bluuebunny/crossref_metadata_2025_split huggingface dataset:
Separation andin vitroculture of goat fetal testicular cells and behaviour of male germ-line stem cells (mGSCs)
Authors: Wu-Zi Dong , Jin-Lian Hua , Shu-Zhen Zhuang , Wen-Zheng Shen , Zhong-Ying Dou 
Published: October 2005

AbstractSeminiferous tubule tissues from goat fetal testis were treated using four methods. One of the treatments, with 0.1% collagenase for 15 min and then with an enzyme mixture (containing 0.25% trypsin, 10 μg/ml DNase I, 0.04% EDTA) for 5 min showed ideal cell dispersal after centrifugation and washing. Thein vitroculture of the cells separated with a different medium system showed that mulberry-shaped male germ-line stem cell (mGSC) clusters and a single layer of Sertoli cells appeared as the original generation in 120 h. The mGSC clusters developed in suspension and were distributed in different locations from the Sertoli cells in the plate. The cells in the marginal area of incompletely digested mGSCs clusters, and the single cells from them, were obviously homogenized after culture for 5 days in a fibre cell feeding layer from fetal mice (MEF feeding layer), whereas identically treated mGSCs clusters co-cultured with Sertoli cells in the same culture system did not displayed clear homogenization in marginal cells of the clusters. Also, the first generation of mGSC clusters co-cultured with Sertoli cells could form the same morphological properties with the original generation; cells in clusters were tight and mGSCs divided more slowly than those in the MEF feeding layer.
Cloning and expression ofSpodoptera lituranucleopolyhedrovirusgp41gene inEscherichia coliand preparation of antibody
Authors: Li-Jing Pan , Zhao-Fei Li , Chong Yin , Lei Lv , Yi Pang 
Published: October 2005

AbstractGP41, a major glycoprotein, identified in the occlusion-derived virions (ODV) of baculoviruses, is required for the egress of nucleocapsids from the nucleus in the pathway of budded virion (BV) synthesis. Using the polymerase chain reaction (PCR), the open reading frame (ORF) ofSpodoptera lituranucleopolyhedrovirus (SpltMNPV)gp41gene was obtained from SpltMNPV genomic DNA. The PCR product was cloned into pMD18-T vector to get the recombinant plasmid (pT-gp41). Thegp41gene was recombinedin vitrowith prokaryotic expression vector pQE30 and transformed intoEscherichia coliM15 [pREP4]. The M15 [pREP4] strain, containinggp41recombinant plasmid, expressed a 37.9 kDa 6×His-tag fusion protein after induction with 1 mmol/l isopropylthio-β-d-galactoside (IPTG). The fusion protein was purified with a nickel-nitrilotriacetic acid (Ni–NTA) resin column and used as the immunogen to raise GP41-specific antibody. Western blotting analysis indicated that the antibody was suitable to be used for further analysis of GP41 protein.
Construction of antagonistic, insecticidal, and endophytic strains of Bacillus spp. by protoplast fusion
Authors: Nian-Long Yan , Si-Xin Qiu , Hong He , Xiong Guan , Fang-Ping Hu 
Published: October 2005

AbstractA multifunctional-engineering strain was constructed by fusing the insecticidal bacterium (Bacillus thuringiensis) with vip3A gene and the endophytic bacterium (B. subtilis) protoplasts. Results of polymerase chain reaction (PCR) showed that the insecticidal vip3A gene of B. thuringiensis was present in the fusant. The fusant's antagonism to Fusarium oxysporum f. sp. cucumerinum, the adjusted mortality to Plutella xylostella and the endophytic characters were detected. The result showed that the antagonism of the fusant was against the F. oxysporum f. sp. cucumerinum; the adjusted mortality to P. xylostella reached 59% and it had insecticidal characteristics.
Bone morphogenetic protein receptor IB as a candidate gene for prolificacy in Small Tail Han and Hu ewes
Authors: Ya-Dong Yan , Ming-Xing Chu , Yong-Qing Zeng , Li Fang , Su-Cheng Ye , Li-Min Wang , Qing-Kun Guo , Dai-Qin Han , Zhao-Xin Zhang , Xi-Jun Wang , Zhao-Xin Zhang 
Published: October 2005

