Proteomic analysis of rat hippocampus exposed to the antidepressant paroxetine.

Antidepressant drugs can exert significant effects on the mood of a patient suffering major depression and other disorders. These drugs generally have pharmacological actions on the uptake or metabolism of the neurotransmitters serotonin, noradrenaline and, to a lesser extent, dopamine. However, there are many aspects of antidepressant action we do not understand. We have applied proteomic analysis in a rat hippocampal model in an attempt to identify relevant molecules that operate in pathways functionally relevant to antidepressant action. Rats were administered either 5 mg/kg daily of the antidepressant paroxetine or vehicle for 12 days, then hippocampal protein was recovered and resolved by 2-D gel electrophoresis. After antidepressant exposure, we observed increased expression or modification of cytochrome c oxidase, subunit Va, cyclin-dependent kinase inhibitor 2A interacting protein, dynein, axonemal, heavy polypeptide 3 and RHO GDP-dissociation inhibitor alpha. Decreased expression or modification was observed for complexin 1 (CPLX1), alpha-synuclein, parvalbumin, ribosomal protein large P2, prohibitin, nerve growth factor, beta subunit (NGFB), peroxiredoxin 6 (PRDX6), 1-acylglycerol-3-phosphate O-acyltransferase 2_predicted, cystatin B (CYTB) and lysosomal membrane glycoprotein 1. CPLX1, the most strongly regulated protein observed, mediates the fusion of cellular transport vesicles with their target membranes and has been implicated in the pathophysiology of mood disorders, as well as antidepressant action. CPLX1 and the other proteins identified may represent links into molecular processes of importance to mood dysregulation and control, and their respective genes may represent novel candidates for studies of antidepressant pharmacogenetics.

Sexual function in women with and without urinary incontinence and/or pelvic organ prolapse.

The sexual function of women with and without urinary incontinence and/or pelvic organ prolapse (UI/POP) was compared using a condition-specific validated questionnaire, the Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire (PISQ). Eighty-three women with UI/POP and 56 without agreed to participate. PISQ scores were significantly lower among women with UI/POP than in those without (P = 0.003). No differences in the stages of sexual excitement were noted between groups. The frequency of intercourse was less with UI/POP than without (P= 0.04). Women with UI/POP restricted sexual activity for fear of losing urine more frequently than did those without (P = 0.005). No differences were reported in patients' or partners' sexual satisfaction. This study found that women with UI/POP have poorer sexual functioning than those without, as measured by the PISQ, and report less frequent sexual activity. In addition, women with UI/POP are more likely to restrict sexual activity for fear of incontinence, although they report similar levels of satisfaction with their sexual relationships as do women without UI/POP.

Patching together the genetics of Gorlin syndrome.

BACKGROUND: Gorlin syndrome is an autosomal dominant disorder characterized by developmental defects and susceptibility to cancer, especially to basal cell carcinomas. The genetic basis of this disorder has recently been elucidated. METHODS: In this article previous studies are reviewed in which loss of heterozygosity analysis of tumours and normal tissue pointed to a region on chromosome 9 as being involved in Gorlin syndrome. In this light, Knudson's two-hit model is discussed. The identification of the involvement of the patched gene in Gorlin syndrome is reviewed. New data on genotype-phenotype correlations in the syndrome are presented. RESULTS: Loss-of-heterozygosity studies, together with standard family studies using linkage analysis, have proved useful in identifying the location of a gene with complex phenotypic expression. CONCLUSION: The application of the two-hit model, as utilized in loss-of-heterozygosity studies, has been very useful in elucidating the genetic basis of Gorlin syndrome. There may be a correlation between certain aspects of the mutations in patched and the clinical presentation of the disorder in families.

Sjögren-Larsson syndrome is caused by a common mutation in northern European and Swedish patients.

Sjögren-Larsson syndrome (SLS) is an autosomal recessive disorder characterized by congenital ichthyosis, mental retardation, and spastic diplegia or tetraplegia. Patients with SLS have deficient activity of fatty aldehyde dehydrogenase (FALDH), an enzyme involved in long-chain fatty alcohol oxidation. The cDNA encoding FALDH has recently been cloned and several different mutations have been found in SLS patients. We have now identified a point mutation (C943 --> T) in 7 of 19 kindreds of European descent, accounting for 24% of the SLS alleles. The C943T mutation was only found in patients of northern European ancestry from Sweden, the Netherlands, Germany, and Belgium. Haplotype analysis suggested that the patients carrying the C943T allele were distantly related. All four Swedish patients were homozygous for C943T, indicating that this mutation is probably the major cause of SLS in the inbred Swedish families. The mutation leads to the substitution of serine for the highly conserved proline 315 in the FALDH protein, and expression studies confirm that it destroys enzymatic activity. The mutation was readily detected with an MnlI restriction enzyme digestion test. The finding that C943T is a common SLS mutation in northern European and Swedish patients affords a rapid simple method for diagnosing SLS by screening patients for this mutation with DNA-based methods.

Genomic organization and expression of the human fatty aldehyde dehydrogenase gene (FALDH).

Mutations in the fatty aldehyde dehydrogenase (FALDH) gene cause Sjögren-Larsson syndrome (SLS)-a disease characterized by mental retardation, spasticity, and congenital ichthyosis. To facilitate mutation analysis in SLS and to study the pathogenesis of FALDH deficiency, we have determined the structural organization and characterized expression of the FALDH (proposed designation ALDH10) gene. The gene consists of 10 exons spanning about 30.5 kb. A TATA-less promoter is associated with the major transcription initiation site found to be 258 bp upstream of the ATG codon. The GC-rich sequences surrounding the transcription initiation site encompassed regulatory elements that interacted with proteins in HeLa nuclear extracts and were able to promote transcription in vitro. FALDH is widely expressed as three transcripts of 2, 3.8, and 4.0 kb, which originate from multiple polyadenylation signals in the 3' UTR. An alternatively spliced mRNA was detected that contains an extra exon and encodes an enzyme that is likely to have altered membrane-binding properties. The FALDH gene lies only 50-85 kb from ALDH3, an aldehyde dehydrogenase gene that has homologous sequence and intron/exon structure.

