bluuebunny/crossref_metadata_2025 huggingface dataset:
Bronchial challenges with aerosolized food in asthmatic, food‐allergic children
Authors: Roberts G. , Golder N. , Lack G.
Published: March 2003

Background:  Allergic asthma is usually considered to be provoked by aeroallergens. However, we have recently recognized a group of children with food allergies who also develop asthma when exposed to the aerosolized form of the food.

Methods:  Between 1997 and 1999 we prospectively identified children with an immunoglobulin (Ig)E‐mediated food allergy who develop asthma on inhalational exposure to the relevant food allergen while it is being cooked. Subjects were exposed for 20 min to the aerosolized form of the allergen and the symptoms and the lung function were monitored. Aerosolization was achieved by cooking the food in a small room. Where possible challenges were double‐blinded.

Results:  We identified 12 children with an IgE‐mediated food allergy who developed asthma on inhalational exposure to food. The implicated foods were fish, chickpea, milk, egg or buckwheat. Nine out of the 12 children consented to undergo a bronchial food challenge. Five challenges were positive with objective clinical features of asthma. Additionally, two children developed late‐phase symptoms with a decrease in lung function. Positive reactions were seen with fish, chickpea and buckwheat. There were no reactions to the seven placebo challenges.

Conclusions:  We have presented a prospective series of children with food allergy who developed symptoms of asthma with exposure to aerosolized food allergens. Our data demonstrates that, as in the case of other aeroallergens, inhaled food allergens can produce both early‐ and late‐phase asthmatic responses. This highlights the importance of considering foods as aeroallergens in children with coexistent food allergy and allergic asthma. For these children, dietary avoidance alone may not be sufficient and further environmental measures may be required to limit exposure to aerosolized food.
Assessment of profilin as an allergen for latex‐sensitized patients
Authors: Nieto A. , Mazón A. , Boquete M. , . Carballada F , Asturias J. A. , Martínez J , Martínez A.
Published: March 2003

Background:  The presence of the actin‐binding protein, profilin, has been demonstrated in natural latex extracts; but the clinical significance of this molecule as an allergen for latex‐allergic patients is not clear. We studied the allergenic relevance of isolated latex natural and recombinant profilin, by in vivo and in vitro techniques, in two populations of spina bifida children (SB) and adults allergic to latex (AL).

Methods: 
Profilin is present in small amounts in latex extracts obtained from low ammoniated (LA) natural latex. Its purification by affinity chromatography resulted difficult due to Heb v 1 unspecific binding. Therefore a method was developed to obtain natural profilin from natural latex, combining affinity chromatography (PLP, poly‐L‐proline Sepharose column) and previous ammonium sulfate fractionation. Alternatively, latex c‐serum containing a low amount of Hev b 1 and a relatively higher profilin content could be used. Recombinant latex profilin isoform (rHev b 8) was cloned by PCR amplification. The entire coding region of Hev b 8 was subcloned into the expression vector pKN172 and a non‐fusion form of Hev b 8 was expressed in
Escherichia coli BL21 (DE3). Purified recombinant protein was obtained after a single passage through PLP‐Sepharose column.

Results:  Natural and recombinant purified Hev b 8 were tested cutaneously by intradermoreaction (ID) in 17 SB and 14 AL patients. They were positive in 15 SB and 14 AL patients. No wheals were produced when tested in nonatopic control patients. Only 42% of sera from latex‐allergic patients revealed specific IgE titers of class 1 or higher by enzyme immunoassay and only 39% of them exhibited IgE binding by SDS‐PAGE immunoblotting with any natural or recombinant Hev b 8 forms.

Conclusion:  It seems that profilin is a relevant allergen for both groups of patients from a frequency point of view, but with scarce presence in natural latex extracts and raw sources, with a subsequent low IgE induction capacity.
Itraconazole treatment of allergic bronchopulmonary aspergillosis in patients with cystic fibrosis
Authors: Skov M , Høiby N , Koch C
Published: March 2003

Background:  Allergic bronchopulmonary aspergillosis (ABPA) in cystic fibrosis (CF) patients is a potentially fatal inflammatory disease due to the dual‐type immune response provoked by the fungal antigens. Despite serious side effects long‐term treatment with corticosteroids is often required. Itraconazole has been reported to be a useful steroid‐sparing agent.

Methods:  In a retrospective follow‐up of 21 CF patients from a total of 250 treated once or twice within a five‐year study period (1994–98), 9 patients were treated with systemic glucocorticosteroids in combination with itraconazole and 12 patients were treated with itraconazole (200–600 mg/day) as monotherapy.

Results: 
During treatment the percentage of
Aspergillus fumigatus
(AF)‐positive sputum cultures significantly reduced (
P
 < 0.05); precipitating antibodies to AF decreased significantly in all patients (
P
 < 0.05); forced expiratory volume (FEV
1) increased to pre‐exacerbation level; total IgE levels decreased in 42% of patients on monotherapy and in 56% on combination therapy. Specific IgE (radioallergosorbant; RAST) level decreased in 6 of 21 patients. Eleven patients had transient increased levels of alanine transaminase (ALAT). One patient had isolated increase in alkaline phosphatase and another in aspartate transaminase (ASAT).

