Translation and Psychometric Testing of the Hägerbäumer Presenteeism Scale in English.

PURPOSE: Interest in presenteeism has increased in research. Presenteeism is a behaviour of going to work despite illness. It has been predominantly measured using single items, which introduce limitations to validity. To overcome these limitations, Hägerbäumer developed a German multi-item presenteeism scale. METHODS: The aim of the study was to provide an English translation and psychometric testing of the scale. This was conducted in two phases with native English-speaking employed adults. Phase 1 includes translation and cognitive debriefing, phase 2 testing construct validity and internal consistency reliability. RESULTS: Cognitive debriefing with 10 employees revealed no problems with understanding or answering the translated items. In total, 487 employed adults participated in the study, of which data from 287 were included in the analysis. For structural validity, the goodness-of-fit indicators all reached their thresholds (TLI = 0.98, CFI = 0.99, RMSEA = 0.07, SRMR = 0.02). The scale does not show differences between sexes and age groups but between sectors (F6,70.95 = 5.53, p < 0.001). The internal consistency reliability was satisfactory with α = 0.89 (CI 95%, 0.87-0.91). CONCLUSION: The translated multidimensional scale for measuring presenteeism at the behavioural level demonstrated good psychometric properties in an initial validation. Further psychometric testing is required before using this scale in cross-national comparison in research and international companies.

Risks and Recommendations in Prenatally Detected De Novo Balanced Chromosomal Rearrangements from Assessment of Long-Term Outcomes.

The 6%-9% risk of an untoward outcome previously established by Warburton for prenatally detected de novo balanced chromosomal rearrangements (BCRs) does not account for long-term morbidity. We performed long-term follow-up (mean 17 years) of a registry-based nationwide cohort of 41 individuals carrying a prenatally detected de novo BCR with normal first trimester screening/ultrasound scan. We observed a significantly higher frequency of neurodevelopmental and/or neuropsychiatric disorders than in a matched control group (19.5% versus 8.3%, p = 0.04), which was increased to 26.8% upon clinical follow-up. Chromosomal microarray of 32 carriers revealed no pathogenic imbalances, illustrating a low prognostic value when fetal ultrasound scan is normal. In contrast, mate-pair sequencing revealed disrupted genes (ARID1B, NPAS3, CELF4), regulatory domains of known developmental genes (ZEB2, HOXC), and complex BCRs associated with adverse outcomes. Seven unmappable autosomal-autosomal BCRs with breakpoints involving pericentromeric/heterochromatic regions may represent a low-risk group. We performed independent phenotype-aware and blinded interpretation, which accurately predicted benign outcomes (specificity = 100%) but demonstrated relatively low sensitivity for prediction of the clinical outcome in affected carriers (sensitivity = 45%-55%). This sensitivity emphasizes the challenges associated with prenatal risk prediction for long-term morbidity in the absence of phenotypic data given the still immature annotation of the morbidity genome and poorly understood long-range regulatory mechanisms. In conclusion, we upwardly revise the previous estimates of Warburton to a morbidity risk of 27% and recommend sequencing of the chromosomal breakpoints as the first-tier diagnostic test in pregnancies with a de novo BCR.

Erwinia amylovora pyrC mutant causes fire blight despite pyrimidine auxotrophy.

