bluuebunny/crossref_metadata_2025 huggingface dataset:
Oilseed rape flour: another allergen causing occupational asthma among farmers
Authors: Alvarez M.J. , Estrada J.L. , Gozalo F. , Fernandez‐Rojo F. , Barber D.
Published: March 2003

Background: Farmers are exposed to a wide variety of sensitizers. Since occupational asthma (OA) can lead to permanent disability, exposure discontinuation is the preferred treatment. When this is not possible, the identification of the causative allergen may allow an alternative therapy.

Methods: We present three farmers diagnosed with OA as a consequence of handling fodder. We carried out skin tests with common and occupational allergens and with oilseed rape (OSR) extract. Total and specific serum IgE levels were measured. The patients underwent the OSR‐bronchial provocation test (OSR‐BPT). The day before and 24 h after the OSR‐BPT, the methacholine (M)‐BPT and induced sputum were performed. Eosinophil percentages and ECP levels were measured in the sputum samples.

Results: OSR sensitization (skin tests and specific serum IgE) was detected in all the patients. The OSR‐BPT elicited early responses in two subjects. Methacholine sensitivity, sputum eosinophils, and sputum ECP levels increased 24 h after the OSR‐BPT in all the patients.

Conclusions: We have demonstrated that inhalation of OSR flour causes bronchoconstriction, induces an eosinophilic inflammatory bronchial response, and increases bronchial hyperresponsiveness in sensitized asthmatics. OSR flour contained in animal fodder should be considered another potential cause of OA among farmers.
Pigeon allergens in indoor environments: a preliminary study
Authors: Curtis L. , Lee B.‐S. , Cai D. , Morozova I. , Fan J.‐L. , Scheff P. , Persky V. , Einoder C. , Diblee S.
Published: March 2003

Background:  Few studies have measured pigeon allergens in non pigeon coop environments. This study was conducted to determine approximate pigeon dropping allergen concentrations in indoor environments.

Methods:  Polyclonal antibody serum was prepared by injecting a rabbit three times with crude wild pigeon dropping extract in 50 mM Tris buffer with Freund's adjuvant. One hundred and fifteen dust samples were collected in a pigeon‐infested school, pigeon coops, homes and hospitals and analyzed by a direct competitive pigeon enzyme‐linked immunosorbent assay (ELISA).

Results:  The highest level of pigeon allergen inhibitory activity were recorded in four samples from pigeon coop bedding samples with a median activity of 11.2% relative to pigeon droppings. The second highest level of pigeon allergens was in a pigeon‐infested high school with a median or 7.4% activity relative to pigeon droppings. At an entrance underneath pigeon roosts, one sample had a relative inhibitory activity of 62.3%. Pigeon allergen inhibitory levels were generally low in the home and hospital samples, but nevertheless 46 out of 89 of these samples were still above detection limit.

Conclusions:  This study suggests that large concentrations of pigeon allergens can be found in buildings without domestic pigeons such as the pigeon‐infested high school.
Polymorphism 4G/5G in the plasminogen activator inhibitor‐1 (PAI‐1) gene is associated with IgE‐mediated allergic diseases and asthma in the Czech population
Authors: Buč D. , Izakovičová Hollá L. , Vácha J.
Published: March 2003

Background: Plasminogen activator inhibitor type 1 (PAI‐1) is a glycoprotein that belongs to the serine protease inhibitor superfamily and has an essential role in tissue remodeling after inflammation. Recently, a single base pair deletion/insertion (4G/5G) polymorphism of the PAI‐1 gene has been associated with an increased risk of asthma in nuclear families from the UK.

Methods: The present study was thus conducted to determine the association of this polymorphism with the development of IgE‐mediated asthma and other allergic diseases in the Czech population. A case‐control approach was used in our study. DNA taken from subjects with clinically manifested asthma and other allergic diseases (n = 207) and from reference ethnically age‐gender‐matched unrelated subjects (n = 186) was examined for base deletions/insertions in the PAI‐1 gene.

Results: A significant association (P = 0.0035) was observed between the PAI‐1 promoter polymorphism and IgE‐mediated allergic diseases altogether. Furthermore, the 4G allele frequency was also significantly higher in the asthmatic patients than in the control group (P = 0.0148).

Conclusions: Our findings support the idea that the 4G allele of the 4G/5G polymorphism in the PAI‐1 gene may be a risk factor for IgE‐mediated asthma and allergic diseases.
Up‐regulation of IL‐18 in allergic rhinitis
Authors: Verhaeghe B. , Gevaert P. , Holtappels G. , Lukat K. F. , Lange B. , Van Cauwenberge P. , Bachert C.
Published: March 2003

Background:  This paper reports a study on the concentrations of the pro‐inflammatory cytokines IL‐18 and IL‐1β in nasal secretions of allergic rhinitis patients in relation to ECP and nasal symptoms.

Methods:  We measured IL‐18 and IL‐1β concentrations using ELISA, and eosinophilic cationic protein (ECP) using the CAP system, in nasal secretions of 15 seasonal allergic rhinitis (SAR) patients at six visits throughout the pollen season. Pollen exposure, nasal and ocular symptoms were monitored daily. Furthermore, we measured IL‐18, IL‐1β and ECP concentrations in nasal secretions of 19 controls and 20 symptomatic persistent allergic rhinitis (PAR) patients with house dust mite allergy.