Conclusions:  High dose itraconazole as monotherapy or in combination with systemic glucocorticosteroids seems effective in CF patients with ABPA. No hepatotoxicity was observed during long‐term therapy.
Sputum eosinophil count in a large population of patients with mild to moderate steroid‐naive asthma: distribution and relationship with methacholine bronchial hyperresponsiveness
Authors: Louis R. , Sele J. , Henket M. , Cataldo D. , Bettiol J. , Seiden L. , Bartsch P.
Published: March 2003

Background: Although airway eosinophilia is seen as a cardinal feature of asthma, data eosinophilia are still lacking on the proportion of the asthma group exhibiting raised airway eosinophilia. This study aimed to assess the distribution of sputum eosinophil count and its relationship with methacholine bronchial hyperresponsiveness in mild to moderate steroid‐naive asthmatic people.

Methods: Sputum was induced by inhalation of hypertonic saline (NaCl 4.5%) in 118 mild to moderate steroid‐naive asthmatic people consecutively recruited from our outpatient clinic, and in 44 healthy people. The asthma group was selected on the basis of an forced expiratory volume in 1 s (FEV1) of ≥ 70% predicted, and a provocative methacholine concentration causing a fall of 20% in FEV1 (PC20 methacholine; PC20M) ≤ 16 mg/ml.

Results: In the asthma group, the median (range) of the percentage and the absolute values of sputum eosinophils were 4.8% (0–75) and 38 103/g (0–14 191), respectively, vs 0% (0–2.3) (P < 0.001) and 0 103/g (0–53) (P < 0.001) in healthy participants. Based on the 95% percentile for normal values calculated from our healthy group, 69% of the asthma group had significantly raised sputum eosinophil count (that is > 2%). In the asthma group, multiple regression analysis followed by a stepwise procedure revealed that sputum eosinophil count was significantly and inversely associated with PC20M accounting for 16% of its total variance (P < 0.001) while neutrophil counts positively related to PC20M accounting for 4% of total variance (P < 0.05). By contrast, no significant relationship was found between either eosinophil or neutrophil counts and the slope of forced vital capacity (FVC) vs FEV1 from the methacholine challenge.

Conclusions: We conclude that two‐thirds of people in the mild to moderate asthma group had increased sputum eosinophilia, which plays a limited role in determining the degree of methacholine airway hyperresponsiveness.
Latex hypersensitivity: relationship with positive prick test and patch test responses among hairdressers
Authors: Nettis E. , Dambra P. , Soccio A. L. , Ferrannini A. , Tursi A.
Published: March 2003

Background: Natural rubber latex is a frequent cause of IgE‐mediated allergy in hairdressers; but a non‐IgE‐mediated allergy to latex proteins can also occur. Sixty‐one hairdressers, reporting latex glove‐related symptoms, were enrolled in the study.

Methods: They were evaluated for latex allergy by assessment of serum specific IgE and skin prick tests. Patients were patch tested to a rubber additive series and to latex. Proportions were compared by one‐tailed Fisher's exact test; mean values of continuous variables were compared by Student's t‐test.

Results: A diagnosis of IgE‐mediated latex allergy was made in 11 (18%) hairdressers; patch tests with rubber‐related antigens were positive in 24 (39.3%) subjects. The most positive patch tests were those with para‐phenylenediamine (33%). Two hairdressers (3.3%) showed delayed reactions to the latex sample. Hairdressers with IgE‐mediated latex allergy were older and presented a personal history of atopy and local hives after exposure to latex; they were more likely to have a positive response to other allergens on prick or patch testing.

Conclusions: Our study demonstrates a certain number of false‐positive and false‐negative results when reported symptoms of possible latex sensitivity were compared to prick and CAP tests to latex, indicating the problems of relying solely on self‐report.
Aspirin‐intolerant asthma: role of cyclo‐oxygenase enzymes
Authors: Picado C.
Published: March 2003

Aspirin‐induced asthma and rhinitis (AIAR) appear to be precipitated by the inhibition of cyclo‐oxygenase (COX). By inhibiting COX pathway aspirin diverts arachidonic acid metabolites to the lipoxygenase pathway. There are two isoforms of COX, namely COX‐1 and COX‐2. Metabolites derived from COX‐1 are involved in cellular housekeeping functions. COX‐2 can be induced in cells exposed to proinflammatory substances and growth factors. Recent studies have reported that patients with AIAR have decreased activity of COX‐2 and lower production of PGE2 in the upper airway and peripheral blood cells. Considering the protective effect of exogenous PGE2 on aspirin‐induced bronchoconstriction and the interdependence of PGE2 and cisteinyl leukotriene production, a reduced PGE2 synthesis may render aspirin‐sensitive patients more susceptible to the inhibitory effect of NSAIDs drugs and also lead to an increase in cysteinyl leukotriene release.
Deciphering the immune pathomechanism of cutaneous drug reactions
Authors: Pichler W. J.
Published: March 2003

The present peptide fragments to T‐cells in the context of MHC molecules. Small chemical compounds are thought to acquire immunogenicity by covalent binding to proteins or peptides. This requires chemical reactivity of the compound or its metabolite. We recently elaborated a new pathway of drug presentation to T‐cells, namely that even chemically nonreactive drugs like sulfamethoxazole (SMX) can be recognized in an MHC dependent way by T‐cell receptors. This concept of ‘noncovalent drug presentation’ is based on the finding that glutaraldehyde fixed cells could present chemically nonreactive drugs like sulfamethoxazole or lidocaine; and that the drug binding to MHC‐peptide complexes is rather labile, as the drug can be washed away – in contrast to covalently binding drugs. Our data show that drugs can bind in a pharmacological way to immune‐receptors like MHC or TCR, and that this labile binding is sufficient to elicit an immune response. This drug‐presentation may occur in the skin and lead to different symptoms, dependent on the function of the reactive T‐cell.
Monoclonal antibodies to amoxicillin express different idiotypes determined by anti‐idiotype antibodies production
Authors: Mayorga C. , Torres M. J. , Romano A. , Moreno F. , Sánchez‐Sabater E. , Juárez C. , Blanca M.
Published: March 2003