Erwinia amylovora bacteria cause fire blight disease, which affects apple and pear production worldwide. The Erw. amylovora pyrC gene encodes a predicted dihydroorotase enzyme involved in pyrimidine biosynthesis. Here, we discovered that the Erw. amylovora pyrC244::Tn5 mutant was a uracil auxotroph. Unexpectedly, the Erw. amylovora pyrC244::Tn5 mutant grew as well as the wild-type in detached immature apple and pear fruits. Fire blight symptoms caused by the pyrC244::Tn5 mutant in immature apple and pear fruits were attenuated compared to those caused by the wild-type. The pyrC244::Tn5 mutant also caused severe fire blight symptoms in apple tree shoots. A plasmid-borne copy of the wild-type pyrC gene restored prototrophy and symptom induction in apple and pear fruit to the pyrC244::Tn5 mutant. These results suggest that Erw. amylovora can obtain sufficient pyrimidine from the host to support bacterial growth and fire blight disease development, although de novo pyrimidine synthesis by Erw. amylovora is required for full symptom development in fruits. Significance and impact of the study: This study provides information about the fire blight host-pathogen interaction. Although the Erwinia amylovora pyrC mutant was strictly auxotrophic for pyrimidine, it grew as well as the wild-type in immature pear and apple fruits and caused severe fire blight disease in apple trees. This suggests that Erw. amylovora can obtain sufficient pyrimidines from host tissue to support growth and fire blight disease development. This situation contrasts with findings in some human bacterial pathogens, which require de novo pyrimidine synthesis for growth in host blood, for example.

First Report of Penicillium crustosum Causing Blue Mold on Stored Apple Fruit in Serbia.

Penicillium crustosum Thom (1930) causes blue mold on pome fruits and is also regularly found on cheese, nuts, and soil (1,3). The fungus produces a wide range of mycotoxins such as penitrem A, roquefortine C, terrestric acid, and cyclopenol, which impact human health (1). In January and February 2013, 20 decayed apples, 'Golden Delicious' and 'Jonagold' (Malus × domestica Borkh.) with blue mold symptoms were collected from cold storages in Svilajnac and Bela Crkva, Serbia. Decayed areas were light to medium brown with blue green sporulation on the surface of the lesion. Decayed tissue was soft and watery with a sharp margin between the diseased and healthy areas. One isolate from each cultivar was designated JP2 ('Golden Delicious') and JBC7 ('Jonagold') and further characterized. Conidiophores of both isolates were terverticillate, stipes were septate with rough walls, and phialides were ampulliform. Conidia were smooth, borne in columns, and were spherical to subglobose. Conidial dimensions for JP2 were 3.2 to 4.56 (3.73) × 2.64 to 4.3 (3.32) µm and for JBC7 were 3.1 to 4.46 (3.65) × 2.81 to 4.27 (3.31) µm (n = 50). The isolates were cultured on Czapek yeast autolysate agar (CYA), malt extract agar (MEA), and yeast extract sucrose agar (YES) media and incubated at 25°C for 7 days. Mycelia were white with heavy sporulation yielding grayish green colonies on all media. Colonies were radially sulcate and velutinous, with clear exudate, and produced a yellow to orange reverse on CYA and YES. On MEA, colonies were plane, low, and mycelia subsurface with conidia having a dry powdery appearance. Crusts of conidial masses formed after 10 or more days. No growth was observed on CYA when these isolates were incubated at 37°C. Both isolates were identified as P. crustosum Thom using morphological characters according to (2) and (1). Species level identification was confirmed by isolating genomic DNA followed by amplification of the ß-tubulin locus using gene specific primers via conventional PCR (4). MegaBLAST analysis of the 2X consensus nucleotide sequences revealed that JP2 and JBC7 (GenBank KJ433984 and 85) were 99% identical to P. crustosum culture collection isolate IBT 21518 (JN112030.1). Koch's postulates were examined using two apple cvs. Idared and Kolacara. Ten fruit per cultivar per isolate were inoculated on two sides of each fruit; 20 fruit were used as water-only inoculated controls. Fruit were washed with soap and water, surface sanitized with 70% ethanol, and placed into polyethylene boxes. Using a finishing nail, 4-mm wounds were created and inoculated with 50 µl of a 3 × 105/ml conidial suspension or Tween-treated sterile distilled water. Boxes with inoculated and control fruit were stored at 25°C for 10 days. The inoculated fruit developed small, soft, watery lesions, which enlarged into decayed areas with defined edges and abundant sporulation on the surface. Symptoms were identical to the original ones, while the control fruit remained symptomless. The fungus was re-isolated from infected tissue and showed the same morphological characteristics as the original isolates, thus completing Koch's postulates. Blue mold occurs during long term storage of apples and is predominantly caused by P. expansum. This is the first report of P. crustosum causing postharvest blue mold decay on apple fruit obtained from storage in Serbia and indicates that P. crustosum is an emerging pathogen for the Serbian pome fruit growing and packing industry. References: (1) J. C. Frisvad and R. A. Samson. Stud. Mycol. 49:1, 2004. (2) J. I. Pitt and A. D. Hocking. Fungi and Food Spoilage, 239. Springer, 2009. (3) P. G. Sanderson and R. A. Spotts. Phytopathology 85:103. 1995. (4) P. L. Sholberg et al. Postharvest Biol. Technol. 36:41, 2005.

