicated that in comparison with normal subjects, glucuronidation of acetaminophen was impaired in subjects with Gilbert's syndrome, a genetically-based impairment of bilirubin glucuronidation. Thus, inter-subject and ethnic differences in acetaminophen disposition have pharmacogenetic and toxicological implications. This study was conceived to explore these differences. Urinary excretion of acetaminophen and its metabolites was observed in 125 Caucasian and 33 Oriental subjects. No appreciable difference was noted in the mean fraction of drug excreted as glucuronide between the two groups (51.5% in Caucasians vs 51.8% in Orientals). However, the data strongly indicated that the excretion of acetaminophen glucuronide was not normally distributed. Bimodality was apparent in both groups, with 20% of Caucasian and 33% of Oriental subjects displaying relatively extensive glucuronidation. In addition, glucuronidation displayed a strong negative correlation with sulfation (r = -0.97), suggesting the existence of a compensatory mechanism between the two metabolic pathways. The mean fractional excretions of cysteine and mercapturate conjugates did show significant differences between Caucasians and Orientals (p < 0.005). In addition, the ratio of mercapturate to total GSH-derived conjugates recovered appeared to be bimodal, indicating possible heterogeneity in the conversion of the cysteine conjugate to mercapturate via N-acetylation.


Pharmacogenetics of alcohol metabolism and alcoholism.

The pharmacogenetic differences among individuals in their capacity to metabolize ingested alcohol are possibly responsible for the large inter-individual and inter-ethnic variations observed in the outcome of alcohol use and misuse. Based on results of adoption, twin, and family studies it is now widely accepted that the vulnerability to alcoholism is determined by genetic factors as well as by environment. There is a constant search for biological markers and specific genes which could identify individuals genetically predisposed to alcohol abuse and alcoholism. Numerous 'candidate genes' for alcoholism have been suggested including the alcohol metabolizing enzymes, alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). Both ADH and ALDH exhibit genetic heterogeneity. An atypical form of ADH (ADH2), which contains a variant beta 2 subunit instead of the usual beta 1 subunit, differs substantially from the usual form in its kinetic properties and is found more frequently among the Japanese, Chinese and other Mongoloid populations than in Caucasoids and Negroids. A widely prevalent genetic polymorphism has been observed for ALDH; about 50% of Japanese and Chinese livers possess an inactive ALDH (ALDH2 isozyme) whereas none of the Caucasian or Negroid populations show this isozyme abnormality. These metabolic polymorphisms seem to contribute to differences in the in vivo elimination rate of ethanol and acetaldehyde, and may explain differences in alcohol-related behaviour and its disease outcome. Taken together, Orientals who possess an atypical ALDH2 gene are more sensitive to acute responses to alcohol, tend to be discouraged from drinking alcohol, and consequently are at lower risk of developing alcohol-related disorders. However, more work is needed to support these findings. Recent advances in molecular genetics have made it possible to analyze directly the human genome. This may help in a better understanding of the complex genetic and environmental factors in alcohol abuse by providing prospects for identification of gene loci which may be responsible for predisposition to, and protection from, alcoholism.


Biotransformation of caffeine, paraxanthine, theobromine and theophylline by cDNA-expressed human CYP1A2 and CYP2E1.

Six human cytochrome P450s expressed in HepG2 cells using vaccinia virus cDNA-directed expression, were used to study the biotransformation of caffeine and its metabolites. CYP1A2 alone was responsible for caffeine 3-demethylation and paraxanthine 7-demethylation; in addition, 1A2 catalysed virtually all reactions related to caffeine and its metabolites. The metabolic profile of caffeine biotransformation by CYP1A2 averaged 81.5% for paraxanthine, 10.8% for theobromine and 5.4% for theophylline formation. It remained quite uniform when caffeine concentrations were varied. The most striking finding was that CYP2E1 (the ethanol-inducible form) had major influences upon caffeine metabolism: in particular, it catalysed the formation of theophylline and theobromine from caffeine. Thus, the in vivo metabolite profiling of caffeine may reveal CYP2E1 activities in addition to the previously documented activities of CYP1A2, polymorphic N-acetyltransferase and xanthine oxidase.


Detection of human DNA mutations with nonradioactive, allele-specific oligonucleotide probes.

We describe a method of detecting human DNA mutations with nonradioactive, biotinylated allele-specific oligonucleotide probes. This method can detect seven different mutations in the butyrylcholinesterase, cystic fibrosis, and N-acetyltransferase genes under identical assay conditions. This indicates that it may be used to detect mutations responsible for a wide variety of genetic diseases and pharmacogenetic conditions. The method involves first amplifying selected DNA fragments by the polymerase chain reaction and dot blotting the amplified DNA in duplicate onto small nitrocellulose squares. Each dot blot is then hybridized in individual wells containing a tetramethylammonium chloride solution with short biotinylated probes specific for either the normal or mutant allele. Successfully hybridized probes are detected by a simple colorimetric reaction using an avidin-alkaline phosphatase conjugate, which yields a very strong, clear signal. DNA from homozygous normal or mutant individuals hybridizes only to the normal- or mutant-specific probes respectively, while DNA from heterozygous individuals hybridizes equally well with both probes. These results can be easily interpreted to assign a genotype to the sample DNA. This method is amenable to automation, and may be useful in clinical laboratories for diagnosis of a wide variety of DNA mutations responsible for unusual reactions to drugs and environmental chemicals.