Penicillins are β‐lactam antibiotics able to generate several antigenic determinants that are recognized by the immune system. To study the differences in the antigen binding site of two monoclonal antibodies (Mab) specific to amoxicillin, polyclonal rabbit anti‐idiotypic antibodies were produced. One Mab, AO3.2 (IgG2a), specific to a structure formed by the acyl‐side chain structure and a part of the nuclear region of amoxicillin. The second one, AO6.2 (IgE), is specific to the side chain of amoxicillin, although it also recognizes the side chain of other penicillins (penicillin G and ampicillin). These antibodies were used to immunize rabbits in order to produce polyclonal anti‐idiotipic antibodies, which were purified in several steps by affinity chromatography. The specificity and cross‐reactivity studies were made by ELISA and ELISA inhibition. The results suggest that the anti‐Id antibodies produced are the internal image of the antigen, since the binding to their specific idiotype is blocked mainly by the original hapten (amoxicillin): in 98% of the cases with anti‐id‐1 (induced against AO3.2) and in 59% with anti‐id‐2 (induced against AO6.2). The absence of cross‐reactivity of each anti‐idiotypic antibody with the different Mabs specific to amoxicillin shows that the idiotypes induced by the same hapten have differences that are reflected by the nonrecognition of these anti‐idiotypes. We conclude that such a small molecule as amoxicillin can present several antigenic determinants that induce a panel of antibody specificities especially directed against the side chain.
Anti‐T‐cell strategies in the treatment of allergic disease
Authors: Larché M.
Published: March 2003

Specific allergen immunotherapy (SIT) has been shown to be effective in modulating allergic responses in diseases such as rhinitis and asthma. However, the ability of whole allergen to cross link mast cell bound IgE, resulting in release of mediators such as histamine, has limited the application of this therapy to carefully selected patients who have failed conventional pharmacotherapy. The use of peptide sequences corresponding to T cell epitopes of the allergen has been postulated as an alternative to SIT in which high molar doses of T cell epitope can be delivered over a shorter time period and with improved safety. Using peptides from the sequence of the major cat allergen, Fel d 1, we have demonstrated the ability to induce transient T cell activation, resulting in isolated late asthmatic reactions, which are followed by prolonged periods of allergen‐specific hyporesponsiveness, both to peptide re‐challenge and to cutaneous challenge with whole allergen. Thus, peptide therapy may prove safe and efficacious in the treatment of allergic diseases.
Chronic urticaria: impact of allergic inflammation
Authors: Hein R.
Published: March 2003

Urticaria is defined by weals (hives), with or without angioedema, that appear and peak in minutes to hours, usually disappear within 24 h, and are accompanied by pruritus that worsens during the night. Urticaria is caused by cutaneous mast cell degranulation, attributed to immunological, nonimmunological, and idiopathic causes. Chronic idiopathic urticaria (CIU) is the diagnosis when the pathophysiological mechanism of persistent urticaria remains unclear; up to half of patients with CIU have functional autoantibodies directed against the high‐affinity receptor for IgE (FcεRI) or against IgE itself, which appear to induce mast cell degranulation. Systemic histamine H1‐receptor antagonists, such as desloratadine, are central to the management of CIU. The efficacy and safety of desloratadine, 5 mg once daily, was studied in a double‐blind, randomized, placebo‐controlled, multicentre trial that included 190 patients, ages 12 and above, with at least a 6‐week history of CIU and experiencing a flare of at least moderate severity. Desloratadine was superior to placebo in controlling pruritus and total symptoms after the first dose, and its superiority was maintained throughout the full 6 weeks of the study. Measures of sleep, daily activity, therapeutic response, and global CIU status were also significantly improved with desloratadine after the first dose and maintained throughout the study.
A brief review of cryptic duplications of 21q as an emerging cause of Down syndrome: practical considerations for accurate detection
Authors: Garcia‐Heras Jaime , Nagesh Rao P
Published: March 2003

We review five cryptic duplications of 21q in patients with Down syndrome (DS) that were inherited from parental balanced translocations. All cases were identified by fluorescence in situ hybridization (FISH) and/or DNA diagnosis because the phenotype was inconsistent with the initial cytogenetic studies. These rearrangements seem to escape detection without expanded testing and are probably more frequent than expected. For this reason we propose a series of steps combining objective clinical diagnostic criteria, FISH and DNA methods to achieve an accurate ascertainment.
A microdeletion syndrome due to a 3‐Mb deletion on 19q13.2 – Diamond–Blackfan anemia associated with macrocephaly, hypotonia, and psychomotor retardation
Authors: Cario H , Bode H , Gustavsson P , Dahl N , Kohne E
Published: March 2003

We report on a boy with congenital pure red blood cell aplasia [Diamond–Blackfan anemia (DBA)] and severe congenital hypotonia, macrocephaly, hypertelorism, a broad and tall forehead, medial epicanthus, and facial hypotonia with mouth‐breathing and drooling, an affable and out‐going personality, and a general psychomotor retardation. These features show similarity to the phenotype of the X‐linked FG syndrome. DBA was diagnosed at the age of 4 months, and the boy underwent treatment with transfusion and with prednisolone. He had a normal 46, XY karyotype, but fluorescence in situ hybridization (FISH) analysis to metaphase chromosomes revealed a 3‐Mb deletion on 19q13.2. This chromosomal region has previously been linked to the DBA phenotype and one 19q13 microdeletion has been identified in a patient with DBA. This deletion coincides with the deletion reported here.