AbstractThe bone morphogenetic protein receptor IB (BMPR-IB) gene, which controls the fecundity of Booroola Merino ewes, was studied as a candidate gene for the prolificacy of Small Tail Han and Hu ewes. A single nucleotide polymorphism of the BMPR-IB gene was detected in both high (Small Tail Han and Hu) and low (Suffolk and Dorset) fecundity sheep breeds by polymerase chain reaction–single-strand conformation polymorphism (PCR-SSCP) analysis. The results indicated the presence of the same mutation (A746G) of the BMPR-IB gene in both Small Tail Han and Hu ewes and in Booroola Merino ewes, but not in both Suffolk and Dorset ewes. In Small Tail Han ewes, frequencies of BB, B+ and ++ genotypes were 0.524, 0.383 and 0.093, respectively. In Hu ewes, these frequencies were 0.882, 0.118 and 0.000. The BMPR-IB genotype distributions were significantly different (P<0.001) among high- and low-fecundity sheep breeds. Small Tail Han ewes with genotype BB had 0.92 (P<0.01) and 1.02 (P<0.01) lambs more than those with genotype ++ in the first and second parity, respectively. These results demonstrated that the BMPR-IB gene is a major gene affecting the prolificacy in both Small Tail Han and Hu ewes, and could be used as a molecular genetic marker to select the litter size in sheep.
Remarks on product 𝑉𝑀𝑂
Authors: Lacey Michael , Terwilleger Erin , Wick Brett 
Published: October 2005

Well known results related to the compactness of Hankel operators of one complex variable are extended to little Hankel operators of two complex variables. Critical to these considerations is the result of Ferguson and Lacey (2002) characterizing the boundedness of the little Hankel operators in terms of the product BMO of S.-Y. Chang and R. Fefferman (1985), (1980).
Characterization of the T cell response to the major hazelnut allergen, Cor a 1.04: evidence for a relevant T cell epitope not cross‐reactive with homologous pollen allergens
Authors: Bohle B. , Radakovics A. , Lüttkopf D. , Jahn‐Schmid B. , Vieths S. , Ebner C. 
Published: October 2005

SummaryBackground
IgE antibodies specific for the major birch‐pollen allergen, Bet v 1, cross‐react with homologous allergens in particular foods, e.g. apples, carrots and hazelnuts. In a high number of tree pollen‐allergic individuals, this cross‐reactivity causes clinical symptoms, commonly known as the ‘birch‐fruit‐syndrome’.Objective
To characterize the T cell response to the Bet v 1‐related major allergen in hazelnuts, Cor a 1.04, and its cellular cross‐reactivity with Bet v 1 and the homologous hazel pollen allergen, Cor a 1.Methods
Using recombinant Cor a 1.04, T cell lines (TCL) and T cell clones (TCC) were established from peripheral blood mononuclear cells of tree pollen‐allergic patients with associated food allergy. T cell epitopes were determined using overlapping synthetic peptides in Cor a 1.04‐reactive TCL and TCC. In parallel, reactivity to Bet v 1 and Cor a 1 was tested.Results
In total, 20 distinct T cell epitopes on the hazelnut allergen were identified. Several Cor a 1.04‐specific TCL and TCC reacted with pollen allergens albeit less pronounced than with the hazelnut allergen. Several Cor a 1.04‐specific TCC did not react with pollen allergens. Interestingly, these clones were found to react with the Bet v 1‐related major allergen in carrots, Dau c 1. The cellular cross‐reactivity between both food allergens could be associated with the most frequently recognized T cell epitope of Cor a 1.04, Cor a 1.04142–153.Conclusions
The major hazelnut allergen cross‐reacts with the major allergens of birch and hazel pollen but apparently contains a relevant T cell epitope not shared with pollen allergens. Our finding may have important implications for the specific immunotherapy of tree pollen‐allergic patients suffering from concomitant hazelnut allergy.
Impairment in quality of life is directly related to the level of allergen exposure and allergic airway inflammation
Authors: Roberts G. , Mylonopoulou M. , Hurley C. , Lack G. 
Published: October 2005

