Gamma-COP, a coat subunit of non-clathrin-coated vesicles with homology to Sec21p.

Constitutive secretory transport in eukaryotes is likely to be mediated by non-clathrin-coated vesicles, which have been isolated and characterized [(1989) Cell 58, 329-336; (1991) Nature 349, 215-220]. They contain a set of coat proteins (COPs) which are also likely to exist in a preformed cytosolic complex named coatomer [(1991) Nature 349, 248-250]. From peptide sequence and cDNA structure comparisons evidence is presented that one of the subunits of coatomer, gamma-COP, is a true constituent of non-clathrin-coated vesicles, and that gamma-COP is related to sec 21, a secretory mutant of the yeast Saccharomyces cervisiae.     

Characterization of cisplatin adducts of oligonucleotides by fast atom bombardment mass spectrometry

The products of the reaction of the antitumor drug cisplatin (cis-diamminedichloroplatinum(II)) with four oligonucleotide tetramers, d(GpCpGpC), d(GpGpCpC), d(TpGpApT), and d(TpGpCpT), were separated by gel permeation chromatography and characterized by negative- and positive-ion fast atom bombardment (FAB) mass spectrometry. Fragment ions indicating the oligonucleotide sequence and the position of cisplatin binding were observed in MS/MS spectra following collisional activation and B/E-linked scanning. Positive-ion FAB MS/MS spectra were characterized by platinum-containing product ions. Nonplatinated sequence ions and internal fragment ions were present primarily in the negative-ion spectra. The most prominent fragment ions containing platinum were [HB2.Pt.B3H]+ and [HB1.Pt.B2H]+, where B1, B2, and B3 were bases in the oligonucleotide tetramer, one of which was usually guanine. Both singly and doubly charged platinum complexes were observed, probably indicating reduction of Pt(II) during the FAB ionization process. The location of the platinum complex bound to each oligonucleotide sequence could be determined, and the binding sites observed by mass spectrometry were similar to those previously determined by other methods. FAB ionization with collisional activation and MS/MS analysis could serve as a new method for structural analysis of platinated oligonucleotides.     

Optimizing the production of transformed pea (Pisum sativum L.) callus using disarmed Agrobacterium tumefaciens strains

For optimization of the transformation procedure with Pisum sativum L. stern explant callus was used to test the effect of disarmed Agrobacterium tumefaciens strains, cocultivation procedures (preconditioning of explants; use of Nicotiana tabacum L. nurse cultures), duration of cocultivation (2, 3 or 4 days), and agents for selection (kanamycin or hygromycin). The succinamopine strain EHA101(pBI1042) produced the highest percentage of transformed calli (77%) when used in conjunction with tobacco nurse culture during four days of cocultivation. Using this strain, kanamycin (76%) and hygromycin (77%) were equally effective selective agents, but for strain LBA4404(pBI1042) percentage of transformed calli was higher for hygromycin (63%) than for kanamycin (17%). The procedures and strains shown to be optimal for transformation of pea callus will now be complemented by a pea regeneration system.     

Interferon-γ activates the cleavage of double-stranded RNA by bovine seminal ribonuclease

Bovine seminal ribonuclease (BS-RNase), a dimeric homologue of RNase A, cleaves both single- and double-stranded RNA and inhibits the growth of tumor cells. Its catalytic activity against double-stranded RNA, either homopolymeric ([³H]polyA/polyU) or mixed sequence, is enhanced by bovine or human recombinant interferon-γ (IFN-γ). Activation is seen with as little as 4–10 interferon units per assay. Enhancing the degradation of double-stranded RNA, an intermediate in the growth cycle of many viruses, could contribute to IFN-γ's ability to control cell growth and induce an antiviral state.     

A flow cytometric study of chromosomes from rat kangaroo and chinese hamster cells

Summary Chromosomes from rat kangaroo (PTK) and chinese hamster (CHV 79) cells have been prepared for quantitative flow-cytometric analysis. The preparation time was optimized down to 30 (PTK) and 40 min (CHV 79). DAPI was used as a AT-sensitive fluorescent dye to stain for monoparameter DNA measurements. Simultaneous two-parameter DNA-protein analysis was carried out with DAPI and SR 101 (as a general protein fluorochrome) in combination. The karyotype of the PTK cells with 13 (14) chromosomes was separated into 10 DNA peaks. The X-chromosome bearing the nucleolus organizer region generates a distinct peak. The karyotype of the CHV 79 cells with 22 chromosomes was separated into 15 peaks. The DNA profile obtained indicates a geometric grading of the chromosomal amount of AT components in the karyotype of this particular cell line. The simultaneous DNA-protein analysis performed show enough sensitivity of the instrument utilizing high power UV excitation illumination to discriminate the two color emission consisting of blue (DAPI) and red (SR 101) fluorescence. Color overlapping could be completely avoided. Additionally, the quality (number, location, and resolution of peaks) of the DNA distribution was not influenced by the simultaneous application of a second fluorescent stain. Fluorescence activated electronic sorting applied on chromosomal fluorescence distributions providing purified fractions of chromosomes for subsequent biochemical and biological determinations is discussed.     

Die Wirkung von Harnmarken auf Artgenossen beim Haushund

An 11 Versuchstagen wurden die Reaktionen von insgesamt 3 Rüden und 2 Hündinnen auf einem rund 2stündigen Spaziergang in nicht oder wenig bekanntem Gebiet beobachtet. Auf einem ersten Rundgang wurden die spontan abgegebenen Harnmarken eines Versuchstieres mit kleinen Objekten markiert, dann ein zweiter Hund an loser Leine den gleichen Weg (meist in umgekehrter Richtung) entlanggeführt und dessen Reaktionen auf die markierten Stellen registriert. Im ganzen wurden bei den ersten Rundgängen 261 Harnmarken abgesetzt. In 47,1 % der Fälle ging der im zweiten Rundgang vorbeigeführte Hund ohne ersichtlichen Grund an der Harnmarke vorbei. Bei Beachtung der Marken zeigten sich 6 verschiedene Reaktionen: 1. Beschnuppern, 2. Harnen über oder neben die Marke, 3. Scharren mit den Hinterpfoten nach Überharnen, 4. Belecken der Marke, 5. Knurren beim Scharren, 6. Zähneklappern nach Beschnuppern und Belecken. Die Marken von feindlichen Rüden und läufigen Hündinnen werden nach Beschnuppern signifikant häufiger als die eigenen bzw. diejenigen einer nichtläufigen Hündin mit einer weiteren Reaktion beantwortet. Die Marken nichtläufiger Hündinnen und die eigenen Marken ergaben keine unterschiedlichen Reaktionen beim Rüden. Eine Hündin reagiert auf die Marken eines (feindl.) Rüden nicht mehr als ein Rüde auf die eigenen Marken. Die Harnmarken vermitteln dem Hunderüden Informationen darüber, ob sie von einer läufigen Hündin, einem fremden Rüden oder von ihm selbst (bzw. einer nicht-läufigen Hündin) stammen.     

Plasmon-Assisted Crystalline Silicon Solar Cell with TiO2 as Anti-Reflective Coating

Plasmonics - The present study focuses on the employment of TiO2 (titanium dioxide) film as an anti-reflective coating (ARC) on thin crystalline silicon (Si)-based solar cells along with the...