Polymorphonuclear leukocyte-mediated bone degradation and influence of blood mononuclear cells on the mouse calvarial model.

OBJECTIVE: To investigate the relationships between degradation of bone and activated blood polymorphonuclear leukocytes (PMNL) and mononuclear leukocytes (ML) as well as their soluble products in vitro. MATERIALS AND METHODS: A neonatal mouse calvarial bone model was used to assess the activity of degradation (by measuring the amount of 45Ca release) by normal human blood leukocytes, separated PMNL and ML following 24-hour incubation. The effects of conditioned culture medium obtained from Staphylococcus aureus-stimulated ML on PMNL-mediated calvarial bone loss were also studied. RESULTS: It was demonstrated that isolated human PMNL rapidly degraded bone in a dose and time dependent manner. The PMNL-mediated bone degradation was enhanced by conditioned medium obtained from Staphylococcus aureus-stimulated ML. CONCLUSION: These findings implicate PMNL as major contributors to early bone loss in infectious diseases such as acute haematogenous osteomyelitis.

Sjögren-Larsson syndrome is caused by mutations in the fatty aldehyde dehydrogenase gene.

Sjögren-Larsson syndrome (SLS) is an inherited neurocutaneous disorder characterized by mental retardation, spasticity and ichthyosis. SLS patients have a profound deficiency in fatty aldehyde dehydrogenase (FALDH) activity. We have now cloned the human FALDH cDNA and show that it maps to the SLS locus on chromosome 17p11.2. Sequence analysis of FALDH amplified from fibroblast mRNA and genomic DNA from 3 unrelated SLS patients reveals distinct mutations, including deletions, an insertion and a point mutation. The cloning of FALDH and the identification of mutations in SLS patients opens up possibilities for developing therapeutic approaches to ameliorate the neurologic and cutaneous symptoms of the disease.

Genetic homogeneity in Sjögren-Larsson syndrome: linkage to chromosome 17p in families of different non-Swedish ethnic origins.

Sjögren-Larsson syndrome (SLS) is a rare, autosomal recessive disorder that is characterized by congenital ichthyosis, mental retardation, and spastic diplegia or tetraplegia. Three United States families, three Egyptian families, and one Israeli Arab family were investigated for linkage of the SLS gene to a region of chromosome 17. Pairwise and multipoint linkage analysis with nine markers mapped the SLS gene to the same region of the genome as that reported in Swedish SLS pedigrees. Examination of recombinants by haplotype analysis showed that the gene lies in the region containing the markers D17S953, D17S805, D17S689, and D17S842. D17S805 is pericentromeric on 17p. Patients in two consanguineous Egyptian families were homozygous at the nine marker loci tested, and another patient from a third family was homozygous for eight of the nine, suggesting that within each of these families the region of chromosome 17 carrying the SLS gene is identical by descent. Linkage of the SLS gene to chromosome 17p in families of Arabic, mixed European, Native American, and Swedish descent provides evidence for a single SLS locus and should prove useful for diagnosis and carrier detection in worldwide cases.

Mutations in the gene for transglutaminase 1 in autosomal recessive lamellar ichthyosis.

We recently mapped the disease locus for severe autosomal recessive lamellar ichthyosis (LI) to chromosome 14q11 and showed complete linkage with TGM1, the gene encoding transglutaminase 1. We have now identified point mutations in TGM1 in two of the multiplex LI families used in the linkage study. Each nucleotide change causes a non-conservative amino acid substitution of histidine for one of two adjacent arginine residues in exon 3 of the gene (Arg141His, Arg142His). Within the transglutaminase family, these arginines are invariant within a conserved region, distant from the catalytic site of the enzyme. We hypothesize that these mutations adversely affect formation of crosslinks essential in production of cornified cell envelopes and a normal stratum corneum layer of the skin.

Polymorphism in two genes for B2 high sulfur proteins of wool.

Variation in the nucleotide sequence of the B2 high-sulfur protein genes has not been reported previously. This paper reports 15 nucleotide substitutions in each of the genes for the B2A and B2C proteins and a length of polymorphism in the B2A gene which translates to the insertion/deletion of one 30-nucleotide repeat sequence. Evidence is presented for gene conversion occurring within the B2 high-sulfur multigene family. These DNA polymorphisms may account for some of the microheterogeneity observed in the B2 high-sulfur proteins and may also be useful genetic markers of the B2 high-sulfur protein gene loci for future use in analysing wool fibre characteristics.

Soluble products of Staphylococcus aureus-stimulated blood mononuclear cells in enhancing polymorphonuclear leukocyte degradation of bone.

Rapid, extensive loss of infected bone implies abnormal localized inflammatory cell activity. We have demonstrated, using a live bone Ca-45 release model, that polymorphonuclear leukocytes degrade bone in a dose dependent manner. Staphylococcus aureus-stimulated blood mononuclear leukocytes release soluble products in vitro that enhance that process. Despite the usually accepted roles of osteoclasts and their blood-borne monocytic precursors in normal bone remodelling, these results indicate that considerable early pathological infected bone loss may be attributable to inflammatory polymorphonuclear leukocytes.