We suggest that the complex phenotype of our patient, including both DBA and the associated features, represent a microdeletion syndrome.
Functional characterization of two low density lipoprotein receptor gene mutations by fluorescence flow cytometric assessment of receptor activity in stimulated human T‐lymphocytes
Authors: Raungaard Bent , Kjærulf Jensen Henrik , Uffe Brorholt‐Petersen Jens , Heath Finn , Færgeman Ole
Published: March 2003

We report a functional characterization of the W23X and W66G low density lipoprotein (LDL) receptor gene mutations. The authors used two‐color fluorescence flow cytometry to measure LDL receptor activity in stimulated T‐lymphocytes, prepared from patients heterozygous for the W23X or W66G mutation, and compared the results with measurements of LDL receptor activity in stimulated T‐lymphocytes prepared from unrelated healthy control subjects. It was found that the W23X mutation significantly reduced LDL receptor expression and LDL binding and internalization, and that the W66G mutation significantly reduced LDL receptor expression and LDL binding. LDL internalization in patients heterozygous for the W66G mutation was not significantly reduced. The data support the concepts that the W23X mutation prevents production of LDL receptors (class I) and that the W66G mutation produces LDL receptors unable to recycle normally in cells (class V).
A germline mutation at the extreme 3′ end of the APC gene results in a severe desmoid phenotype and is associated with overexpression of beta‐catenin in the desmoid tumor
Authors: Couture Jean , Mitri Angela , Lagace Real , Smits Ron , Berk Terri , Bouchard Henri‐Louis , Fodde Riccardo , Alman Benjamin , Bapat Bharati
Published: March 2003

Desmoid tumors arise sporadically or as part of the extraintestinal manifestations of familial adenomatous polyposis (FAP). In FAP, two distinct clinical presentations of the desmoid phenotype are seen: 1) one or a few desmoid tumors present predominantly in the abdominal wall or the abdomen; 2) a florid proliferation of tumors early in life, mostly near the axial skeleton or extremities. These different phenotypes have been associated with different sites of germline mutations in the adenomatous polyposis coli gene (APC gene).  We present a large, French‐Canadian kindred with a florid desmoid tumor phenotype caused by a germline mutation at codon 2643‐2644 of the APC gene. The phenotype was characterized by the early onset of multiple tumors, arising near the axial skeleton and in proximal extremities. The penetrance of desmoid tumors was near 100% in this kindred. However, the expression of the disease was variable amongst the different affected relatives. Many gene carriers had cutaneous cysts. Polyposis of the colon was rarely observed in the affected individuals and we did not document upper gastro‐intestinal polyps. The mutant APC allele did not express a stable truncated protein in vivo. Molecular analysis of the proband's tumor DNA revealed a somatic inactivating mutation of the wild‐type allele. Immunohistochemistry on the tumor also demonstrated elevated levels of beta‐catenin.  The present study demonstrates that this extreme 3′APC mutation is associated with a severely penetrant desmoid phenotype and attenuated polyposis coli. It also suggests the involvement of the beta‐catenin pathway in the development of desmoid tumors in FAP. The natural history of the disease is variable between individuals, and surgical interventions have to be timed appropriately due to the frequent recurrences.
Perceptions of Ashkenazi Jewish breast cancer patients on genetic testing for mutations in BRCA1 and BRCA2
Authors: Phillips Kelly‐Anne , Warner Ellen , Meschino Wendy S , Hunter Jon , Abdolell Mohamed , Glendon Gordon , Andrulis Irene L , Goodwin Pamela J
Published: March 2003

The perceived benefits and risks of genetic testing may vary between groups of individuals with different cultural, demographic, and family history features. This multicentre study examined the factors that influenced the decision to undergo genetic testing for BRCA1 and BRCA2 in Canadian Jewish women with breast cancer. A self‐administered questionnaire was developed and distributed to 134 individuals enrolled in a research‐based testing program for Ashkenazi women. The questionnaire assessed demographic, social, and family history parameters, and the influence of medical, family, social, psychological, and cultural/religious factors on decision making about genetic testing. Seventy‐six percent of women completed the questionnaire. Forty‐one percent of study participants had no family history of breast or ovarian cancer. The most important factors influencing the decision to undergo testing were a desire to contribute to research, potential benefit to other family members, curiosity, and the potential for relief if not found to be a carrier (endorsed by 87, 78, 70, and 60% of participants, respectively). The main perceived risks of undergoing genetic testing related to insurance discrimination, confidentiality, accuracy and interpretability of results, potential impact on marriage prospects for family members, and focus on the Jewish community (endorsed by 28, 24, 30, 17, and 14% of participants, respectively). This study provides novel information on the motivating factors for BRCA1 and BRCA2 mutation testing in Canadian women of Ashkenazi Jewish descent. The focus on altruistic factors and those related to perceived psychological benefits of testing is notable.
Sporadic cardiac and skeletal myopathy caused by a de novo desmin mutation
Authors: Park Kye‐Yoon , Dalakas Marinos C , Semino‐Mora Christina , Lee Hee‐Suk , Litvak Svetlana , Takeda Kazuyo , Ferrans Victor J , Goldfarb Lev G
Published: March 2003