Results:  In SAR, the increase of IL‐18, IL‐1β and ECP paralleled the pollen flight with a time delay. IL‐18 and IL‐1β significantly increased during the pollen season compared to baseline, and differently from ECP, remained elevated until 4 weeks after the season. In PAR, the concentrations of IL‐18 and ECP, but not IL‐1β, were significantly higher compared to controls, with IL‐18 concentrations also being significantly higher than in SAR.

Conclusion:  This is the first study to demonstrate the up‐regulation of IL‐18 in nasal secretions in allergic rhinitis. The persistence of elevated IL‐18 concentrations until after the season and the high concentrations in PAR compared to SAR suggests its role in persistent allergic inflammation.
Treatment of asthmatic patients with a cysteinyl leukotriene receptor‐1 antagonist montelukast (Singulair), decreases the eosinophil survival‐enhancing activity produced by peripheral blood mononuclear leukocytes in vitro
Authors: Becler K. , Håkansson L. , Rak S.
Published: March 2003

Background: Montelukast (Singulair, MSD) has been shown to have a beneficial effect on the clinical symptoms of asthma. We aimed to investigate the effect of montelukast treatment on the production of eosinophil survival‐enhancing cytokines by peripheral blood mononuclear cells (PBMNC).

Methods: PBMNC obtained from 15 grass‐allergic patients (7 treated with montelukast and 8 with a placebo) were cultured for 72 h. Eosinophils from allergic patients were cultured with MNC supernatants alone or with addition of neutralizing antibodies, and the proportion of living cells was assessed by flow cytometry. In another experiment PBMNC from 6 allergic patients were cultured in vitro in the presence of montelukast or vehicle. Following stimulation the production of GM‐CSF in monocytes was assessed.

Results: Eosinophil survival in the MNC supernatants from the placebo‐treated patients was significantly (P < 0.05) higher than in supernatants from montelukast‐treated patients. GM‐CSF was the predominant cytokine responsible for the eosinophil survival‐enhancing activity (ESEA). In vitro production of GM‐CSF by allergen‐stimulated monocytes was significantly suppressed by addition of montelukast.

Conclusion: Treatment of patients with montelukast decreased the production of MNC‐derived cytokines, particularly GM‐CSF. We suggest that cysteinyl leukotriene receptor‐1 (CysLT‐R1) antagonists may act, at least partially, by diminishing the production of GM‐CSF from PBMNCs.
Rapid method for arrayed investigation of IgE‐reactivity profiles using natural and recombinant allergens
Authors: Suck R. , Nandy A. , Weber B. , Stock M. , Fiebig H. , Cromwell O.
Published: March 2003

Background: The availability of increasing numbers of purified natural and recombinant allergens offer the possibility for component‐resolved characterization of IgE binding. To make use of this potential, fast and simple methods with high capacity have to be developed.

Methods: A laboratory multiscreen device was used in an innovative two‐dimensional approach. In the first step, natural and recombinant allergens were immobilized onto the membrane using the sample chambers as application mask and, after blocking and rotating the membrane through 90°, the same device was used to apply and incubate sera of allergic patients. Enzyme‐linked immunoassay (ELISA) quantification of specific IgE was performed for purposes of comparison.

Results: Proteins were most efficiently bound onto nitrocellulose in 20 mM sodium hydroxide (NaOH). Up to 45 proteins or extracts could be investigated with a maximum of 45 sera in a single application, resulting in a resolution of 2025 spots on one membrane with a size comparable to a standard Western blot. A high correlation for IgE‐binding between natural and recombinant allergens was observed. Development of the membrane resulted in very evenly distributed square patterns. The results corresponded with the conventional ELISA measurements of specific IgE.

Conclusions: The innovative usage of a standard incubation device for both application of proteins as well as screening of sera provides a simple high throughput method for the characterization of IgE binding to allergens. The results are important for component resolved diagnosis of allergy by means of fast monitoring of IgE‐ and IgG‐reactivity spectra. Recombinant allergens may be used as targets for these purposes.
Nasal provocation test (NPT) with aspirin: a sensitive and safe method to diagnose aspirin‐induced asthma (AIA)
Authors: Alonso‐Llamazares A. , Martinez‐Cócera C. , Domínguez‐Ortega J. , Robledo‐Echarren T. , Cimarra‐Alvarez M. , Mesa del Castillo M.
Published: March 2003

Background: We have aimed to determine the sensitivity and specificity of a simpler technique with less risk than oral provocation to diagnose aspirin‐induced asthma (AIA).

Methods: We studied a group of 20 AIA patients compared to a control group with 40 aspirin‐tolerant patients (confirmed by oral provocation test): 10 asthmatic patients and 30 healthy subjects. A nasal provocation test (NPT) with lysine acetylsalicylic acid (L‐ASA) was carried out. Nasal and pulmonary functions were monitored with anterior active rhinomanometry (AAR) and spirometry. An L‐ASA solution (900 mg/ml L‐ASA, equivalent to 500 mg/ml acetylsalicylic acid) was diluted with saline solution. We administered four increasing doses: 5, 25, 50 and 100 mg/ml acetylsalicylic acid (ASA) with saline solution control. Nasal and pulmonary functions were monitored with rhinomanometry and spirometry. The patients were controlled for nasal inspiratory peak flow and expiratory peak flow.

Results: The results showed high sensitivity and specificity: 80% and 92.5%, respectively, with an 84.2% positive predictive value and an 89.2% negative predictive value. The patients had no bronchial or systemic symptoms, and no decreases over 20% were recorded in the FEV1.

