bluuebunny/crossref_metadata_2025 huggingface dataset:
Syntenic assignment of dopamine tautomerase (DCT) to bovine chromosome 12
Authors: Sun H S , Womack J E , Kirkpatrick B W
Published: July 2010

Heterologous primers were used to amplify an exon and intron‐containing segment of the bovine homologue of the human dopachrome tautomerase gene. After confirmation of homo‐logy by sequence analysis (exon sequence similarity greater than 90%), bovine‐specific primers were developed for synteny mapping purposes. The dopachrome tautomerase gene was assigned to bovine chromosome 12 (BTA12) with 97% concordance to the coagulation factor 10 locus. Together with previous synteny mapping of bovine chromosome 12 genes, fms‐related tyrosine kinase, esterase D and 5‐hydroxytryptamine receptor 2, this assignment further indicates conservation between human chromosome 13q and bovine chromosome 12.
Optimization of single‐strand conformation polymorphism and sequence analysis of the mitochondrial control region in Pagellus bogaraveo (Sparidae, Teleostei): rationalized tools in fish population biology
Authors: Ostellari L. , Bargelloni L. , Penzo E. , Patarnello P. , Patarnello T.
Published: July 2010

We report the isolation and sequencing of 400–550 base pairs (bp) of the mitochondrial DNA (mtDNA) control region of eight species of Sparidae (Perciformes, Teleostei). This sequence information allowed us to design specific primers to one of these species (Pagellus bogaraveo). The new set of primers was used to test a rationalized approach to study the mtDNA nucleotide variability at the intraspecific level. The single‐strand conformation polymorphism (SSCP) technique was applied to detect sequence variation in two non‐overlapping fragments of the control region of 32 individuals of P. bogaraveo. To assess the sensitivity of the method, the nucleotide sequence of the analysed region was determined for all the specimens. The results showed that, for one of the two fragments, SSCP analysis was able to detect 100% of the underlying genetic variability. In sharp contrast, nucleotide variation of the second DNA fragment was completely unresolved by SSCP under different experimental conditions. This suggests that the resolution power of SSCP is crucially dependent on the nature of the fragment subjected to the analysis; therefore, a preliminary test of the sensitivity of the method should be performed on each specific DNA fragment before starting a large‐scale survey. A rationalized approach, combining the SSCP technique and a simplified sequencing procedure, is proposed for studying intraspecific polymorphism at the mtDNA control region in fish.
Cloning and characterization of 414 polymorphic porcine microsatellites
Authors: Alexander L J , Rohrer G A , Beattie C W
Published: July 2010

We report the sequences, sizes, and number of alleles of 414 new porcine microsatellites that were cloned in our laboratory and 21 micro‐satellites derived from GenBank DNA sequences. We also confirm the usefulness of porcine microsatellite primer pairs derived from short interdispersed elements.
Transfer of bovine microsatellites to the cervine (Cervus elaphus)
Authors: Kühn R , Anastassiadis C , Pirchner F
Published: July 2010

Bovine microsatellites were used to amplify DNA of red deer (Cervus elaphus). Fourteen of 27 bovine systems (52%) displayed polymorphism, while no (CA)n‐repeat was detected in seven systems and six systems gave no amplificates in red deer. The allele number ranged from 2 to 7, the polymorphism information content between 0.24 and 0.76. The results demonstrate that transfer of microsatellite systems between families of the same order (artiodactyla) is possible. Molecular genetic research will help to clarify the differentiation and ecology of wild animals and will contribute to define criteria needed for the preservation of endangered species.
The porcine TTR locus maps to chromosome 6q
Authors: Archibald A L , Couperwhite S , Jiang Z H
Published: July 2010

The sequence of a cDNA clone encoding porcine transthyretin (prealbumin) was used to develop polymorphic markers for the TTR locus. The single‐strand conformation polymorphism (SSCP) detected is caused by a silent AIT mutation in the penultimate coding codon and can also be revealed as a SacI restriction fragment length polymorphism (RFLP). The TTR locus was mapped to chromosome 6q by segregation and linkage analysis with these polymorphisms. This assignment confirms the predictions of homology between human chromosome 18 and pig chromosome 6q2.5‐2.6.
Alignment of the PiGMaP and USDA linkage maps of porcine chromosomes 3 and 9
Authors: Groenen M A M , de Vries B J , van der Poel J J
Published: July 2010

The European Pig Gene Mapping Project (PiGMaP) and United States Department of Agriculture (USDA) porcine linkage maps for chromosomes 3 and 9 have been aligned by typing three USDA microsatellites from chromosome 3 and five from chromosome 9 on the PiGMaP reference families. Using the crimap linkage analysis package, revised multipoint linkage maps were constructed for chromosome 3 and 9. Inclusion of these USDA markers in the multipoint analysis resulted in an increase in length of 47% and 33% respectively for these two PiGMaP linkage groups. This increase in size is mainly the result of extension of the ends of both linkage groups.
Variation in the control region sequence of the sheep mitochondrial genome
Authors: Wood N. J. , Phua S. H.
Published: July 2010