Desmin myopathy is a familial or sporadic disorder characterized by intracytoplasmic accumulation of desmin in the muscle cells. We and others have previously identified desmin gene mutations in patients with familial myopathy, but close to 45% of the patients do not report previous family history of the disease. The present study was conducted to determine the cause of desmin myopathy in a sporadic patient presenting with symmetrical muscle weakness and atrophy combined with atrioventricular conduction block requiring a permanent pacemaker. A novel heterozygous R406W mutation in the desmin gene was identified by sequencing cDNA and genomic DNA. Expression of a construct containing the patient's mutant desmin cDNA in SW13 (vim‐) cells demonstrated a high pathogenic potential of the R406W mutation. This mutation was not found in the patient's father, mother or sister by sequencing and restriction analysis. Testing with five microsatellite markers and four intragenic single nucleotide polymorphisms excluded alternative paternity. Haplotype analysis indicates that the patient's father was germ‐line mosaic for the desmin mutation. We conclude that de novo mutations in the desmin gene may be the cause of sporadic forms of desmin‐related cardiac and skeletal myopathy.
Short arm rearrangements of sex chromosomes with haploinsufficiency of the SHOX gene are associated with Leri–Weill dyschondrosteosis
Authors: Palka G , Stuppia L , Guanciali Franchi P , Chiarelli F , Fischetto R , Borrelli P , Giannotti A , Fioretti G , Rinaldi Mm , Mingarelli R , Rappold Ga , Calabrese G
Published: March 2003

Twelve patients with different features of Turner syndrome, and with Xp and Yp rearrangements involving the pseudoautosomal region (PAR1) are described. In all patients, FISH analysis showed loss of one copy of the Short Stature Homeobox (SHOX)‐containing gene. Ten patients had short stature and one disproportionate (mesomelic) normal stature, while the last one had normal stature. Skeletal abnormalities, including shortened ulna, were detected in nine subjects, and in six of them Madelung deformity was observed. These clinical data indicated a genotype–phenotype correlation between haploinsufficiency of SHOX, and short stature and skeletal abnormalities.
Low density lipoprotein receptor‐related protein gene polymorphisms and risk for late‐onset Alzheimer's disease in a Japanese population
Authors: Hatanaka Y , Kamino K , Fukuo K , Mitsuda N , Nishiwaki‐Ueda Y , Sato N , Satoh T , Yamamoto H , Yoneda H , Imagawa M , Miki T , Ohta S , Ogihara T
Published: March 2003

Low density lipoprotein (LDL) receptor‐related protein (LRP) gene polymorphisms located in the 5′ region and in exon 3, and the apolipoprotein E (APOE) genotype were determined in 100 Japanese patients affected by late‐onset Alzheimer's disease (AD). We matched 246 controls for age and found no association between the polymorphism located in the 5′ region of the LRP gene. The distribution of LRP exon 3 genotypes and alleles did not differ between AD and the control groups. However, the frequency of T allele in the Alzheimer's group having APOE‐ɛ4 was lower than that in the control group having APOE‐ɛ4, but it was only marginally significant (p=0.022). Age of onset was significantly younger in the patients with CC genotype than those carrying the T allele (p=0.03), and this trend was more evident among non‐APOE‐ɛ4 carriers (p=0.008). These results support the possibility that ApoE and LRP may contribute to the development of AD.
Sudden death caused by pulmonary thromboembolism in Proteus syndrome
Authors: Slavotinek AM , Vacha SJ , Peters KF , Biesecker LG
Published: March 2003

We report 3 patients with Proteus syndrome (PS) who died suddenly from pulmonary embolism (PE). The first patient was a male diagnosed with PS at 12 years who had varicose veins, portal vein thrombosis, right iliac vein occlusion and recurrent PE. At age 25 years, he was admitted to the hospital with a severe headache. Despite therapeutic doses of warfarin, investigations for an acute episode of breathlessness showed PE and he was unable to be resuscitated. The second case was a 9‐year‐old male with PS who collapsed at home and could not be revived. Autopsy revealed that the cause of death was a PE associated with thrombosis of the deep veins (DVT). The third patient was a 17‐year‐old female undergoing inpatient treatment for sinusitis when she unexpectedly arrested. She could not be revived and a full autopsy revealed a large PE with no identified DVT.  We conclude that PE is a serious complication of PS and recommend vigilance concerning the signs and symptoms of thrombosis and PE in individuals with PS, including children. Aggressive evaluation and treatment should be considered urgently in patients with PS and signs or symptoms of DVT.
Influence of 699C→T and 1080C→T polymorphisms of the cystathionine β‐synthase gene on plasma homocysteine levels
Authors: Aras Ö , Hanson NQ , Yang F , Tsai MY
Published: March 2003

The association of moderately elevated total homocysteine (tHcy) levels with coronary artery disease is increasingly being recognized. However, the role of genetic influence on plasma tHcy levels is not completely understood. We studied 1 055 individuals with respect to the effect of two silent polymorphisms, the 699C→T and the 1080C→T, of the cystathionine β‐synthase (CBS) gene on plasma tHcy levels. Individuals who were heterozygous or homozygous for the T699 allele had lower post‐methionine load (PML) tHcy levels when compared to individuals with the C/C genotype. This association was statistically significant (p=0.005) for the T/T genotype compared to the C/C genotype and became even more significant (p=0.000002) when individuals carrying the 68‐bp insertion (844ins68) and the T1080 allele were excluded from the analysis. With regard to the 1080C→T polymorphism, the T1080 allele was associated with significantly lower PML tHcy levels only when individuals carrying the 844ins68 and T699 allele were excluded from the study (p=0.01 for 1080T/T genotype compared to 1080C/C genotype). We speculate that the 699C→T and 1080C→T polymorphisms may be in linkage disequilibrium with regulatory elements that upregulate CBS gene transcription.
Recombinant Down syndrome: a case report and literature review
Authors: Lazzaro SJ , Speevak MD , Farrell SA
Published: March 2003