SummaryBackground
Allergic disease has been shown to impair health‐related quality of life (HRQL). The relationship between HRQL and either allergen exposure or allergic inflammation has not been previously assessed.Objective
To assess the relationship between HRQL and both grass pollen exposure and airway inflammation using the Paediatric Allergic Disease Quality of Life Questionnaire (PADQLQ). This is a novel questionnaire previously developed to assess the multi‐system aspects of allergic disease.Methods
Eighty‐four subjects, aged 6–17 years, with seasonal allergic rhinoconjunctivitis, asthma and/or cutaneous manifestations were assessed before and during the grass pollen season. They were assessed with the PADQLQ, a visual analogue scale (VAS) to assess quality of life, symptom diary and exhaled nitric oxide (FENO).Results
HRQL, as measured by the PADQLQ, significantly correlated with the average pollen count in the previous week (regression coefficient 0.038, 95% confidence interval (CI) 0.027–0.049, P<0.001). The PADQLQ score was also found to be significantly associated with airway inflammation as measured by FENO (regression coefficient 0.410, 95% CI 0.175–0.646, P=0.001). Additionally, PADQLQ showed a high degree of correlation with symptom scores and quality of life as measured by a VAS, good within‐subject reliability and a small minimal important difference (0.20, 95% CI −0.09 to 0.49 on a seven‐point scale).Conclusion
HRQL is related to both allergen load and allergic inflammation and the PADQLQ has excellent cross‐sectional and longitudinal validity with respect to quality of life and symptoms.
Occupational sensitization to ribosome‐inactivating proteins in researchers
Authors: Szalai K. , Schöll I. , Förster‐Waldl E. , Polito L. , Bolognesi A. , Untersmayr E. , Riemer A. B. , Boltz‐Nitulescu G. , Stirpe F. , Jensen‐Jarolim E. 
Published: October 2005

SummaryBackground
Ribosome‐inactivating proteins (RIPs) are expressed in many plants. Because of their anti‐infectious and anti‐proliferative effects, intensive research is going on for applying these toxins in therapy against viral infections or malignancies. Recently, we demonstrated that type I allergy against RIPs from elderberry can occur.Objective
Stimulated by our study, a group of RIP researchers reported that some of the employees had suspected allergy to RIPs.Methods and Results
We tested their sera in ELISA on natural RIPs. Specific IgE in four subjects were found against dianthin30, gelonin, momordin, PAP‐S, saporin, ricin and volkensin. In contrast, asparin and lychnin did not show any IgE binding. When separating extracts of plants containing the toxins in SDS‐PAGE, RIPs appeared to be the predominant constituents. Interestingly, among the other plant proteins, they were exclusively recognized by IgE in immunoblot. RIPs derived from close botanical families share high sequence homologies. Nevertheless, in IgE inhibition experiments with human sera, cross‐reactivity between RIPs also derived from non‐related plants could be demonstrated.Conclusion
We conclude that sensitization and IgE induction to RIPs may occur upon exposure. This has to be considered when applying them in therapy against malignancies or viral infections.
Multi‐dimensional phenotyping: towards a new taxonomy for airway disease
Authors: Wardlaw A. J. , Silverman M. , Siva R. , Pavord I. D. , Green R. 
Published: October 2005