Evidence for a dissociation in the control of sparteine, debrisoquine and metoprolol metabolism in Nigerians.

The 0-8 hour urinary distributions of the metabolic ratios of sparteine (100 mg), debrisoquine (10 mg) and metoprolol (100 mg) were measured in 165 healthy, unrelated, black Nigerian medical students. There was a weak correlation (rs = 0.51, p < 0.001; n = 82) between the metoprolol/alpha-hydroxymetoprolol (M/HM) and the sparteine/total (2- + 5-) dehydrosparteine (S/DHS) ratios. No significant correlations were found between the debrisoquine/4-hydroxydebrisoquine (D/HD) and M/HM ratios (rs = 0.16, n = 33) and between the D/HD and S/DHS ratios (rs = 0.31, n = 38). Both visual inspection and kernel density analysis of the data suggested the presence of two phenotypic groups for sparteine oxidation, with 4% of the population studied being putative poor metabolizers. In contrast biomodality was not apparent in the distribution of the log10M/HM and log10D/HD ratios. These findings provide evidence for a dissociation in the control of metoprolol, sparteine and debrisoquine oxidation in Nigerians and highlight the difficulties in the interpretation of data from pharmacogenetic studies in different ethnic groups.


Characterization of a shoot-specific, GA3- and ABA-regulated gene from tomato.

A tomato (Lycopersicon esculentum) gene (GAST1) that encodes an RNA whose abundance increases > 20-fold in shoots of the GA-deficient gib1 mutant following spraying with GA3 has been characterized. An increase in GAST1 RNA levels is detectable 2 h after treatment and levels continue to increase for at least an additional 10 h. Between 12 and 24 h following treatment, the amount of GAST1 RNA begins to decline and at 48 h the level is nearly equivalent to that of water-treated control plants. Nuclear runoff analysis indicates that 8 h after treatment with GA3, transcription of the GAST1 gene has increased only threefold, suggesting that GA acts both transcriptionally and post-transcriptionally. ABA partially inhibits the GA-mediated increase in GAST1 RNA abundance while ethephon, kinetin, and 2,4-D have little effect. GAST1 RNA is detectable in untreated leaves, stems, petioles and flowers, but not in roots. The GAST1 gene encodes a 0.7 kb transcript. The sequence of the GAST1 cDNA and genomic clones indicates that the gene is interrupted by three introns and potentially encodes a 112 amino acid protein of unknown function.


Vicilin with carboxy-terminal KDEL is retained in the endoplasmic reticulum and accumulates to high levels in the leaves of transgenic plants.

Gene constructs were designed to test the effect of the endoplasmic reticulum (ER)-targeting signal, KDEL, on the level of accumulation of a foreign protein in transgenic plants. The gene for the pea seed protein vicilin was modified by the addition of a sequence coding for this tetrapeptide at its carboxyl terminus. The altered gene was placed under the control of a CaMV 35S promoter and its expression in the leaves of both tobacco and lucerne (alfalfa) was compared with that of an equivalent vicilin construct lacking the KDEL-coding sequence. The presence of the ER-targeting signal led to a greatly enhanced accumulation of the heterologous protein. In lucerne and tobacco leaves, the level of vicilin-KDEL protein was 20 and 100 times greater than that of the unmodified vicilin, respectively. These differences in expression level could not be explained by corresponding differences in the steady-state levels or the translatability of the mRNAs. However, when the stability of vicilin and vicilin-KDEL proteins was compared in their respective transgenic hosts, unmodified vicilin was found to be degraded with a half-life of 4.5 h while vicilin-KDEL was much more stable with a half-life of more than 48 h. Immunogold labelling of leaf tissues from transgenic lucerne and tobacco showed the presence of vicilin associated with large aggregates within the ER lumen of vicilin-KDEL plants. No such aggregates were detected in transgenic plants expressing wild-type vicilin. It is concluded that the carboxy-terminal KDEL caused the retention of the modified vicilin in the ER, and that this retention led to the increased stability and higher level of accumulation of vicilin-KDEL in leaves of transgenic plants.


Characterization of cDNAs encoding two isoforms of granule-bound starch synthase which show differential expression in developing storage organs of pea and potato.

We have isolated cDNA clones to two isoforms of granule-bound starch synthase (GBSS) from pea embryos and potato tubers. The sequences of both isoforms are related to that of glycogen synthase from E. coli and one, GBSSI, is very similar to the waxy protein of maize and other species. In pea, GBSSII carries a novel 203-amino-acid domain at its N-terminus. Genes encoding both proteins are expressed during pea embryo development, but GBSSII is most highly expressed earlier in development than GBSSI. Similarly, GBSSI and GBSSII are differentially expressed in developing potato tubers. Expression of both isoforms is much lower in other organs of pea than in embryos. GBSSII is expressed in every organ tested while GBSSI is not expressed in roots, stipules or flowers. The possible consequences of this differential use of GBSS isoforms are discussed.


A nuclear protein fraction binding to dA/dT-rich sequences upstream from the radish rDNA promoter [corrected].