First Report of Botryosphaeria dothidea Causing White Rot on Apple Fruit in Maryland.

Botryosphaeria dothidea (Moug.:Fr.) Ces. De Not. causes perennial cankers on apple trees and causes white rot on apple fruit in the field and during storage (1). Prolonged periods of warm wet weather favor rapid disease outbreaks that result in severe losses, which range from 25 to 50% for the southeastern United States (3). A B. dothidea isolate was obtained from decayed 'Fuji' apple fruit exhibiting white rot symptoms from a local farm market in Beltsville, MD, in May 2010. The fruit had characteristic large dark brown lesions with irregular margins and decay expanded unevenly toward the core and the tissue was soft. The pathogen was isolated from symptomatic tissue by spraying the lesion surface with 70% ethanol. The skin with aseptically removed with a scalpel and small pieces of tissue were placed on potato dextrose agar (PDA) and incubated at 20°C. Once fungal growth was evident, the cultures were hyphal-tip transferred to individual PDA plates and incubated at 20°C. The B. dothidea isolate produced black aerial mycelium with a white margin on PDA and had a black reverse. Conidiomata were evident after 10 to 14 days at 20°C only on oatmeal agar. Conidia were hyaline, smooth and straight, fusiform with an subobtuse apex and a truncate base 20 to 26 (24.33) × 4 to 7 (5) µm (n = 50). Genomic DNA was isolated from the fungus and amplified with gene specific primers (ITS 4 and 5) for the ribosomal DNA internal transcribed spacer region ITSI-5.8S-ITS2 as described by White et al. (4). Both forward and reverse strands of the 542-bp amplicon were sequenced and assembled into a contig. The nucleotide sequence (GenBank Accession No. KC473852) indicated 99% identity to B. dothidea isolate CMM3938 (JX513645.1) and to voucher specimens CMW 25686, 25696, and 25222 (FM955381.1, FM955379.1, and FM955377,1). Koch's postulates were conducted using three 'Golden Delicious' apple fruit that were wound-inoculated with 50 µl of a mycelial suspension of the fungus, obtained from aseptically scraping a 7-day-old PDA culture, and was also repeated using 'Fuji' apple fruit. Large, brown, slightly sunken, soft lesions with undefined edges developed 5 days after inoculation at 20°C and water-only inoculated fruit were symptomless. The fungus was reisolated from infected tissue and was morphologically identical to the original isolate from decayed apple fruit. To determine if the B. dothidea isolate was resistant to postharvest fungicides, the minimum inhibitory concentration (MIC) was conducted using the 96 well plate method with a mycelial suspension of the fungus as described by Pianzzola et al. (2). The MIC for the isolate was >1 ppm for Mertect and Scholar and 50 ppm for Penbotec, which are well below the labeled rates for these postharvest fungicides and the experiment was repeated. To our knowledge, this is the first report of B. dothidea causing white rot on apple fruit in Maryland. References: (1) A. R. Biggs and S. S. Miller. HortScience 38:400, 2003. (2) M. J. Pianzzola et al. Plant Dis. 88:23, 2004. (3) T. B. Sutton. White rot and black rot. Pages 16-20 in: Compendium of Apple and Pear Diseases, A. L. Jones and H. S. Aldwinckle, eds. The American Phytopathological Society, St Paul, MN, 1991. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Application. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

First Report of Neofusicoccum ribis Causing Postharvest Decay of Apple Fruit from Cold Storage in Pennsylvania.