Conclusion: NPT has a high sensitivity and specificity in the diagnosis of AIA. An oral provocation should be performed to confirm the result whenever the clinical situation of the patient permits it.
In vitro kinetics of allergen‐ and microbe‐induced IL‐4 and IFN‐γ mRNA expression in PBMC of pollen‐allergic patients
Authors: Laaksonen K. , Waris M. , Mäkelä M.J. , Terho E. O. , Savolainen J.
Published: March 2003

Background: According to a hypothesis allergens induce Th2 responses in allergic patients, and microbes induce Th1 responses. We studied the kinetics of in vitro allergen‐, tuberculin (PPD)‐ and tetanus toxin (TT)‐induced IFN‐γ and IL‐4 mRNA expression in peripheral blood mononuclear cell (PBMC) cultures of pollen‐allergic patients and healthy controls.

Methods: PBMC of 10 birch or timothy pollen‐allergic patients and of 13 healthy controls were stimulated in vitro with allergen (birch or timothy), PPD or TT. Pellets and supernatants were collected at 24, 48, 72 and 96 h after stimulation. IFN‐γ and IL‐4 production was measured by enzyme linked immunosorbent assay and mRNA expression using RT‐PCR and time‐resolved fluorometry.

Results: Allergen induced IFN‐γ production and mRNA expression in PBMC more in allergic patients than in healthy controls. Also allergen induced IL‐4 mRNA expression more in allergic patients than in healthy controls. PPD induced IFN‐γ mRNA expression both in allergic patients and healthy controls, whereas IFN‐γ production was induced only in healthy controls and IL‐4 was not induced at all. TT induced IFN‐γ mRNA expression in both groups, IFN‐γ production in allergic patients, and IL‐4 mRNA expression in both allergic patients and healthy controls.

Conclusions: In vitro stimulation with allergen induced both IFN‐γ and IL‐4 mRNA expression of PBMC in allergic patients. These observations challenge the clearcut division of microbe‐specific Th1 and allergen‐specific Th2 responses in peripheral blood.
Relationship of skin‐prick reactivity to aeroallergens and hyperresponsiveness to challenges with methacholine and adenosine monophosphate
Authors: Fowler Stephen J. , Lipworth Brian J.
Published: March 2003

Background: Clarification of the relationship between atopy and bronchial hyperresponsiveness (BHR), both key features of asthma, is critical to our understanding of the disease. We therefore investigated the putative relationship between skin‐prick reactivity to aeroallergens and BHR to direct and indirect stimuli.

Methods: We performed a retrospective analysis of data from 332 patients presenting with a diagnosis of asthma. Patients were characterized by skin prick tests (SPT), spirometry and bronchial challenge with methacholine and adenosine monophosphate (AMP).

Results: For patients who had BHR to methacholine but not AMP, the presence of atopy was associated with a lower PD20 (the provocative dose of methacholine producing a fall in FEV1 of 20%), amounting to a geometric mean (95% confidence interval (CI)) of 2.3‐fold (1.4–4.0) difference. Furthermore, the number of skin‐prick positive (SPP) responses was related to methacholine reactivity: 0–1 SPP, PD20 = 69.9 µg; 2–4 SPP, PD20 = 47.8 µg; 5–8 SPP, PD20 = 35.6 µg. There was a 2.0‐ fold (1.1–3.6) difference between the groups with a low (0–1 SPP) and high (5–8 SPP) degree of skin‐prick reactivity. A similar pattern was seen when data were analyzed including only perennial allergens. Spirometry was unrelated to the degree of skin‐prick reactivity.

Discussion: These results suggest that skin‐prick reactivity to aeroallergens is associated with BHR to methacholine.
Nuclear factor‐κB activity is down‐regulated in nasal polyps from aspirin‐sensitive asthmatics
Authors: Picado C. , Bioque G. , Roca‐Ferrer J. , Pujols L. , Mullol J. , Benitez P. , Bulbena O.
Published: March 2003

Background: We examined whether a decreased activity of nuclear factor(NF)‐κB), a transcriptional regulator of cyclooxygenase‐2 (COX‐2), could account for down‐regulation of COX‐2 in nasal polyps of aspirin‐sensitive asthmatics.

Methods: Nasal polyps were obtained from 17 aspirin‐intolerant asthma/rhinitis patients (AIAR; 7 men, mean age 48 ± 12 years) and 23 aspirin‐tolerant asthma/rhinitis patients (ATAR; 12 men, mean age 65 ± 11 years). COX‐2 mRNA expression was measured using semiquantitative reverse transcriptase competitive polymerase chain reaction (RT‐PCR), and the results were expressed as mean ± standard error of 106 molecules of mRNA/µg of total RNA. NF‐κB binding was measured with 32P‐labeled oligonucleotides and electrophoretic mobility shift assay (EMSA), and the results were expressed as a percentage with respect to the mean EMSA obtained in 19 healthy nasal mucosa.

Results: The mean levels of COX‐2 mRNA expression (0.25 ± 0.06) and NF‐κB activity (89 ± 13) in nasal polyps from AIAR were significantly lower than in polyps from ATAR (COX‐2 = 1.58 ± 0.50, and NF‐κB = 143 ± 12, P < 0.01 and P < 0.05, respectively). Levels of COX‐2 mRNA and NF‐κB activity in polyps from patients on corticosteroid therapy did not differ statistically from those who were not on this therapy before polypectomy.

