Identification of novel translocation between short arm of chromosome 4 and long arm of chromosome 6 in an infertile man using Interphase Chromosome Profiling (ICP).

Conventional cytogenetics has always been a favourite to detect chromosomal aberrations. Carriers of chromosomal translocation are often phenotypically normal but are infertile. Couples are often advised to go for karyotyping, but culture failure or improper metaphase spread with poor banding often makes the analysis difficult. We report here a novel translocation between short arm of chromosome 4 and long arm of chromosome 6 in an infertile man using an advanced molecular cytogenetic technique of Interphase Chromosome Profiling (ICP).

Minor allergen patterns in birch pollen allergen products-A question of pollen?

BACKGROUND: Contrary to the scientific differentiation between major and minor allergens, the regulatory framework controlling allergen products in the EU distinguishes relevant and non-relevant allergens. Given the lack of knowledge on their clinical relevance, minor allergens are usually not controlled by allergen product specifications. Especially, in birch pollen (BP) allergen products, minor allergens are commonly disregarded. OBJECTIVES: To quantify three minor allergens in BP allergen products from different manufacturers and to assess the influence of the utilized BP on minor allergen patterns. METHODS: Apart from common quality parameters such as Bet v 1 content, Bet v 4, Bet v 6 and Bet v 7 were quantified in 70 BP allergen product batches from six manufacturers, using ELISA systems developed in-house. Batch-to-batch variability was checked for agreement with a variability margin of 50%-200% from mean of the given batches for individual allergen content. Subsequently, minor allergen patterns were generated via multidimensional scaling and related to information on the pollen lots used in production of the respective product batches. RESULTS: Like the already established Bet v 4 ELISA, the ELISA systems for quantification of Bet v 6 and Bet v 7 were successfully validated. Differences in minor allergen content between products and batch-to-batch consistency were observed. Correlations between minor and major allergen content were low to moderate. About 20% of batches exceeded the variability margin for at least one minor allergen. Interestingly, these fluctuations could not in all cases be linked to the use of certain BP lots. CONCLUSIONS AND CLINICAL RELEVANCE: The impact of the observed minor allergen variability on safety and efficacy of BP allergen products can currently not be estimated. As the described differences could only in few cases be related to the used pollen lots, it is evident that additional factors influence minor allergens in BP allergen products.

Allergen exposure chambers: harmonizing current concepts and projecting the needs for the future - an EAACI Position Paper.

BACKGROUND: Allergen exposure chambers (AECs) are clinical facilities allowing for controlled exposure of subjects to allergens in an enclosed environment. AECs have contributed towards characterizing the pathophysiology of respiratory allergic diseases and the pharmacological properties of new therapies. In addition, they are complementary to and offer some advantages over traditional multicentre field trials for evaluation of novel therapeutics. To date, AEC studies conducted have been monocentric and have followed protocols unique to each centre. Because there are technical differences among AECs, it may be necessary to define parameters to standardize the AECs so that studies may be extrapolated for driving basic immunological research and for marketing authorization purposes by regulatory authorities. METHODS: For this task force initiative of the European Academy of Allergy and Clinical Immunology (EAACI), experts from academia and regulatory agencies met with chamber operators to list technical, clinical and regulatory unmet needs as well as the prerequisites for clinical validation. RESULTS: The latter covered the validation process, standardization of challenges and outcomes, intra- and interchamber variability and reproducibility, in addition to comparability with field trials and specifics of paediatric trials and regulatory issues. CONCLUSION: This EAACI Position Paper aims to harmonize current concepts in AECs and to project unmet needs with the intent to enhance progress towards use of these facilities in determining safety and efficacy of new therapeutics in the future.

Effects of work-family conflict and job insecurity on psychological distress.

