bluuebunny/crossref_metadata_2025 huggingface dataset:
Linkage mapping of the retinoic acid receptor‐γ gene to porcine chromosome 5
Authors: Messer L , Wang L , Yelich J , Pomp D , Geisert R , Rothschild M F
Published: July 2010

The porcine retinoic acid receptor‐γ gene (RARG) has been mapped by restriction fragment length polymorphism analysis to porcine chromosome 5. The placement of RARG distal to the diacylglycerol kinase gene increases the length Of the existing map (PiGMaP) and adds a fifth type‐I marker to this sparsely mapped chromosome. This augments the homology of pig chromosome 5 and human chromosome 12 established by previous comparative mapping.
Allele frequencies of the major milk proteins in the Finnish Ayrshire and detection of a new K‐casein variant
Authors: Ikonen T , Ruottinen O , Erhardt G , Ojala M
Published: July 2010

A total of 20990 Finnish Ayrshire cows were phenotyped for the major milk proteins by isoelectric focusing in polyacrylamide gels. The predominant alleles in the Finnish Ayrshire were αS1‐casein B (0.999), αS2‐casein A (0.991), β‐casein A1 (0.509) and α2 (0.490), α22‐casein A (0.612) and β‐lactoglobulin B (0.716). The K‐casein E allele (0.307) was also rather common in the Finnish Ayrshire. A new K‐casein variant (K‐casein F) was demonstrated in two Finnish Ayrshire cows, a dam and a daughter.
Restriction fragment length polymorphism in amplification products of the bovine butyrophilin gene: assignment of bovine butyronhilin to bovine chromosome 23
Authors: Taylor C , Everest M , Smith C
Published: July 2010

A polymorphism was identified in the bovine butyrophilin (BTN) gene by digesting poly‐merase chain reaction products with the restriction enzyme HaeIII. This polymorphism was segregating in a Holstein‐Friesian sire selected as part of an ongoing study directed towards the identification of quantitative trait loci affecting milk composition. Screening of a half‐sib family established for the heterozygous sire allowed the localization of BTN to bovine chromosome 23 (BTA23).
Characterization and linkage mapping often sheep microsatellite markers derived from a sheep x hamster cell hybrid
Authors: Lumsden J M , Lord E A , Montgomery G W
Published: July 2010

A cosmid library has been constructed from a sheep x hamster cell hybrid containing sheep chromosome t1, rob (6;24). Clones containing sheep DNA were identified by hybridizing to a total sheep genomic DNA probe. Small fragments (<500 bp) containing (AC)n microsatellites were subcloned and sequenced. Ten microsatellite markers were characterized and six were mapped back to chromosomes 6 and 24. The remaining microsatellites mapped to chromosome 26, which was shown to be present in a small proportion of cells of the cell line.
The porcine skeletal muscle ryanodine receptor gene structure coding region 1 to 10 614 harbouring 71 exons
Authors: Wen G , Leeb T , Reinhart B , Brenig B
Published: July 2010

The skeletal muscle ryanodine receptor (RYR1) belongs to a family of calcium release channels that are expressed in different tissues. The RYR1 gene is one of the largest genes characterized, so far, containing a 15 253 nucleotide ORF in swine. To study the genomic organization of the porcine skeletal muscle ryanodine receptor gene we have isolated seven genomic fragments spanning 72.7kb of chromosomal DNA of chromosome 6q12. This region harbours exons 1 to 71 coding for 3538 amino acids (69.6%) of the ryanodine receptor 1.
Polymorphism at the ovine major histocompatibility complex class II loci
Authors: Escayg A P , Hickford J G H , Bullock D W , Montgomery G W , Dodds K G
Published: July 2010

Southern hybridization analysis of the ovine major histocompatibility complex (MHC) (MhcOvar) class II region, using sheep‐specific probes for the DQA1, DQA2, DQB and DRA loci, has revealed extensive polymorphism. DQA1 and DQAP had eight and 16 alleles respectively, DQB had six and DRA had three alleles. Little information was derived from the DRB locus owing to extensive cross‐hybridization between the DRB probe and the DQB locus. Differences in allele frequency between breeds were revealed. At the DQA1 locus a null allele (DQA1‐N) was observed with a frequency of between 27% and 45%, making this the most common DQA1 allele in all breeds examined. The frequency of DQA1‐N homozygotes was between 11% and 18%, raising questions as to the functional significance of the DQA1 gene. Linkage analysis between the DQA1, DQA2, DQB and DRA loci did not reveal any recombination.
Characterization of isogenic carp (Cyprinus carpio L.) lines with a genetically determined high or low antibody production
Authors: Wiegertjes G F , Voorthuis P , Groeneveld A , Van Muiswinkel W B , Stet R J M , Bongers A B J , Doulabi B Zandieh
Published: July 2010

Antibody production to dinitrophenyl‐keyhole limpet haemocyanin (DNP‐KLH) served as the immune parameter to divergently select carp (Cyprinus carpio L.) to produce high‐ and low‐responder F, hybrid lines. Antibody production to trinitrophenyl‐lipopolysaccharide [TNP‐LPS) and to DNP‐KLH were similar in magnitude. By contrast, some high‐responder lines were low responders to DNP‐human serum albumin, and vice versa. Low‐responder carp were relatively susceptible to infection with the parasite Trypanoplasma borreli. This suggested that at least one gene with a major influence on resistance differed between the two homozygous parents (69, 85) used to generate the high‐ and low‐responder homozygous families, respectively. The isogenic lines showed no within‐line variation in DNA fingerprints, but differed with respect to their MhcCyca‐DAB genes.
Conservation of the RD‐BF‐C2 organization in the pig MHC class‐III region: mapping and cloning of the pig RD gene
Authors: Peelman L. J. , Mattheeuws M. , VAN Zeveren A. , van de Weghe A. , Bouquet Y.
Published: July 2010

