Padlock probes reveal single-nucleotide differences, parent of origin and in situ distribution of centromeric sequences in human chromosomes 13 and 21.

Chromosome centromeres, composed of repeated DNA sequences, orchestrate the correct segregation of chromatids in cell division. We have examined the centromeres of human chromosomes 13 and 21 by studying the distribution, in situ, of two alpha satellite sequences that differ in a single nucleotide position. This was possible using padlock probes, oligo-nucleotides that can be ligated into circles upon target recognition. The segregation of individual 13 and 21 homologues in a family was followed by monitoring of the signals from two differentially labelled probes, specific for either sequence variant. A characteristic arrangement of the repeat motifs in three separate spots, oriented transverse to the length axis of the metaphase chromosomes and bilaterally symmetric, indicates that only parts of the detected regions are involved in the centromeric region, joining the sister chromatids before anaphase.

Diamond-Blackfan anaemia: genetic homogeneity for a gene on chromosome 19q13 restricted to 1.8 Mb.

Diamond-Blackfan anaemia (DBA; MIM#205900) is a rare disorder manifested as a pure red-cell aplasia in the neonatal period or in infancy. The clinical hallmark of DBA is a selective decrease in erythroid precursors and anaemia. Other lineages are usually normal and the peripheral white blood cell count is normal. In approximately one-third of cases, the disease is associated with a wide variety of congenital anomalies and malformations. Most cases are sporadic, but 10-20% of them follow a recessive or a dominant inheritance pattern. A female with DBA and a chromosomal translocation involving chromosome 19q was recently identified. We undertook a linkage analysis with chromosome 19 markers in multiplex DBA families of Swedish, French, Dutch, Arabic and Italian origin. Significant linkage to chromosome 19q13 was established for dominant and recessive inherited DBA with a peak lod score at D19S197 (Zmax = 7.08, theta = 0.00). Within this region, a submicroscopic de novo deletion of 3.3 Mb was identified in a patient with DBA. The deletion coincides with the translocation break-point and, together with key recombinations, restricts the DBA gene to a 1.8-Mb region. The results suggest that, despite its clinical heterogeneity, DBA is genetically homogeneous for a gene in 19q13.

The gene encoding ribosomal protein S19 is mutated in Diamond-Blackfan anaemia.

Diamond-Blackfan anaemia (DBA) is a constitutional erythroblastopenia characterized by absent or decreased erythroid precursors. The disease, previously mapped to human chromosome 19q13, is frequently associated with a variety of malformations. To identify the gene involved in DBA, we cloned the chromosome 19q13 breakpoint in a patient with a reciprocal X;19 chromosome translocation. The breakpoint occurred in the gene encoding ribosomal protein S19. Furthermore, we identified mutations in RPS19 in 10 of 40 unrelated DBA patients, including nonsense, frameshift, splice site and missense mutations, as well as two intragenic deletions. These mutations are associated with clinical features that suggest a function for RPS19 in erythropoiesis and embryogenesis.

Occupational exposure to whole body vibrations and birth outcomes - A nationwide cohort study of Swedish women.

BACKGROUND: More women in reproductive ages are entering occupations where exposure to whole body vibrations (WBV) is common (e.g. in transportation and construction). Previous studies based on self-assessed exposure suggest increased risks of adverse birth outcomes, but it is unclear at what exposure levels and if the current exposure guidelines are appropriate during pregnancy. OBJECTIVES: To investigate whether occupational WBV-exposure increases the risk of preterm birth, low birth weight, and/or small-for-gestational age, in a large, nationwide, prospective, cohort study. DESIGN: The Fetal Air Pollution Exposure cohort (FAIR) was formed by merging data from multiple, national registers, and the present study includes singletons born 1994-2014 to working women in Sweden (n = 1,091,080 births). WBV-exposure was assessed quantitatively using a job-exposure matrix based on measurements, and calculated odds ratios were adjusted for potential confounders such as smoking and BMI, and other occupational exposures like noise, combustion particles, and physically and psychologically strenuous work. Data on absence from work (full-/part time, sick leave, parental leave, etc.) was also used. RESULTS: Exposure to WBV during pregnancy, among women with low absence from work (n = 476,419), was associated with an increased risk of preterm birth, below the occupational exposure limit (1.15 m/s2). Compared to unexposed mothers, the OR was 1.38 (95% CI: 1.05, 1.83) for exposure ≥0.5 m/s2, corresponding to an increase from 47/1000 cases to 65/1000 cases. No increased risk was found for small-for-gestational age. CONCLUSIONS: Exposure to WBV was associated with an increased risk of preterm birth. The results suggest that the current permissible exposure and action levels for WBV-exposure do not adequately protect pregnant women with continuous exposure.