Conclusion: This study shows that the low expression of COX‐2 mRNA in nasal polyps from aspirin‐sensitive patients is associated with a down‐regulation of NF‐κB activity.
I1307K APC and hMLH1 mutations in a non‐Jewish family with hereditary non‐polyposis colorectal cancer
Authors: Yuan Zhi Qiang , Kasprzak Lidia , Gordon Philip H , Pinsky Leonard , Foulkes William D
Published: March 2003

We describe a French Canadian hereditary non‐polyposis colorectal cancer (HNPCC) kindred which carries a novel truncating mutation in hMLH1. Interestingly, the I1307K APC polymorphism, associated with an increased risk of colorectal cancer, is also present in this family. The I1307K polymorphism has previously only been identified in individuals of self‐reported Ashkenazi Jewish origins. In addition, in this family, there appears to be no relationship between the I1307K polymorphism and the presence or absence of cancer.
Tissue‐specific methylation differences in a fragile X premutation carrier
Authors: Tassone Flora , Longshore John , Zunich Janice , Steinbach Peter , Salat Ulrike , Taylor Annette K
Published: March 2003

Methylation of a premutation was found in a small percentage of blood cells in a male premutation carrier for the FMR1 mutation. To investigate the inter‐tissue heterogeneity and possible clinical implications of this finding, fibroblast cells from the subject were also studied. Although the premutation size was found to be the same in leukocytes and fibroblasts, the methylation pattern was different. In cultured fibroblasts, the premutation was completely unmethylated, as is typical of premutations, whereas methylation of the premutation was detected in a small percentage of lymphocytes. However, the change in methylation did not affect the FMR1 protein (FMRP) expression, as immunocytochemical analysis of FMRP performed on cultured skin fibroblasts and a blood smear revealed normal levels of expression in both tissues.
Cloning of translocation breakpoints associated with Shwachman syndrome and identification of a candidate gene
Authors: Ikegawa Shiro , Masuno Mitsuo , Kumano Yoshiharu , Okawa Akihiko , Isomura Minoru , Koyama Kumiko , Okui Keiko , Makita Yoshio , Sasaki Michiko , Kohdera Urara , Okuda Masumi , Koyama Hirofumi , Ohashi Hirofumi , Tajiri Hitoshi , Imaizumi Kiyoshi , Nakamura Yusuke
Published: March 2003

Shwachman syndrome is an autosomal‐recessive disorder characterized by exocrine pancreatic insufficiency, bone‐marrow dysfunction, and metaphyseal chondrodysplasia. A de novo balanced translocation was recently documented in a patient with this disease. Toward isolating the gene(s) responsible for Shwachman syndrome, we cloned and sequenced the translocation breakpoints in the DNA of this patient. The nucleotide sequences around the breakpoints contained neither repetitive elements nor motifs reported to be implicated in recombination events, although we did detect gains or losses of oligonucleotides at the translocation junctions. By large‐scale genomic sequencing and in silico gene trapping, we identified two novel transcripts in the vicinity of the breakpoints that might represent candidate genes for Shwachman syndrome, one on chromosome 6 and the other on chromosome 12. The gene on chromosome 12 was actually disrupted by the translocation.
Inheritance of two different alleles of the low‐density lipoprotein (LDL)‐receptor gene carrying the recurrent Pro664Leu mutation in a patient with homozygous familial hypercholesterolaemia
Authors: Bourbon Mafalda , Fowler A. Michelle , Sun Xi‐Ming , Soutar Anne K.
Published: March 2003

Familial hypercholesterolaemia (FH) is caused by mutations in the low‐density lipoprotein (LDL)‐receptor gene that result in impaired clearance of plasma LDL and increased risk of coronary heart disease. Numerous different mutations have been found in FH patients world‐wide, the majority of which are infrequent in out‐bred populations and account for 2% or less of patients with the disorder in large cohorts. Thus, it was surprising to find that two homozygous FH patients referred to a single hospital in the UK were both apparently homozygous for the Pro664Leu mutation. One, an Asian patient, was a true homozygote. The other, of English origin, had inherited two different alleles of the LDL‐receptor gene with the same mutation from unrelated parents, as inferred from the haplotype of polymorphic markers. A third, clinically homozygous FH patient, despite being the offspring of first cousins, had inherited one ‘Asian’ Pro664Leu allele, but an allele with a 1‐bp deletion in exon 5 from the other parent. The Pro664Leu mutation in the LDL‐receptor gene has now been described in heterozygous patients of very different ethnic origin and is associated with different haplotypes, suggesting that the same base change at a CpG may have recurred as many as six times.
Cortical dysgenesis in 2 patients with chromosome 22q11 deletion
Authors: Bird Lynne M. , Scambler Peter
Published: March 2003

Two patients with chromosome 22q11 deletion and cortical dysgenesis (gyral abnormalities) are reported in this study. One had unilateral clubfoot in addition to multiple features suggestive of the Di George syndrome (DGS), and the other presented with leg asymmetry and seizures, with subsequent recognition of the velo‐cardio‐facial syndrome (VCFS). In each patient, gyral abnormalities were identified in the hemisphere contralateral to the limb abnormality. A wide range of central nervous system abnormalities have been reported in DGS and VCFS, including three prior reports of gyral abnormalities (lissencephaly, microgyria). The 2 patients reported herein strengthen the association between the 22q11 deletion spectrum and cortical dysgenesis, but the underlying pathogenetic mechanism (primary neural migration vs. vascular disruption) remains unclear.
Prevalence of the C282Y mutation for haemochromatosis on the Island of Majorca
Authors: Guix P , Picornell A , Parera M , Tomás C , Muncunill J , Castro Ja , Rossell J , Vaquer P , Ramon Mm , Obrador A
Published: March 2003

