A novel nuclear antigen

The human oestrogen receptor (hER) mediates some effects of the steroid hormone oestrogen and functions as a ligand-dependent transcription factor in the nuclei of oestrogen-sensitive cells. The measurement of hER levels in breast cancer biopsies provides useful clinical information regarding therapy and prognosis. The current study describes a monoclonal antibody raised against hER aa 497-507 which recognises a novel nuclear antigen. Monoclonal antibodies were raised by immunising mice with a synthetic fragment of the hER (aa 497-507) conjugated to keyhole lymphocyte haemocyanin. Thirty antibody secreting hybridomas were identified. Hybridoma supernatants were characterised by enzyme-linked immunosorbent assay (ELISA), immunological staining using MCF-7 cells, binding studies, and SDS-PAGE Western blotting. One supernatant (15F6) displayed nuclear staining in fixed MCF -7 cells. Staining could be abolished by pre-incubation of the supernatant with the aa 497-507 peptide and peptide conjugates, but not with an unrelated hER peptide (aa 256-275). This antibody also stained the nuclei of hER negative breast cell lines MDA-MB-231 and MDA-MB-330, the breast cell line T47D, and liver cell line HepG2. Immunological staining of human tissue sections reveal the antigen to be present in the nuclei of keratocytes in skin and tubule and luminal endothelial cells of the kidney. The antibody identified a 120 kD band on Western blots with cytosols prepared from human breast cell lines and in solubilised cells. The antibody does not precipitate 16a-iodooestradiol-labelled ER from MCF-7 cells. Expression-linked screening of the MCF-7 cDNA library with antibody 15F6 identified nine positive clones. Antibody staining could be blocked by pre-incubating the antibody with hER aa 497-507-BSA conjugate, but not with an unrelated hER peptide conjugate. The (260 bp) clones were found to be identical. Submission of sequence to BLAST protein and nucleotide databases revealed a lack of homology to known proteins and genes. Sequence was matched to expressed sequence tags (ESTs) from brain, liver/spleen, uterus, ovary, colon, heart, and placenta. To further define the epitope of antibody 15F6, the sequence was translated and three peptides containing potential epitopes, comparable to the hER aa 497-507 region, were synthesised and tested by ELISA. The putative epitope was shown to be contained within one of these peptides

Оценка выбросов в атмосферу отходящих дымовых газов ТЭЦ и технологий их переработки

Сравнительная оценка выбросов в атмосферу отходящих дымовых газов, при сжигании органических топлив и способы их переработки.Comparative assessment of emissions of flue gases into the atmosphere, during combustion of organic fuels and methods of their processing

Совершенствование системы вознаграждения персонала бюджетного учреждения на основе комплексной оценки его деятельности

Объектом исследования является МБОУ 슫СОШ № 18슻 Цель работы - является оценка системы вознаграждения труда персонала бюджетного учреждения, ее оптимизации на основе комплексной оценке его деятельности В процессе исследования проводились способы группировки и сравнения, графические и аналитические методы. В результате исследования были выявлены недостатки и предложен комплекс мер по их устранению. Степень внедрения: сделать систему оплаты труда взаимосвязанной с конкретным результатом каждого работника и учреждения в целом. Область применения: бюджетные учреждения. Экономическая эффективность работы заключается в объективной оценке оплаты труда в бюджетном учреждении и в разработке практических рекомендаций по совершенствованию системы оплаты труда в анализируемом учреждении.The object of study is MBOU "SOSH № 18" Purpose - is to assess the remuneration system of the personnel budget of the institution, its optimization on the basis of a comprehensive assessment of its activities In the process of investigation the methods of grouping and comparison of graphical and analytical methods. The study revealed shortcomings and proposed the complex of measures on their elimination. Level of implementation: to make the wage system is interconnected with the concrete result of each employee and the institution as a whole. Scope: budget companies. Economic efficiency is an objective assessment of remuneration in budgetary institutions and in the development of practical recommendations for improving the system of remuneration in the analyzed institution

The membrane distal half of gp 130 is responsible for the formation of a ternary complex with IL-6 and the IL-6 receptor

AbstractGp130 is the signal transducing subunit of the interleukin-6 receptor. Signaling is initiated by the complex formation of gp130 with IL-6 bound to the IL-6 receptor (IL-6R). We have subdivided the extracellular domain of gp130 in two parts and expressed the mutant proteins as soluble IgG fusion proteins in COS-7 cells. By studying the formation of the ternary complex we show that the membrane distal half of gp130 which contains a cytokine receptor domain is responsible for the interaction with the IL-6/IL-6R complex. Interestingly this is the same region which is believed to be involved in specific recognition of the related cytokines LIF, OM, and probably also of CNTF and IL-11