The RD gene, named after the arginine (R) and aspartic acid (D) repeat in the central part of its protein, was initially mapped in the mouse H‐2S subregion between C4 and BF. It was later mapped in the same position in the human MHC and here we show it is also conserved in the pig MHC class III region, close to the complement BF gene. A pig RD genomic clone was isolated from a γ‐phage library. Hybridizations on genomic DNA separated with pulsed field gel electrophoresis identified common 220kb Nrul, 130 kb EagI and 200 kb Mlul bands for RD, BF and C2. The RD gene has also a 17 kb Kpnl and 11 kb Sad fragment in common with BFbut not with C2. The close linkage of the RD and BF genes was further established by hybridization of BF to a genomic γ‐phage clone also containing the RD gene. This genomic RD clone overlaps with a γ ‐phage clone previously isolated and containing the complete BF gene and the 3′ part of C2. The distance between RD and BF is about 6 kb. The junction between the two complement genes BF and C2 was sequenced and the BF 5′ promoter region, overlapping the 3′ noncoding region of C2, was compared with that of the human BF promoter. The overall homology was about 80% and all but one identified promoter elements were found in the same position in both genes. The results obtained demonstrate the RD‐BF‐C2 organization is strongly conserved between human, mouse and pig. No polymorphisms were detected in either the RD gene or in the BF promoter region using polymerase chain reaction and restriction fragment polymorphism analysis.
Genetic mapping of quantitative trait loci affecting susceptibility in chicken to develop pulmonary hypertension syndrome
Authors: Rabie T. S. K. M. , Crooijmans R. P. M. A. , Bovenhuis H. , Vereijken A. L. J. , Veenendaal T. , van der Poel J. J. , Van Arendonk J. A. M. , Pakdel A. , Groenen M. A. M.
Published: July 2010

Pulmonary hypertension syndrome (PHS), also referred to as ascites syndrome, is a growth‐related disorder of chickens frequently observed in fast‐growing broilers with insufficient pulmonary vascular capacity at low temperature and/or at high altitude. A cross between two genetically different broiler dam lines that originated from the White Plymouth Rock breed was used to produce a three‐generation population. This population was used for the detection and localization of quantitative trait loci (QTL) affecting PHS‐related traits. Ten full‐sib families consisting of 456 G2 birds were typed with 420 microsatellite markers covering 24 autosomal chromosomes. Phenotypic observations were collected on 4202 G3 birds and a full‐sib across family regression interval mapping approach was used to identify QTL. There was statistical evidence for QTL on chicken chromosome 2 (GGA2), GGA4 and GGA6. Suggestive QTL were found on chromosomes 5, 8, 10, 27 and 28. The most significant QTL were located on GGA2 for right and total ventricular weight as percentage of body weight (%RV and %TV respectively). A related trait, the ratio of right ventricular weight as percentage to total ventricular weight (RATIO), reached the suggestive threshold on this chromosome. All three QTL effects identified on GGA2 had their maximum test statistic in the region flanked by markers MCW0185 and MCW0245 (335–421 cM).
Subsurface sediment remobilization and fluid flow in sedimentary basins: an overview
Authors: Huuse Mads , Jackson Christopher A.‐L. , Van Rensbergen Pieter , Davies Richard J. , Flemings Peter B. , Dixon Richard J.
Published: July 2010

Subsurface sediment remobilization and fluid flow processes and their products are increasingly being recognized as significant dynamic components of sedimentary basins. The geological structures formed by these processes have traditionally been grouped into mud volcano systems, fluid flow pipes and sandstone intrusion complexes. But the boundaries between these groups are not always distinct because there can be similarities in their geometries and the causal geological processes. For instance, the process model for both mud and sand remobilization and injection involves a source of fluid that can be separate from the source of sediment, and diapirism is now largely discarded as a deformation mechanism for both lithologies. Both mud and sand form dykes and sills in the subsurface and extrusive edifices when intersecting the sediment surface, although the relative proportions of intrusive and extrusive components are very different, with mud volcano systems being largely extrusive and sand injectite systems being mainly intrusive. Focused fluid flow pipes may transfer fluids over hundreds of metres of vertical section for millions of years and may develop into mud volcano feeder systems under conditions of sufficiently voluminous and rapid fluid ascent associated with deeper focus points and overpressured aquifers. Both mud and sand remobilization is facilitated by overpressure and generally will be activated by an external trigger such as an earthquake, although some mud volcano systems may be driven by the re‐charge dynamics of their fluid source. Future research should aim to provide spatio‐temporal ‘injectite’ stratigraphies to help constrain sediment remobilization processes in their basinal context and identify and study outcrop analogues of mud volcano feeders and pipes, which are virtually unknown at present. Further data‐driven research would be significantly boosted by numerical and analogue process modelling to constrain the mechanics of deep subsurface sediment remobilization as these processes can not be readily observed, unlike many conventional sediment transport phenomena.
A literature review of interprofessional working and intermediate care in the UK
Authors: Rout Amelia , Ashby Sue , Maslin‐Prothero Sian , Masterson Abigail , Priest Helena , Beach Mike , Harrison Gina , Mann Phillip , Sturdy Deborah
Published: July 2010

Aims and objectives.  This systematic literature review aimed at addressing two questions: first, what evidence exists regarding intermediate care in the UK; and what interventions have been used to develop interprofessional working in intermediate care in the UK? A systematic review of the literature from 2000–2006 resulted in a total of 104 full‐text articles describing research into intermediate care in the UK.