The C282Y mutation of the HFE gene has been reported to be present in most of the patients with hereditary haemochromatosis (HH) of Northern European ancestry. HH affects approximately 1/300 individuals, but it is not evenly distributed in the different European countries. In the present study, polymerase chain reaction (PCR) and restriction‐enzyme digestion were used to analyse the frequency of the most important mutation in haemochromatosis (C282Y) in subjects from Majorca (Balearic Islands, Spain) and patients with haemochromatosis. The results were compared with other studies from Spain and Europe. A total of 420 Majorcan chromosomes were analysed and the C282Y mutation was observed at a frequency of 2.62%±0.8 (11 heterozygotes: eight men and three women). In the group of hereditary haemochromatosis probands, 13 out of 14 were homozygous for the C282Y mutation. In the distribution of the C282Y mutation, a north‐west to south‐east cline was detected, supporting the Celtic origin of this mutation.
Resolution of a mispaired secondary structure intermediate could account for a novel micro‐insertion/deletion (387 insA/del 8 bp) in the PYGM gene causing McArdle's disease
Authors: Martín MA , Rubio JC , García A , Fernández MA , Campos Y , Krawczak M , Cooper DN , Arenas J
Published: March 2003

We report two siblings with McArdle's disease who are both compound heterozygotes for two non‐identical frameshift mutations in the PYGM gene; a previously reported 753 delA in exon 18 and a novel 387 insA/del 8 bp in exon 10. The novel mutation is predicted to result in premature termination of translation 33 amino acids downstream of the site of mutation, potentially encoding a severely truncated protein of 419 amino acids instead of 841 amino acids. The complete lack of myophosphorylase activity observed in muscle derived from one sibling suggests that this mutation has deleterious functional consequences. The underlying mechanism of mutagenesis may have been slipped mispairing mediated by the formation of a Moebius loop‐like secondary intermediate.
Analysis of myocilin gene mutations in Japanese patients with normal tension glaucoma and primary open‐angle glaucoma
Authors: Mabuchi F , Yamagata Z , Kashiwagi K , Tang S , Iijima H , Tsukahara S
Published: March 2003

The myocilin gene was identified as a gene (MYOC) that caused primary open‐angle glaucoma (POAG). Although a normal tension glaucoma (NTG) patient with the myocilin gene mutation was previously reported, no study using large numbers of patients with NTG has been reported. Single‐strand conformation polymorphism analysis and subsequent sequence analysis were performed for genotyping the myocilin gene in 114 unrelated Japanese patients with NTG. One hundred and nineteen patients with POAG and 100 control subjects without glaucoma were studied as reference subjects. Five amino acid sequence changes of the myocilin were identified: Arg46Stop (one NTG), Arg76Lys (four NTG, 10 POAG, seven control), Arg158Gln (one NTG, one POAG, one control) found in only Japanese, Asp208Glu (four NTG, three POAG, one control), Pro481Ser (one control). Pro481Ser was novel. Arg76Lys always occurred with 1–83 from G to A in the promoter as it was reported in Chinese. Although some Japanese patients with NTG had sequence changes of the myocilin gene, there were no apparent specific mutations in patients with NTG.
W44C mutation in the connexin 26 gene associated with dominant non‐syndromic deafness
Authors: Tekin M , Arnos KS , Xia XJ , Oelrich MK , Liu XZ , Nance WE , Pandya A
Published: March 2003

Although more than 50% of recessive non‐syndromic deafness is attributed to mutations in the connexin 26 (Cx26) gene, only a few reported families have shown dominant transmission of the trait. The W44C mutation was originally reported in two families from the same geographic region of France, which exhibited dominant non‐syndromic hearing loss. In this report, we describe a third family with early‐onset severe‐to‐profound non‐syndromic hearing loss segregating with the W44C mutation. Our observation places W44C among recurrent mutations in the Cx26 gene and emphasizes the importance of screening for this as well as other Cx26 mutations in autosomal dominant families.
The VACTERL association: lessons from the Sonic hedgehog pathway
Authors: Kim JH , Kim PCW , Hui C‐C
Published: March 2003

VACTERL represents a non‐random association of congenital anomalies in humans of poorly known etiology and pathogenesis. From our mutant analysis of Gli genes, which encode transcription factors mediating Sonic hedgehog (Shh) signal transduction, we observed that defective Shh signaling leads to a spectrum of developmental anomalies in mice strikingly similar to those of VACTERL. In this review, we will discuss the function of the three Gli transcription factors in Shh signaling and mammalian development. We propose that VACTERL could be caused by defective Shh signaling during human embryogenesis and suggest that the Gli mutant mice can serve as useful models for studying the pathogenesis of VACTERL.
The risk for congenital heart defects in offspring of individuals with congenital heart defects
Authors: Romano‐Zelekha O , Hirsh R , Blieden L , Green MS , Shohat T
Published: March 2003