Improved structural characterization of chromosomal breakpoints using high resolution custom array-CGH.

Array-CGH is a powerful tool for the rapid detection of genomic imbalances. By customizing the array it is possible to increase the resolution in a targeted genomic region of interest and determine the structure of the breakpoints with high accuracy, as well as to detect very small imbalances. We have used targeted custom arrays to zoom in on 38 chromosomal breakpoints from 12 different patients carrying both balanced and unbalanced rearrangements. We show that it is possible to characterize unbalanced breakpoints within 17-20,000 bp, depending on the structure of the genome. All of the deletion and duplication breakpoints were further refined and potential underlying molecular mechanisms of formation are discussed. In one of seven carriers of apparently balanced reciprocal translocations we detected a small deletion of 200 bp within the previously FISH-defined breakpoint, and in another patient, a large deletion of 11 Mb was identified on a chromosome not involved in the translocation. Targeted custom oligonucleotide arrays make it possible to perform fine mapping of breakpoints with a resolution within the breakpoint region much higher compared to commercially available array platforms. In addition, identification of small deletions or duplications in apparently balanced rearrangements may contribute to the identification of new disease causing genes.

Occupational exposure to UV light and mortality from multiple sclerosis.

BACKGROUND: The etiology of multiple sclerosis (MS) is largely unknown; low exposure to ultraviolet (UV) light has been a suggested risk factor. The aim of this study was to investigate whether occupational exposure to UV light reduces the risk of death from MS. METHODS: The cohort was based on all individuals in the Swedish census in 1980. All MS-related deaths were identified in the national registry of causes of death. A job-exposure matrix was developed to classify the occupational exposure to UV light. RESULTS: MS was recorded as a cause of the death for 839 individuals. The risk of MS-related death decreased with increasing occupational exposure to UV light. The relative risk adjusted for age, sex, and socioeconomic status was 0.48 (95% CI 0.28-0.80) in the high-exposure group and 0.88 (95% CI 0.73-1.06) in the intermediate-exposure group. CONCLUSIONS: Occupational exposure to UV light was associated with a reduced risk of MS. Our findings are corroborated by previous observations that UV light has a preventive role in the development of MS, although the possibility of reversed causality cannot be completely ruled out.

Exposure to Particles and Nitrogen Dioxide Among Workers in the Stockholm Underground Train System.

OBJECTIVES: Exposure to fine particles in urban air has been associated with a number of negative health effects. High levels of fine particles have been detected at underground stations in big cities. We investigated the exposure conditions in four occupational groups in the Stockholm underground train system to identify high-exposed groups and study variations in exposure. METHODS: PM1 and PM2.5 were measured during three full work shifts on 44 underground workers. Fluctuations in exposure were monitored by a real-time particle monitoring instrument, pDR, DataRAM. Qualitative analysis of particle content was performed using inductively coupled plasma mass spectrometry. Nitrogen dioxide was measured using passive monitors. RESULTS: For all underground workers, the geometric mean (GM) of PM1 was 18 µg/m3 and of PM2.5 was 37 µg/m3. The particle exposure was highest for cleaners/platform workers, and the GM of PM1 was 31.6 µg/m3 [geometric standard deviation (GSD), 1.6] and of PM2.5 was 76.5 µg/m3 (GSD, 1.3); the particle exposure was lowest for ticket sellers, and the GM of PM1  was 4.9 µg/m3 (GSD, 2.1) and of PM2.5 was 9.3 µg/m3 (GSD, 1.5). The PM1 and PM2.5 levels were five times higher in the underground system than at the street level, and the particles in the underground had high iron content. The train driver's nitrogen dioxide exposure level was 64.1 µg/m3 (GSD, 1.5). CONCLUSIONS: Cleaners and other platform workers were statistically significantly more exposed to particles than train drivers or ticket sellers. Particle concentrations (PM2.5) in the Stockholm underground system were within the same range as in the New York underground system but were much lower than in several older underground systems around the world.