Background.  The review was the first stage of a large, national project evaluating and developing interprofessional working among health and social care staff, particularly in relation to the intermediate care of older people.

Design.  Systematic literature review.

Methods.  All the literature was reviewed by one reviewer, and a second review was carried out by a team of reviewers to ensure each article was reviewed twice, independently. One article was reviewed by all the reviewers to ensure inter‐rater reliability; finally, all the reviews were amalgamated, which resulted in one summary per article.

Results.  The main findings drawn from this systematic literature review are that research carried out on intermediate care in the UK has a diverse set of aims, for example economic evaluations, delivery of intermediate care and exploring the views and perceptions of those involved in intermediate care.

Conclusions.  Although several articles include discussions about the importance of interprofessional working in intermediate care, no article specifically focused on the interprofessional focus of intermediate care, and there was no research about interventions used to develop interprofessional working.

Relevance to clinical practice.  Intermediate care as a policy has been interpreted very differently across the four countries of the UK; there is no one preferred or consistent interpretation to its delivery.
Diabetes and oral health: doctors' knowledge, perception and practices
Authors: Al‐Habashneh Rola , Barghout Nicola , Humbert Lewis , Khader Yousef , Alwaeli Hayder
Published: July 2010

Objectives  To assess Jordanian doctors' knowledge of the connection between diabetes and oral health and assess their willingness to advise their diabetic patients to seek dental treatment and determine the associated factors.

Methods  Data were collected from 164 doctors practising in Jordan using a structured questionnaire. Chi‐squared test and regression analyses were conducted to reveal factors influencing the awareness, perception and knowledge of health care professionals regarding diabetes and oral health.

Results  Of the respondents, 70% had heard of the link between diabetes and oral health. The majority agreed that diabetes increased the tendency to have periodontitis but only half advised their diabetic patients to consult a dentist concerning their oral health. Only a third of doctors agreed that oral health was an issue in controlling diabetes. Books, magazines and pamphlets were the main source of information with the rate of 58%, medical journals and medical curriculum were the second and third sources, respectively. General medical practitioners were less informed than specialized doctors about the relationship between oral health and diabetes. Factors that significantly predicted doctors would advise dental visits were: (1) being a specialist (P = 0.037); (2) having positive knowledge about the association between diabetes and oral health (P = 0.02, P = 0.007 and P = 0.004, respectively).

Conclusion  There is limited knowledge of the relationships between oral health and diabetes. The more knowledgeable doctors are, the more likely they are to make dental referrals. Screening and referral by health professionals may benefit diabetic patients by improving access to dental care. Therefore, there is a need to educate doctors about oral health and diabetes.
Are COX‐2 inhibitors preferable to combined NSAID and PPI in countries with moderate health service expenditures?
Authors: Perić Aneta , Toskić‐Radojičić Marija , Dobrić Silva , Damjanov Nemanja , Miljković Branislava , Antunović Mirjana , Vezmar Sandra
Published: July 2010

Rationale  In developed countries, cyclooxygenase 2 (COX‐2) inhibitors were shown to be less costly than the combination of non‐steroidal anti‐inflammatory drugs (NSAIDs) and proton pump inhibitors (PPIs) in treatment of patients with high risk of serious gastrointestinal (GI) adverse effects. It is questionable if such results apply to developing countries where health service costs are lower and there is high discrepancy between generic and patent protected drug prices.

We analysed the direct cost of treatment with generic NSAIDs in combination with PPIs versus branded COX‐2 inhibitors in patients with high risk of serious GI adverse effects from the perspective of the public health service in Serbia.

Methods   Total cost of treatment of serious GI complications and the use of NSAID+PPI versus COX‐2 inhibitors were calculated. A model for estimation of cost of treatment of NSAID+PPI versus COX‐2 inhibitors which included the probability of developing serious GI adverse effects was developed.

Results   Total cost of treatment of serious GI adverse effects resulted in an average of $814/patient. Considering the relative risk of such adverse effects for patients with four or more risk factors, the least costly treatment over 6 months was the use of celecoxib ($487). Compared with diclofenac+omeprazole, cost savings were estimated at $59 and $22 per patient with celecoxib and etoricoxib, respectively.

Conclusion   Cost savings may be achieved by using COX‐2 inhibitors in patients at high risk of GI adverse effects even in countries with moderate health care service expenditures. Such possibility requires further investigation.
EHR adoption among doctors who treat the elderly
Authors: Yeager Valerie A. , Menachemi Nir , Brooks Robert G.
Published: July 2010

Objectives  The purpose of this study is to examine Electronic Health Record (EHR) adoption among Florida doctors who treat the elderly. This analysis contributes to the EHR adoption literature by determining if doctors who disproportionately treat the elderly differ from their counterparts with respect to the utilization of an important quality‐enhancing health information technology application.

Methods  This study is based on a primary survey of a large, statewide sample of doctors practising in outpatient settings in Florida. Logistic regression analysis was used to determine whether doctors who treat a high volume of elderly (HVE) patients were different with respect to EHR adoption.

Results  Our analyses included responses from 1724 doctors. In multivariate analyses controlling for doctor age, training, computer sophistication, practice size and practice setting, HVE doctors were significantly less likely to adopt EHR. Specifically, compared with their counterparts, HVE doctors were observed to be 26.7% less likely to be utilizing an EHR system (OR = 0.733, 95% CI 0.547–0.982). We also found that doctor age is negatively related to EHR adoption, and practice size and doctor computer savvy‐ness is positively associated.