The DNA sequences of the control region of the mitochondrial genome of fifty unrelated sheep were determined in order to ascertain the extent and distribution of its variability. A consensus sequence was derived, and 1081 differences from it were observed amongst the fifty animals. Some constant groups of differences were observed that were held in common by a number of animals, which thus fell into two main groups, although neither group was typical of anyof the breeds sampled. The consensus sequence also allowed comparison between the control region sequences of sheep and other mammals. The sequence contains four tandem repeats of a 75 base‐pair motif that accounts for the difference in its size from the cattle control region, to which it is otherwise very similar. Comparison with the cattle sequence allowed the determination of the homologues of various functionally important sites. The homologues of the transcription promoters, the origin of replication and the central conserved sequence block were all identified by this method.
Chicken red blood cells as a substrate for direct polymerase chain reaction
Authors: Khatib H. , Gruenbaum Y.
Published: July 2010

A protocol is presented for direct polymerase chain reaction (PCR) amplification of DNA from chicken nucleated red blood cells. Chicken blood in EDTA was found to have a strong inhibitory effect on the PCR. Consequently, PCR using this protocol should be performed only on a narrow range of blood volumes, from 0125 to 025 |ll.
Cytogenetic mapping of immunity‐related genes in the domestic horse
Authors: Musilova P. , Kubickova S. , Vychodilova‐Krenkova L. , Kralik P. , Matiasovic J. , Hubertova D. , Rubes J. , Horin P.
Published: July 2010

Chromosomal locations of 19 horse immunity‐related loci (CASP1, CD14, EIF5A, FCER1A, IFNG, IL12A, IL12B, IL12RB2, IL1A, IL23A, IL4, IL6, MMP7, MS4A2, MYD88, NOS2A, PTGS2, TFRC and TLR2) were determined by fluorescence in situ hybridization. For IFNG and PTGS2, this study is a confirmation of their previously reported position. In addition, microsatellite (HMBr1) was localized in the same region as IFNG. All genes were assigned to regions of conserved synteny and the data obtained in this study enhance the comparative human–horse map. Cytogenetic localization of IL6 to ECA4q14–q21.1 suggested a new breakage point that changes the order of loci compared with HSA7. The map assignments of these loci serve as anchors for other loci and will aid in the search for candidate genes associated with traits in the horse.
Application of bovine microsatellite markers for genetic diversity analysis of Swiss yak (Poephagus grunniens)
Authors: Nguyen T. T. , Genini S. , Ménétrey F. , Malek M. , Vögeli P. , Goe M. R. , Stranzinger G.
Published: July 2010

In order to assess the applicability of bovine microsatellite markers for population genetic studies in Swiss yak, 131 bovine microsatellite markers were tested on a panel of 10 animals. Efficient amplification was observed for 124 markers (94.6%) with a total of 476 alleles, of which 117 markers (94.3%) were polymorphic. The number of alleles per locus among the polymorphic markers ranged from two to nine. Seven loci (ILSTS005, BMS424B, BMS1825, BMS672, BM1314, ETH123 and BM6017) failed to amplify yak genomic DNA. Two cattle Y‐chromosome specific microsatellite markers (INRA126 and BM861) amplified genomic DNA from both male and female yaks. However, two additional markers on cattle Y‐chromosome (INRA124 and INRA189) amplified DNA from only males. Of the polymorphic markers, 24 microsatellites proposed by CaDBase for within‐ and cross‐species comparisons and two additional highly polymorphic markers (MHCII and TGLA73) were used to investigate the genetic variability and the population structure of a Swiss yak herd that included 51 additional animals. The polymorphic information content ranged from 0.355 to 0.752, while observed heterozygosity (HO) ranged from 0.348 to 0.823. Furthermore, a set of 13 markers, organized into three multiplex polymerase chain reactions, was evaluated for routine parentage testing. This set provided an exclusion probability in a family of four yaks (both parents and two offspring) of 0.995. These microsatellites serve as useful tools for genetic characterization of the yak, which continues to be an important domestic livestock species.
Diverse products of near‐surface sediment mobilization in an ancient eolianite: outcrop features of the early Jurassic Navajo Sandstone
Authors: Bryant Gerald , Miall Andrew
Published: July 2010

The Navajo Sandstone of the Colorado Plateau, USA, displays a wide range of soft‐sediment deformation (SSD) features, including decametre‐scale features that have not been found in any modern desert environment. Laboratory simulations and partial analogues from other depositional environments suggest that these features derived from episodic liquefaction and fluidization of unconsolidated dune deposits. Outcrop details at many locations preserve the effects of fluid‐escape dynamics through porous, permeable, well‐sorted sand, which was partitioned by subtle textural changes at depositional boundaries between successive dune deposits and, less commonly, by distinct lithofacies changes marking the interface between wet and dry depositional environments. Extreme deformation and turbulent sediment flow have effaced primary structures in some zones of deformation; but other sites preserve the ductile modification of primary structures. Some outcrops preserve evidence of dramatic alterations in topography and sedimentation patterns due to localized compaction and large, subsurface, sediment displacements. Particularly notable among these extraordinary features are those representing the foundering of active dunes, sediment eruptions, and the subsidence of interdune surfaces in the ancient erg. The distinctive patterns of deformation in the Navajo Sandstone, interpreted in the context of its primary sedimentary architecture, provide unique insights into environmental conditions in the region during the Early Jurassic. Widespread ductile deformation in the Navajo indicates extraordinarily wet conditions for an active dune environment. The truncation of successive deformation features by deflation surfaces establishes the episodic nature of deformation in the unit and also suggests climatic variability. The widespread, episodic occurrence of SSD features, viewed within the seismic context provided by palaeotectonic reconstructions, strongly suggests that earthquake triggering of localized liquefaction controlled the distribution of deformation in this unit.
Three‐dimensional seismic analysis of the morphology and spatial distribution of chimneys beneath the Nyegga pockmark field, offshore mid‐Norway
Authors: Hustoft Steinar , Bünz Stefan , Mienert Jürgen
Published: July 2010

