Characterization of allelic variants at chromosome 15q14 in schizophrenia.

Evidence of genetic linkage for schizophrenia at chromosome 15q14 has been reported in nine independent studies, but the molecular variants responsible for transmission of genetic risk are unknown. National Institute of Mental Health Schizophrenia Genetics Initiative families were genotyped for single nucleotide polymorphisms (SNPs) and dinucleotide repeat markers in the 15q14 linkage region and analyzed based on the presence of particular alleles of the dinucleotide repeat marker D15S165 in the 15q14 region. Two alleles showed both familial transmission disequilibrium and population-wide association with schizophrenia. The two groups identified by these two D15S165 alleles differ in age of onset, number of hospitalizations and intensity of nicotine abuse, as well as in predominant ethnicity. Variations in the frequency of SNPs in CHRNA7, the alpha-7-nicotinic acetylcholine receptor subunit gene at 15q14, were found in each group. Further sequencing in these two groups may yield more definitive identification of the molecular pathology.

A novel immunocytochemistry technique to measure hsp70 induction in L929 cells exposed to cadmium chloride.

The heat shock response has been suggested as a potential biomarker in toxicology. A vast amount of stimuli have been shown to induce heat shock proteins and new techniques to measure the response are constantly being assessed. In this study we use a novel immunocytochemistry technique to measure heat shock protein 70 (hsp70) induction in L929 cells exposed to cadmium chloride. Hsp70 induction was quantifiably measured using a soluble coloured substrate and qualitatively measured using a coloured substrate that precipitated at the location of hsp70. Using the insoluble coloured substrate hsp70 was identified predominantly within the cytoplasm of control cells. At intermediate cadmium concentrations hsp70 was observed to translocate to the nucleus. At these intermediate concentrations a heterogeneous heat shock response was observed. At lethal concentrations a strong heat shock response was observed with a widespread cellular response. This study demonstrates the potential of this immunocytochemistry technique to measure toxicological effects in cells by identifying the response quantitatively and qualitatively.

Linkage disequilibrium for schizophrenia at the chromosome 15q13-14 locus of the alpha7-nicotinic acetylcholine receptor subunit gene (CHRNA7).

The transmission/disequilibrium test was used for fine mapping of the linkage of schizophrenia to the chromosome 15q13-14 region, the site of a candidate gene, the alpha7 nicotinic acetylcholine receptor subunit gene (CHRNA7), in parent-child triads from the NIMH Schizophrenia Genetics Initiative families. This candidate gene was identified from neurobiological studies of deficits in schizophrenics of the inhibitory gating of the P50 auditory evoked potential. The neurobiological deficit was also used as a phenotype for subsequent linkage analysis. In the present study, significant genotype-wise disequilibrium (P < 0.007) was found at D15S165, a polymorphic simple sequence marker physically located within 1 megabase of both CHRNA7 and a partially duplicated, expressed sequence that includes exons 5-10 of CHRNA7. Replication of this result was found in an additional set of families. The results support this region as a chromosomal location involved in the genetic transmission of schizophrenia.

Smoking and mental illness.

Patients with mental illness have a higher incidence of smoking than the general population and are the major consumers of tobacco products. This population includes subjects with schizophrenia, manic depression, depression, posttraumatic stress disorder (PTSD), attention-deficit disorder (ADD), and several other less common diseases. Smoking cessation treatment in this group of patients is difficult, often leading to profound depression. Several recent findings suggest that increased smoking in the mentally ill may have an underlying biological etiology. The mental illness schizophrenia has been most thoroughly studied in this regard. Nicotine administration normalizes several sensory-processing deficits seen in this disease. Animal models of sensory deficits have been used to identify specific nicotinic receptor subunits that are involved in these brain pathways, indicating that the alpha 7 nicotinic receptor subunit may play a role. Genetic linkage in schizophrenic families also supports a role for the alpha 7 subunit with linkage at the alpha 7 locus on chromosome 15. Bipolar disorder has some phenotypes in common with schizophrenia and also exhibits genetic linkage to the alpha 7 locus, suggesting that these two disorders may share a gene defect. The alpha 7 receptor is decreased in expression in schizophrenia. [(3)H]-Nicotine binding studies in postmortem brain indicate that high-affinity nicotinic receptors may also be affected in schizophrenia.