The external spacer (ES) of rRNA nuclear genes (rDNA) contains the sequences that control rDNA transcription initiation and enhancement. The ES is also characterized in most species by the presence of multiple repeated elements. In higher plants very few data are available on the cis- and trans-acting elements which control rDNA transcription. Using electrophoretic mobility shift assays (EMSA) it is shown that nuclear extracts from young radish leaves (NER) contain a protein fraction which binds to specific sequences in the radish ES. DNase I footprinting analysis allows mapping of the NER protein binding to dA/dT homopolymer stretches and to a 13-bp dA/dT-rich short repeat, found both in the seven approximately 100 bp repeat regions (located -1077 to -740 from transcription initiation site) and the region (-120 to -55) containing the putative promoter for rDNA transcription initiation. Whether this ES binding is due to a single or several different proteins is not known. So far, protein(s) binding to dA/dT-rich regions of a plant rDNA ES has not yet been described. Whether it is a specific RNA polymerase I transcription factor(s) or plays a more general role in genome expression remains to be elucidated.


Maize C4 photosynthesis involves differential regulation of phosphoenolpyruvate carboxylase genes.

Maize as a C4 plant partitions CO2 fixation in two consecutive, spatially separated steps, thus eliminating photorespiration. The crucial enzyme for primary CO2 fixation is a C4-specific phosphoenolpyruvate carboxylase (PEPC). The differential expression of the unique C4-specific gene pepcZm1 and two non-C4-specific genes, pepcZm2A and pepcZm3B, in leaf, root, and stem is reported here. It is shown, in a transient homologous system, that this tissue-specific regulation is mainly controlled by their distinct promoters. The light induction of the C4-specific pepcZm1 in illuminated etiolated (greening) leaves probably relies on light-dependent developmental changes instead of an immediate responsiveness found for other maize genes. Analyses of deleted, mutated, and hybrid promoters revealed the redundant nature of a 14mer which is repeated four times and a decisive function of the TATA box-like motif, TATTT, and the sequences directly preceding it. No consensus sequences to other photosynthetic gene promoters were uncovered. Although light induces the expression of C4 PEPC and other photosynthetic genes in maize, this co-ordination is apparently mediated through different signal transduction pathways and distinct regulatory elements. This study indicates that the acquisition of a new promoter is at least partially responsible for the C4-specific expression of pepcZm1 essential for C4 photosynthesis.


Late embryogenesis-abundant genes encoding proteins with different numbers of hydrophilic repeats are regulated differentially by abscisic acid and osmotic stress.

The late embryogenesis-abundant (Lea) genes, which are suggested to act as desiccation protectants during seed desiccation and in water-stressed seedlings, can be induced by abscisic acid (ABA) and various kinds of water-related stress. Using cotton Lea cDNAs as probes it was found that several of the Lea genes are conserved at the mRNA level in dicots and monocots. By screening a barley cDNA library with a cotton Lea D19 cDNA a family of three members was isolated. The putative B19 proteins have strong similarities to the Em protein in wheat and to LEA proteins from several dicots. However, the middle part of the B19 proteins consists of a 20-amino acid motif repeated three and four times in B19.3 and B19.4, respectively, but only once in B19.1. The gene products are strongly hydrophilic, the internal 20-amino acid motif being the most hydrophilic part. This motif is found once in cotton Lea D19 but is repeated twice in cotton Lea D132, indicating that the repeats are universal among monocot and dicot B19-like genes. The B19 genes are regulated similarly during embryo development, but to very different levels. In contrast, they are differentially regulated by ABA and various types of osmotic stress. In immature embryos all three genes are responsive to ABA and mannitol. However, B19.1 is also responsive to salt. Cold stress does not induce B19 mRNAs; only a stabilization of the transcript levels is seen. These results suggest that the responses to salt stress and exogenous ABA operate through different pathways.


[The physiology of erection: study of the intrapenile tissue].

The mechanisms which control erection depend on the integrity of the intrapenile smooth muscle fibres, the endothelium which lines them, and the thick membrane, the albuginea, which surrounds them. These dynamic structures are subject to neurological control which stimulates the release of neurotransmitters and neuromodulators. These substances exert synergistic and antagonistic effects and act via receptors which have been identified by means of pharmacological studies. The objective of this paper is to simply present the current state of knowledge concerning the physiology of the erectile tissue of the penis.


[Role and contribution of karyotyping in male infertility].

In the context of the aetiological investigation of male infertility, the authors stress the place and the contribution of blood karyotype testing in the light of their personal experience based on 1,612 subjects. This examination has an important place, as about 15% of azoospermic subjects and 6 to 7% of subjects with oligospermia less than 10 million spermatozoa per ml, either alone or in combination with other abnormalities of the semen examination, present a congenital chromosomal abnormality. A remarkable constancy of the results was observed according to identical recruitment criteria. The contribution of this examination is also important: the medical and psychological value of detecting the cause of azoospermia, genetic counselling and antenatal chromosomal diagnosis for non-azoospermic subjects with an equilibrated structural abnormality, in whom treatment allows a chance of procreation, genetic counselling for the family of these subjects in order to prevent the appearance of a chromosomally abnormal infant. In conclusion, the authors argue in favour of the routine use of this test in all infertile subjects with at least isolated oligospermia less than 10 million spermatozoa per ml.


[Quantification of nucleolar organizers (NORs) in renal carcinomas. Comparison with the nuclear grade].

Twenty three renal tumours: 6 clear cell carcinomas, 3 granular cell carcinomas, 8 mixed cell carcinomas, 3 tubulopapillary tumours, 2 spindle cell carcinomas and 1 oncocytoma, were retrospectively submitted to a staining technique which reveals argyrophilic proteins related to Nucleolar Organizer Regions (NORs). The semiautomated quantification of the number and mean surface area of the argyrophilic granules was performed by means of a quantimeter. The results were compared to the nuclear grade, which constitutes the reference histological prognostic factor. A very good correlation was observed between NORs and nuclear grade. The mean number of argyrophilic granules per nucleus (n) and more particularly their surface area (s) were very significantly increased in high-grade carcinomas (n = 1.96), s = 3.72) in comparison with low-grade carcinomas (n = 1.50, s = 1.35). Quantification of NORs therefore allowed objective differentiation between a low nuclear grade and a high nuclear grade. The quantification method used emphasised the value of not only measuring the number but also the surface area of the argyrophilic granules.