Three‐dimensional (3D) seismic data analysis has identified 415 seismic chimneys in the Nyegga area, on the mid‐Norwegian continental margin. The majority of the seismic chimneys connect with pockmarks at the modern seafloor, suggesting that the seismic chimneys represent the acoustic imprint of highly focused, cross‐stratal fluid flow. A semi‐quantitative spatial analysis of the seismic chimney complex, measuring parameters describing the dimension, geometry and root zone, provides constraints on the geological controls on chimney formation. Most of the seismic chimneys in the Nyegga region descriptively fall into the categories of ‘blowout chimneys’, because they diagnostically link to topography‐controlled leakage locations within strata showing acoustic indications of free gas. We find that approximately 60% of the fluid‐escape chimneys emanate from free gas layers below the gas hydrate stability zone. The timing of highly focused fluid expulsion through chimneys and the formation of the Nyegga pockmark field postdate the last glacial maximum at 25 000 years before present.
How do we know when peanut and tree nut allergy have resolved, and how do we keep it resolved?
Authors: Byrne A. M. , Malka‐Rais J. , Burks A. W. , Fleischer D. M.
Published: July 2010

Over the last two decades, the prevalence of peanut and tree nut allergy has increased throughout the western world. Adverse reactions to these foods account for over 50% of all deaths resulting from food‐related anaphylaxis. Until recently, evidence suggested that all peanut and tree nut allergy were permanent. It is now known that about 20% and 10%, respectively, of young patients outgrow peanut and tree nut allergies. Achieving tolerance is associated with increasing circulating T regulatory cells and reduced production of allergen‐specific IgE. Reliable predictors of resolution are not yet available. A direct correlation between skin test weal size and allergen‐specific IgE, at the time of diagnosis and likelihood of resolution, has been reported. Resolution of peanut or tree nut allergy cannot be determined conclusively by either allergen‐specific IgE analysis or by skin prick testing. Oral food challenge is the gold standard for determining resolution of food allergy. Food challenges should only be undertaken in a clinical setting fully equipped to deal with a potential severe adverse reaction. Approximately 8% of patients who outgrow peanut allergy may suffer a recurrence, but recurrent tree nut allergy has not been reported to date. Infrequent ingestion of peanut may be related to the re‐emergence of allergy. Induction of tolerance through oral immunotherapy or sublingual immunotherapy is now being actively studied, but remains experimental. Studies have reported short‐term desensitization to peanut, but ongoing follow‐up will determine whether tolerance is achieved long term.

Cite this as: A. M. Byrne, J. Malka‐Rais, A. W. Burks and D. M. Fleischer, Clinical & Experimental Allergy, 2010 (40) 1303–1311.
Hereditary haemorrhagic telangiectasia
Authors: SHARATHKUMAR A. A. , SHAPIRO A.
Published: July 2010

Summary.  Hereditary haemorrhagic telangiectasia (also known as Osler‐Weber‐Rendu syndrome) is a relatively common, under‐recognized autosomal‐dominant disorder that results from multisystem vascular dysplasia. It is characterized by telangiectases and arteriovenous malformations of skin, mucosa and viscera. This article summarizes the clinical manifestations and the management of this disorder and its management. This review underscores an urgent need to conduct prospective multicentre studies to develop evidence‐based management guidelines for this disease.
Diagnosis and treatment of inherited factor X deficiency
Authors: BROWN D. L. , KOUIDES P. A.
Published: July 2010

Summary.  Factor X is a vitamin K‐dependent, liver‐produced serine protease that serves a pivotal role in coagulation as the first enzyme in the common pathway to fibrin formation. Inherited factor X deficiency is a rare autosomal recessive bleeding disorder that is estimated to occur in 1:1 000 000 individuals up to 1:500 carriers. Several international registries of FX‐deficient patients have greatly expanded the knowledge of clinical phenotype. A proposed classification of severity is based on FX:C activity measurements: an FX:C measurement <1% is severe, an FX:C measurement of 1–5% is moderate and an FX:C measurement of 6–10% is mild. Levels above 20% are infrequently associated with bleeding and heterozygotes are usually asymptomatic. Among patients with FX:C levels <10%, unlike moderate or severe haemophilia A and B, mucocutaneous bleeding symptoms such as epistaxis and menorrhagia occur in the majority. In addition, patients with moderate–severe deficiency may have symptoms similar to that of haemophilia A and B, including haemarthrosis, intracranial haemorrhage, and gastrointestinal bleeding. Genotype characterization may offer important clues about clinical prognosis. More than 80 mutations of the F10 gene have been identified, most of which are missense mutations. There is no specific FX replacement product yet readily available, but fresh frozen plasma and prothrombin complex concentrates can be used for treatment of bleeding symptoms and preparation for surgery.
Original article: Extraction of lipid from scallop (Patinopecten yessoensis) viscera by enzyme‐assisted solvent and supercritical carbon dioxide methods
Authors: Zhou Da‐Yong , Zhu Bei‐Wei , Tong Lei , Wu Hai‐Tao , Qin Lei , Tan Hui , Chi Ya‐Li , Qu Jing‐Ya , Murata Yoshiyuki
Published: July 2010