Conclusions  Despite the fact that EHR adoption has improved in recent years, doctors in Florida who serve the elderly are less likely to adopt EHRs. As long as HVE doctors are adopting EHR systems at slower rates, the elderly patients treated by these doctors will be at a disadvantage with respect to potential benefits offered by this technology.
Different, difficult or distinct? Mothers' and fathers' perceptions of temperament in children with and without intellectual disabilities
Authors: Boström P. , Broberg M. , Hwang C. P.
Published: July 2010

Background  Can ratings of temperament be a way of identifying young children with intellectual disabilities (ID) who are at risk for being experienced as difficult? We aimed to explore parents' reports of temperament in their young children with or without ID, as well as positive and negative impact of the child on parents.

Method  Mothers and fathers of 55 children recently diagnosed with ID and 183 age‐matched typically developing (TD) children completed the EASI Temperamental Survey and two scales of the Family Impact Questionnaire measuring positive and negative impact of the child on parents.

Results  Parents rated children with mixed ID/DD (developmental delay) as shyer and more impulsive, and less active and sociable when compared with TD children. Children with mixed ID/DD were also reported to have more negative and less positive impact on the family compared with the TD group. In subgroup analyses, children with Down syndrome and cerebral palsy/motor impairment were described as having less negative impact on parents and were described as low in negative emotionality. Children with autism spectrum disorder (ASD), ID/DD nos and other less common diagnoses had a similar pattern of temperament with high emotionality, shyness and impulsivity, and low activity and sociability. Parents of children with ASD and ID/DD reported the highest level of negative impact.

Conclusions  Temperamental characteristics such as high negative emotionality and impulsivity, which can be identified earlier than behavioural problems, could be indicators of negative impact on parents of young children with ID. Despite great variability in temperament among children with mixed ID/DD, results indicated common temperamental characteristics among children with ASD, ID/DD and other diagnosis.
Cloning and functional analysis of chicory root fructan1‐exohydrolase I (1‐FEH I): a vacuolar enzyme derivedfrom a cell‐wall invertase ancestor? Mass fingerprint of the 1‐FEH I enzyme
Authors: Van den Ende Wim , Michiels An , De Roover Joke , Verhaert Peter , Van Laere André
Published: July 2010

This paper describes the cloning and functional analysis of chicory (Cichorium intybus L.) fructan 1‐exohydrolase I cDNA (1‐FEH I). To our knowledge it is the first plant FEH cloned. Full‐length cDNA was obtained by a combination of RT–PCR, 5′ and 3′ RACE using primers based on N‐terminal and conserved amino acid sequences. Electrophoretically purified 1‐FEH I enzyme was further analyzed by in‐gel trypsin digestion followed by matrix‐assisted laser desorption ionization and electrospray time‐of‐flight tandem mass spectrometry. Functionality of the cDNA was demonstrated by heterologous expression in potato tubers. 1‐FEH I takes a new, distinct position in the phylogenetic tree of plant glycosyl hydrolases being more homologous to cell‐wall invertases (44–53%) than to vacuolar invertases (38–41%) and fructosyl transferases (33–38%). The 1‐FEH I enzyme could not be purified from the apoplastic fluid at significantly higher levels than can be explained by cellular leakage. These and other data suggest a vacuolar localization for 1‐FEH I. Also, the pI of the enzyme (6.5) is lower than expected from a typical cell‐wall invertase. Unlike plant fructosyl transferases that are believed to have evolved from a vacuolar invertase, 1‐FEH I might have evolved from a cell‐wall invertase‐like ancestor gene that later obtained a vacuolar targeting signal. 1‐FEH I mRNA quantities increase in the roots throughout autumn, and especially when roots are stored at low temperature.
Antibody blocking of MHC II on human activated regulatory T cells abrogates their suppressive potential
Authors: Peiser M. , Becht A. , Wanner R.
Published: July 2010

Natural regulatory CD4+CD25+Foxp3+ T cells control peripheral immune responses. Freshly isolated regulatory T‐cell populations are regarded as being unable to suppress the proliferation of strongly stimulated effector T cells. We now provide evidence that it is not the strength of the proliferative signal to effector T cells but activation and accessibility of regulatory T cells that determine whether suppression may occur. Human regulatory T cells were initially cocultured with allogeneic monocyte‐derived dendritic cells for a short time and were then rendered accessible for effector T cells by removal of the dendritic cells. That way activated regulatory T cells effectively suppressed the proliferation of autologous effector T cells which was strongly driven by cell‐sized Dynabeads coated with CD3/CD28 antibodies. Although regulatory T cells are known to display MHC II molecules and to upregulate their expression along with activation, a role of MHC II molecules in forming the contact to effector T cells was not yet envisaged. However, blocking of MHC II on activated regulatory T cells abrogated their suppressive potential. It should not be excluded that self‐MHC molecules on physically accessible activated regulatory T cells arrange the contact to effector T cells.
Diagnostic accuracy of the atopy patch test in children with food allergy‐related gastrointestinal symptoms
Authors: Canani R. Berni , Ruotolo S. , Auricchio L. , Caldore M. , Porcaro F. , Manguso F. , Terrin G. , Troncone R.
Published: July 2010

Background:  Little is known about the diagnostic accuracy of atopy patch tests (APT) in the clinical practice of pediatric gastroenterology. Moreover, APTs containing purified food extracts have recently become available, but their diagnostic accuracy is largely undefined.

Patients and methods:  We evaluated the diagnostic accuracy of food challenge, skin prick test (SPT), serum specific IgE determination, and APT using fresh food and commercial food extracts in parallel in children referred for suspected food allergy‐related gastrointestinal symptoms.