The use of stress proteins as a biomarker of sub-lethal toxicity: induction of heat shock protein 70 by 2-isobutyl piperidine and transition metals at sub-lethal concentrations.

The stress response is a highly conserved reaction to various physical, chemical and biological stimuli. The ubiquity of the response occurring across taxonomic classes has identified heat shock proteins as potential biomarkers. In this study using the neutral red assay, silver stained one-dimensional SDS-PAGE, Western blotting and ELISA, the use of heat shock proteins as biomarkers of sub-lethal toxicity was examined. Hsp70 was induced in the mouse connective tissue cell line (L929) at sub-lethal concentrations for three transition metals (cadmium, mercury and copper) and for 2-isobutyl piperidine, a novel compound whose chemical structure is similar to a toxin found in the Colorado potato beetle (Leptinotarsa decemlineata). Hsp70 induction was found to increase in a dose-dependent fashion. Expression of other potentially interfering proteins was found to decrease with increasing toxin concentration. The induction of hsp70 at sub-lethal concentrations by the transition metals and 2-isobutyl piperidine demonstrates the potential of hsp70 as a biomarker of sub-lethal toxicity.

Inhibitory neurophysiological deficit as a phenotype for genetic investigation of schizophrenia.

Many investigators have proposed that biological endophenotypes might facilitate the genetic analysis of schizophrenia. A deficit in the inhibition of the P50 evoked response to repeated auditory stimuli has been characterized as a neurobiological deficit in schizophrenia. This deficit is linked to a candidate gene locus, the locus of the alpha7-nicotinic cholinergic receptor subunit gene on chromosome 15q14. Supportive evidence has been found by other investigators, including: 1) linkage of schizophrenia to the same locus; 2) linkage of bipolar disorder to the locus; and 3) replication of the existence of this neurobiological deficit and its relation to broader neuropsychological deficits in schizophrenia. It is certain that there are many genetic factors in schizophrenia and bipolar disorder; what is needed is a complete and precise description of the contribution of each individual factor to the pathophysiology of these illnesses.

Smoking and schizophrenia: abnormal nicotinic receptor expression.

Biological and genetic evidence suggests a role for the neuronal nicotinic receptors in the neuropathophysiology of schizophrenia. Nicotine normalizes an auditory evoked potential deficit seen in subjects who suffer from the disease. Nicotinic receptors with both high and low affinity for nicotine are decreased in postmortem brain of schizophrenics compared to control subjects. The chromosomal locus of the human alpha-7 gene (15q14) is linked to the gating deficit with a lod of 5.3, and antagonists of the alpha-7 receptor (alpha-bungarotoxin and methyllycaconitine) induce a loss of gating in rodents. We have cloned the human alpha-7 gene and found it to be partially duplicated proximal to the full-length gene. The duplication is expressed in both the brain and in peripheral blood cells of normal subjects, but is missing in some schizophrenic subjects. The results of these studies suggest the presence of abnormal expression and function of the neuronal nicotinic receptor gene family in schizophrenia.

Genomic organization and partial duplication of the human alpha7 neuronal nicotinic acetylcholine receptor gene (CHRNA7).

The human alpha7 neuronal nicotinic acetylcholine receptor gene (HGMW-approved symbol CHRNA7) has been characterized from genomic clones. The gene is similar in structure to the chick alpha7 gene with 10 exons and conserved splice junction positions. The size of the human gene is estimated to be larger than 75 kb. A putative promoter 5' of the translation start in exon 1 has been cloned and sequenced. The promoter region lacks a TATA box and has a high GC content (77%). Consensus Sp1, AP-2, Egr-1, and CREB transcription factor binding sites appear to be conserved between bovine and human genes. The alpha7 nAChR gene was found to be partially duplicated, with both loci mapping to the chromosome 15q13 region. A yeast artificial chromosome contig was constructed over a genetic distance of 5 cM that includes both alpha7 loci and the region between them. Four novel exons are described, located in genomic clones containing the partially duplicated gene. The duplicated sequences, including the novel exons, are expressed in human brain.