Neofusicoccum ribis (Slippers, Crous & M.J. Wingf.), previously known as Botryosphaeria ribis (Grossenb. & Duggar), is an aggressive fungal plant pathogen that is part of the N. ribis/N. parvum species complex that causes stem cankers on a variety of woody plant species (2). An isolate of N. ribis was obtained from decayed 'Honeycrisp' apple fruit from a commercial cold storage facility located in Pennsylvania in October of 2011. The decayed apple fruit sample had a brownish lesion that was soft, dry, and leathery on the surface while sporulation was not evident. To conduct Koch's postulates, three 'Golden Delicious' apple fruits were wound-inoculated with a 50-µl mycelial suspension, obtained from aseptically scraping a 7-day-old potato dextrose agar (PDA) culture of the fungus, and was repeated using 'Fuji' apple fruit. The inoculated fruit developed lesions, while water-inoculated fruit were symptomless after 5 days at 20°C. N. ribis was reisolated from infected tissue and was morphologically identical to the original isolate. Genomic DNA was isolated, a portion of the ß-tubulin gene was amplified with the gene specific primers, and the amplicon was sequenced and analyzed using BLAST (1). The nucleotide sequence (GenBank Accession No. KC47853) had 99% identity with N. ribis SEGA8 isolate (JN607146.1). The N. ribis isolate produced a grayish-white mycelium with abundant aerial hyphae on PDA and had an olive-colored reverse. Microscopic investigation revealed septate mycelia with right angle branching and conidiomata were not evident on PDA, V8, oatmeal agar (OMA), or water agar (WA). Growth on WA was sparse and transparent, and aerial mycelial growth was not produced. Growth rate analyses were conducted on PDA, V8, and OMA and were 10.1 (±1.39), 20.4 (±1.15), and 17.6 (±0.70) mm/day at 20°C and the experiment was repeated. The minimum inhibitory concentrations (MIC) for the N. ribis isolate was carried out for three postharvest fungicides as described by Pianzzola et al. (3). Briefly, 96 well plates were filled with PDA alone (0 ppm) and PDA amended with 10 fungicide concentrations ranging from 1 to 1,200 ppm for thiabendazole (Mertect), and 1 to 1,000 ppm for fludioxonil (Scholar) and pyrimethanil (Penbotec). A mycelial suspension of the fungus was obtained from pure culture, 50 µl of the mycelial suspension was pipetted into each well, and allowed to grow for 72 h at 25°C. The experiment was conducted twice. The N. ribis isolate displayed MIC values of >1 ppm thiabendazole (Mertect), >1 ppm fludioxonil (Scholar), and 50 ppm pyrimethanil (Penbotec), which are all well below the labeled application rates for these postharvest fungicides. To our knowledge, this is the first report of N. ribis causing postharvest decay on apple fruit obtained from a commercial storage facility in Pennsylvania. References: (1) S. F. Altschul et al. J. Mol. Biol. 215:403, 1990. (2) D. Pavlic et al. Mycologia 101:636, 2009. (3) M. J. Pianzzola et al. Plant Dis. 88:23, 2004.

First Report of Penicillium carneum Causing Blue Mold on Stored Apples in Pennsylvania.