In the present study, lipid was extracted from scallop (Patinopecten yessoensis) viscera by using the enzyme‐assisted solvent method and the supercritical carbon dioxide (SC‐CO2) method. Soxhlet extraction with ethyl ether produced a yield of 23.7 ± 0.6 g of lipid 100 g−1 of dry matter. Enzyme‐assisted solvent extraction allowed recovering 60.6 ± 1.5% of P. yessoensis viscera lipid from the samples treated with papain, whereas a lipid recovery rate of 78.3 ± 0.6% was achieved by SC‐CO2 extraction. The lipid extracted was divided into the unsaponifiable fraction (sterol) and the saponifiable fraction (fatty acid) and analysed by gas chromatography mass spectrometry. Results indicated that the fatty acid composition and sterol composition for lipids extracted by different methods were slightly different. Eicosapentaenoic acid and docosahexaenoic acid were dominant polyunsaturated fatty acids accounting for 35–40% of the total fatty acid.
Bisphosphonate‐associated osteonecrosis of the jaw. Knowledge and attitudes of dentists and dental students: a preliminary study
Authors: López‐Jornet Pia , Camacho‐Alonso Fabio , Molina‐Miñano Francisco , Gomez‐Garcia Francisco
Published: July 2010

Objective  The objective was to determine the knowledge of dentists and dental students of bisphosphates and osteonecrosis of the jaw (ONJ) in the Autonomous Community of Murcia, Spain.

Material and method  A structured questionnaire was elaborated on knowledge, attitude and practice regarding aspects of the aetiology, diagnosis and prevention of bisphosphonate‐associated ONJ. The questionnaire was administered to two groups: group I with 60 dental students and group II, 60 dentists with well‐established professional activity.

Results  As regards the subjects’ knowledge of bisphosphonate‐associated osteonecrosis, 30 (50%) students and 41 (68.36%) dentists (P = 0.0041) had up to date knowledge. Most correctly identified risk factors involved. Only eight (13.33%) students and 20 (33.33%) dentists (P = 0.010) knew how to treat osteonecrosis once established.

Conclusions  Training strategies need to be established as regards bisphosphonate‐associated ONJ.
Does systematically organized care improve outcomes for women with diabetes?
Authors: Lowe Julia , Byles Julie , Dolja‐Gore Xenia , Young Anne
Published: July 2010

Objective  To investigate whether financial reimbursement for a bundle of diabetes care items self‐reported by general practitioners (GPs) leads to improved outcomes for women with diabetes.

Methods  Longitudinal cohort study of women in the Australian Longitudinal Study on Women's Health aged 45–50 and 70–75 years when recruited in 1996.

Outcomes  Short Form 36‐item (SF‐36), Medicare and pharmaceutical benefits costs 2002–2005, uptake of annual cycle of care for diabetes (ACC).

Results  Annual cycle of care claims were identified for 23% of 388 mid‐age, and 40% of 616 older women with diabetes. ACC was not associated with statistically significantly higher costs in either group. Women for whom the GP had received an ACC fee were more likely to have been overweight, had more GP visits, more medications, and more ‘no cost’ visits. Unlike older women, mid‐age women for whom the GP had received an ACC fee were more likely to have difficulty managing on their income and tended to have worse physical and social function scores prior to the time the ACC was introduced and compared with other women with diabetes continued to have poorer scores at subsequent surveys.

There was no association between ACC, co‐morbidities or country of birth. Women who developed diabetes after the first survey (incident cases) tended to have better SF‐36 health profile scores and lower costs than those who reported diabetes on the first survey (prevalent cases).

Conclusions  General practitioners of women with diabetes, who have more health care encounters and poorer health‐related quality of life, have adopted ACC with little impact on the decline in quality of life of the women nor on health care costs.
Discharge management for patients in Flemish psychiatric hospitals
Authors: Desplenter Franciska , Laekeman Gert , Moons Philip , Simoens Steven
Published: July 2010

Rationale and aims  At the end of the 1990s, a case management service called ‘discharge management’ was implemented in Belgian psychiatric hospitals. This study aimed to describe the profile of patients receiving discharge management in Flemish psychiatric hospitals as well as to analyse the discharge management indicators of these hospitals on micro and meso level.

Methods  Ten Flemish psychiatric hospitals participated. A descriptive analysis of the profile of patients receiving discharge management (gender, age, length of stay, family situation, assistance at home, living environment and Global Assessment of Functioning) and of the indicators of discharge management (screening, interdisciplinary patient file, interdisciplinary meeting, timely announcement of discharge date, transfer of discharge documents, readmission and institutionalization) were carried out.

Results  Of the 1306 patients included in the database, one‐fourth received discharge management. In general, patients (54% were male) were about 45 years old, stayed for 55 days in hospital, were single and had no aid at home. Most of them came from and returned to their own home. On the micro level of discharge management, nearly all patients were screened and half of them were positively screened. Half of these patients received discharge management. Of the discharged patients who received discharge management (meso level), 13.5% were institutionalized after discharge, 37.6% had been previously admitted and 27.2% received discharge management during that previous admission. Differences between diagnostic groups occurred.

Conclusion  Hospitals have made efforts to support and prepare patients for discharge, although a number of improvements are suggested for different indicators.
Hypoglycaemia with pioglitazone: analysis of data from the Prescription‐Event Monitoring study
Authors: Vlckova Veronika , Cornelius Victoria , Kasliwal Rachna , Wilton Lynda , Shakir Saad
Published: July 2010

Aims and objectives  To investigate the relationship between patients' characteristics, use of concomitant anti‐diabetic therapies and the incidence of hypoglycaemia, an acute complication of the treatment of diabetes mellitus reported by general practitioners (GPs) during the first 9 months of the treatment with pioglitazone.