Further investigation of a chromosome 15 locus in schizophrenia: analysis of affected sibpairs from the NIMH Genetics Initiative.

Linkage of a neurophysiological deficit associated with schizophrenia, i.e., the failure to inhibit the auditory P50 response, was previously reported at chromosome 15q14. The marker with the highest pairwise lod score, D15S1360, was isolated from a yeast artificial chromosome containing a candidate gene, the alpha7-nicotinic acetylcholine receptor gene. In the present study, this linkage was further investigated in a subset of the NIMH Genetics Initiative schizophrenia families. These families have not been studied neurophysiologically, as were the families in the original report. Therefore, the DSMIII-R diagnosis of schizophrenia was used as the affected phenotype. Twenty families fulfilled the criteria of at least one sibpair concordant for schizophrenia, along with their two parents or another affected relative outside the nuclear family, available for genotyping. Sibpair analysis showed a significant proportion of D15S1360 alleles shared identical-by-descent (0.58; P < 0.0024). The results further support the involvement of this chromosomal locus in the genetic transmission of schizophrenia.

Nicotinic receptors, smoking and schizophrenia.

Neuronal nicotinic acetylcholine receptor expression was examined in schizophrenia. The incidence of smoking in schizophrenia is remarkably high and nicotine has been found to normalize an auditory evoked potential deficit seen in most subjects who suffer from this disease. Antagonists and agonists of a specific subset of this receptor family, the alpha7 nicotinic receptor, were found to regulate the gating of filtering of auditory information in both humans and in an animal model. The alpha7 gene has been cloned and a polymorphic dinucleotide repeat near the gene was used for linkage analysis, showing the alpha7 locus to be linked to the P50 deficit. Expression of the alpha7 receptor, which binds nicotine with low affinity, is reduced in the hippocampus of schizophrenics. [3H]-nicotine binding, a measure of the high affinity nicotinic receptors, was also decreased in schizophrenics and does not increase in response to tobacco use, as is seen in control subjects. The results of these studies suggest the presence of abnormal expression and function of the neuronal nicotinic receptor gene family in schizophrenia.

Indications for penetrating keratoplasty and associated procedures, 1989-1995.

PURPOSE: To identify changing trends in indications for penetrating keratoplasty and associated surgical procedures. METHODS: Review of charts from all patients who underwent penetrating keratoplasty at Wills Eye Hospital from January 1, 1989 through December 31, 1995. RESULTS: A total of 2,442 corneal transplants were performed in 2,186 patients. The leading indication for penetrating keratoplasty was pseudophakic corneal edema, accounting for 634 cases (26.0%); 54.7% of them were associated with anterior chamber intraocular lenses, 36.4% with posterior chamber intraocular lenses, and 3.1% with iris-fixated intraocular lenses. Regraft (17.8%), Fuchs' dystrophy (15.7%), and keratoconus (13.2%) followed pseudophakic corneal edema in frequency. Cataract extraction, with or without intraocular lens implantation, was combined with penetrating keratoplasty in 439 cases of 1,264 phakic eyes (34.7%). Intraocular lens exchange was performed in 285 of the 634 cases of pseudophakic corneal edema (44.9%). CONCLUSION: Pseudophakic corneal edema was the leading indication for penetrating keratoplasty, with an increasing number of cases associated with posterior chamber intraocular lenses during the study period (p = 0.001). The number of regrafts steadily increased between 1989 and 1995 (p = 0.001), being the second most common indication for corneal transplantation since 1992.

Food as a motivator in a client with Prader Willi syndrome.