Blue mold decay occurs during long term storage of apples and is predominantly caused by Penicillium expansum Link. Apples harvested in 2010 were stored in a controlled atmosphere at a commercial Pennsylvania apple packing and storage facility, and were examined for occurrence of decay in May 2011. Several decayed apples from different cultivars, exhibiting blue mold symptoms with a sporulating fungus were collected. One isolate recovered from a decayed 'Golden Delicious' apple fruit was identified as P. carneum Frisvad. Genomic DNA was isolated, 800 bp of the 3' end of the ß-tubulin locus was amplified using gene specific primers and sequenced (4). The recovered nucleotide sequence (GenBank Accession No. JX127312) indicated 99% sequence identity with P. carneum strain IBT 3472 (GenBank Accession No. JF302650) (3). The P. carneum colonies strongly sporulated and had a blue green color on potato dextrose agar (PDA), Czapek yeast autolysate agar (CYA), malt extract agar (MEA), and yeast extract sucrose agar (YES) media at 25°C after 7 days. The colonies also had a beige color on plate reverse on CYA and YES media. The species tested positive for the production of alkaloids, as indicated by a violet reaction for the Ehrlich test, and grew on CYA at 30°C and on Czapek with 1,000 ppm propionic acid agar at 25°C; all of which are diagnostic characters of this species (2). The conidiophores were hyaline and tetraverticillate with a finely rough stipe. Conida were produced in long columns, blue green, globose, and averaged 2.9 µm in diameter. To prove pathogenicity, Koch's postulates were conducted using 20 'Golden Delicious' apple fruits. Fruits were washed, surface sterilized with 70% ethanol, and placed onto fruit trays. Using a nail, 3-mm wounds were created and inoculated with 50 µl of a 106/ml conidial suspension or water only as a negative control. The fruit trays were placed into boxes and were stored in the laboratory at 20°C for 7 days. The inoculated fruit developed soft watery lesions, with hard defined edges 37 ± 4 mm in diameter. The sporulating fungus was reisolated from infected tissue of all conidia inoculated apples and confirmed to be P. carneum by polymerase chain reaction (PCR) using the ß-tubulin locus as described. Water inoculated control apples were symptomless. Originally grouped with P. roqueforti, P. carneum was reclassified in 1996 as a separate species (1). P. carneum is typically associated with meat products, beverages, and bread spoilage and produces patulin, which is not produced by P. roqueforti (1,2). Our isolate of P. carneum was susceptible to the thiabendazole (TBZ) fungicide at 250 ppm, which is below the recommended labeled application rate of 600 ppm. The susceptibility to TBZ suggests that this P. carneum isolate has been recently introduced because resistance to TBZ has evolved rapidly in P. expansum (4). To the best of our knowledge, P. carneum has not previously been described on apple, and this is the first report of P. carneum causing postharvest decay on apple fruits obtained from storage in Pennsylvania. References: (1) M. Boyson et al. Microbiology 142:541, 1996. (2) J. C. Frisvad and R. A. Samson. Stud. Mycol. 49:1, 2004. (3) B. G. Hansen et al. BMC Microbiology 11:202, 2011. (4) P. L. Sholberg et al. Postharvest Biol. Technol. 36:41, 2005.

Recurrence of congenital heart defects in families.

BACKGROUND: Knowledge of the familial contribution to congenital heart diseases (CHD) on an individual and population level is sparse. We estimated an individual's risk of CHD given a family history of CHD, as well as the contribution of CHD family history to the total number of CHD cases in the population. METHODS AND RESULTS: In a national cohort study, we linked all Danish residents to the National Patient Register, the Causes of Death Register, the Danish Central Cytogenetic Register, and the Danish Family Relations Database, yielding 1 763 591 persons born in Denmark between 1977 and 2005, of whom 18 708 had CHD. Individuals with CHD were classified by phenotype. We estimated recurrence risk ratios and population-attributable risk. Among first-degree relatives, the recurrence risk ratio was 79.1 (95% confidence interval [CI] 32.9 to 190) for heterotaxia, 11.7 (95% CI, 8.0 to 17.0) for conotruncal defects, 24.3 (95% CI,12.2 to 48.7) for atrioventricular septal defect, 12.9 (95% CI, 7.48 to 22.2) for left ventricular outflow tract obstruction, 48.6 (95% CI, 27.5 to 85.6) for right ventricular outflow tract obstruction, 7.1 (95% CI, 4.5 to 11.1) for isolated atrial septal defect, and 3.4 (95% CI, 2.2 to 5.3) for isolated ventricular septal defect. The overall recurrence risk ratio for the same defect was 8.15 (95% CI, 6.95 to 9.55), whereas it was 2.68 (95% CI, 2.43 to 2.97) for different heart defects. Only 2.2% of heart defect cases in the population (4.2% after the exclusion of chromosomal aberrations) were attributed to CHD family history in first-degree relatives. CONCLUSIONS: Specific CHDs showed highly variable but strong familial clustering in first-degree relatives, ranging from 3-fold to 80-fold compared with the population prevalence, whereas the crossover risks between dissimilar cases of CHD were weaker. Family history of any CHD among first-degree relatives accounted for a small proportion of CHD cases in the population.

