Cell-Free Synthesis of Selenoproteins in High Yield and Purity for Selective Protein Tagging

The selenol group of selenocysteine is much more nucleophilic than the thiol group of cysteine. Selenocysteine residues in proteins thus offer reactive points for rapid post-translational modification. Herein, we show that selenoproteins can be expressed in high yield and purity by cell-free protein synthesis by global substitution of cysteine by selenocysteine. Complete alkylation of solvent-exposed selenocysteine residues was achieved in 10 minutes with 4-chloromethylene dipicolinic acid (4Cl-MDPA) under conditions that left cysteine residues unchanged even after overnight incubation. Gd^III−Gd^III distances measured by double electron–electron resonance (DEER) experiments of maltose binding protein (MBP) containing two selenocysteine residues tagged with 4Cl-MDPA-Gd^III were indistinguishable from Gd^III−Gd^III distances measured of MBP containing cysteine reacted with 4Br-MDPA tags.     

Use of stem cells from mismatched related donors

Mismatched haploidentical bone marrow transplantations from a related donor have been the topic of clinical and laboratory research for more than 20 years. During that time, new treatment strategies have been designed based on animal experiments, and, since our group introduced the megadose inoculum which combines T-cell-depleted bone marrow cells with a large number of granulocyte colony-stimulating factor-mobilized peripheral blood stem cells and a more intensive conditioning regimen, have done much to overcome the problems of graft-versus-host disease and graft rejection. As most patients have a full haplotype mismatched relative available, this technique means that a far greater number of patients with hematologic malignancies can be offered a T-cell-depleted transplantation as curative therapy.     

Immunocytochemical evaluation of protein kinase C translocation to the inner nuclear matrix in 3T3 mouse fibroblasts after IGF-I treatment

The complex pathway which links the agonist-cell membrane receptor binding to the response at the genome level involves, among other elements, protein kinase C (PKC). Agonists acting at the cell membrane can affect an autonomous nuclear polyphosphoinositide signaling system inducing an activation of nuclear phosphoinositidase activity and a subsequent translocation of PKC to the nuclear region. The fine localization of PKC has been investigated by means of electron microscopy quantitative immunogold labeling in 3T3 mouse fibroblasts, mitogenically stimulated by IGF-I. The enzyme, which in untreated cells is present in the cytoplasm, except for the organelles, and in the nucleoplasm, after IGF-I treatment is reduced in the cytoplasm and almost doubled in the nucleus. The PKC isoform translocated to the nucleus is the alpha isozyme, which is found not only associated with the nuclear envelope but mainly with the interchromatin domains. By using in situ matrix preparations, PKC appears to be retained at the nuclear matrix level, both at the nuclear lamina and at the inner nuclear matrix, suggesting a direct involvement in the phosphorylation of nuclear proteins which are responsible for the regulation of DNA replication.     

Effects of sympathetic stimulation on microcirculatory dynamics in patients with essential acrocyanosis. A study using mental stress

Cytokines are central to the development of effective immunity against microbial pathogens and their beneficial effects in the control of a variety of infections of man and animals are well-documented. However, it appears that cytokines may also have detrimental effects in infections by actually enhancing microbial growth. These observations emphasise the fact that many successful pathogens possess mechanisms enabling them to evade the immune response and that caution needs to be exercised in using cytokines as therapeutic agents to control infections in various human diseases.     

Examination of coping resources and family adaptation in mothers and daughters of incestuous versus nonclinical families.

Mother–daughter dyads from incestuous vs nonclinical families were compared in terms of (1) family life stress, (2) problem-solving appraisal, (3) direct coping behaviors, (4) mother–daughter communication, and (5) family adaptation and cohesion. 15 mother–daughter pairs from an incestuous family involving an adult male caretaker were matched on several variables to 16 mother–daughter pairs from a nonclinical family. The results revealed that mothers from the incest group (compared with the nonclinical group) reported greater amounts of family life stress, had more negative perceptions of their problem-solving appraisal, and reported more communication difficulties. The daughters from the incest group appeared to be quite similar to the nonclinical group daughters, although they did acknowledge more family stress. Implications of the results for therapeutic interventions as well as future research were discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Analysis of the validity of vaginal-cervical cytologic examination in 162 pre- and post-menopausal patients

The authors examine 162 cervico-vaginal smears of women in peri-menopause presenting cellular alterations (ASCUS). These patients were treated with a local topic therapy as the persistence of these alterations in 23 cases (14.2%) second level analyses were performed: gynecological visit, colposcopy and biopsy. In addition to inflammatory aspects 2 in situ carcinomas and 2 moderate dysplasias were recorded. We believe therefore that cytology could be a valid aid also in pre-menopausal women in the cervico-vaginal pathology.     

Unusual essential oils with aromatic properties

Summary The essential oil obtained by steam distillation of cinchona bark (Cinchona spp.,Rubiaceae family) — a plant well known for its pharmacological properties, also employed in the food industry, especially in making liqueurs - was investigated. The oil, of completely unknown chemical composition, was analyzed by GLC and GLC/MS after separation on a silica gel column, and one hundred components were identified. The presence of 2-hexyl-3-methylmaleic anhydride is rather unusual.

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Ungewöhnliche ätherische Öle mit aromatischen Eigenschaften. Die flüchtigen Aromastoffe der China-Rinde

Zusammenfassung Es wurde ätherisches Öl analysiert, das durch Wasserdampfdestillation von Chinarinde (Cinchona spp. derRubiaceae-Familie) gewonnen wurde. Diese Pflanze ist aufgrund ihrer pharmakologischen Eigenschaften bekannt und wird auch in der Lebensmittelindustrie verwendet, ganz besonders für die Likör- und Magenbitterherstellung. Die chemische Zusammensetzung dieses Öls war unbekannt. Nach Abtrennung auf einer Kieselgelsäule wurden im Öl mit GLC und GLC/MS etwa 100 Verbindungen identifiziert. Das Vorhandensein von 2-Hexyl-3-methyl-maleinsäureanhydrid ist ziemlich ungewöhnlich.

     

Chitosan particles agglomerated scaffolds for cartilage and osteochondral tissue engineering approaches with adipose tissue derived stem cells

In the XML file of the original article, H. Redl’s affiliation is incorrect. It is listed correctly in both the paper and PDF versions of the article, and can be found below: The online version of the original article can be found at