Blood markers of inflammation and coagulation and exposure to airborne particles in employees in the Stockholm underground.

OBJECTIVES: Although associations have been found between levels of ambient airborne particles and cardiovascular disease (CVD) in the general population, little is known about possible cardiovascular effects from high exposure to particles in underground railway systems. This study investigates risk markers for CVD in employees exposed to particles in the Stockholm underground system. METHODS: 79 workers (54 men and 25 women) in the Stockholm underground were investigated between November 2004 and March 2005. All were non-smokers aged 25-50 years. Three exposure groups were delineated: 29 platform workers with high exposure to particles, 29 train drivers with medium exposure and 21 ticket sellers with low exposure (control group). A baseline blood sample was taken after 2 non-working days, and a second sample after 2 working days, for analysis of levels of plasminogen activator inhibitor-1 (PAI-1), high-sensitivity C-reactive protein (hs-CRP), interleukin-6, fibrinogen, von Willebrand factor and factor VII. The study investigated changes in plasma concentrations between sample 1 and sample 2, and differences in average concentrations between the groups. RESULTS: No changes between sample 1 and 2 were found that could be attributed to particle exposure. However, the highly exposed platform workers were found to have higher plasma concentrations of PAI-1 and hs-CRP than the ticket sellers and train drivers. This suggests that particle exposure could have a long-term inflammatory effect. These differences remained for PAI-1 in the comparison between platform workers and ticket sellers after adjusting for body mass index. CONCLUSIONS: Employees who were highly exposed to airborne particles in the Stockholm underground tended to have elevated levels of risk markers for CVD relative to employees with low exposure. However, the differences observed cannot definitely be linked to particle exposure as such.

Assessing informal caregivers' experiences: a qualitative and psychometric evaluation of the Caregiver Reaction Assessment Scale.

With the aim to evaluate the Swedish version of the Caregiver Reaction Assessment Scale (CRA), informal caregivers (n = 209) to individuals with a malignant disease, dementia or a physical impairment were recruited. The CRA was developed in the USA and is a self-rating questionnaire consisting of five subscales, measuring family members' reactions to the experience of caring for a relative with mental or physical illnesses. Data were analysed using psychometric and qualitative methods. Findings indicated good internal consistency, and a factor analysis confirmed the structure with five subscales; however, an overlap of items between the subscales was found. A content analysis of respondents' comments indicated that there were problems due to presuppositions inherent in the questions and with the meaning of words. Problems regarding inclusion/exclusion aspects in some concepts were also found. Two aspects of caregiver reactions were found to be missing or only partly covered: worrying and positive experiences. Results from this study suggest that the Swedish version is useful for assessment of caregiver reactions but needs further refinement.

Arc and resistance welding and tumours of the endocrine glands: a Swedish case-control study with focus on extremely low frequency magnetic fields.

BACKGROUND: Mechanisms for potential effects of extremely low frequency (ELF) magnetic fields on carcinogenesis have not been identified. A potential pathway could be an interaction with the endocrine system. AIMS: To analyse occupational exposure to ELF magnetic fields from welding, and tumours of the endocrine glands. METHODS: This case-control study was based on a cohort with an increased prevalence of high exposed individuals. A total of 174 incident cases of tumours of the endocrine glands, 1985-94, were identified and data were obtained from 140 (80%) of these cases; 1692 controls frequency matched on sex and age were selected, and information on 1306 (77%) individuals was obtained. A short questionnaire was sent to a work administrator at the workplaces of the cases and controls. The exposure assessment was based on questions about job tasks, exposure to different types of welding, and exposure to solvents. RESULTS: There was an overall increased risk for all tumours of the endocrine glands for individuals who had been welding sometime during the follow up. The increased risk was attributable to arc welding; for resistance welding there was no clear evidence of an association. We found an increased risk for the adrenal glands in relation to arc welding, and for the parathyroid glands in relation to both arc welding and resistance welding. An imprecise increase in risk was also noted for tumours of the pituitary gland for arc welding. No confounding effect was found for solvent exposure, and there was no sign of biological interaction. CONCLUSION: The increased risks of endocrine gland tumours related to welding might be explained by exposure to high levels of ELF magnetic fields.