National time trends in congenital heart defects, Denmark, 1977-2005.

BACKGROUND: Time trends in congenital heart defects (CHD) by specific phenotype and with long follow-up time are rarely available for an entire population. We present trends in national CHD prevalences over the past 3 decades. METHODS: We linked information from the National Patient Register, the Causes of Death Register, and the Danish Cytogenetic Central Register for all persons born in Denmark, 1977 to 2005, and registered in the Civil Registration System, yielding a cohort of 1,763,591 persons-18,207 with CHD. Individuals with CHDs were classified by phenotype (heterotaxia, conotruncal defect, atrioventricular septal defect, anomalous pulmonary venous return, left and right ventricular outflow tract obstructions, septal defects, complex defects, associations, patent ductus arteriosus, unspecified, and other specified) by combining International Classification of Diseases codes using a hierarchical approach. RESULTS: From 1977 to 2005, the overall CHD birth prevalence increased from 73 to 113 per 10,000 live births. Generally, prevalence increased for defects diagnosed in infancy, until 1996-1997, and then stabilized. For each 5-year interval, isolated septal defects and severe defects increased by 22% (95% CI, 20%-25%) and 5% (95% CI, 4%-7%), respectively. Among the severe defects, conotruncal defects and atrioventricular septal defect showed the largest prevalence increases. Women had a lower prevalence of severe defects during the 1980s. The CHD prevalence increase was unchanged when persons with extracardiac defects or chromosomal aberrations were excluded. CONCLUSIONS: CHD birth prevalence increased from the beginning of the 1980s but stabilized in the late 1990s.

Systematic re-examination of carriers of balanced reciprocal translocations: a strategy to search for candidate regions for common and complex diseases.

Balanced reciprocal translocations associated with genetic disorders have facilitated the identification of a variety of genes for early-onset monogenic disorders, but only rarely the genes associated with common and complex disorders. To assess the potential of chromosomal breakpoints associated with common/ complex disorders, we investigated the full spectrum of diseases in 731 carriers of balanced reciprocal translocations without known early-onset disorders in a nation-wide questionnaire-based re-examination. In 42 families, one of the breakpoints at the cytogenetic level concurred with known linkage data and/or the translocation co-segregated with the reported phenotype, for example, we found a significant linkage (lod score=2.1) of dyslexia and a co-segregating translocation with a breakpoint in a previously confirmed locus for dyslexia. Furthermore, we identified 441 instances of at least two unrelated carriers with concordant breakpoints and traits. If applied to other populations, re-examination of translocation carriers may identify additional genotype-phenotype associations, some of which may be novel and others that may coincide with and provide additional support of data presented here.

Low frequency of large genomic rearrangements of BRCA1 and BRCA2 in western Denmark.