Summary
All the real knowledge which we possess, depends on methods by which we distinguish the similar from the dissimilar. The greater the number of natural distinctions this method comprehends the clearer becomes our idea of things. The more numerous the objects which employ our attention the more difficult it becomes to form such a method and the more necessary. [1].
Classification is a fundamental part of medicine. Diseases are often categorized according to pre‐20th century descriptions and concepts of disease based on symptoms, signs and functional abnormalities rather than on underlying pathogenesis. Where the aetiology of disease has been revealed (for example in the infectious diseases) a more precise classification has become possible, but in the chronic inflammatory diseases, and in the inflammatory airway diseases in particular, where pathogenesis has been stubbornly difficult to elucidate, we still use broad descriptive terms such as asthma and chronic obstructive pulmonary disease, which defy precise definition because they encompass a wide spectrum of presentations and physiological and cellular abnormalities. It is our contention that these broad‐brush terms have outlived their usefulness and that we should be looking to create a new taxonomy of airway disease—a taxonomy that more closely reflects the spectrum of phenotypes that are encompassed within the term airway inflammatory diseases, and that gives full recognition to late 20th and 21st century insights into the disordered physiology and cell biology that characterizes these conditions in the expectation that these will map more closely to both aetiology and response to treatment. Development of this taxonomy will require a much more complete and sophisticated correlation of the many variables that make up a condition than has been usual to employ in an approach that encompasses multi‐dimensional phenotyping and uses complex statistical tools such as cluster analysis.
Comparison of purified Dermatophagoides pteronyssinus allergens and extract by two‐dimensional immunoblotting and quantitative immunoglobulin E inhibitions
Authors: Weghofer M. , Thomas W. R. , Pittner G. , Horak F. , Valenta R. , Vrtala S. 
Published: October 2005

SummaryBackground
The allergens of the house dust mite (Dermatophagoides pteronyssinus, Der p), one of the most important indoor allergen sources, occur as isoallergens that differ in their amino acid sequence. These variations may influence allergenic activity and thus may have impact on diagnostic tests and specific immunotherapy.Objective
We investigated whether single purified recombinant mite allergens contain the IgE epitopes of the natural Der p isoallergens.Methods
A panel of purified recombinant (rDer p 2, 5, 7, 8, 10 and 14) and two natural (nDer p 1 and 4) mite allergens were used to establish IgE reactivity profiles of Der p allergic patients and to inhibit IgE reactivity to two‐dimensionally separated Der p isoallergens. In addition, we determined the percentage of Der p extract‐specific IgE which could be preadsorbed with a mixture of purified mite allergens (nDer p 1, rDer p 2, 5, 7, 8 and 10) from sera of mite‐allergic patients (n=18) in a non‐denaturing RAST‐based inhibition.Results
We demonstrate that single recombinant mite allergens inhibit IgE reactivity to the corresponding natural isoallergens. A mixture of purified mite allergens (nDer p 1, rDer p 2, 5, 7, 8 and 10) bound on an average 76% of Der p‐specific IgE antibodies.Conclusion
The studied recombinant and natural mite allergens contain a large portion of Der p‐specific IgE and may be used for diagnostic tests and therapy of Der p allergy.
Effects of (R)‐ and (S)‐isomers of β‐adrenergic agonists on eosinophil response to interleukin‐5
Authors: Volcheck G. W. , Kelkar P. , Bartemes K. R. , Gleich G. J. , Kita H. 
Published: October 2005