Results:  Eighty‐nine food challenges were performed in 60 patients (38 boys, median age 23 months, range 3–48 months): 31 tested positive for cow's milk (CM), 19 for hen's egg (HE), and two for wheat. Specific immunoglobulin E (IgE) determination, and SPT, respectively, were positive in 7/31 and 14/31 of patients with cow's milk allergy (CMA), and in 7/19 and 7/19 with HE allergy. The results of APT with fresh food vs a commercial assay were (1) CM: sensitivity: 64.5%vs 6.4%, specificity 95.8%vs 95.6%, positive predictive value (PPV) 95.2%vs 66.6% and negative predictive value (NPV) 67.6%vs 43.1%; (2) HE: sensitivity 84.2%vs 5.2%, specificity 100%vs 100%, PPV 100%vs 100% and NPV 75.0%vs 33.3%.

Conclusions:  Atopy patch test is a useful tool in the diagnostic work up of children with food‐allergy‐related gastrointestinal symptoms. The diagnostic accuracy of ATP was higher with fresh food than with commercial food extracts.
Sublingual immunotherapy with a standardized cat dander extract: evaluation of efficacy in a double blind placebo controlled study
Authors: Álvarez‐Cuesta E. , Berges‐Gimeno P. , Mancebo E. G. , Fernández‐Caldas E. , Cuesta‐Herranz J. , Casanovas M.
Published: July 2010

Background:  Little information is available on the clinical efficacy of sublingual immunotherapy (SLIT) using extracts derived from mammalian epithelia.

Objectives:  To assess clinical efficacy of cat SLIT based on natural exposure challenge test (NCT).

Material and methods:  Fifty cat allergic patients with rhinoconjunctivitis with or without asthma were included in a randomized double blind placebo controlled clinical trial of cat SLIT during 1 year. Twenty‐five patients received active treatment and 25 placebo. Sublingual immunotherapy efficacy was assessed by natural exposure challenge to a cat in a cat‐room and by skin tests. Airborne Fel d 1 levels, symptom scores and peak expiratory flow (PEF) values were monitored.

Results:  Thirty‐three (66%) out of 50 patients completed the treatment. Fel d 1 content of the maximum concentration was 0.51 μg per ml. During the build up phase, the accumulated dose was 1.7 μg of Fel d 1 and during the entire length of the study was 17.1. No adverse reports were reported. The active group showed a marked reduction (62%) in symptoms during the NCT (P < 0.001) with no changes in placebo group. Active group also showed a reduced PEF response to cat exposure (P < 0.05), and an improvement in skin test reactivity to a standardized cat extract (P < 0.05), without significant changes in placebo group. Mean Fel d 1 exposure during the NCT was 6.2 ± 2.21 ng/m3.

Conclusions:  The results suggest that the cat SLIT used in this study was able to improve cat allergy based on natural exposure challenge.
Latex allergy: the sum quantity of four major allergens shows the allergenic potential of medical gloves
Authors: Palosuo T. , Reinikka‐Railo H. , Kautiainen H. , Alenius H. , Kalkkinen N. , Kulomaa M. , Reunala T. , Turjanmaa K.
Published: July 2010

Background:  Assessment of allergenic potential of medical devices made of natural rubber latex (NRL) requires the measurement of concentrations of specific allergenic proteins or polypeptides eluting from rubber.

Methods:  Four NRL allergens (Hev b 1, 3, 5, and 6.02) were quantified in all medical glove brands marketed in Finland in 1999, 2001, and 2003 (n = 208) by a capture enzyme immunoassay. The results were compared with those obtained from previous nationwide market surveys, using a skin prick test‐validated human IgE‐based ELISA‐inhibition method.

Results:  A high overall correlation (r = 0.87, 95% CI 0.83–0.90) emerged between the sum values of the four allergens(μg/g glove) and IgE‐ELISA inhibition (allergen units, AU/ml, 1 : 5 diluted glove extract). The sum of four allergens when set at 0.15 μg/g discriminated ‘low allergenic’ (<10 AU/ml) from ‘moderate‐ to high‐allergenic’ (≥10 AU/ml) gloves at a sensitivity of 0.93 (95% CI 0.85–0.98) and specificity of 0.90 (95% CI 0.83–0.94). When the sum was below the detection limit (0.03 μg/g) all gloves belonged to the previously defined low‐allergen category.

Conclusions:  By comparing the sum concentration of four selected NRL allergens with results obtained in human IgE‐ELISA inhibition, it was possible set a cut‐off level (0.15 μg/g) below which virtually all gloves contain low or insignificant amounts of allergens, and can be considered as low allergenic. At different cut‐off‐points, one could calculate the likelihood of a given glove to belong to the previously defined low, moderate or high allergen categories.
Variable p‐CREB expression depicts different asthma phenotypes
Authors: Chiappara G , Chanez P , Bruno A , Pace E , Pompeo F , Bousquet J , Bonsignore G , Gjomarkaj M
Published: July 2010

Background:  Chromatin modification may play a role in inflammatory gene regulation in asthma. Cyclic adenosine mono‐phosphate response element‐binding protein (CREB), with the specific co‐activator, the CREB‐binding protein (CBP), contributes to the acetylation of chromatin and to the transcription of pro‐inflammatory genes.

Objectives:  To evaluate the expression of CBP and of phospho‐CREB (p‐CREB) in bronchial biopsies and in peripheral blood mononuclear cells (PBMC) of controls (C), untreated (UA), inhaled steroid treated (ICS) and steroid‐dependent asthmatic (SDA) patients.