Methods  We used data collected for the Prescription‐Event Monitoring (PEM) study conducted by the Drug Safety Research Unit for patients prescribed pioglitazone between November 2000 and June 2001 by their GP in England. A Cox proportional‐hazards regression model was used to assess this relationship.

Results  The original pioglitazone PEM cohort included 12 772 patients (mean age 60.9 years); 53% (6777) were male. A total of 77 patients experienced at least one hypoglycaemic episode (9.64 per 1000 patient‐years). Women were estimated to have twice the hazard of having a hypoglycaemic event compared with men [hazard ratio (HR) 2.05; confidence interval (CI) 1.24, 3.41]. Patients taking combination therapy with sulfonylurea or insulin were estimated to have approximately three and four times the hazard of having an event compared with those who were not taking these adjunctive therapies [HR = 3.11 (CI 1.64, 5.88); HR = 4.15 (CI 1.74, 9.91), respectively]. Patients treated with adjunctive metformin were 25% less likely to experience hypoglycaemia than those who did not take concomitant metformin (HR = 0.75; CI 0.44, 1.27).

Conclusions  This study has shown that the treatment with pioglitazone was associated with a low incidence of hypoglycaemia. The factors possibly increasing the risk of hypoglycaemia were concomitant therapy with sulfonylurea or insulin and female gender.
New aspects of HbA1c as a risk factor for cardiovascular diseases in type 2 diabetes: an observational study from the Swedish National Diabetes Register (NDR)
Authors: Eeg‐Olofsson K. , Cederholm J. , Nilsson P. M. , Zethelius B. , Svensson A.‐M. , Gudbjörnsdóttir S. , Eliasson B.
Published: July 2010

Abstract.  Eeg‐Olofsson K, Cederholm J, Nilsson PM, Zethelius B, Svensson A‐M, Gudbjörnsdóttir S, Eliasson B (Institute of Medicine, Sahlgrenska University Hospital, University of Gothenburg, Gothenburg; Department of Public Health and Caring Sciences/Family Medicine and Clinical Epidemiology, Uppsala University, Uppsala; Department of Clinical Sciences, Lund University, University Hospital, Malmö; Department of Public Health and Caring Sciences/Geriatrics, Uppsala University, Uppsala; and Center of Registers in Region Västra Götaland, Gothenburg, Sweden) New aspects of HbA1c as a risk factor for cardiovascular diseases in type 2 diabetes: an observational study from the Swedish National Diabetes Register (NDR). J Intern Med 2010; 268: 471–482.

Aims.  To analyse the association between glycosylated haemoglobin A1c (HbA1c) and cardiovascular disease (CVD) in patients with type 2 diabetes in the Swedish National Diabetes Register (NDR).

Methods.  An observational study of 18 334 patients (age 30–79 years, previous CVD in 18%, baseline HbA1c 5.0–10.9%) who were followed for 6 years (mean 5.6 years) from 1997/1998 until 2003.

Results.  Hazard ratios per 1% unit increase in baseline or updated mean HbA1c for fatal/nonfatal coronary heart disease (CHD), CVD and total mortality were 1.11–1.13, 1.10–1.11 and 1.09–1.10, respectively (all P < 0.001), adjusted for several risk factors and clinical characteristics in Cox regression. Adjusted 6‐year event rates increased with higher baseline or updated mean HbA1c with no J‐shaped risk curves, in all patients and also when subgrouping by shorter (mean 3 years) or longer (mean 14 years) diabetes duration, by presence or absence of previous CVD, or by treatment with oral hypoglycaemic agents (OHAs) or insulin. Risk reductions of 20% for CHD and 16% for CVD (P < 0.001) were found in patients with a baseline mean HbA1c of 6.5%, compared to those with a mean level of 7.5%. Compared to OHA‐treated patients, insulin‐treated patients had an increased risk of total mortality, due almost exclusively to an increased risk of non‐CVD mortality, and due less to a weakly significant increased risk of fatal CVD. HbA1c was not associated with non‐CVD mortality.

Conclusions.  This observational study showed progressively increasing risks of CHD, CVD and total mortality with higher HbA1c, and no risk increase at low HbA1c levels even with longer diabetes duration, previous CVD or treatment with either insulin or OHAs. Patients achieving HbA1c <7% showed benefits for risk reduction.
Specificity of IgE antibodies to sequential epitopes of hen's egg ovomucoid as a marker for persistence of egg allergy
Authors: Järvinen K.‐M. , Beyer K. , Vila L. , Bardina L. , Mishoe M. , Sampson H. A.
Published: July 2010

Background:  Approximately two‐thirds of egg‐allergic infants become tolerant within the first 5 years of life.

Objective:  We sought (1) to compare the recognition of sequential (linear) and conformational binding sites of ovomucoid, ovalbumin and ovotransferrin, by IgE antibodies of children with persistent and transient egg allergy, (2) to identify immunodominant IgE‐and IgG‐binding epitopes of ovomucoid, and (3) to compare epitope‐specificity of IgE antibodies between patients with differing natural histories of egg allergy.

Methods:  Using immunodot‐blots or ImmunoCAPs, IgE‐antibodies against conformational (native) and sequential (reduced and alkylated) egg proteins were determined at the time of clinical reactivity in patients who retained their allergy and in those who developed clinical tolerance. IgE‐ and IgG‐binding epitopes were mapped for ovomucoid using overlapping decapeptides on SPOTs membranes. Recognition of the major IgE‐binding epitopes were compared between patients with differing natural histories of egg allergy.