SummaryBackground
Racemic β2‐adrenergic receptor agonists (β2‐agonists) are used frequently to treat patients with asthma. Potential differences in the biological activities and clinical efficacies among racemic β2‐agonists and their isomers are controversial, and research into these possible differences is limited.Objective
We hypothesized that the (S)‐ and the (R)‐isomers of β2‐agonists have opposing effects on the activation of inflammatory cells.Methods
Isolated human eosinophils were pretreated with 1 : 1 racemic (R,S)‐, (R)‐ or (S)‐albuterol, isobutyl methylxanthine (IBMX), and stimulated with IL‐5. The kinetics of superoxide production were examined by reduction of cytochrome c, and the effects of pharmacological agents on superoxide production were monitored for 180 min.Results
(R,S)‐albuterol inhibited IL‐5‐induced superoxide production. This inhibition was enhanced by a cyclic adenosine monophosphate (cAMP) phosphodiesterase inhibitor, IBMX, and was reversed by the selective β2‐adrenergic receptor antagonist, ICI 118, 551, verifying the involvement of both cAMP and the β2‐adrenergic receptor. In addition, (R)‐albuterol alone, similarly to (R,S)‐albuterol, significantly inhibited IL‐5‐induced superoxide production up to 60 min (P<0.05, n=4), but the inhibition was lost with longer incubation. In contrast, (S)‐albuterol with IBMX did not inhibit IL‐5‐induced superoxide production before 60 min, but it significantly enhanced IL‐5‐mediated superoxide production after 60 min (P<0.05, n=4). When both were present as racemic (R,S)‐albuterol, the inhibitory effect of (R)‐albuterol was not affected by (S)‐albuterol.Conclusion
When incubated with IL‐5‐activated eosinophils, (R)‐albuterol shows anti‐inflammatory effects and (S)‐albuterol shows pro‐inflammatory effects in the presence of IBMX. The kinetics of these effects are different, and when used simultaneously, (R)‐albuterol predominates. When marked usage of the (S)‐isomer is anticipated, racemic (R,S)‐albuterol should be used clinically with caution.
Atopic dermatitis in young children is associated with impaired interleukin‐10 and interferon‐γ responses to allergens, vaccines and colonizing skin and gut bacteria
Authors: Dunstan J. A. , Hale J. , Breckler L. , Lehmann H. , Weston S. , Richmond P. , Prescott S. L. 
Published: October 2005

SummaryBackground
A significant proportion of children with food allergy and more severe forms of atopic dermatis (AD) go on to develop persistent forms of allergic disease such asthma. Defining immune dysregulation in these children will be of great value in understanding disease pathogenesis.Objective
In this study we characterized the immune responses of young infants (6–18 months of age) with moderate‐to‐severe AD (a modified SCORAD25) and compared these (n=53) with responses of non‐allergic children with no history of dermatitis or sensitization of the same age (n=20).Methods
Mononuclear cell cytokine responses to allergens (egg ovalbumin (OVA), β‐lactoglobulin (BLG), house dust mite (HDM)), vaccines (tetanus toxoid (TT), diphtheria toxoid (DT)), intestinal flora (heat‐killed Lactobacillus species (HKLB)), heat‐killed Staphylococcus aureus (HKSA), S. aureus enterotoxin B (SEB) and mitogen (phytohaemaglutinin (PHA)) were compared in children with AD with unaffected children.Results
Children with AD had significantly lower spontaneous (unstimulated) production of regulatory cytokine IL‐10 (P<0.001), as well as IFN‐γ (P<0.001) and TNF‐α (P<0.001) compared with the unaffected children. After allowing for differences in baseline levels IL‐10 responses to virtually all stimuli (food allergens (P=0.003), vaccines P=0.01, intestinal flora (heat‐killed Lactobacillus species (HKLB), P=0.005) and skin flora (heat‐killed Staphylococcus aureus (HKSA), P=0.003)) were also significantly attenuated in children with AD. The only exception was HDM, to which responses were stronger in children with AD [P=0.05]. Although there were no significant correlations between HDM IgE and HDM cytokine responses at this age, T‐helper type 2 (Th2) IL‐5 (P=0.014) and IL‐13 (P=0.004) responses to HDM were significantly more frequent in the children with AD. However, while children with AD showed significantly attenuated Th1 IFN‐γ responses to food allergens (OVA, P=0.007 and BLG, P<0.001) and vaccines (DT, P=0.008 and TT, P<0.001), these children showed no difference in Th1 IFN‐γ responses to HDM or microbial agents (HKSA and HKLB).Conclusion
A increase in propensity for Th2 responses to aeroallergens in children with AD is associated with early impaired production of IL‐10 regulatory cytokine to a broad range of environmental stimuli including foods, intestinal flora, S. aureus, and vaccines.
The squamous cell carcinoma antigens as relevant biomarkers of atopic dermatitis
Authors: Mitsuishi K. , Nakamura T. , Sakata Y. , Yuyama N. , Arima K. , Sugita Y. , Suto H. , Izuhara K. , Ogawa H. 
Published: October 2005