Familial hypercalcemia and hypercalciuria caused by a novel mutation in the cytoplasmic tail of the calcium receptor.

Familial hyperparathyroidism (HPT), characterized by hypercalcemia and hypercalciuria, and familial benign hypocalciuric hypercalcemia (FHH) are the most common causes of hereditary hypercalcemia. The calcium-sensing receptor (CaR) regulates PTH secretion and renal calcium excretion. Heterozygous inactivating mutations of the gene cause FHH, whereas CaR gene mutations have not been demonstrated in HPT. In a kindred with 20 affected individuals, the hypercalcemic disorder segregated with inappropriately higher serum PTH and magnesium levels and urinary calcium levels than in unaffected members. Subtotal parathyroidectomy revealed parathyroid gland hyperplasia/adenoma and corrected the biochemical signs of the disorder in seven of nine individuals. Linkage analysis mapped the condition to markers flanking the CaR gene on chromosome 3q. Sequence analysis revealed a mutation changing phenylalanine to leucine at codon 881 of the CaR gene, representing the first identified point mutation located within the cytoplasmic tail of the CaR. A construct of the mutant receptor (F881L) was expressed in human embryonic kidney cells (HEK 293), and demonstrated a right-shifted dose-response relationship between the extracellular and intracellular calcium concentrations. The hypercalcemic disorder of the present family is caused by an inactivating point mutation in the cytoplasmic tail of the CaR and displays clinical characteristics atypical of FHH and primary HPT.

Memory effects of clomipramine treatment: relationship to CSF monoamine metabolites and drug concentrations in plasma.

Performance on tasks tapping automatic and voluntary aspects of memory, attention, and motor speed was examined in 14 patients with major depressive disorder, before and after 3 weeks of treatment with clomipramine (150 mg/day), a potent serotonin and noradrenaline uptake blocker with anticholinergic side effects. Performance on tasks requiring frontal functions improved or did not change, whereas verbal learning and retention, where hippocampal functioning is critical, were impaired. The latter tasks were negatively related to cerebrospinal fluid (CSF) 5-HIAA levels and plasma concentration of clomipramine. The results provide further support for the regulatory role of monoaminergic systems in cognition. Furthermore, we found the automatic-voluntary capacity distinction less heuristically useful. Physiological mechanisms regulating different aspects of cognition and memory appeared to be more closely related to the type of task used than to its capacity-demanding properties.

Deletion including the oligophrenin-1 gene associated with enlarged cerebral ventricles, cerebellar hypoplasia, seizures and ataxia.

Non-specific X-linked mental retardation is a heterogeneous group of disorders with an incidence of approximately 1 in 500 males. A recently identified gene in Xq12, encoding a Rho-GTPase-activating protein, was found to be mutated in individuals with mental retardation. We describe here two sisters with a 46,XY karyotype and a microdeletion of the oligophrenin-1 gene and 1.1 Mb of flanking DNA. We have characterised the molecular interval defining this microdeletion syndrome with the fibre-FISH technique. A visual physical map of 1.2 Mb was constructed which spans the oligophrenin-1 gene and the androgen receptor gene. The analysis of the patients revealed a deletion which extended from the 5' end of the AR gene to a region approximately 80 kb proximal to the EPLG2 gene. The clinical manifestations of the two sisters include psychomotor retardation, seizures, ataxia, hypotonia and complete androgen insensitivity. Cranial MRI scans show enlargement of the cerebral ventricles and cerebellar hypoplasia. Our findings give further support for the involvement of the oligophrenin-1 gene in specific morphological abnormalities of the brain which is of importance in the investigation of male patients presenting with mental retardation. In combination with our results from physical mapping we suggest that a region around the oligophrenin-1 locus is relatively bereft of vital genes.