We report a case of a child with features of Down syndrome (DS) but with an atypical karyotype. Initial chromosome analysis was 46,XX,dup(21q).ish 21(wcp21+). The father's chromosomes were normal. However, the mother was found to have mosaicism for a pericentric inversion of chromosome 21 (19/30 cells). The revised chromosome result of the child was 46,XX,rec(21)dup(21q)inv(21)(p12q21.1)mat. A literature review of similar cases (hereafter referred to as rec dup(21q)) was conducted to aid counselling about recurrence risks and the prognosis for this child. All previous reports of rec dup(21q) were secondary to a maternal pericentric inversion. Male carriers did not seem to be at risk of having offspring with the rec dup(21q), although the number of male carriers was limited. In those with rec dup(21q), the risk of congenital heart disease was similar to that of trisomy 21. In reported cases, the facial appearance was suggestive of Down syndrome but perhaps less striking. Although the data are limited, there is an indication the developmental disabilities and short stature are milder in those with rec dup(21q) compared to trisomy 21. These observations promote the concept that the region of chromosome 21 proximal to the duplication contains genetic information contributing to the expression of some features of Down syndrome.
Familial aggregation of QT‐interval variability in a general population: results from the NHLBI Family Heart Study
Authors: Hong Y , Rautaharju Pm , Hopkins Pn , Arnett Dk , Djoussé L , Pankow Js , Sholinsky P , Rao Dc , Province Ma
Published: March 2003

QT‐interval prolongation is associated with increased risk of cardiac death. Although information on genetics and molecular mechanisms of the congenital long QT syndrome is mounting, limited data are available on the genetics of QT interval in the general population. Heart rate adjusted QT intervals (Bazett's QTc, and QT index (QTI)) were assessed by electrocardiography in 2399 members aged 25–91 years of 468 randomly selected families participating in the NHLBI Family Heart Study. Familial correlation and segregation analyses were performed to evaluate the genetics of the variability of QT interval in this population. The parent–offspring (0.14±0.03) and sibling (0.18±0.03) correlations for age and sex‐adjusted QTc were moderate, while the spouse correlation was close to zero (0.09±0.06). This suggests that there are familial/genetic influences on QT‐interval variability. Segregation analysis results suggest that there is a major effect in addition to heritable multifactorial effects (h2=0.34), but the major effect did not follow Mendelian inheritance. Further adjustments of QTc for other major cardiovascular risk factors did not significantly change the results. Similar results were found for QTI.  The QT‐interval variation in the general population is influenced by moderate heritable multifactorial effects in addition to a major effect. A major gene effect is not directly supported.
Identification of novel C253Y missense and Y864X nonsense mutations in the insulin receptor gene in type A insulin‐resistant patients
Authors: Osawa H , Nishimiya T , Ochi M , Niiya T , Onuma H , Kitamuro F , Kaino Y , Kida K , Makino H
Published: March 2003

Mutations in the insulin receptor gene have been reported in cases of type A insulin resistance. We report herein two cases of type A insulin resistance, which involve some novel mutations. Case 1 is a heterozygote of the C253Y missense mutation and case 2 is a heterozygote of the Y864X nonsense mutation. In the C253Y missense mutation in exon 3, a cysteine residue is replaced with tyrosine in the cysteine‐rich domain of the α subunit. The Y864X in exon 13 results in a truncated receptor, which is devoid of most of the β subunit. This mutant receptor could not be expressed on a cell membrane since the transmembrane domain is missing. Other significant mutations were not found for the entire coding regions and splice/donor sites.
Genetic analysis of early onset cerebellar ataxia with retained tendon reflexes in four Tunisian families
Authors: Marzouki N , Belal S , Benhamida C , Benlemlih M , Hentati F
Published: March 2003

Spinocerebellar ataxias comprise a poorly understood group of inherited degenerative neurological diseases. Attempts to classify hereditary ataxias on the basis of the neurological features or specific clinical signs such as tendon reflex changes have proven to be unsatisfactory.  Early onset cerebellar ataxia (EOCA) is generally inherited as an autosomal‐recessive trait. Thus far, we do not have accurate answers to several questions about its classification. However, significant clinical heterogeneity observed in four Tunisian families with typical EOCA clinical features reinforces the hypothesis of genetic heterogeneity underlying this phenotype.  We have demonstrated that three of the four families studied were not linked to Friedreich's ataxia (FA), vitamin E deficiency ataxia (AVED), and autosomal dominant cerebellar ataxia (ADCA) loci. The fourth family showed homozygosity for a large pathological expansion of GAA repeat in all patients, the parents being heterozygous for this mutation. We have also noted, in the case of the family studied, that there was instability in the transmission of the mutation, along with a phenomenon of anticipation comparable to that observed in dominant triplet repeat diseases.  EOCA is thus clinically indistinguishable from FA, yet genetically independent of all known candidate genes. Genetic mapping is required for research into the causal gene and an understanding of the disease's physiopathologic mechanisms.
Cryptic familial t(11;18)(q25;q23) incidentally detected by interphase FISH
Authors: Schultz Ln , Schmidt P , Tabor A , Bryndorf T , Christensen B , Lundsteen C
Published: March 2003