Background: Congenital heart defects (CHDs) occur in approximately 1% of all live births. Although most CHDs are of unknown etiology, a family history of CHDs is a known risk factor, and offspring of individuals with CHDs are at a higher risk of having CHDs. The aim of this study was to investigate the relative risk for CHDs to offspring of individuals with CHDs.  Methods: The prevalence rates of CHDs in offspring of 203 individuals with CHDs and 282 individuals without CHDs were investigated. The study participants completed a questionnaire that included information on medical and reproductive history, lifestyle indicators, and family history of CHDs and other congenital malformations. The prevalence rates of CHDs in offspring were calculated.  Results: The prevalence of CHDs was 3.1% (18/575) in offspring of individuals with CHDs and 1.3% (8/589) in offspring of individuals without CHDs. The adjusted odds ratio for CHDs to offspring of parents with CHDs was 1.73 (95% confidence interval [95% CI] 0.89–2.44, p=0.02). The estimated relative risk for offspring to females with CHD was higher than for males [2.3 (95% CI 1.1–4.7, p=0.03) versus 1.31 (95% CI 0.48–4.30, p=0.66), respectively]. There was no suggestion of association between CHDs and maternal smoking, alcohol consumption, and use of medication during pregnancy.  Conclusions: Offspring of parents with CHDs are at a higher risk for CHDs compared with the general population. Couples where one member is affected with CHD should receive pre‐conceptional or pre‐natal genetic counseling and should be informed about the magnitude of the potential risk of CHDs to the offspring.
Fundus albipunctatus and retinitis punctata albescens in a pedigree with an R150Q mutation in RLBP1
Authors: Katsanis N , Shroyer NF , Lewis RA , Cavender JC , Al‐Rajhi AA , Jabak M , Lupski JR
Published: March 2003

Fundus albipunctatus (FA; OMIM 136880) is a rare form of apparently stationary night blindness characterized by the presence of myriad symmetrical round white dots in the fundus with a greater concentration in the midperiphery. A distantly similar but distinct clinical entity, retinitis punctata albescens (RPA), is also characterized by aggregation of irregular white flecks but is progressive and evolves to generalized atrophy of the retina. We studied 4 consanguineous kindreds diagnosed with FA from Saudi Arabia. Given the substantial phenotypic variation and overlap between different flecked retinal dystrophies, we evaluated all known genes associated with such conditions by both genetic analysis and direct sequencing. In one kindred, KKESH‐099, we identified a homozygous R150Q alteration in RLBP1, the gene encoding the cellular retinaldehyde binding protein, associated previously with both recessive retinitis pigmentosa (arRP) and RPA. Examination of several patients aged 3–20 years over a 9‐year period presented no evidence for either RP or RPA. In contrast, clinical examination of individuals with the same mutation in their fourth and fifth decade revealed signs consistent with RPA. The data suggest that the R150Q mutation in RLBP1 may result in RPA with slow progression. More importantly, younger individuals diagnosed with the milder disorder FA thought to be stationary may evolve to a more devastating and progressive phenotype.
The additive effect of neurotransmitter genes in pathological gambling
Authors: Comings DE , Gade‐Andavolu R , Gonzalez N , Wu S , Muhleman D , Chen C , Koh P , Farwell K , Blake H , Dietz G , MacMurray JP , Lesieur HR , Rugle LJ , Rosenthal RJ
Published: March 2003

As access to gambling increases there is a corresponding increase in the frequency of addiction to gambling, known as pathological gambling. Studies have shown that a number of different neurotransmitters are affected in pathological gamblers and that genetic factors play a role. Polymorphisms at 31 different genes involved in dopamine, serotonin, norepinephrine, GABA and neurotransmitters were genotyped in 139 pathological gamblers and 139 age, race, and sex‐matched controls. Multivariate regression analysis was used with the presence or absence of pathological gambling as the dependent variable, and the 31 coded genes as the independent variables. Fifteen genes were included in the regression equation. The most significant were theDRD2,DRD4,DAT1,TPH,ADRA2C,NMDA1, andPS1genes. The r2or fraction of the variance was less than 0.02 for most genes. Dopamine, serotonin, and norepinephrine genes contributed approximately equally to the risk for pathological gambling. These results indicate that genes influencing a range of brain functions play an additive role as risk factors for pathological gambling. Multi‐gene profiles in specific individuals may be of assistance in choosing the appropriate treatment.
Goals of genetic counseling
Authors: Biesecker BB
Published: March 2003

The goals of genetic counseling have differed over the past three decades. Two schools of thought are prominent in reviewing past literature. One upholds the goal of preventing birth defects and genetic disorders while the other promotes a goal of improved psychological well‐being in client adaptation to a genetic condition or risk. Both types of goals emphasize that clients should make their own reproductive decisions; however, the former relies on clients making decisions that will reduce the impact of genetic disorders. The differences in the types of goals may be due to the training and orientation of genetics health care providers, socio‐cultural views, or priorities of health care settings. Regardless, there are ample reasons to dismiss the prevention of birth defects as a goal. This mini‐review recommends use of genetic counseling sub‐specialties as a framework for considering different client needs and thus different counseling goals and specific aims in the reproductive, pediatric/adult, and common disease settings. Given the extent of new genetic information, technologies, and the need to evaluate genetic counseling practice, genetics health care providers should work toward arriving at consensus on the goals of genetic counseling, and in doing so, the needs of clients should be considered.
The E326K mutation and Gaucher disease: mutation or polymorphism?
Authors: Park JK , Tayebi N , Stubblefield BK , LaMarca ME , MacKenzie JJ , Stone DL , Sidransky E
Published: March 2003