Incidence of testicular cancer and occupation among Swedish men gainfully employed in 1970.

PURPOSE: To estimate occupation-specific risk of seminomas and nonseminoma subtypes of testicular cancer among Swedish men gainfully employed in 1970 over the period 1971-1989. METHODS: Age-period standardized incidence ratios were computed in a dataset linking cancer diagnoses from the Swedish national cancer register to occupational and demographical data obtained in the census in 1970. Log-linear Poisson models were fitted, allowing for geographical area and town size. Taking occupational sector as a proxy for socioeconomic status, occupational risks were recalculated using intra-sector analyses, where the reference group comprised other occupations in the same sector only. Risk estimators per occupation were also computed for men reporting the same occupation in 1960 and 1970, a more specifically exposed group. RESULTS: Seminomas and nonseminomas showed a substantial geographical variation. The association between germ-cell testicular tumors and high socioeconomic groups was found mainly for nonseminomas. Positive associations with particular occupations were more evident for seminomas, for which railway stationmasters, metal annealers and temperers, precision toolmakers, watchmakers, construction smiths, and typographers and lithographers exhibited a risk excess. Concrete and construction worker was the only occupation consistently associated with nonseminomas. CONCLUSIONS: Among the many occupations studied, our results corroborate the previously reported increased risk among metal workers, specifically related with seminomatous tumors in this study. Our results confirm the geographical and socioeconomical differences in the incidence of testicular tumors. These factors should be accounted for in occupational studies. The different pattern of occupations related with seminomas and nonseminomas support the need to study these tumors separately.

Using geographic information systems to assess individual historical exposure to air pollution from traffic and house heating in Stockholm.

A specific aim of a population-based case-control study of lung cancer in Stockholm, Sweden, was to use emission data, dispersion models, and geographic information systems (GIS) to assess historical exposure to several components of ambient air pollution. Data collected for 1,042 lung cancer cases and 2,364 population controls included information on residence from 1955 to the end of follow-up for each individual, 1990-1995. We assessed ambient air concentrations of pollutants from road traffic and heating throughout the study area for three points in time (1960, 1970, and 1980) using reconstructed emission data for the index pollutants nitrogen oxides (NO(x)/NO(2)) and sulfur dioxide together with dispersion modeling. NO(2) estimates for 1980 compared well with actual measurements, but no independently measured (study-external) data were available for SO(2), precluding similar validation. Subsequently, we used linear intra- and extrapolation to obtain estimates for all other years 1955-1990. Eleven thousand individual addresses were transformed into geographic coordinates through automatic and manual procedures, with an estimated error of < 100 m for 90% of the addresses. Finally, we linked annual air pollution estimates to annual residence coordinates, yielding long-term residential exposure indices for each individual. There was a wide range of individual long-term average exposure, with an 11-fold interindividual difference in NO(2) and an 18-fold difference in SO(2). The 30-year average for all study subjects was 20 microg/m(3) NO(2) from traffic and 53 microg/m(3) SO(2) from heating. The results indicate that GIS can be useful for exposure assessment in environmental epidemiology studies, provided that detailed geographically related exposure data are available for relevant time periods.

Monosomy 18q syndrome and atypical Rett syndrome in a girl with an interstitial deletion (18)(q21.1q22.3).

We describe a 6 1/2-year-old girl with an interstitial deletion of chromosome arm 18q (18q21.1q22.3). Her clinical manifestations are a combination of those found in monosomy 18q syndrome and those of Rett syndrome. Cytogenetic analysis demonstrated a deletion of the long arm of chromosome 18, defined by molecular analysis with polymorphic markers as a de novo interstitial deletion, paternally derived. The findings typical of the 18q- syndrome included mental retardation, midface hypoplasia, and hypoplasia of labia majora, and those typical of Rett syndrome were severe mental retardation, autistic behavior, inappropriate hand-washing movements, epilepsy, attacks of sighing and hyperventilation, and progressive scoliosis since the age of 5 years. She did not have microcephaly, and the mental delay was obvious from an early age without a period of normal development, which makes the diagnosis of Rett syndrome atypical. Previously, a girl with mosaicism for a monosomy 18q associated with Rett syndrome has been described. That girl had a terminal deletion of chromosome 18q, which seems to coincide in part with that in the present girl. It is possible that genes in the distal region of 18q are involved in the etiology of Rett syndrome.