[Tumor ablation with focalized ultrasound. In vivo experiment with prostatic adenocarcinoma R3327 Mat-Ly-Lu].

Ultrasound can induce tissue lesions by a combination of thermal and mechanical effects related to the tissue absorption of the energy emitted at the focal point of the transducer. The effects of focused ultrasound were studied in vivo in Fisher/Copenhagen hybrid rats bearing Dunning R3327 experimental prostatic carcinoma. The experimental tumour was transplanted by subcutaneous injection into the abdomen of 20 mg of tumour tissue derived from the Mat-Ly-Lu strain. Treatment was performed under general anaesthesia. The animal, maintained in a sarcophage exposing the tumour, was immersed in degassed water ensuring the interface between the tumour and the 1 MHz transducer. The displacements of the transducer were guided by computerised ultrasound screening, allowing irradiation of the entire tumour. The energy was supplied by a 7.5 kW amplifier in the form of series of impulses of variable duration. 77 rats were treated and the tumour growth was compared to that of non-irradiated control rats. Comparative series demonstrated the following results: 1. Immediate tumour destruction was obtained with a very high acoustic intensity (9,000 Watts/cm2) and a brief exposure time. 2. A transient slowing of the tumour growth rate was observed for an acoustic intensity of between 3,500 and 5,500 Watts/cm2. 3. Partial or total necrosis of the tumour was obtained with intensities of between 300 and 2,750 Watts/cm2 and a long exposure time. Total tumour destruction was obtained in 30 of the 49 rats treated under these conditions. 14 animals developed a local recurrence, 9 animals did not develop a local recurrence but developed metastases and 7 animals obtained long-term survival without local recurrence or metastasis. Under certain experimental conditions, focused ultrasound, without any adjuvant treatment, was able to destroy the Dunning R3327 Mat-Ly-Lu strain experimental tumour and, in certain cases, induced complete cure of this experimental cancer.


[Is the lower urinary tract sensation different in patients with bladder instability?].

The author examined the sensory innervation of the lower urinary tract in two ways: by evaluation perception of bladder fullness and by determining the perception threshold to electrical stimulation of the urethral and bladder wall. The results obtained in 95 patients with detrusor instability were compared to those of 312 patients with no neurological disease and a stable detrusor. Patients with detrusor instability reported sensations of fullness at statistically significantly lower volumes than patients with a stable detrusor. In contrast, the threshold of perception of electrical stimulation was the same in the two groups, which indicates that the sensory system involved in the perception of electrical stimulation is independent of motor function, while the sensation of fullness is influenced by detrusor motor activity. These results constitute additional evidence that perception of fullness and perception of electrical stimulation are related to two different pathways of afferent innervation.


[Cyproterone acetate: cardiovascular tolerance].

Cyproterone acetate, characterised by its antiandrogen and antigonadotrophin actions, is indicated in the treatment of advanced prostatic cancer. It can be used alone or in combination with surgical or chemical castration. This paper reviews the literature concerning the cardiovascular safety of this drug. Metabolic studies evaluating the impact of cyproterone acetate on lipids and certain clotting parameters indicate the absence of any harmful effects on these cardiovascular risk factors. The two trials conducted by the EORTC (30761 and 30 762) demonstrated that the cardiovascular safety of cyproterone acetate was better than that of diethylstilboestrol. Cardiovascular adverse effects were reported in less than 5% of cases. This figure was confirmed by a large number of published studies. A comparable incidence of cardiovascular complications was also observed in series of patients treated by orchidectomy, LHRH agonists and/or pure antiandrogens. It is difficult to formally demonstrate the responsibility of antihormonal treatment in this elderly population in which ischaemic heart disease and cerebral vascular accidents are frequent causes of death.


[Value of tomodensitometry with injection and late images for the diagnosis of urinary leaks in kidney transplants].

Between May 1985 and May 1990, 263 renal transplantations were performed in 254 patients, with 4 kidney-pancreas double grafts. 12 urinomas were observed, i.e. 4.56%. In one half of cases, clinical examination alone was able to establish the diagnosis of urinoma, but in the other 6 cases, complementary investigations were required. Post-contrast computed tomography with late images (between 6 and 24 hours after the injection) would appear to be the most useful investigation. Performed in 6 cases, this examination established the diagnosis of urinoma in 5 cases and also suggested the mechanism of the leak, which was always confirmed at operation. The positive diagnosis is based on extravasation of contrast agent which is clearly visible, even in the presence of low concentrations due to impaired renal function. This extravasation occurred early in 3 cases, but was delayed in 2 cases, emphasising the importance of late images. Other investigations appear to be less useful either because of a lack of sensitivity or because of imprecise anatomical information. The severity of urine leaks and the need for rapid treatment justify post-contrast computed tomography with late images in any case of suspected urinoma.


[Vesico-ureteral reimplantation in kidney transplantation from living relative donor: extravesical or transvesical? Urologic complications and long-term results evaluation].