Germline mutations in BRCA1 and BRCA2 predispose female carriers to breast and ovarian cancer. The majority of mutations identified are small deletions or insertions or are nonsense mutations. Large genomic rearrangements in BRCA1 are found with varying frequencies in different populations, but BRCA2 rearrangements have not been investigated thoroughly. The objective in this study was to determine the frequency of large genomic rearrangements in BRCA1 and BRCA2 in a large group of Danish families with increased risk of breast and ovarian cancer. A total of 617 families previously tested negative for mutations involving few bases were screened with multiplex ligation-dependent probe amplification (MLPA). Two deletions in BRCA1 were identified in three families; no large rearrangements were detected in BRCA2. The large deletions constitute 3.8% of the BRCA1 mutations identified, which is low compared to several other populations.

A missense mutation in exon 13 in BRCA2, c.7235G>A, results in skipping of exon 13.

We report here the functional characterisation of a missense mutation c.7235G>A in BRCA2. By reverse transcriptase polymerase chain reaction the mutation is demonstrated to cause skipping of exon 13. We conclude that the mutation is most likely deleterious.

[Monogenic hereditary diseases]. / Monogent arvelige sygdomme.

Monogenic disorders are caused by the inheritance of single gene mutations; alternatively, a monogenic disorder arises as a consequence of a de novo mutation in either the paternal or maternal germ line. The exponential increase in our understanding of the human genome has resulted in the localisation and cloning of a vast number of disease genes which has enabled the precise characterization of the underlying molecular defect in many of these disorders. Single nucleotide substitutions and microdeletions are the major types of disease-related mutations, but more complex mutations have also been described.

Recurrence of discordant congenital heart defects in families.

BACKGROUND: Variation within a single gene might produce different congenital heart defects (CHDs) within a family, which could explain the previously reported familial aggregation of discordant CHDs. We investigated whether certain groups of discordant CHDs are more common in families than others. METHODS AND RESULTS: Using Danish national population and health registers, we identified CHDs among all singletons born in Denmark during 1977-2005 and their first-degree relatives. In a cohort of 1 711 641 persons, 16 777 had CHDs, which we classified into 14 phenotypes. We estimated relative risks of discordant CHDs by history of specific CHDs in first-degree relatives. The relative risk of any dissimilar CHD given the specified CHD in first-degree relatives was as follows: heterotaxia, 2.00 (95% CI, 0.96 to 4.17); conotruncal defects, 2.78 (95% CI, 2.12 to 3.66); atrioventricular septal defects, 2.25 (95% CI, 1.39 to 3.66); anomalous pulmonary venous return, 1.76 (95% CI, 0.66 to 4.64); left- and right-ventricular outflow tract obstruction, 2.55 (95% CI, 1.87 to 3.48) and 3.09 (95% CI, 2.03 to 4.71), respectively; isolated atrial septal defects, 2.76 (95% CI, 2.11 to 3.61); isolated ventricular septal defects, 2.27 (95% CI, 1.75 to 2.94); persistent ductus arteriosus, 1.92 (95% CI, 1.32 to 2.79); other specified CHDs, 3.29 (95% CI, 2.51 to 4.32); and unspecified CHDs, 2.30 (95% CI, 1.76 to 3.00). Relative risks for all pairwise combinations of discordant CHD phenotypes gave no indications that certain constellations of CHDs cluster more in families than others. CONCLUSIONS: We documented strong familial aggregation of discordant CHD phenotypes. However, we observed no excess clustering of specific CHD phenotypes among the first-degree relatives.

No signature of Y chromosomal resemblance between possible descendants of the Cimbri in Denmark and Northern Italy.