Methods:  We used immunohistochemistry in bronchial biopsies and western blot analysis and immunocytochemistry in PBMC.

Results:  Cyclic adenosine mono‐phosphate response element‐binding protein expression, in the epithelium was similar in all groups, while p‐CREB expression was increased in UA and in SDA in comparison with ICS and C subjects (C vs UA P = 0.002, C vs SDA P = 0.007), (ICS vs SDA P = 0.005), (ICS vs UA P = 0.001). Interestingly, also in the submucosa, p‐CREB was increased in UA and SDA in comparison with ICS and C subjects (C vs UA P = 0.0004) (C vs SDA P < 0.0001) (ICS vs UA P = 0.002) (ICS vs SDA P < 0.0001) and positively correlated with leukocyte infiltration within the bronchi (CD45RB+ cells). Similar results were obtained with PBMC isolated from the same patient groups. Incubation of PBMC in vitro, with fluticasone propionate, decreased the p‐CREB expression induced by cytokine activation (interferon‐γ, tumor necrosis factor‐α).

Conclusions:  This study demonstrates that the expression of p‐CREB is related, in asthma, to the persistent inflammation according to the disease severity. p‐CREB expression can be modulated by glucocorticoids in responsive patients.
Estimation of Membrane Potential and pH Difference across the Cristae Membrane of Rat Liver Mitochondria
Authors: Mitchell P. , Moyle J.
Published: July 2010

The cristae membrane or M phase of resting or respiring rat liver mitochondria becomes relatively permeable to K+ ions in presence of valinomycin. The equilibrium distribution of K+ ions across the membrane can therefore be used to estimate the membrane potential ΔΨ provided that precautions are taken to minimise swelling of the valinomycin‐treated mitochondria. The pH difference ΔpH across the M phase of anaerobic mitochondria has been estimated from the buffering powers of the inner and outer phases and from the change of pH observed on lysing the mitochondria with Triton X‐100. When the anaerobic mitochondria (State 5), in presence of β‐hydroxybutyrate, are brought to a state of steady respiration either in absence (State 4) or in presence (State 3) of phosphate acceptor, the changes of pH and of pK of the medium recorded with H+ ion‐sensitive and K+ ion‐sensitive electrodes can be used to estimate the changes of ΔΨ and ΔpH across the M phase. The absolute values of ΔΨ and ΔpH have been estimated from the values determined in State 5 and the change of these values in the transition from State 5 to States 4 and 3.

The total protonmotive force Δp =ΔΨ– 59 ΔpH across the M phase of the mitochondria oxidising β‐hydroxybutyrate in State 4 at 25° in a 250 mM sucrose medium near pH 7 is estimated to be about 230 mV, of which the major component is ΔΨ when the effect of translocation of K+ ions across the M phase is minimised. Under conditions permitting accumulation of a relatively large quantity of cation (State 6), Δp is not significantly different from that in State 4, but the major component is −ZΔpH.

The effects of changing Δp in mitochondrial suspensions in State 4 with uncoupling agent and with pulses of acid, alkali, calcium salt and ADP have been found to be in accord with the chemiosmotic hypothesis. In particular Δp in State 3 (ADP and Pi present) is estimated to be about 30 mV less than in State 4.
Combining Sites of IgG and IgM Antibodies of Poly‐d‐Alanyl Specificity
Authors: Haimovich J. , Schechter I. , Sela M.
Published: July 2010

Goats and rabbits were immunized with poly‐d‐alanyl ribonuclease. Anti‐poly‐d‐alanyl antibodies were isolated from the goat and rabbit sera by means of a water‐insoluble immuno‐adsorbent prepared from poly‐d‐alanyl rabbit serum albumin and bromoacetyl cellulose. The isolated antibodies and sera were separated into IgM and IgG fractions. The ability of the antibodies and of the serum fractions to precipitate with poly‐d‐alanyl human serum albumin and to inactivate poly‐d‐alanyl bacteriophage T4 was inhibited with alanine peptides of increasing size. In all cases the efficiency of inhibition as a function of the size of the inhibitory peptide increased up to the tetrapeptide. The penta‐d‐alanine, on the other hand, showed an extent of inhibition similar to the tetrapeptide. These results suggest that the antigen‐combining sites of both IgG and IgM antibodies are similar in size and capable of accomodating a peptide composed of four alanine residues.
The Δ5‐3β‐Hydroxysteroid Dehydrogenase of Rat Ovarian Tissue
Authors: Sulimovici S. , Boyd G. S.
Published: July 2010

The enzymic conversion of pregnenolone to progesterone catalysed by a Δ5‐3β‐hydroxysteroid dehydrogenase and Δ5‐3‐ketosteroid isomerase has been studied in ovaries of immature rats pretreated with pregnant mare's serum gonadotrophin. The enzymes were found to be located in both particulate fractions (mitochondria and microsomes). The Km and the Vmax values of the mitochondrial dehydrogenase were found to be not significantly different from those of the microsomal dehydrogenase.

The mitochondrial dehydrogenase was found to be mainly NAD+ dependent while the microsomal enzyme can utilise NADP+ as well as NAD+ as cofactor.

The reaction catalysed by the‐Δ5‐3β‐hydroxysteroid dehydrogenase was found to be reversible in both subcellular fractions.

2α‐Cyano,4,4,17α‐trimethyl‐17β‐hydroxy‐5‐androsten‐3‐one (cyanoketone), a synthetic androstane added to the incubation medium in very small amounts inhibited both dehydrogenases.