During a prospective prenatal study of numerical abnormalities of chromosomes 13, 18, 21, X and Y using locus‐specific probes, we incidentally found a case with only one signal for chromosome 18 per cell in a chorionic villus sampling (CVS) associated with an otherwise apparently normal G‐banded karyotype. This led us to discover a cryptic t(11;18) segregating in a four‐generation family. The CVS was performed because of mental retardation in the brother to the father of the fetus. A subtelomeric chromosome 18 probe revealed one signal on 18qter and one on 11qter of the father. Thus the father had a balanced reciprocal t(11;18) in spite of the apparently normal G‐banded karyotype. Using the same probes, we found an unbalanced translocation 46,XX,‐18,+der (18)t(11;18)‐(q25;q23)pat in the fetus. Further investigation of the family showed the translocation in balanced and unbalanced form in four generations in mentally normal and retarded individuals, respectively. The study emphasizes the need for a follow‐up with molecular cytogenetic techniques in dysmorphic and retarded children.
APOC3, CETP, fibrinogen, and MTHFR are genetic determinants of carotid intima‐media thickness in healthy men (the Stanislas Cohort)
Authors: Pallaud C , Sass C , Zannad F , Siest G , Visvikis S
Published: March 2003

The purpose of this study was to examine the relationship between carotid intima‐media thickness (CIMT) inter‐individual variability and 16 polymorphisms of 11 genes associated with cardiovascular risk factors (genes among lipid and homocysteine metabolisms, blood viscosity, platelet aggregation, leukocyte adhesion and renin‐angiotensin system).  CIMT was measured by high resolution B‐mode ultrasonography in an healthy population of 77 men and 84 women, aged 35–54 years and selected from a French Cohort: the Stanislas Cohort.  The polymorphisms studied were genotyped by a multilocus approach. Statistical analyses were carried out by ANOVA, after adjustment of CIMT for age, body mass index, and smoking, and by multiple regression analyses. No association was found with APOB Thr71Ile, APOC3 −482C/T, −455T/C, GpIIIa P1A, AT1R 1166A/C, AGT Met235Thr, CBS Ile278Thr, SELE 98G/T, and SELE Ser128Arg, polymorphisms neither in men nor in women. Although, in women we did not find any association for APOC3 3206T/G, 3175C/G, 1100C/T, CETP Ile405Val, MTHFR 677C/T and fibrinogen –455G/A polymorphisms; in men these polymorphisms were associated with CIMT variability (p≤0.01; p≤0.05). The most interesting finding was that altogether these genes in men were able to explain a considerable part, 20.6%, of CIMT variability. Therefore, our study gives a new opportunity to understand CIMT variability.
The DSL domain in mutant JAG1 ligand is essential for the severity of the liver defect in Alagille syndrome
Authors: Yuan Zr , Okaniwa M , Nagata I , Tazawa Y , Ito M , Kawarazaki H , Inomata Y , Okano S , Yoshida T , Kobayashi N , Kohsaka T
Published: March 2003

Alagille syndrome (AGS) is a congenital multi‐system anomaly mainly characterized by paucity of intrahepatic bile ducts caused by haploinsufficiency of the Jagged 1 gene (JAG1). To explore the relationship between genotype and phenotype, we analyzed the JAG1 gene in 25 Japanese AGS families at the genomic DNA level and identified 15 point mutations and one large deletion. Analysis of the genotype and phenotype strongly indicated that the Delta/Serrate/Lag‐2 (DSL) domain in JAG1 protein played an essential role in determining the severity of the liver disorder. In four sporadic cases, missing an entire DSL domain in mutant JAG1 resulted in progressive liver failure and all 4 patients needed a liver transplant at a very young age. This correlation was further confirmed by statistical analysis (χ2=9.143, p<0.001). Our finding demonstrated that the DSL domain in JAG1 appears to be essential for normal liver development and function.
LDL‐receptor gene mutations and the hypocholesterolemic response to statin therapy
Authors: Brorholt‐Petersen JU , Jensen HK , Raungaard B , Gregersen N , Faergeman O
Published: March 2003

Studies of the cholesterol lowering effect of statin therapy as a function of low‐density lipoprotein (LDL)‐receptor mutation type have not produced a clear picture, possibly because they included patients with several different kinds of LDL‐receptor mutations. We studied the response to treatment with fluvastatin in 28 patients with heterozygous familial hypercholesterolemia as a result of a receptor‐negative mutation (Trp23‐stop) and in 30 patients with a receptor‐binding defective mutation (Trp66‐Gly) to test the hypothesis that response to treatment depends on the type of mutation. Patients were randomized to 12 weeks of treatment with fluvastatin 40 mg daily and 12 weeks of placebo treatment, preceded by a placebo run‐in period of 8 weeks in a double‐blind, cross‐over design. Untreated plasma concentrations of lipids and lipoproteins were similar in the two groups of patients. Plasma cholesterol and LDL cholesterol response to therapy tended to be less marked in receptor‐binding defective patients, but the differences were not statistically significant. A tabulation of the results of the present and earlier studies suggests that differences in treatment response as an apparent function of LDL‐receptor gene mutational type occur mainly in populations with recent genetic admixture (<400 years). In such populations, persons with the same mutation in the LDL‐receptor gene are more likely to share other but undetermined genetic variations affecting the pharmacology of statins.
Flow cytometry: an ‘old’ tool for novel applications in medical genetics
Authors: Wedemeyer N , Pötter T
Published: March 2003