Screening for CGG trinucleotide repeat expansion in the fragile X mental retardation 1 gene in schizophrenic patients.

Patients diagnosed using DSM-III-R criteria as having schizophrenia and other related disorders (n = 128) were assessed for CGG trinucleotide repeat expansion in the fragile X mental retardation 1 (FMR-1) gene. One subject, a woman with schizophreniform disorder, was found to have a premutation of the gene. Her case report is given. The present investigation supports the view that mutation or premutation of the FMR-1 gene is not of importance for the aetiology of the vast majority of schizophrenic patients.

Field potentials evoked in rat primary somatosensory cortex (SI) by impulses in cutaneous A beta- and C-fibres.

A projection of cutaneous C-afferent fibres to the contralateral primary somatosensory cortex (SI) was examined in the halothane-anaesthetized rat. Field potentials evoked by electrical stimulation of the right sural nerve were recorded at the surface of an intracortically within the left SI cortex. A late surface positive potential (latency 110-190 ms, mean 136 ms) was evoked by sural stimulation at a strength that activated A- and C-fibres. A selective anodal block of impulse conduction in A-fibres proximal to the stimulating electrodes showed that impulses in C-fibres generate the late potential also in the absence of a preceding A-fibre input. Stimulation of the sural nerve at two sites caused a shift in latency of the late potential corresponding to a conduction velocity of the primary afferent fibres of less than 1.0 m/s. The distribution of the C-fibre-evoked potential in SI was similar to that of the A beta-fibre evoked 'primary' potential suggesting that the investigated projection of cutaneous C-fibres to SI has a somatotopic organization.

Clinical characteristics and biological parameters in temperamental clusters of suicide attempters.

A sample of 215 suicide attempters was categorized in a cluster analysis into four groups according to temperamental trails. Monoamine metabolites in the cerebrospinal fluid were analysed (n = 106). Dexamethasone suppression tests (DST) were performed (n = 154) and the activity of the enzyme monoamine oxidase in platelets (pl-MAO) was assessed (n = 103). Patients belonging to the two clusters with the most deviant temperament profiles (nos 2 and 3) were young and scored high on the Beck Hopelessness Scale and the Suicide Assessment Scale. "Cluster 3" ("neurotic, impulsive, aggressive") patients often had dysthymia and axis II, cluster B diagnoses (e.g. borderline or histrionic personality). "Cluster 2" ("neurotic and introverted") patients often had major depression. The "Cluster 1", with on the whole a normal temperament profile, had significantly higher levels of post-DST cortisol than the other clusters. The "Cluster 4" had a normal temperament profile. Adjustment disorders were most common in "Cluster 1" and "Cluster 4". The monoamine metabolite levels did not differ between the clusters, and the differences in pl-MAO activity disappeared after adjusting for age and gender. The results suggest that temperament profiles in suicide attempters are related to psychiatric diagnoses, suicidality, hopelessness, and post-DST cortisol, but are not predictive of completed suicide.

Continuous superporous agarose beds in radial flow columns.

Continuous superporous agarose beds constitute a new support material for chromatography, biocatalysis and electrophoresis. The bed consists of a single piece of agarose gel, homogeneously transected by flow-carrying pores, which easily can be varied in the range of 10-100 microm. In this work, large diameter beds (60 mm) were prepared and used in specially designed radial flow columns. The basic chromatographic properties of the beds were investigated by size-exclusion chromatography experiments. In an affinity chromatography application one bed was derivatized with Cibacron Blue 3GA and used for the purification of lactate dehydrogenase from a crude bovine heart extract. In a biotransformation application one bed was provided with immobilized beta-galactosidase and used in the production of lactose-free milk.