Results of two techniques of ureteroneocystostomy were evaluated in a series of 400 consecutive living renal transplantations. In the first 150 patients (group A) PAQUIN ureteroneocystostomy was utilized and in the second 250 (group B) DE CAMPOS-FREIRE technique was performed. Significant urological complications were seen in 13 (8.6%) patients of group A and in 10 (4%) of group B. Bladder leak occurred in 5 patients of group A and in only 1 of group B (P < 0.05). Complications related to ureteral stenosis and ureteral necrosis were comparable in the two groups. High pressure vesicoureteral reflux was demonstrated in 2 of 85 patients of group A and in 15 of 160 of group B (P < 0.05). Abnormalities of intravenous urography were significantly higher in group A. The incidence of immediate and delayed post-transplant urinary infection was not statistically different among the two groups. We advocate the use of DE CAMPOS-FREIRE approach over the PAQUIN reimplantation, not only because of the marginal differences in the incidence of urological complications, but also because of the technical advantages offered by the former approach.


[Bladder pheochromocytoma (report of a new case)].

Bladder pheochromocytoma is a rare tumor. The authors report a new case and imaging modalities for localizing extrasurrenal pheochromocytomas are reviewed.


[A case of bladder pheochromocytoma].

Phaeochromocytoma is an endocrine neuroectodermal tumour usually located in the adrenal gland, but occasionally arising in other organs. The authors report a case of a vesical phaeochromocytoma presenting with haematuria. Radiological and endoscopic investigations revealed a right laterovesical submucosal tumour. Partial cystectomy was performed. The normality of the postoperative endocrine survey suggested disappearance of the tumour and absence of any other sites.


[Cancer of the kidney mimicking renal multilocular cyst].

Two cases of multilocular cystic adenocarcinoma of the kidney are presented. Although ultrasonography (US) and x-ray computed tomography (CT) are generally considered to be excellent diagnostic methods for renal cystic lesions, in the 2 cases reported here they failed to give the correct diagnosis. These unusual cases altered us to the possibility that the capabilities of modern diagnostic imaging techniques can lead to a false sense of security regarding the prognosis. To our knowledge, there are no reliable clinical features which distinguish multilocular cystic adenocarcinoma from multilocular cystic nephroma.


[Leiomyosarcoma of the kidney].

Leiomyosarcoma of the kidney is a rare neoplasm. Diagnosis is obtained by CT-Scan and immunohistochemical study. Radical nephrectomy is the only therapeutic way whose efficacity is known. The authors report a case of two localisations (kidney and bladder) of leiomyosarcoma.


[A case of hemangiopericytoma of the pelvis].

Haemangiopericytoma is an uncommon tumour of vascular origin. The authors report a case of one of the rarer sites of this tumour in man: paravesical pelvic haemangiopericytoma. Although modern imaging techniques have provided useful information concerning the hypervascular and clearly demarcated appearance of this tumour which displaces but does not invade adjacent organs, its diagnosis can only be established by histology. Its degree of malignancy and its invasive potential are unclear. The risk of local recurrence and metastases in more than one half of cases justifies wide surgical excision, possibly combined with adjuvant radiotherapy, and long-term follow-up.


[Sarcoma of the spermatic cord. Report of 2 cases. Review of the literature].

Illustrated by a case of rapidly fatal rhabdomyosarcoma in a child and a leiomyosarcoma in an adult, the authors differentiate the various histological types of sarcomas affecting the spermatic cord. Based on a review of the literature, they stress the great variety and various clinical courses related to the histological type. The predominant role of the histologist is stressed. The most widely used classifications are reviewed, leading to treatments in which primary surgery should be as complete as possible and combined with lymph node dissection or radiotherapy and chemotherapy. At the present time, the use of chemotherapy appears to be the decisive step in the improvement of the survival rates. The lymphophilic nature of rhabdomyosarcomas, which are embryonal in 95% of cases in children, can be schematically contrasted with the haematogenous nature of other sarcomas. The current approach to sarcomas of the spermatic cord, a minor site in comparison with the frequency of other soft tissue sarcomas, consists of an aggressive multidisciplinary treatment.


[Transplantation of a horse-shoe kidney. Report of a case and review of the literature].

The authors report a case of transplantation of a horseshoe kidney in two recipients. Based on this case and a review of the literature, they emphasise the technical precautions required for collection and transplantation of these malformed kidneys.


[Isolated urethral injury following coital faux pas. Report of a case with no associated injury of the corpora cavernosa].

An urethral lesion due to coital faux pas is usually associated with a lesion of the corpora cavernosa, but may occasionally be isolated. Suspected clinically due to the presence of urethral bleeding, the diagnosis of ruptured urethra is based on urethrography and, more importantly, urethroscopy, which appears to be more reliable for defining the complete or partial nature and the site of the rupture. Even in incomplete forms, surgical suture of the lesion appears to give better results than urinary diversion alone.


[Portal hypertension: a rare cause of hematuria].

A patient with undiagnosed cirrhosis presented with massive haematuria 16 years after augmentation colonoplasty. Portal hypertension (PHT) was responsible for a portocaval collateral circulation with vesical varices detected by low pressure endoscopy performed between episodes of haematuria. A very similar case was published in 1991. Surgical haemostatis with partial or total resection of the colonoplasty was only temporarily effective in our patient, who subsequently underwent liver transplantation due to repeated episodes of hepatic encephalopathy. No recurrence of haematuria was observed six years after the diversion. This rare cause of haematuria must be considered in view of the widespread use of intestinal conduits in urology: a dozen cases have been reported after transileal cutaneous ureterostomy, occurring an average of 3 years after the operation. Direct treatment of the varices, either by surgical devascularisation or resection or be endoscopic laser sclerosis or coagulation, is only temporarily effective. Treatment of the PHT appears to be the most effective measure in the long term: beta-blockers administered in the absence of episodes of haematuria or portal by-pass surgery in the case of failure.