The action of adenosine 3′,5′‐cyclic‐monophosphoric acid and other nucleotides (AMP, ADP, ATP, adenosine‐5′‐diphosphoribose and adenosine‐2′,3′‐cyclic‐monophosphoric acid) on the dehydrogenase was tested. It was found that adenosine‐3′,5′‐cyclic‐monophosphoric acid markedly inhibited both dehydrogenases while the other nucleotides had virtually no effect on the enzyme system which converts pregnenolone to progesterone. The adenosine‐3′,5′‐cyclic‐monophosphoric acid was apparently a non‐competitive inhibitor of the Δ5‐3β‐hydroxysteroid dehydrogenase and Δ5‐3‐ketosteroid isomerase in ovarian mitochondria. The inhibition of the microsomal Δ5‐3β‐hydroxysteroid dehydrogenase system by the adenosine‐3′,5′‐cyclic‐monophosphoric acid was released by increasing the concentration of exogenous NAD+ suggesting a competitive inhibitory effect.
Fatty Acid Degradation in Escherichia coli
Authors: Overath P. , Pauli G. , Schairer H. U.
Published: July 2010

An enzymatic activity is described in oleate‐grown cells of E. coli, which has the cofactor requirements of a long chain acid: CoA ligase (AMP). This enzyme is at least partially membrane bound and activates saturated and mono‐ and polyunsaturated fatty acids. It is inactive with butyrate and hexanoate. The enzyme is induced by oleate under similar conditions, but at different relative rates, as three other enzymes of fatty acid degradation (thiolase, crotonase, hydroxyacyl‐CoA dehydrogenase) which are under coordinate control.

A mutant lacking acyl‐CoA synthetase synthesizes the other three enzymes at the basal rate of the wild type in the presence or absence of oleate in the medium. This suggests that the fatty acyl‐CoA derivatives rather than the fatty acids serve as inducers. The same mutant is unable to incorporate fatty acids into bacterial lipids. The acyl‐CoA synthetase is therefore required for lipid synthesis from exogenous fatty acids in vivo.

The isolation and properties of mutants unable to degrade oleate are described. A pleiotropic mutant lacking thiolase, crotonase and hydroxyacyl‐CoA dehydrogenase still contains the inducible acyl‐CoA synthetase. Mutants lacking thiolase and hydroxyacyl‐CoA dehydrogenase have inducible wild type levels of the other enzymes.

Mutants affecting the synthesis of thiolase, hydroxyacyl‐CoA dehydrogenase or crotonase map between the loci metE and rha on the E. coli‐chromosome and are probably closely linked. The mutant lacking acyl‐CoA synthetase is not linked to the other mutants and maps near the his‐locus.

Only fatty acids with more than twelve carbon atoms serve as inducers or can be used as sole carbon source by the wild type. This specificity is therefore typical for the abolition of repression by the interaction with the fatty acids.

Spontaneous mutants can be isolated which grow on decanoate. These are constitutive for the synthesis of all four enzymes investigated and can only grow on fatty acids with more than eight carbon atoms. Thus, after destruction of the repression mechanism the growth behavior is a reflection of the substrate specificity of the acyl‐CoA synthetase.

Starting with the constitutive mutants, spontaneous mutants have been isolated which can grow on butyrate. These fall into two classes: (a) One class grows on butyrate and poorly on long chain fatty acids. It makes all four enzymes constitutively but has twenty times more thiolase activity than the parent. This thiolase activity is not due to a gene linked to metE. (b) The other class can grow on butyrate and fatty acids with more than eight carbon atoms. It synthesizes the enzymes constitutively.

The data suggest that the expression of the linked structural genes of crotonase, hydroxy‐acyl‐CoA dehydrogenase and thiolase and the unlinked gene of the acyl‐CoA synthetase from a regulon which is controlled by a common regulator gene.
The Effect of Vitamin A Deficiency on Myelination in the Central Nervous System of the Rat
Authors: Clausen J.
Published: July 2010

The age‐dependent deposition of sulfatide, glycolipids and phospholipids in the whole brain during the period of myelination in the normal rat has been estimated and expressed as μg lipid stored in the tissue per week per dry weight, per mg total protein or per mg DNA.

Deficiency in vitamin A during the period of myelination in the rat gives rise to a statistically significant decrease in the deposits of sulfatide per week in the whole brain.

The decreased weekly deposits of sulfatide in the vitamin A deficient rat brain is associated with a slight depression in the weekly storage of phospholipids in the whole brain.

The myelin fraction, isolated by gradient ultracentrifugation, showed a significantly lower sulfatide content in the deficient rats than in the group of rats fed and raised on an optimal diet. Expressed per dry weight of the myelin, the total content of the phospholipids was found to be increased in the vitamin A deficient group when compared to the myelin from the normal animals.

The vitamin A deficiency from birth resulted in half of the normal serum value of vitamin A within five weeks. However, no absolute lack of the vitamin could be induced by the deficiency. This is in agreement with previous studies, showing it impossible to deplete the liver completely of vitamin A. This lack of complete depletion may explain the decrease, but not the absolute lack, of sulfatide formation in the rat brain.

The formation of sulfatide from galactocerebroside and inorganic 35S‐labelled sulfate in vitro was found to be highest in the sediment of 100,000 ×g ultracentrifugation both in the normal and in the vitamin A deficient animals.

The vitamin A deficiency in the period of myelination heavily depressed the formation of 35S‐labelled active sulfate and sulfatide in vitro compared to the activities of normal rats with an identical age (two months).
Chelation of Iron and Zinc by Protoporphyrin Catalyzed by Mitochondrial Preparations
Authors: Mazanowska A. , Dancewicz A. M. , Malinowska T. , Kowalski E.
Published: July 2010

Sonication, freezing and thawing, treatment of mitochondria with Tween‐Span surfactants and other procedures applied to isolate ferrochelatase from the liver mitochondria showed that the most active in releasing the enzyme was the combined action of Tween 20 at a final concentration of 15 mg/ml and sonic irradiation on the mitochondrial suspensions, previously subjected to freezing and thawing.