Results:  The patients with long‐lasting egg allergy had a higher concentrations of IgE antibodies against sequential and native ovomucoid and ovalbumin than the children who subsequently gained tolerance (P < 0.01). Four major IgE‐binding epitopes were identified in ovomucoid at amino acid 1–10, 9–20, 47–56, and 113–124. IgE antibodies of all seven patients with persistent egg allergy recognized these epitopes whereas none of the 11 children who outgrew their egg allergy did so.

Conclusions:  Patients with persistent egg allergy develop IgE antibodies against more sequential and conformational epitopes of ovomucoid and ovalbumin. The presence of serum IgE antibodies to specific sequential epitopes of ovomucoid may be used as a screening instrument for persistent egg allergy.
Putative association of a TLR9 promoter polymorphism with atopic eczema
Authors: Novak N. , Yu C.‐F. , Bussmann C. , Maintz L. , Peng W.‐M. , Hart J. , Hagemann T. , Diaz‐Lacava A. , Baurecht H.‐J. , Klopp N. , Wagenpfeil S. , Behrendt H. , Bieber T. , Ring J. , Illig T. , Weidinger S.
Published: July 2010

Background:  Toll‐like receptors (TLR) play a pivotal role in the induction of first‐line defense mechanisms of the innate immune system and trigger adaptive immune responses to microbial pathogens. Genetic variations in innate immunity genes have been reported to be associated with a range of inflammatory disorders. Deficiencies on the level of immunity receptors such as pathogen‐recognition receptors are suspected to affect the maturation of our immune system and to avail thereby the high prevalence of atopic diseases and susceptibility of atopic patients to microbial infections.

Aims of the study:  We evaluated TLR9 as susceptibility gene for atopic eczema (AE).

Methods:  Analyses of four tag single‐nucleotide polymorphisms in two panels of families containing a total of 483 parent‐affected offspring trios as well as a cohort of 274 unrelated adult AE cases and 252 hypernormal population‐based controls have been performed.

Results:  In both family cohorts, polymorphism C‐1237T, which is located within the promoter region of the TLR9 gene, was significantly associated with AE, in particular the intrinsic subtype of AE. No associations were seen in the case–control cohort. Luciferase reporter gene assays revealed significantly higher promoter activity of the TT allelic variant at this single nucleotide polymorphism site.

Conclusion:  These observations suggest that the TLR9 promoter polymorphism C‐1237T might affect AE susceptibility in particular in patients with the intrinsic variant of AE.
Facial thermography is a sensitive and specific method for assessing food challenge outcome
Authors: Clark A. T. , Mangat J. S. , Tay S. S. , King Y. , Monk C. J. , White P. A. , Ewan P. W.
Published: July 2010

Background:  Oral challenge is widely used for diagnosing food allergy but variable interpretation of subjective symptoms may cause error. Facial thermography was evaluated as a novel, objective and sensitive indicator of challenge outcome.

Methods:  A total of 24 children with a history of egg allergy underwent oral challenge, which were scored positive when objective symptoms occurred or negative after all doses were consumed without reaction. Facial temperatures were recorded at baseline and 10‐min intervals. The difference between mean and baseline temperature (ΔT), maximum ΔT during challenge (ΔTmax) and area under curve of ΔT against time (ΔTAUC) were calculated for predefined nasal, oral and forehead areas, and related to objective challenge outcome.

Results:  There were 13 positive and 11 negative challenges. Median nasal ΔTAUC and ΔTmax were greater in positive compared with negative challenges (231‐ and 5‐fold, respectively; P < 0.05). In positive challenges, nasal temperatures showed an early transient rise at 20 min, preceding objective symptoms at median 67 min. There was a sustained temperature increase from 60 min, which was reduced by antihistamines. A cut‐off for nasal ΔTmax of 0.8°C occurring within 20 min of the start of the challenge predicted outcome with 91% sensitivity (positive predictive value [PPV] 100%) and 100% specificity (negative predictive value [NPV] 93%). Subjective symptoms occurred in four of 13 positive and three of 11 negative challenges.

Conclusions:  Facial thermography consistently detects a significant early rise in nasal temperature during positive compared with negative food challenges, which is evident before objective symptoms occur. Thermography may therefore provide a sensitive method to determine outcome of food challenges and investigate the pathophysiology of food allergic reactions.
Diagnosis and natural course of allergy to cooked potatoes in children
Authors: De Swert L. F. A. , Cadot P. , Ceuppens J. L.
Published: July 2010

Background:  Allergy to heat stable potato proteins can cause severe allergic disease in children.

Objective:  To study diagnostic criteria for allergy to cooked potatoes and to describe its clinical characteristics and natural history.

Methods:  Thirty‐six children, aged 4–36 months, with atopic symptoms and having a positive potato‐CAP and/or skin prick test (SPT) were included. Potato allergy was documented by means of provocation, or elimination and reintroduction or an unequivocal clinical history. Potato‐CAP and SPT with a commercial extract were evaluated for diagnostic performance.