Flow cytometry was originally established as an automated method for measuring optical or fluorescence characteristics of cells or particles in suspension. In the meantime, flow cytometers have become user‐friendlier, less expensive instruments with an increasing importance in clinical diagnostics. Besides the classical fields of application, such as immunophenotyping blood cells or analyzing the cell cycle status by measuring the DNA content, novel flow cytometric methods have been developed to identify and to quantify disease‐related gene sequences. Here we give an overview of current and future applications, including the detection of viral sequences via microsphere‐based PCR assays and the analysis of single nucleotide polymorphisms, reflecting individual phenotypic traits. Furthermore, flow cytometry allows the quantification of gene expression changes as well as the isolation of differentially expressed gene sequences.  Flow cytometry is also convenient for multiplex analyses, e.g. when hybridizing DNA samples to a mixture of various microsphere populations each coated with different DNA probes. Last but not least, the use of magnetic beads in combination with flow cytometers coupled with automated devices enables molecular diagnostics on a large scale. Overall, this review demonstrates flow cytometry as a rapid, sensitive, and reproducible tool applicable to a wide range of medical genetic approaches.
Familial resemblance for glucose and insulin metabolism indices derived from an intravenous glucose tolerance test in Blacks and Whites of the HERITAGE Family Study
Authors: Hong Y , Weisnagel SJ , Rice T , Sun G , Mandel SA , Gu C , Rankinen T , Gagnon J , Leon AS , Skinner JS , Wilmore JH , Bergman RN , Bouchard C , Rao DC
Published: March 2003

Type 2 diabetes mellitus (T2DM), characterized by hyperglycemia, is a complex disease primarily caused by impairment in insulin sensitivity (SI) and insulin secretion. While a strong genetic component for T2DM is well established, there are few reports on racial differences in the magnitude of the genetic effects of T2DM and indices of glucose and insulin metabolism. We report here on the familial resemblance for traits related to glucose metabolism at pre‐exercise training levels in 492 members from 99 sedentary White families and 259 members from 108 Black families participating in the multicenter HERITAGE Family Study. All these traits were obtained from the frequently sampled intravenous glucose tolerance test (IVGTT). They include glucose disappearance index (Kg), an overall index for glucose tolerance, acute insulin response to glucose (AIRGlucose) which is an index for insulin secretion, and those derived from the minimal model including SIand the disposition index (DI). DI, derived as the product of SIand AIRGlucose, is a measure of the activity of the B‐cells adjusted for insulin resistance. After adjustment for age, sex, and body mass index, the maximal heritability estimates in Blacks (Whites) are 48±14% (25±8%) for Kg, 44±14% (46±8%) for AIRGlucose, 38±12% (44±8%) for SIand 32±14% (24±8%) for DI. Interestingly, Blacks have higher heritability for overall glucose tolerance than Whites but there is no race difference in heritability estimates for insulin sensitivity or insulin secretion.
Haplotype analysis of the USH1D locus and genotype–phenotype correlations
Authors: Liu X‐Z , Blanton SH , Bitner‐Glindzicz M , Pandya A , Landa B , MacArdle B , Rajput K , Bellman S , Webb BT , Ping X , Smith RJH , Nance WE
Published: March 2003

Usher syndrome (USH) is characterised by hearing impairment and progressive pigmentary retinopathy. USH can be divided into three subtypes based on the severity and progression of the major clinical findings. These subtypes are genetically heterogeneous, with at least six loci for USH1, three for USH2 and one for USH3. In the present study, five unrelated consanguineous families with USH1 were analysed for linkage to markers flanking the six USH1 loci. Two of these families, one Pakistani and one Turkish, demonstrated linkage to the USH1D locus. In another family, haplotype segregation was consistent with linkage to USH1C. The remaining families were not linked to any of the six USH1 loci, providing support for the existence of at least one additional USH1 locus. Analysis of these two new USH1D families allowed us to narrow the USH1D candidate region to a 7.3‐cM interval with a telomeric flanking marker at D10S1752. Comparison of the affected haplotypes in our Pakistani family with the original Pakistani USH1D family yielded no evidence for a founder effect. The identification of two additional affected families suggests that the USH1D may be a more common form of USH1 than originally suspected. The USH1D (CDH23) gene has recently been cloned. Mutation analysis has shown two different CDH23 mutations in the two Pakistani USH1D families studied, which confirmed our finding that there was no evidence for a founder effect by haplotype analysis. The interesting correlations between genotype and phenotype in CDH23 are also summarised.
Association of a single nucleotide polymorphism in the TIGR/MYOCILIN gene promoter with the severity of primary open‐angle glaucoma
Authors: Colomb E , Nguyen TD , Béchetoille A , Dascotte J‐C , Valtot F , Brézin AP , Berkani M , Copin B , Gomez L , Polansky JR , Garchon H‐J
Published: March 2003

Primary open‐angle glaucoma (POAG) is a highly prevalent optic neuropathy and a major cause of irreversible blindness, with elevation of intraocular pressure (IOP) being a primary risk factor. The trabecular meshwork‐inducible glucocorticoid response (TIGR)/MYOCILIN (MYOC) gene coding region is mutated in 3–4% of POAG patients. Here, in a retrospective study of 142 POAG patients, we evaluated the influence on glaucoma phenotype of a novel biallelic polymorphism (−1000C/G) located in the upstream region of the MYOC gene. Allele frequencies were similar among patients and controls. However, the G allele (frequency 17.6%), also designated as MYOC.mt1, was associated with an increased IOP (+4.9 mmHg, p=0.0004) and a more damaged visual field (p=0.02). Both effects were predominant in females. Moreover, whereas IOP in MYOC.mt1 noncarriers decreased very markedly to the normal range between diagnosis and inclusion in the study (p=3×10−5 in both males and females), reflecting successful therapy, it decreased less noticeably in MYOC.mt1+ male patients (p=0.005) and not at all in MYOC.mt1+ female patients. MYOC.mt1 appears therefore to be an indicator of poor IOP control and greater visual field damage in diagnosed POAG patients, potentially due to a lack of response to therapeutic intervention. Its typing might help in the selection of treatment paradigms for the management of POAG patients.