A 20–40 fold purification of the enzyme was achieved by ammonium sulphate fractionation and chromatography on dextran gels of “solubilized” ferrochelatase.

Variations of the yield of the release of ferrochelatase from the mitochondria suggest that the localization of the enzyme within the mitochondrial structure is heterogeneous.
Peptide Mapping of Cornea Collagens from Chick Embryos by Dodecylsulfate/Polyacrylamide Gel Electrophoresis
Authors: KAO Winston Whei‐Yang , FOREMAN Carol A.
Published: July 2010

Using differential salt fractionation, different collagen types were obtained from corneas of 17‐day‐old chick embryos. The collagen precipitated by 1.7 M and 2.5 M NaCl consisted of type I collagen, having an α1:α2 ratio of 2. The collagen precipitated by 5 M NaCl contained αA, αB, α1 and α2 chains having an αA:αB ratio of 1 and an α1:α2 ratio of 10. The α1 and α2 chains isolated from corneas were found to have higher mobility in polyacrylamide gel electrophoresis in sodium dodecylsulfate as compared with that of their counterparts isolated from embryonic chick tendons. When the α1 and α2 chains isolated by polyacrylamide gel electrophoresis in sodium dodecylsulfate were subjected to limited protease degradations, the peptide maps obtained from corneal α chains were not identical to those of tendon α chains. It is suggested that the type I collagen in cornea is different from type I collagen in tendon. Also the results indicated that the α1 chain present in the 5 M NaCl precipitate is a type I α1 chain.
Structural Studies on Heparan Sulphates
Authors: FRANSSON Lars‐Åke , SJÖBERG Ingrid , HAVSMARK Birgitta
Published: July 2010

Three heparan sulphate fractions were subjected to degradation by periodate oxidation and alkaline elimination. The starting materials were one low‐sulphated fraction rich in glucuronic acid and N‐acetylglucosamine (I), one fraction (IV) that was medium‐suphated, contained similar proportions of N‐acetyl and N‐sulphate as well as of glucuronic and iduronic acid and one fraction (V) that was oversulphated, contained an excess of N‐sulphate and was rich in iduronic acid. Selective periodate oxidation of glucuronic acid residues within N‐acetylated regions followed by scission in alkali yielded three categories of fragments that were isolated by gel and ion‐exchange chromatography. (a) N‐Acetylglucosamine‐R compounds where R is the remnant of an oxidised and degraded glucuronic acid residue. (b) Medium‐sized oligosaccharides, of the general structure glucosamine‐(glycuronic acid‐glucosamine) n‐R where n= 1–4, contained largely glucuronic acid associated with glucosamines that were N‐sulphated, N‐acetylated or unsubstituted. In saccharides where n= 4 indications of an alternating arrangement of N‐sulphate and N‐acetyl groups as well as of iduronic and glucuronic acid residues were obtained. The glucuronic acid residues of the oligosaccharide fragments were susceptible to reoxidation with periodate. The smaller saccharides were not depolymerised by HNO2 despite the presence of N‐sulphate groups. (c) Longer oligosaccharide fragments (n≥ 5) that were highly sulphated and contained heparin‐like repeating units, i. e. iduronic acid – glucosamine‐N‐sulphate with ester‐sulphate on both sugars. These oligosaccharides were susceptible to deaminative cleavage.
Catabolism of Thyroliberin by Rat Adenohypophyseal Tissue Extract
Authors: BAUER Karl , KLEINKAUF Horst
Published: July 2010

Rapid fragmentation of thyroliberin (< Glu‐His‐Pro‐NH2) by rat adenohypophyseal tissue enzymes could be demonstrated. Based on the identification of the metabolic products and by the demonstration that the individual enzymatic reactions can be preferentially blocked by enzyme inhibitors, specific and sensitive biochemical tests could be developed in order to monitor the enzymatic activities after gel chromatographic fractionation of the tissue extracts. These findings are in agreement with the interpretation that the observed degradation of thyroliberin by hypophyseal tissue extracts may follow the proposed pathways. The primary enzymatic cleavage of thyroliberin is either initiated by the action of a ‘thyroliberin‐deamidating enzyme’ (thyroliberin → < Glu‐His‐Pro‐OH → NH3), or by the action of a pyroglutamate aminopeptidase (thyroliberin → <Glu + His‐Pro‐NH2). While the pyroglutamate aminopeptidase also catayzes the subsequent degradation of deamidated thyroliberin (<Glu‐His‐Pro‐OH → <Glu + His‐Pro‐OH), the enzymatic deamidation of His‐Pro‐NH2 is not catalyzed by the ‘thyroliberin‐deamidating enzyme’ but by a post‐proline dipeptidyl aminopeptidase. Hydrolysis of the common intermediary metabolite His‐Pro‐OH to the free amino acids is apparently catalyzed by a proline dipeptidase. In addition to these enzymatic events rapid cyclization of His‐Pro‐NH2 to histidyl‐proline‐diketopiperazine () could be observed. This reaction however is mainly due to the non‐enzymatic intramolecular condensation reaction which is characteristic for proline‐containing dipeptide derivatives. An enzymatic activity which catalyzes this reaction could not be observed when the enzyme fractions were tested. Enzymatic degradation of , by hypophyseal tissue extracts could also not be observed.