A 34-year-old man with Prader Willi syndrome and an associated learning disability had had ulcers on both legs for 20 years because the healing process was interrupted by self-harm. This multi-disciplinary team used behaviour modification techniques with food being the motivator to heal both legs. Such a motivator would not normally be advocated because people with this syndrome often have excessive appetites but in this case it was felt that the possible gains would outweigh the risks.

Repair of Descemet's membrane detachment with intracameral injection of 20% sulfur hexafluoride gas.

Detachment of Descemet's membrane is a rare complication of cataract surgery that can cause permanent corneal decompensation. We report five patients with Descemet's membrane detachments who were successfully treated with intracameral injection of 20% sulfur hexafluoride gas with resultant resolution of their corneal edema.

A 20-yr longitudinal study of Olympic oarsmen.

Nine 1972 silver-medalist oarsmen were studied before the Olympic Games and 10 and 20 yr later. Peak power, metabolic responses, and heart rate were recorded during rowing ergometry; blood lactate was measured following exercise. The skinfold equation yielded percent body fat. The average change (multiple analysis of variance) among measurements from 1972 to 1992 was 37.5 +/- 3% (P < 0.01). Average changes between 1972 and 1982 and between 1982 and 1992 were similar, 17 and 18%, respectively (P < 0.01). The most significant change between 1972 and 1992 was decreased peak blood lactate (106%). Decreases in peak power, VE, and VO2 (ml.kg-1.min-1) were all similar, approximately 40%, and were significant. Body fat increased (from 12.3 to 15.6%), and absolute VO2 and relative VO2 (lean body mass) decreased 30% (P < 0.01). Only body weight, heart rate, and O2 pulse showed smaller changes, but these changes were still significant (P < 0.05). Relative peak VO2 decreased from 65.5 to 46.8 ml.kg-1.min-1 from 1972 to 1992 and at a rate of 10%.decade-1. The most significant changes between 1972 and 1982 were increases in percent body fat (from 12.3 to 16.3%) and decreases in VO2 values (P < 0.01). There was less change in body fat between 1982 and 1992, but lactate significantly decreased (P < 0.01), as did peak power and absolute and relative VO2 and VE. Although fitness levels in former elite oarsmen decreased each decade, these declines were somewhat arrested by regular aerobic training. Body fat increased and metabolic capacity decreased rapidly during the first decade, whereas anaerobic capacity decreased more significantly in the second decade. Anaerobic capacity diminished at a significantly greater rate than aerobic capacity, probably as a result of the aging process and emphasis on aerobic training in post-competitive years.

Ocular injuries related to air bag inflation.

BACKGROUND AND OBJECTIVES: The effectiveness of air bags as a safety device in decreasing fatalities and reducing morbidity in frontal impact motor vehicle accidents has been well established. However, case reports have surfaced documenting ocular injuries related to air bag inflation. The purpose of this study was to identify and analyze ocular injuries sustained during air bag inflation. PATIENTS AND METHODS: Hospital emergency room records were reviewed retrospectively for a period of 4 years for patients who had sustained ocular injuries in motor vehicle accidents. Those injuries associated with air bag inflation were analyzed as to the nature of the injury, type of vehicle, speed, and visual morbidity. RESULTS: Fourteen cases of ocular injury related to air bag inflation were identified. Hyphema (nine cases) and cornea abrasion (eight cases) were the most common. Three cases, all patients who had been wearing glasses, sustained serious and permanent ocular damage. CONCLUSION: While air bags are an unequivocal safety feature, ocular injuries do occur that are related to air bag inflation. Eyeglass wear may present an additional risk factor.

Ocular injuries associated with eyeglass wear and airbag inflation.