Results:  Presenting symptoms in children with proven potato allergy (n = 17) were eczema (16 of 17), gastrointestinal complaints (eight of 17), urticaria and/or angioedema (five of 17), wheezing/rhinitis (three of 17) and anaphylaxis (two of 17). Fifteen children had previously diagnosed cow's milk protein allergy and were egg‐sensitized. Potato‐CAP at cut‐off >2 kU/l provided a 100% sensitivity and a 62.5% specificity for diagnosis of potato allergy, while a SPT score ≥ 3 had a 100% sensitivity and a score ≥ 4 had a 100% specificity. Tolerance to cooked potato was achieved in 80% of subjects at age 16–102 months. Of 12 subjects having reached the age of 3 years during the study, 10 were re‐evaluated at age 3–6 years: seven of 10 subjects had developed clinical pollen allergy, compared with four of 18 atopic controls (P < 0.05).

Conclusions:  Potato‐CAP and SPT at specific cut‐off are valuable tools in the diagnosis of allergy to cooked potato. Most children with potato allergy develop tolerance at mean age of 4 years. Allergy to cooked potatoes is a risk factor for the development of pollen allergy.
The prevalence, cost and basis of food allergy across Europe
Authors: Mills E. N. C. , Mackie A. R. , Burney P. , Beyer K. , Frewer L. , Madsen C. , Botjes E. , Crevel R. W. R. , Van Ree R.
Published: July 2010

The development of effective management strategies to optimize the quality of life for allergic patients is currently hampered by a lack of good quality information. Estimates of how many individuals suffer from food allergy and the major foods involved vary widely and inadequacies of in vitro diagnostics make food challenges the only reliable means of diagnosis in many instances. The EuroPrevall project brings together a multidisciplinary partnership to address these issues. Cohorts spanning the main climatic regions of Europe are being developed in infants through a birth cohort, community surveys in school‐age children and adults and an outpatient clinic study. Confirmatory double‐blind placebo‐controlled food challenge diagnosis is being undertaken using foods as they are eaten with titrated doses to allow no‐effect and lowest‐observable effect levels for allergenic foods to be determined. The cohorts will also facilitate validation of novel in vitro diagnostics through the development of the EuroPrevall Serum Bank. Complementary studies in Ghana, western Siberia, India and China will allow us to gain insights into how different dietary patterns and exposure to microorganisms affect food allergies. New instruments to assess the socioeconomic impact of food allergy are being developed in the project and their application in the clinical cohorts will allow, for the first time, an assessment to be made of the burden this disease places on allergy sufferers and their communities.
Eczematous reactions to food in atopic eczema: position paper of the EAACI and GA2LEN
Authors: Werfel T. , Ballmer‐Weber B. , Eigenmann P. A. , Niggemann B. , Rancé F. , Turjanmaa K. , Worm M.
Published: July 2010

Food allergy and atopic eczema (AE) may occur in the same patient. Besides typical immediate types of allergic reactions (i.e. noneczematous reactions) which are observed in patients suffering from AE, it is clear that foods, such as cow’s milk and hen’s eggs, can directly provoke flares of AE, particularly in sensitized infants. In general, inhaled allergens and pollen‐related foods are of greater importance in older children, adolescents and adults. Clinical studies have revealed that more than 50% of affected children with AE that can be exacerbated by certain foods will react with a worsening of skin eczema either alone or in addition to immediate symptoms. Adolescents and adults may also react to foods, but reactions to ‘classical’ food allergens, such as hen’s eggs and cow’s milk, are not as common as in childhood. Some patients with AE do react to pollen‐associated foods. Food‐induced eczema should not be neglected by the allergologist: On the one hand, food can be a relevant trigger factor of persistent moderate‐to‐severe AE; on the other hand, unnecessary diets which are not based on a proper diagnosis may lead to malnutrition and additional psychological stress on patients suffering from AE. Eczematous reactions to food can only be diagnosed by a thorough diagnostic procedure, taking into account the patient’s history, the degree of sensitization and the clinical relevance of the sensitization. The latter has often to be proven by oral food challenges. Upon oral food challenge it is most important to evaluate the status of the skin with an established score (e.g. SCORAD, EASI) after 24 h and later because otherwise worsening of eczema will be missed.
The sodium‐dependent glucose cotransporter SLC5A11 as an autoimmune modifier gene in SLE
Authors: Tsai L.‐J. , Hsiao S.‐H. , Tsai L.‐M. , Lin C.‐Y. , Tsai J.‐J. , Liou D.‐M. , Lan J.‐L.
Published: July 2010

Genetic studies in several human autoimmune diseases suggest that the pericentromeric region of chromosome 16 might harbor an autoimmune modifier gene. We hypothesized that the sodium‐dependent glucose cotransporter gene SLC5A11 is such a gene, and so might interact with immune‐related genes. Herein, this hypothesis was tested in a genetic evaluation of the multiple gene effect in systemic lupus erythematosus (SLE). We used the case–control candidate gene association approach. Eight immune‐related genes involved in inflammation and autoantibody generation and clear‐up [interleukin 1 receptor antagonist (IL1RN), interleukin 1‐beta (IL1‐β), tumor necrosis factor‐alpha (TNF‐α), lymphotoxin‐alpha (LTA), tumor necrosis factor ligand superfamily, member 6 (TNFSF6), programmed cell death 1 (PDCD1), C2, and complement component 4 (C4)] were selected for study. Frequency of each candidate’s genotype and allele between case and control were compared. Results were stratified by reanalyzing genotype data with relevant symptoms. Finally, improved computational data mining was used to analyze the phenotypes in a large data set. In the frequency analysis, only IL1‐β was significantly associated with SLE. Stratification analysis showed a significant association with SLE symptoms between SLC5A11 and the other immune‐related genes, with the exceptions of TNFSF6 and C4. SLC5A11 was significantly associated with low C4 (as was TNF‐α), anti‐Smith antibody (anti‐Sm) (as was C2), serositis, and alopecia. Finally, SLC5A11 interacted with PDCD1, TNF‐α, LTA, and C4. After our study, we concluded that SLC5A11 is involved with some immune effects and interacts with immune‐related gene(s), consistent with its function as an autoimmune modifier gene. Furthermore, SLC5A11 might induce apoptosis through the TNF‐α, PDCD1 pathway. The present genotype–phenotype mapping approach should be applicable to genetic study of other complex diseases.
The Formation of Triple‐Helical Collagen Molecules from α1 or α2 Polypeptide Chains
Authors: Tkocz C. , Kühn K.
Published: July 2010