[Posterior epispadias with unknown complete duplicated urethra].

The authors report a case of posterior epispadias associated with complete ventral urethra only discovered during the urethroplasty phase of the operation. This malformation is a rare form of urethral duplication and should be considered in any case of apparently isolated posterior epispadias.


[Reconstruction of the penile skin in Fourier's gangrene: the use of a helical skin graft].

The emergency treatment of Fournier's and perineal gangrene is now well defined and is based on excision of the necrotic tissues and broad spectrum antibiotics combined with hyperbarid oxygen therapy and symptomatic resuscitation measures. However, several procedures have been described for the reconstruction of the excised zones. In the light of one case of Fournier's gangrene, the authors evaluate reconstruction of the penile skin by means of a helical split-skin graft. With a follow-up of one year, the morphological and functional result was excellent. The patient has normal erection and the suture line does not contain any retractile adhesions. The authors therefore consider that this simple, but little known procedure should be part of the urologist's therapeutic arsenal for reconstruction of the penile skin.


[Ureteral replacement with ileal graft with anti-reflux valve].

The authors describe an ureteroileoplasty with a new anti-refluxing technique. A subtotal ureteroplasty is carried out, anti-refluxing proceeding is made. This one is based on Benchekroun's hydraulic valve principle. A transversal ileostomy is carried out 8 cm below the uretero-ileal anastomosis, which is invaginated and fixed to the proximal layer of the ileostomy with Dexon 3/0, the ileostomy is then closed using Dexon 2/0. The hydraulic valve may then ensure anti-reflux.


[New findings on the physiology of erection or the concept of the active sponge].

Until these last years, erection was mainly explained by neurovascular mechanisms. The erectile bodies were assimilated as a simple sponge passively filled or emptied by an unbalance between the arterial and venous flows. In fact, the erectile bodies themselves control the erection thanks to their potent smooth musculature. By contracting, they induce both flaccidity and detumescence, since their relaxation releases the erection. The erectile bodies work really as a kind of active sponge having the property to be autonomous and distortionable. However, the contractility of the erectile bodies cannot alone explain all the erection. This complex hydraulic process requires also complementary vascular mechanisms (cavernous outflow blockade, arteriodilatation) and tissular mechanisms (extrinsic compression of erectile striated muscles, specific distortion of each erectile body) under both hormonal and neuro-psychic control. By pointing up the fact that the active and passive tissue mechanisms play a major role for erection, this active sponge concept underlines the necessity for investigating the erectile tissues and allows to reconsider both physiopathology and treatment of dyserections.


[Contribution of the ambulatory urodynamic monitoring to the study of micturition disorders].

An outpatient urodynamic study allows simultaneous recording of bladder, urethral and abdominal pressures over a long duration with physiological bladder filling and under outpatient conditions. One hundred and eighteen outpatient urodynamic recordings were performed in patients presenting with disturbances of micturition such as urinary incontinence or dysuria. In the group of incontinent patients, the outpatient urodynamic recording was superior to the standard urodynamic assessment for the diagnosis of unstable bladder (p < 0.001). In the group of dysuric patients, the outpatient urodynamic recording allowed a precise evaluation of the detrusor activity and a simplified assessment of micturition. Its use combined with a urecholine test allows analysis of the onset, duration and intensity of action of the drug in comparison with a standard test.


[Cutaneous trans-jejunal ureterostomy: an original technique used in 29 patients].

The authors report an original technique of transintestinal cutaneous diversion using the jejunum A 10 to 15 cm segment of jejunum is raised from the second loop and is attached to the skin transperitoneally. The stoma lies below the ureterojejunal anastomosis and the two ureters, sutured according to Wallace's technique, are implanted end-to-end into the proximal and extraperitonealised end at the height of the sacral promontory. 29 patients have been treated by this technique for various indications: 21 bladder tumors, 5 neurogenic bladders, 1 bladder exstrophy, 1 cervical cancer, 1 prostatic cancer. 15 patients (52%) had previously received pelvic irradiation. The mean follow-up was 47 months (2-192 months) and the mortality was zero. 4 patients (13.8% developed early complications: 2 cases of urinary tract obstruction and 2 intestinal obstructions requiring 3 operations. 6 patients developed 7 minor late complications (24.1%): 2 cases of pyelonephritis, 4 cases of renal stones, only one of which required an operation, and 1 case of prolapsed stoma. None of the patients developed any alteration in renal function or dilatation of the upper urinary tract after the operation. The addition of sodium bicarbonate was found to be useless, as none of the patients developed any metabolic disorders. Transjejunal cutaneous ureterostomy achieves excellent drainage by means of a short graft with a stoma situated below the level of the ureteric implantation, as reflected by the absence of any long-term deterioration in renal function despite full lumen ureterojejunal implantation without an antireflux device. This technique is simple to perform and is indicated in all situations, particularly after pelvic irradiation. It is associated with low morbidity, no mortality and ensures an excellent long-term result.


[Cholelithiasis and ileo-ceco-cystoplasty (study of 39 patients)].