OBJECTIVE: The purpose of this report is to detail three cases of serious ocular injury associated with eyeglass wear during airbag inflation following motor vehicle crashes. DESIGN: Although there were too few cases to provide valid statistics, an attempt was made to determine if there were characteristics common to the glasses or accidents that would provide preventive information. MATERIALS AND METHODS: The emergency room records were reviewed for a 4-year period and cases were extracted involving ocular injuries related to airbag inflation. Fourteen cases were found, and three patients were wearing glasses. These cases were studied in detail. MEASUREMENTS AND MAIN RESULTS: All three patients sustained serious, vision-compromising ocular damage. CONCLUSIONS: Eyeglass wear presents an additional risk factor for ocular injury during airbag inflation. Frame damage and lens shatter provide potential for both blunt and penetrating types of ocular injury.

Noise removal from multiple MRI images.

We introduce a novel technique for magnetic resonance image (MRI) restoration, using a physical model (spin equation). We determine a set of three basis images (proton density and nuclear relaxation times) from the MRI data using a nonlinear optimization method, and use those images to obtain restorations of the original image. MRIs depend nonlinearly on proton density, two nuclear relaxation times, T1 and T2, and two control parameters, echo time (TE) and relaxation time (TR). We model images as Markov random fields and introduce a maximum a posteriori restoration method, based on nonlinear optimization, which reduces noise while preserving resolution.

Detection of de novo mutations and analysis of their origin in families with X linked hypohidrotic ectodermal dysplasia.

Hypohidrotic ectodermal dysplasia (EDA) has been localised to the q12-q13.1 region of the X chromosome by both physical and genetic mapping methods. Although linkage analysis using closely linked flanking markers can clarify the carrier status for many females at risk for the disorder, knowledge of the origin of the mutation in instances of possible de novo mutation is critical for accurate genetic counselling of families. Two methods have been used to confirm de novo mutation in families with EDA and to trace their origin. Direct detection of three de novo molecular deletions, one arising during oogenesis and the other two during spermatogenesis, was achieved by Southern analyses using cosmids isolated from the EDA region as probes. Seven de novo mutations arising during spermatogenesis, and two possible de novo mutations during oogenesis, were identified by an analysis of the cosegregation of the disorder with polymorphic markers closely linked to and flanking the EDA locus. The confirmation and analysis of the origin of the 10 de novo mutations greatly assisted genetic counselling in these families. The apparent 3.5:1 excess of male to female origin of mutation in families studied with unidentified types of mutation is similar to other studies of X linked disorders, and suggests that the majority of these mutations may involve single base pair substitutions.

Characterisation of molecular DNA rearrangements within the Xq12-q13.1 region, in three patients with X-linked hypohidrotic ectodermal dysplasia (EDA).

A panel of somatic cell hybrids and X-linked hypohidrotic ectodermal dysplasia (EDA) patient-derived cell lines, containing different rearranged X chromosomes, have been used to refine the physical map of the Xq12-q13.1 region. The patient-derived material included genomic DNA from an EDA male (EDA family 1015) with an interstitial deletion, and a cell line GM0705A, obtained from an isolated female patient with a de novo balanced (X;9) translocation, and the somatic hybrid, AnLy, derived from this cell line. This map subdivides the region into at least 6 mapping-intervals. DNA probes from DXS732 and DXS453, identified as the closest flanking marker loci to the EDA locus, were used to identify homologous Yeast Artificial Chromosome (YAC) clones. Two of the DXS732-specific YACs were shown by fluorescent in situ hybridisation (FISH) analysis to bridge the (X;9) translocation breakpoint. These two YACs were also screened against the ICRF human X chromosome cosmid library and identified 36 cosmid clones. Direct cosmid-cosmid hybridisation analysis placed subsets of these clones within four different cosmid contigs. Mapping of anchor clones from each contig, against the mapping panel, localised all these contigs within the Xq12-q13.1 region. One cosmid, ICRFc104C03.184, identified potential junctional-fragments in several restriction digests of AnLy hybrid DNA. This was confirmed by FISH analysis of the GM0705A cell line with total cosmid ICRFc104C03.184, in which both chromosomal elements of the (X;9) translocation were identified. A single-copy probe pC03.184E2, derived from this cosmid, also identified the der(9)-derived junctional fragment when hybridised against AnLy DNA.(ABSTRACT TRUNCATED AT 250 WORDS)