BACKGROUND: Work-family conflict (WFC) and job insecurity are important determinants of workers' mental health. AIMS: To examine the relationship between WFC and psychological distress, and the co-occurring effects of WFC and job insecurity on distress in US working adults. METHODS: This study used cross-sectional data from the 2010 National Health Interview Survey (NHIS) for adults aged 18-64 years. The 2010 NHIS included occupational data from the National Institute for Occupational Safety and Health (NIOSH) sponsored Occupational Health Supplement. Logistic regression models were used to examine the independent and co-occurring effects of WFC and job insecurity on distress. RESULTS: The study group consisted of 12059 participants. In the model fully adjusted for relevant occupational, behavioural, sociodemographic and health covariates, WFC and job insecurity were independently significantly associated with increased odds of psychological distress. Relative to participants reporting WFC only, participants reporting no WFC and no job insecurity had lower odds of moderate and severe distress. Co-occurring WFC and job insecurity was associated with significantly higher odds of both moderate [odds ratio (OR) = 1.55; 95% confidence interval (CI) 1.25-1.9] and severe (OR = 3.57; 95% CI 2.66-4.79) distress. CONCLUSIONS: Rates of WFC and job insecurity were influenced by differing factors in working adults; however, both significantly increased risk of adverse mental health outcomes, particularly when experienced jointly. Future studies should explore the temporal association between co-occurring WFC and job insecurity and psychological distress.

Standardization of allergen products: 3. Validation of candidate European Pharmacopoeia standard methods for quantification of major birch allergen Bet v 1.

BACKGROUND: The BSP090 project aims at establishing European Pharmacopoeia Reference Substances in combination with the corresponding ELISA methods for the quantification of major allergens in allergen products. Two sandwich ELISAs proved suitable for quantification of Bet v 1, the major birch pollen allergen, in preceding phases of BSP090. METHODS: Two Bet v 1-specific ELISA systems were compared with respect to accuracy and precision in a ring trial including 13 laboratories. Model samples containing recombinant rBet v 1.0101 as well as native birch pollen extracts were measured independently at least three times in each facility. The assessment was completed with a comparative quantification of Bet v 1 in 30 marketed birch allergen products in one laboratory, simulating the future use as reference method. RESULTS: In the collaborative study, both candidate ELISAs confirmed their suitability to quantify recombinant and native Bet v 1. ELISA-A showed higher precision and lower interlaboratory variability, yet ELISA-B exhibited slightly higher accuracy. Subsequent parallel measurement of Bet v 1 in a panel of 'real-life' birch allergen products indicated better repeatability of ELISA-B. Both systems detected substantial differences in Bet v 1 content between allergen products, but the effect was more pronounced using ELISA-B due to persistently higher values compared to ELISA-A. CONCLUSIONS: In the collaborative study, no deciding differences were observed between the two candidate ELISAs. Further comparison under conditions simulating the intended use combined with the criterion of long-term availability enabled the selection of one Bet v 1-specific ELISA for proposal as European Pharmacopoeia standard method.

Identification of the predicted 5s-4f level crossing optical lines with applications to metrology and searches for the variation of fundamental constants.

We measure optical spectra of Nd-like W, Re, Os, Ir, and Pt ions of particular interest for studies of a possibly varying fine-structure constant. Exploiting characteristic energy scalings we identify the strongest lines, confirm the predicted 5s-4f level crossing, and benchmark advanced calculations. We infer two possible values for optical M2/E3 and E1 transitions in Ir^{17+} that have the highest predicted sensitivity to a variation of the fine-structure constant among stable atomic systems. Furthermore, we determine the energies of proposed frequency standards in Hf^{12+} and W^{14+}.

Molecular characterization of repetitive DNA sequences from B chromosome in Plantago lagopus L.

In Plantagolagopus the B chromosome has been reported by our laboratory to be the result of massive amplification of ribosomal DNA sequences. However, the presence of transposable elements and other repetitive DNA sequences together with rDNA could not be ruled out. The present study clearly demonstrated the presence of transposable elements in the B chromosomes. These transposable elements were characterized and their nature assessed. Physical mapping using double fluorescent in situ hybridization was performed using DNA probes for 5S and copia elements. The results clearly proved that the B chromosome is a mix of 5S, 45S rDNA and transposable elements. Based on the FISH and Fiber-FISH patterns, it can be concluded that while 45S rDNA sequences are restricted to the subtelomeric regions, the 5S rDNA sequences are interspersed with transposons in the body of the B chromosome. These results have further enhanced our understanding of the organization and evolution of B chromosomes.

Magnetic irreversibility and magnetization processes in Ni5Al3/NiO core/shell nanoparticle system.