SummaryBackground
Although it is thought that both Th1‐ and Th2‐type inflammations are involved in the pathogenesis of atopic dermatitis (AD), it is controversial which immune response is more involved in regulating the clinical severity of AD. We recently found that the squamous cell carcinoma antigens 1 (SCCA1) and SCCA2 are novel biomarkers of bronchial asthma, downstream of IL‐4 and IL‐13.Objective
We examined whether SCCA1 and SCCA2 could also serve as biomarkers of AD, reflecting its Th2‐type immune responses, and whether the expression level of SCCA was correlated with clinical severity of AD.Method
We compared the expression of SCCA1 and SCCA2 at the mRNA and protein levels in both involved and uninvolved skin of AD patients and in normal control skin. We next analysed induction of SCCA by IL‐4 or IL‐13 in keratinocytes. Finally, we compared the serum level of SCCA with laboratory parameters reflecting Th2‐type inflammation and clinical severity in AD patients.Results
SCCA1 and SCCA2 were highly expressed in involved skin of AD patients, compared with their uninvolved skin, at both mRNA and protein levels. SCCA protein was dominantly expressed in suprabasal keratinocytes in the epidermis of AD patients. Either IL‐4 or IL‐13, but not IFN‐γ or TNF, induced production of SCCA in keratinocytes. These result suggest that SCCA is induced in AD skin, probably due to direct actions of IL‐4 and/or IL‐13 on keratinocytes. Serum levels of SCCA were well correlated with eosinophil numbers and serum lactate dehydrogenase levels, and weakly with serum IgE levels, in AD patients. Furthermore, serum levels of SCCA were strongly correlated with clinical severity.Conclusions
Th2‐type inflammation dominantly regulates the clinical severity of AD, and SCCA is a relevant biomarker of AD, reflecting both Th2‐type inflammation and clinical severity.
Dysfunction of the mitotic:meiotic switch as a potential cause of neoplastic conversion of primordial germ cells
Authors: ADAMAH D. J. B. , GOKHALE P. J. , EASTWOOD D. J. , RAJPERT DE‐MEYTS E. , GOEPEL J. , WALSH J. R. , MOORE H. D. , ANDREWS P. W. 
Published: October 2005

SummaryGerm cell tumours (GCT) are thought to arise as the result of a defect in early development, probably shortly after arrival of the migrating primordial germ cells (PGC) in the genital ridge when, if in a male genital ridge, the germ cells arrest in mitosis, but in a female genital ridge they enter meiosis. We suggest that dysfunction of the mitotic:meiotic switch, with cells aberrantly co‐expressing functions pertinent to both states, might provide the genetic instability that could initiate tumour development. If this hypothesis is correct, GCT could arise because of disruption in the function of any one of a number of different genes involved in controlling mitosis and meiosis, rather than being dependent upon a single prominent susceptibility gene. The Notch signalling system is one candidate system for controlling the switch and we have identified expression of Notch2 and Notch4 in seminomas and carcinoma in situ. Thus those two members of the Notch family are candidates for proto‐oncogenes that could play a role in GCT development. We have also identified a human homologue of the synaptonemal complex protein, SCP3, and have found its apparently aberrant expression in some established EC cell lines. One possibility is that abnormal regulation of such proteins involved in the synaptonemal complex could also lead to genetic instability in PGC and so also initiate tumour development.