The triple‐helical collagen molecule of vertebrates consists of two or three non‐identical polypeptide chains: [(α1)2(α2)] or [(α1)(α2)(α3)]. We have attempted to form molecules from three identical chains: (α1)3 or (α2)3. The yield of (α1)3‐molecules obtained after renaturation of α1 from calf or rat skin collagen and the yield of [(α1)2(α2)]molecules after renaturation of unfractionated collagen from both species is almost the same. (α1)3‐molecules are able to form fibrils of the native type and long‐spacing segments. Their melting temperature of approximately 34° is only slightly less than that of native molecules. In contrast the (α2)3‐molecules is much less stable with a melting range of 20–24°. Therefore, (α2)3‐molecules are formed at lower temperatures and with a lower yield than (α1)3‐ and [(α1)2(α2)]molecules. Renaturation experiments with a mixture of α1 and α2 (ratio 2:1) shows that the reformation of the [(α1)2(α2)]molecule is favoured over the (α1)3‐molecule, in spite of the fact that both molecule have almost the same stability. Long‐spacing segments from (α1)3, (α2)3 or native [(α1)2(α2)]molecules exhibit a similar if not identical crossstriation pattern in the electron microscope. Therefore, the conclusion can be drawn that the α1 and α2 chains are homologous.
Untersuchungen über den Abbaumechanismus einer gereinigten Polygalakturonase aus Aspergillus niger
Authors: Koller A. , Neukom H.
Published: July 2010

Using an Aspergillus niger enzyme preparation, a pure polygalacturonase has been isolated by chromatography on cellulose phosphate.

The alkali‐labile enzyme has an isoelectric point of 4.5 and a pH optimum of 5.5; its activation energy is 8.3 kcal/mole.

The enzyme hydrolyzes 48% of the glycosidic bonds in pectic acid. Besides di‐ and monogalacturonic acid, the hydrolysate mostly contains trigalacturonic acid.

Depending on the degree of esterification, the methyl ester of pectic acid is degraded with varying intensity. If the degree of esterification exceeds 75%, pectin is no longer degraded. Acetylation of the secondary alcohol groups, up to 70%, has little effect.

Di‐ and trigalacturonic acid are not hydrolyzed. Tetragalacturonic acid is split into mono‐and trigalacturonic acid, penta‐ and hexagalacturonic acid into tri‐, di‐ and monogalacturonic acid.

The isolated enzyme is an endopolygalacturonase. The study of the degradation products strongly suggests that two carboxyl groups of pectic acid become attached to the binding sites of the enzyme molecule. These two groups are separated from the active center by three and six galacturonic acid units respectively.

Blocking of the binding sites at higher substrate concentrations causes a decrease in enzyme activity.
Membrane Adenosine Triphosphatase of Micrococcus lysodeikticus
Authors: Muñoz E. , Salton M. R. J. , Ng M. H. , Schor M. T.
Published: July 2010

A protein component which accounts for 10% of the total protein of isolated cell membranes of Micrococcus lysodeikticus has been partially characterized. The protein can be released into solution from the membrane complex and in this “soluble” state it possesses Ca++‐dependent adenosine triphosphatase activity. This enzyme has been purified about 50‐fold, by a procedure based on its association‐dissociation properties with the membrane. The progressive purification and degree of homogeneity have been demonstrated by examining the various stages in the soluble ATPase preparation by disc electrophoresis and by gel diffusion against membrane antiserum. On treatment with 8 M urea or 1% sodium dodecyl sulfate followed by disc electrophoresis, clear evidence of dissociation was obtained, indicating the existence of subunits. The purified protein has a sedimentation coefficient of 14–15 S as determined by sucrose gradient centrifugation or analytical ultracentrifugation. A sedimentation coefficient of 3.5 S has been estimated for the main product of the protein dissociated by sodium dodecyl sulfate again an indication of the dissociation of the protein into subunits, probably identical on the basis of molecular weight. The “soluble” enzyme was activated by Ca++, optimum activation being observed at ATP/Ca++ ratio of 1/1 and pH 7.5 under which conditions 50–60 μmoles of ATP were hydrolyzed per mg protein per 10 min at 37°. The ATPase was quite stable at 4°. The Ca++‐dependent ATPase activity was inhibited by ADP, sodium azide, and Mg++.

When the membrane‐ATPase complex remains stable, no hydrolytic activity for ATP is shown unless trypsin activation. Our system shows similarities with the “allotopic” properties described for some mitochondrial enzymes. From this and previous work in our laboratory a close relationship between the “stalked particles” seen in M. lysodeikticus cell membranes by electron microscopy and ATPase appears to be evident.