Gaucher disease is caused by mutations in the gene for human glucocerebrosidase, a lysosomal enzyme involved in the intracellular hydrolysis of glucosylceramide. While over 150 different glucocerebrosidase mutations have been identified in patients with Gaucher disease, not all reported mutations have been fully characterized as being causative. One such mutation is the E326K mutation, which results from a G to A nucleotide substitution at genomic position 6195 and has been identified in patients with type 1, type 2 and type 3 Gaucher disease. However, in each instance, the E326K mutation was found on the same allele with another glucocerebrosidase mutation. Utilizing polymerase chain reaction (PCR) screening and restriction digestions of both patients with Gaucher disease and normal controls, we identified the E326K allele in both groups. Of the 310 alleles screened from patients with Gaucher disease, the E326K mutation was detected in four alleles (1.3%). In addition, screening for the E326K mutation among normal controls from a random population revealed that three alleles among 316 screened (0.9%) also carried the E326K mutation. In the normal controls with the E326K allele, the glucocerebrosidase gene was completely sequenced, but no additional mutations were found. Because the E326K mutation may be a polymorphism, we caution that a careful examination of any allele with this mutation should be performed to check for the presence of other glucocerebrosidase mutations.
Participation rates of Ashkenazi Jews in a colon cancer community‐based screening/prevention study
Authors: Cappelli M , Hunter AGW , Stern H , Humphreys L , Van Houten L , O'Rourke K , Viertelhausen S , Perras H , Lagarde AE
Published: March 2003

In a recent colon cancer risk study, genetic assessment and colonoscopy were offered to virtually all of the adult Ashkenazi Jews in an urban community. The present study was designed to examine factors influencing participation and response in the initial study and to suggest strategies for improving participation in future health promotion programs.

The study comprised a random sample of three groups of individuals who had been targeted for participation in the previous study: those who had (a) agreed to participate (n = 234); (b) declined participation (n = 179); and (c) failed to respond to a mailed recruitment package (n = 128). All participants completed a brief telephone survey.

Key multivariate predictors of both response and participation were individuals' perceptions of the drawbacks of participating in colon cancer screening research and the degree of decisional conflict they experienced. Response was further predicted by the influence of spouses, family history of colon cancer, past knowledge of genetic testing for colon cancer, and education level. Participation was predicted by awareness that the study was supported by the Ashkenazi Jewish community, past experience with genetic testing, individuals' perceptions of the benefits of participating, and whether or not they had children.

The degree to which individuals understand the purpose and nature of genetic screening research, along with their levels of decisional conflict and other psychosocial factors, may influence the likelihood of their participation in such research. Results of this study suggest a number of possible strategies for improving participation and response rates in disease prevention and detection studies.
Cognitive function in Coffin–Lowry syndrome
Authors: Simensen RJ , Abidi F , Collins JS , Schwartz CE , Stevenson RE
Published: March 2003

Coffin–Lowry syndrome (CLS) is an X‐linked disorder associated with mental retardation, distinctive facies and hands, hypotonia, and skeletal abnormalities. The syndrome results from mutations in the RSK2 gene located in Xp22.2. Although the syndrome has been elucidated clinically, few, if any, studies have focused on the cognitive deficits of the affected males or carrier females. The subjects of the present study were selected from two African‐American families who have the same missense mutation (C340T) in RSK2. The subjects included six affected males, seven carrier females, three normal males and three non‐carrier (normal) females. Normal family members served as contrast/comparison cohorts to control for socio‐economic, sociocultural and genetic variables which would impinge on intellectual abilities. Analysis of cognitive function, as measured by the Stanford‐Binet Intelligence Scale, 4th edn, demonstrated a distinct hierarchy of abilities from normal to carrier to affected patients. The mean composite IQs of the cohorts were 90.8, 65.0 and 43.2 for normal, carrier and affected individuals, respectively. These findings lend support to the clinical concept of negative intellectual effects in carriers of certain X‐linked mental retardation conditions. X‐inactivation studies showed that carrier females had mild to significant skewing. Normal females in the family did not demonstrate skewing. The correlation coefficient between IQ and X‐inactivation status among carriers was not significant.
Development and diseases of the pancreas
Authors: Johansson KA , Grapin‐Botton A
Published: March 2003

The pancreas is a vital gland of exocrine and endocrine function. It is the target of two main affections: diabetes and pancreatic cancer. We describe the tissue interactions, signaling pathways and intracellular targets that are involved in the emergence of the pancreas primordium and its proliferation, morphogenesis and differentiation. It appears that several genes of developmental relevance have an adult function and are involved in pancreas affections. Embryological experimentation in animals contributed to provide candidate genes for human disease and holds promise for future treatments.
A comparison of combinatorial partitioning and linear regression for the detection of epistatic effects of theACE I/DandPAI‐1 4G/5Gpolymorphisms on plasma PAI‐1 levels
Authors: Moore JH , Lamb JM , Brown NJ , Vaughan DE
Published: March 2003

The detection and characterization of epistasis or non‐additive gene–gene interactions remains a statistical challenge in genetic epidemiology. The recently developed combinatorial partitioning method (CPM) may overcome some of the limitations of linear regression for the exploratory analysis of non‐additive epistatic effects. The goal of this study was to compare CPM with linear regression analysis for the exploratory analysis of non‐additive interactive effects of theangiotensin converting enzyme (ACE) insertion/deletion (I/D)andplasminogen activator inhibitor 1 (PAI‐1) 4G/5Gpolymorphisms on plasma PAI‐1 levels in a sample of 50 unrelated African Americans and 117 unrelated Caucasians. Using linear regression, we documented the additive effects of theACEandPAI‐1genes on plasma PAI‐1 levels in African American females (R2 = 0.10), African American males (R2 = 0.16), Caucasian females (R2 = 0.11), and Caucasian males (R2 = 0.09). Using CPM, we found evidence for non‐additive effects of theACEandPAI‐1genes in both African American females (R2 = 0.22) and African American males (R2 = 0.24) but not in Caucasian females (R2 = 0.10) or Caucasian males (R2 = 0.11). The results of this exploratory data analysis support previous experimental, clinical, and epidemiological studies that have proposed as a working hypothesis that theACEgene mediates interaction effects of the fibrinolytic and renin‐angiotensin systems on plasma levels of PAI‐1.
Sibship stability of genotype and phenotype in myotonic dystrophy
Authors: Brisson D , Tremblay M , Prévost C , Laberge C , Puymirat J , Mathieu J
Published: March 2003