Two European populations are believed to be related to the ancient Germanic tribe Cimbri: one living in Northern Italy, the other living in Jutland, Denmark. The people called Cimbri are documented in the ancient Roman historical record. Arriving from the far north their movements can be tracked from successive battles with the Romans. The Cimbri finally entered Italy from the northeast and were defeated at Vercellae (present day Vercelli) in 101 BC by Gaius Marius and his professional legions. Classical sources from the first centuries AD relate the homeland of the Cimbri to the coasts around the Elb estuary (northern Germany) or specifically towards the north (Himmerland in northern Jutland). In the alpine parts of Veneto, northeast of the historical battlefield, local traditions dating back to late medieval time, identify a local population as Cimbri living in Terra dei Cimbri. They are considered the descendents of the Germanic combatants that fled the battlefield at Vercelli. As the defeated Cimbri that possibly fled to the mountains of Northern Italy most likely would have been male (warriors), the present study investigated the possible Y chromosomal diversity of the two present populations using microsatellite markers and single nucleotide polymorphisms. While Cimbri from Himmerland resembled their geographical neighbors from Denmark for the Y-chromosome markers, Cimbri from Italy were significantly differentiated both from Cimbri from Himmerland and from Danes. Therefore, we were not able to show any biological relationship for uniparentally transmitted markers.

The eye of Eila Hopper Ross.

In 1936, when Eila Hopper Ross (nee Hopper) was putting the final stroke of paint on her six-foot oil canvas of a life model at the Ontario College of Art, she had not dreamed that in the year to follow she would embark upon a life's career of anatomical study and attention to intricate detail in her own work as a medical illustrator. Though she had early aspirations herself of going into medicine--having been raised in a medical environment with her father, Dr. David Alexander Hopper, regularly receiving pharmaceutical literature with impressive illustrations--Hopper Ross was not aware of medical illustration as a profession. It was Maria Wishart, then director of the Medical Art Service Unit at the University of Toronto, who saw Hopper Ross' final exhibition at the Ontario College of Art and recommended she consider a career in medical art. As a result, Hopper Ross studied for two years (1937-1939) in the Department of Art as Applied to Medicine at Johns Hopkins University in Baltimore, Maryland under the direction of Professor Max Brödel.

Functional testing of keratin 14 mutant proteins associated with the three major subtypes of epidermolysis bullosa simplex.

Epidermolysis bullosa simplex (EBS) is a group of autosomal dominantly inherited skin disorders characterized by the development of intra-epidermal skin blisters on mild mechanical trauma. The three major clinical subtypes (Weber-Cockayne, Koebner and Dowling-Meara) are all caused by mutations in either the keratin 5 (KRT5) or keratin 14 (KRT14) gene. Previously, we identified three novel KRT14 missense mutations in Danish EBS patients associated with the three different forms of EBS (1). The identified KRT14 mutations represent the full spectrum of the classical EBS subtypes. In the present study we investigated these mutations in a cellular expression system in order to analyse their effects on the keratin cytoskeleton. KRT14 expression vectors were constructed by fusing the nucleotide sequence encoding the FLAG reporter peptide to the 3' end of the KRT14 cDNA sequences. The expression vectors were transiently transfected into normal human primary keratinocytes (NHK), HaCaT or HeLa cells in order to analyze the ability of the mutant K14 proteins to integrate into the existing endogenous keratin filament network (KFN). No effect on the keratin cytoskeleton was observed upon transfection of NHK with the various K14 constructs neither with nor without a subsequently induced heat-stress. In contrast, all constructs, including wild-type K14, caused collapse of the endogenous KFN in a small fraction of the transfected HeLa and HaCaT cells. However, overexpression of the mutation associated with the most severe form of the disease, EBS Dowling-Meara, resulted in a higher number of transfected HaCaT cells with KFN collapse (P < 0.001). Thus, although a background KFN perturbance was observed upon transfection with the wild-type K14 construct, the mutant protein associated with the most severe form of EBS worsened the KFN perturbation significantly compared with the mutant proteins associated with the milder forms of the disease and the normal K14 protein. This shows that the clinical severity of disease-associated mutations identified in patients can be tested using this expression system, although it can not at present be used to discriminate between the milder forms. Assessment of the endogenous K14 protein expression in NHK and HaCaT cells indicated that the higher level of endogenous keratin expression in NHK might make these cells more resistant to perturbation of the keratin cytoskeleton by overexpressed K14 protein than HaCaT cells.