This work brings out many interesting facets of magnetism in the Ni5Al3/NiO core/shell nanoparticle system. Theweakandstrongmagnetic irreversibility lines (TWI(H)andTSI(H)) reproduce the previously reportedH - Tphase diagram at fieldsH⩽30 Oe, but strong departures occur forH > 30 Oe. Comparison with the theoretically predictedH - Tphase diagram allows us to identifyTWIwithTCG+SG, where the paramagnetic (PM)-chiral glass (CG) and PM-spin glass (SG) phase transitions occursimultaneously, andTSIwithTSG, the temperature at which transition to the replica symmetry breakingSGstate takes place. TheTSI(H)transition line abruptly ends at the point (H≃30 Oe,T≃90K). AsHexceeds 30 Oe, a new transition appears which gets completely suppressed at fieldsH>1 kOewhere the magnetic irreversibility ceases to exist. Nointrinsiclong-range ferromagnetic ordering exists but fields as low as 3 kOe suffice to induce long-range ferromagnetic order. At fixed temperatures, the magnetocrystalline anisotropy fluctuations essentially govern the 'approach-to-saturation' in magnetization for fields in the range 3 - 70 kOe. The present nanocrystalline system behaves as an isotropic system with random easy axis in which the magnetization reversal occurs through the coherent rotation of the magnetizations of weakly-interacting single-domain Ni5Al3particles. Saturation magnetization, likeM(T) atH⩾2 kOe, exhibits an anomalous upturn at temperatures below ≈ 30 K. This upturn is associated with the anomalous softening of spin-wave modes which results in the thermal excitation of a large number of non-equilibrium (finite lifetime) magnons. At sub-Kelvin temperatures, these magnons undergo Bose-Einstein condensation.

Insights into microRNA-mediated interaction and regulation of metabolites in tomato.

microRNAs direct regulation of various metabolic pathways in plants and animals. miRNAs may be useful in developing novel/elite genotypes, with enhanced metabolites and disease resistance. We examined miRNAs in tomato. In tomato, miRNAs in the carotenoid pathway have not been fully elucidated. We examined the potential role of miRNAs in biosynthesis of carotenoids, transcript profiling of miRNAs and their possible targets (genes and transcription factors) at different development stages of tomato using stem-loop PCR and RT-qPCR. We also identified miRNAs targeting key flavonoid genes, such as chalcone isomerase (CHI), and dihydroflavonol-4-reductase (DFR). Distinct expression profiles of miRNAs and their targets were found in fruits of three tomato accessions, suggesting carotenoid regulation by miRNAs at various stages of fruit development. This was also confirmed using HPLC of the carotenoids. The present study may help in understanding possible regulation of carotenoid biosynthesis. The identified miRNAs can be exploited to enhance biosynthesis of different carotenoids in plants.

[The therapy allergens ordinance ("Therapieallergene-Verordnung"). Background and effects]. / Die Therapieallergene-Verordnung. Hintergrund und Auswirkungen.

Medicinal products for specific immunotherapy as causal treatment of allergies exist in Germany as authorized medicinal products manufactured batchwise in advance and as named patient products (NPPs) which are exempted from the authorization procedure. With the therapy allergens ordinance ("Therapieallergene-Verordnung (TAV)") which has been in effect since 14 November 2008, this exemption was restricted to therapy allergens indicated for the treatment of rare allergies. NPPs containing at least one of the therapy allergens listed in the annex of the TAV had to be notified to the Paul-Ehrlich-Institut (PEI) by 14 May 2009 to retain their marketability. It had to be stated whether applications for marketing authorization will be submitted for the respective NPPs or if they will be sold off by 14 November 2011. The bulks which are used for manufacturing of the NPPs have been subject to official batch release by PEI since October 2009. Nearly 7,000 NPPs of 10 pharmaceutical entrepreneurs were notified. Marketing authorization applications were submitted for 123 NPPs. This illustrates that, although there are authorized therapy allergens available for all allergens listed in the annex of the TAV, a large number of NPPs with unknown quality, safety, and efficacy have been marketed.

Is the Management of Rectal Cancer Using a Watch and Wait Approach Feasible, Safe and Effective in a Publicly Funded General Hospital?