Myotonic dystrophy (DM1) is caused by an unstable CTG repeat expansion. Despite the evidence of birth order effect in congenital DM1, the expansion's dynamics among sibships is still unknown. The objective of this study was to determine phenotype and CTG repeat size variability in DM1 sibships, and to investigate their predictive values. We compared 86 sib pairs for CTG repeat, 61 for age at onset and 89 for DM1 phenotype. CTG repeats remained stable for 66 of the 86 sib pairs, including 25 of 27 maternal transmissions and 30 of 42 paternal transmissions. Variations of less than 10 years in the age at onset were observed in 44 of 61 sib pairs, including 16 of 18 maternal transmissions and 19 of 28 paternal transmissions. The same phenotypic severity or a variation of only one class was observed among 86 of the 89 sib pairs, including all of the 35 maternal transmissions and 30 of the 33 paternal transmissions. Birth order, intergenesic interval, oldest sib's CTG repeat or parental age and CTG repeat did not exert any significant influence. These results suggest that genotype and phenotype remained stable among sibs, although the paternal origin of the mutation seemed to reduce the predictability of the severity.
Comparison of two different protocols of neonatal screening for cystic fibrosis
Authors: Narzi L , Lucarelli M , Lelli A , Grandoni F , Lo Cicero S , Ferraro A , Matarazzo P , Delaroche I , Quattrucci S , Strom R , Antonelli M
Published: March 2003

The results of two different protocols of neonatal cystic fibrosis (CF) screening in the Lazio region of Italy are reported. The first study, conducted from 1992 to 2000 on about 200,000 newborns, consisted of an immunoreactive trypsin (IRT) protocol without mutation analysis of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, referred to as the IRT/IRT protocol. Approximately 5% of the newborns with a positive first IRT test were also positive at the second test; approximately 57% of the newborns with a high IRT level at the second test were subsequently found to be affected by CF. In September 1998, a second protocol that included mutation analysis (IRT/DNA/IRT protocol) was started. Comparison of the two different screening protocols in terms of sensitivity in detecting CF patients demonstrated that the IRT/DNA/IRT protocol is more effective because it is able to detect a higher number of CF patients than the IRT/IRT protocol. In the same period, in addition to the overall diagnosis performed on a screening basis, 64 other subjects were identified as being affected by CF on the basis of symptomatic findings. The overall incidence of CF (screening + symptoms) was 1 : 2982, while that for carriers was 1 : 27. The sensitivity of the screening program increased over the period from 1992 to 2000, with the enhanced sensitivity in the past 2 years being due to the introduction of the IRT/DNA/IRT protocol.
Analysis of the two common alpha‐1‐antitrypsin deficiency alleles PiMS and PiMZ as modifiers of Pseudomonas aeruginosa susceptibility in cystic fibrosis
Authors: Meyer P , Braun A , Roscher AA
Published: March 2003

Lung disease is the direct cause of death in more than 90% of cystic fibrosis (CF) patients. Proteinase‐antiproteinase imbalances are common in CF and alpha‐1‐antitrypsin (AAT) deficiency. We investigated the hypothesis that the AAT deficiency alleles PiS and PiZ contribute to pulmonary prognosis in CF. Two hundred and sixty‐nine CF patients from Southern Germany were included in this study. The serum concentrations of AAT and C‐reactive protein (CRP) were determined by nephelometry, and patients were screened by polymerase chain reaction (PCR) and restriction enzyme digest for the common AAT deficiency alleles PiS and PiZ. The onset of chronic bacterial colonization by Pseudomonas aeruginosa (Pae) was correlated with the AAT phenotypes PiMM, PiMS and PiMZ. Only three out of nine CF patients (33%) diagnosed with either PiMS or PiMZ had developed chronic Pae lung infection earlier in their lives. The remaining six patients showing a PiMS or PiMZ phenotype showed a later onset of chronic Pae lung infection. Our results indicate that PiMS and PiMZ are not associated with worse pulmonary prognosis in CF. These data need to be confirmed in studies with a much larger number of cases.
The changing survival profile of people with Down's syndrome: implications for genetic counselling
Authors: Glasson EJ , Sullivan SG , Hussain R , Petterson BA , Montgomery PD , Bittles AH
Published: March 2003

Cohort studies have indicated that the survival of individuals with Down's syndrome has dramatically increased over the past 50 years. Early childhood survival in particular has shown major improvement, due largely to advances in cardiac surgery and in general health management. The present study was based on a continuous cohort of 1332 people with Down's syndrome in Western Australia, registered for intellectual disability services between 1953 and 2000. Their life expectancy was 58.6 years, 25% lived to 62.9 years, and the oldest living person is 73 years of age. Life expectancy for males was greater than females by 3.3 years. The substantial increase in survival across the study period means that the life expectancy of people with Down's syndrome is approaching that of the general population, but accompanied by a range of significant mid‐life health problems. The findings are of relevance to all developed countries and have considerable implications in terms of the counselling information provided to families at risk of having a child with Down's syndrome.