Limited data have been reported concerning the gallstone complications of ileocaecocystoplasty. A retrospective study of gallbladder ultrasonography looking for gallstones was performed in 35 of 39 patients undergoing ileocaecocystoplasty between July 1987 and July 1991. The surgical technique used in 33 men and 2 women with a mean age of 64.5 years was that described by Thuroff (Mainz Pouch). Gallbladder ultrasonography was performed a fortnight before the operation and then regularly during follow-up, with a mean follow-up of 24 months. Eight patients, an average of 13 months after ileocaecocystoplasty, developed ultrasonographically detectable gallstones, whose prevalence in the male study population was four times higher than that of the general male population. This study raises the question of whether cholecystectomy should be performed routinely during ileocaecocystoplasty and further studies should be conducted to clarify this problem.


[Recurrence of lithiasis after extracorporeal lithotripsy, percutaneous surgery, and open surgery for calculi of the upper urinary tract].

The safety and short-term effectiveness of percutaneous nephrolithotomy (PCNL) and extracorporeal shock wave lithotripsy (ESWL) have already been well established. However, long-term follow-up studies are essential to prove that the recurrence rate after PCNL and/or ESWL is equal or even better than that of surgery. We reviewed 57 patients treated with ESWL monotherapy, 45 patients treated by PCNL (or by PCNL combined with ESWL for complete staghorn stones) and 59 patients treated by open surgery who all had at least three years of follow-up. The overall recurrence rate for ESWL was 37% for PCNL and for open surgery 39%. ESWL, PCNL and open surgery all showed a higher rate of recurrence and persistent bacteriuria when associated with residual stones. Therefore every effort should be made to remove residual fragments following stone treatment. We conclude that because of its efficacy and low morbidity ESWL should be the treatment of choice for most of renal calculi. Complete staghorn calculi are best treated with open surgery because the complete elimination of all calculous material and the eradication of infection are achieved at a higher rate than by PCNL and ESWL.


[Piezoelectric extracorporeal lithotripsy in children: first results and reflections after 18 sessions].

From 1988 to 1990, we treated 13 children between the ages of 2 and 15 years with stones of the upper urinary tract. These stones were treated by means of 18 sessions of piezoelectric extracorporeal lithotripsy: 14 under general anaesthesia and 4 without even any analgesia. We obtained an 84.5% success rate with no particular adverse effects. These results are similar to those reported for hydroelectric or electromagnetic shock wave lithotripsy, which are associated with a higher morbidity. The efficacy of extracorporeal lithotripsy in paediatrics is remarkable, but clinical studies in adults as well as animal experimentations have demonstrated severe renal contusion and do not formally exclude the possibility of delayed complications. At the present time, this technique should therefore be used very cautiously and must be accompanied by systematic prevention of recurrent stones.


[Does magnetic resonance imaging allow the assessment of the loco-regional extension of cancer of the prostate? Report of 27 anatomo-radiologic comparisons].

27 patients with histologically proven prostatic carcinoma were investigated by magnetic resonance imaging (MRI) (0.5 Tesla) in order to determine the local spread of the cancer. These patients then underwent ilio-obturator lymph node dissection with frozen section examination, followed by radical prostatectomy. Histological examination of the resection specimens was performed on slides prepared from large transverse sections every 5 mn. The MRI examination was especially designed to confirm the diagnosis of capsular effraction based on the signals of the periprostatic fat (PF), periprostatic venous plexuses (PVP) and seminal vesicles (SV). An abnormality of at least one of these structures was considered to indicate the diagnosis of capsular effraction. The MRI data were compared to the histological findings. The results demonstrated a sensitivity of 62% for the PF signal, 52% for the PVP signal and 40% for the SV signal. The overall MRI-Histology correlation was found to be exact in 23 out of 27 cases (Accuracy = 85%). All of our four errors represented understaging (Sensitivity = 81%). When a rigorous methodology is respected, the high accuracy and sensitivity of MRI makes this modality an investigation of choice for the study of the prostatic capsule, as part of the routine staging of prostatic cancer. It should allow a better selection of patients with intracapsular cancer (T1-2), who constitute candidates for radical treatment.


[Color doppler echography in the exploration of vasculogenic impotence].

102 patients with suspected vasculogenic impotence were evaluated with color doppler sonography. Measurement of normal systolic and diastolic velocities were obtained from the cavernosal arteries of patients responding by a full erection after intra-cavernosal injection of 20 mg of Papaverine. A correlation with cavernosometry was obtained in 61 patients and with selective internal pudendal arteriography in 11. The 10 patients with abnormal arteriograms had a systolic velocity < 25 cm/sec. 13 out of the 15 patients with an end diastolic velocity > 5 cm/sec had a venous leak defined by a maintenance flow rate of erection during cavernosometry > 25 ml/mn. End diastolic velocity is an excellent index of the function of the veno-occlusive system, provided the systolic velocity remains at a normal value. In case of arterial insufficiency, a diastolic flow < 5 cm/sec is of no value and cavernometry is mandatory to detect a mixed arterio-venous impotence. The addition of color doppler sonography permitted a more rapid detection of vessels and an easily reproducible measurement of velocities which makes color doppler sonography an excellent screening test for examining patients with potential vasculogenic impotence.


[Electrophoretic study of the protein composition of human seminal vesicle secretion].