AIMS: Although there is emerging evidence to suggest equivalent oncological outcomes using a watch and wait approach compared with primary total mesorectal excision surgery, there is a paucity of evidence about the safety and efficacy of this approach in routine clinical practice. Here we report the long-term outcomes and quality of life from patients managed with watch and wait following a clinical complete response (cCR) to neoadjuvant therapy. MATERIALS AND METHODS: Patients with adenocarcinoma of the rectum with cCR following neoadjuvant therapy managed using watch and wait were retrospectively identified. Demographic data, performance status, pretreatment staging information, oncological and surgical outcomes were obtained from routinely collected clinical data. Quality of life was measured by trained clinicians during telephone interviews. RESULTS: Over a 7-year period, 506 patients were treated for rectal cancer, 276 had neoadjuvant therapy and 72 had a cCR (26.1%). Sixty-three were managed with watch and wait. Thirteen patients had mucosal regrowth. There was no significant difference in the incidence of metastatic disease between the surgical and watch and wait cohorts (P = 0.38). The 13 patients with mucosal regrowth underwent salvage surgery. Eleven of the patients who underwent surgical resection had R0 resections. There was also a statistically and clinically significant improvement in the Functional Assessment of Cancer Therapy - Colorectal (FACT-C) trial outcome index (P = 0.022). CONCLUSION: This study shows that watch and wait is safe and effective outside of tertiary referral centres. It suggests that an opportunistic cCR is durable and when mucosal regrowth occurs it can be salvaged. Finally, we have shown that quality of life is probably improved if a watch and wait approach is adopted.

Validation of an ELISA method for quantification of the major Timothy grass pollen allergen Phl p 5a (BSP090).

Progress towards standardisation of allergen products has been made in recent years. Nevertheless, no standardised test method to quantify the allergen content of grass pollen allergen products is available at present. One aim of the BSP090 project was to validate a quantitative assay for a major Timothy grass (Phleum pratense) pollen allergen, Phl p 5. Qualification of a candidate ELISA system was performed with regard to range, robustness and cross-reactivity in preliminary studies. The assay specifically detected Phl p 5 with a quantification range from 3.9 ng/mL to 62.5 ng/mL. Suitability to quantify recombinant and natural Phl p 5 was further assessed in a collaborative study including 14 laboratories in Europe and the USA. Precision and accuracy of the assay was satisfactory with 93% of calculated Phl p 5 concentrations and 100% of total recoveries being within the ± 30% acceptance range. Similar results were obtained for spike recoveries, with exclusion of the lowest concentration spike, showing spike recoveries exceeding the acceptance range for six laboratories. Inter-assay (repeatability) and inter-laboratory (reproducibility) variability were satisfactory, in the format used in the present study. Robustness towards different statistical methods for data analysis was demonstrated. In conclusion, the assay can easily be established in routine testing and results of the preliminary testing and collaborative study support the proposal of the assessed Phl p 5-specific ELISA as a European Pharmacopoeia general method.

Magnons as a Bose-Einstein condensate in nanocrystalline gadolinium.

The recent observation [S. P. Mathew et al., J. Phys. Conf. Ser. 200, 072047 (2010)] of the anomalous softening of spin-wave modes at low temperatures in nanocrystalline gadolinium is interpreted as a Bose-Einstein condensation (BEC) of magnons. A self-consistent calculation, based on the BEC picture, is shown to closely reproduce the observed temperature variations of magnetization and specific heat at constant magnetic fields.

Regulatory environment for allergen-specific immunotherapy.

Products for specific immunotherapy (SIT) are medicinal products according to the European Regulations. To obtain a marketing authorization (MA) within the European Community, the quality, safety and efficacy have to be proven. During the development phase of a medicinal product, applicants have the opportunity to apply for scientific advice by national competent authorities or the European Medicines Agency (EMA) to compile a suitable development plan for the examination of quality and performance of nonclinical and clinical trials. Moreover, a paediatric investigation plan has to be submitted to the Paediatric Committee of the EMA and has to be approved before submission of an application for MA. Several regulatory procedures exist for obtaining a MA in the European Community. The national procedure leads only to marketability in one country whereas the Mutual Recognition, the Decentralized and Centralized Procedures (CP) are intended for MA in several or all member states of the European Union. The CP is mandatory for certain medicinal products, for example for drug substances derived by biotechnological processes such as recombinant allergens. Named Patient Products for SIT are a specialty because they are manufactured on the basis of an individual prescription and marketed without a MA.