The SDS-PAGE and two-dimensional reductived electrophoretic study of the liquid aspirated from human seminal vesicles showed that this medium contains high molecular weight proteins (greater than 150 kDa). In addition to these high molecular weight proteins, native seminal vesicle secretion is characterised by the presence of a large 47.4 kDa protein band corresponding to seminogelin, described by Lilja. The high molecular weight proteins actually correspond to associations of three types of protein subunits with molecular weights of 75-76, 63-64 and 47-48 kDa, which represent the structural elements of the coagulum; one of these subunits consists of seminogelin. Seminal vesicle secretion contains very little or no proteolytic enzymes, so that its protein composition remains stable even after incubation for 24 hours.


[Urinoma: a complication of celiac plexus infiltration].

A case is described in which a major complication resulted from this technique. We have found no previous record of urinoma due to infiltration of a caustic agent.


[Endo-urologic treatment of ureteral fistulas occurring after pelvic surgery for carcinoma: report of 5 cases].

The authors report 5 cases of ureteric fistula after radical pelvic cancer surgery. Three patients had a recurrent tumour and 3 had received radiotherapy. Endourological methods achieved 4 immediately satisfactory results, but only one complete long-term success. Isolated percutaneous drainage of the pyelocaliceal cavities effectively dried up the urine leak, but carried a high risk of secondary stenosis of the excretory tract in the absence of concomitant intubation of the fistula zone. However, endourological treatment, combined with urinary diversion and catheterisation of the pathological ureter, represented a reliable alternative to conventional surgery.


[Postoperative rectoprostatic fistulas. Report of 2 cases. Review of the literature].

The authors report two cases of acquired prostatorectal fistulae after TURP, which were treated by a Kraske approach. They review the literature of iatrogenic fistulae and all of the techniques of approaching the prostatorectal region. If the bladder drainage is always necessary and may be sufficient in as much as 34% of the patients, the need for a colostomy and the choice of the approach is discussed.


[Pyeloureteral lieberkuhn adenocarcinoma after trans-intestinal ureterostomy for bladder exstrophy].

The authors report a case of pyeloureteric adenocarcinoma in a urinary diversion inserted for bladder exstrophy and in a context of chronic pyelonephritis secondary to renal stones. The various aetiopathogenic hypotheses are discussed. Renal stones and chronic inflammation very probably played an important role in the development of this type of adenocarcinoma in the urothelium.


[Burkitt's lymphoma: an exceptional cause of testicular tumor].

Based on a case of Burkitt's lymphoma presenting in the form of a testicular mass in a 66 year old man, the authors stress the importance of making the diagnosis as early as possible. This malignant lesion has one of the greatest capacities for proliferation and any delay in treatment can decrease the chances of curability of the tumour.


[A rare psychogenic anejaculation. Report of a case of prolactin adenoma].

The authors report a case of prolactinoma discovered during assessment of a case of ejaculation failure attributed for more than eight years to a psychogenic cause. Because of the size of the tumour, surgical resection of the lesion left a persistent hyperprolactinaemia, which only returned to normal after treatment with bromocriptine. The persistence of sexual disorders justified adjuvant treatment with exogenous testosterone. Despite the unusual mode of presentation of hyperprolactinaemia, these data correspond to those reported in the literature. Prolactin assay should therefore be performed in any case of male sexual dysfunction to allow the early diagnosis of prolactinoma and to improve the functional prognosis.


[Anterior sacral meningocele with urologic manifestations. Report of 3 cases].

Anterior sacral meningocele is defined as a spinal fluid-filled thecal sac in the pelvis communicating with the spinal subarachnoid space through a defect in the anterior sacral wall. Since scimitar shape of the sacrum is characteristic, diagnosis is easily confirmed by echography and myelography. The sac very often contains benign tumors and a thickened filum terminale which can achieve a tethered cord syndrome. This congenital malformation, whose autosomal inherited condition has been proposed, has usually few characteristic symptoms but can present itself as a neurogenic bladder from tethered cord origin. Neurosurgical treatment can prevent rupture of the meningocele with meningitis. In addition to symptomatic treatment of the neurogenic bladder, the urologist must advise neurosurgical operation to preserve potency and cure specific bladder dysfunction secondary to tethered cord syndrome.


[The Bourne's test: a relevant contribution to the diagnosis of vesico-enteric fistulas].

Diagnostic verification of vesicoenteric fistulae is necessary in order to establish the indication for surgical therapy, but this is frequently difficult. This is because uroradiological and endoscopic methods are unreliable and the symptoms are often nonspecific. The Bourne test is a simple noninvasive screening method. If necessary, it can be supplemented by computer tomography. The problems are discussed with reference to a case report.


[Microsurgical vaso-vasostomy: a reliable method of vasectomy reversal].

An original microsurgical technique has been used since 1971 to reverse the vasectomies of 2,110 men. Both sides were anastomosed if possible using a meticulous two layered technique with further reinforcing sutures to prevent strain on the join. Other important points include testing the fluid emerging from the testicular end, checking patency of the prostatic end and insisting on good scrotal support in the recovery phase, particularly the important first few days. The series returned a 95% baby rate, with a return of sperm to the ejaculate in 93% of cases.


[Complications of prostatic biopsies. Analysis of a series of 4,764 biopsies].

A multicentre, retrospective series of 4,764 prostatic biopsies revealed 89 complications (1.86%). The commonest complications were infectious (including 10 cases of septicaemia) and antibiotic prophylaxis does not appear to be able to totally prevent them. Thirty three complications occurred after negative biopsies. Many minor complications are probably treated at home by the local doctor and would therefore not be reported to the urologist.