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91.
A simplified and efficient procedure has been developed for performing hybridization reactions and analyses of the percentage of hybridization on glass plates. This method is as accurate as conventional methods, while considerably reducing the time and costs to perform the hybridization in titration hybridization experiments. 相似文献
92.
Ute Gröschel-Stewart Susi Ceurremans Irmtraut Lehr Christine Mahlmeister Eltriede Paar 《Histochemistry and cell biology》1977,50(4):271-279
Summary The injection of rabbits with insoluble or soluble G-actin from chicken smooth or striated muscle will produce antibodies that are equally reactive, and species and tissue non-specific in immunoprecipitation, immunofluorescence and actin-activated Mg2+-ATPase** inhibition test. These antibodies have been used for the identification of actin-containing fibrils in a variety of tissues. When G-actins from chicken smooth or striated muscle are immobilized by chemical linkage to Affi-Gel 702 microbeads, their immunogenicity is increased, but the antibodies obtained against them are species-specific and will only react with actin and actin-containing structures from chicken and are therefore limited in use. It is concluded from this work that insoluble G-actin is the preferable immunogen to obtain precipitating antibodies for wide use.Abbreviations ATPase
Adenosinetriphosphatase
- FITC
Fluoresceinisothiocyanate
- SDS
Sodiumdodecylsulfate
This paper is dedicated to Dr. Dorothy M. Needham, University of Cambridge, England, in honour of her eightieth birthday 相似文献
93.
Summary The thickness of ectoplasmic walls decreases at the contraction phase and increases at the expansion phase of each pulsation cycle. In average, 29% of material forming the walls of expanded veins is evacuated during contraction with the endoplasm streaming. 相似文献
94.
Summary A heme-nonapeptide (H-9-P)1, applicable to electron microscopic cytochemistry via peroxidase-like activity, was prepared by passing horse heart cytochrome c through a column with Sepharose and covalently attached trypsin. After purification by column chromatography (Sephadex G50 Superfine, Biogel P-2) a maximal yield of 50% and purity of >99% was achieved. A concise schedule allows for inexpensive preparation of H-9-P with standard laboratory equipment. H-9-P has the following properties: Its structure is (14) Cys-Ala-Gln-Cys-His-Thr-Val-Glu-Lys (22) with heme attached to Cys (14) and (17). MW=1630, pI=4.95,
E(max)
pH 7
= 397.5 nm,
22 °C, pH 7
397.5 nm
= 1.11 × 105 [Liter/Mole x cm]. With the use of a diaminobenzidine-H2O2-medium — as applied for cytochemistry — we determined spectrophotometrically a pHopt=12.5 and an apparent K5 = 3.14 × 10– 3 [M]. Glutardialdehyde leads to considerable de-activation and, according to SDS-polyacrylamide-gel-electrophoresis, to diffuse crosslinking accompanied by a shift of the active pH-region towards neutral pH values. An attempt was made to optimize the cytochemical assay. The peroxidase-like activity of H-9-P is well comparable to that of other heme-tracers; only horseradish peroxidase has a higher turnover number. When injected to mice or added to cell suspensions, even high concentrations of H-9-P did not entail any signs of toxicity.Abbreviations AAA
amino acid analysis
- AHC
ammoniumhydrogencarbonate
- BSA
bovine serum albumin
- Cyt c
cytochrome c
- DAB
5,3-diaminobenzidine
- GA
glutardialdehyde
- H-8-P
heme-octapeptide
- H-9-P
heme-nonapeptide
- H-11-P
heme-undecapeptide
- HR-POX
horseradish peroxidase
- MW
molecular weight
- PAGE
polyacrylamid-gel-electrophoresis
- pI
isoelectric point
- SDS
sodiumdodecylsulphate
- SG-TLC
silicagel-thin-layer-chromatography
This work was supported by the Österreichische Forschungsfonds 相似文献
95.
Zusammenfassung Kernpolyeder vonMamestra brassicae (L.) wurden auf Nebenwirkungen an zwei Entomophagen-Arten in der Basis- Konzentration (LC100 für den natürlichen Wirt) sowie der 5- und 10 fachen Konzentration geprüft. Schadwirkung durch Kontakt mit frisch angetrocknetem
Belag bzw. durch Verfüttern eines kontaminierten N?hrsubstrates auf den EiparasitenTrichogramma cacoeciae
Marchal konnte nicht nachgewiesen werden. Ebenso wurde die Entwicklung vonT. cacoeciae in parasitierten Wirtseiern durch eine Tauchbehandlung mit der Kernpolyeder-Suspension in verschiedenen Intervallen nach
der Parasitierung nicht gest?rt. Im Wahl- und Nichtwahlversuch konnte keine Repellentwirkung von Wirtseiern festgstellt werden,
die in eine Kernpolyeder-Suspension getaucht worden waren.
Die r?uberischen Larven vonChrysopa carnea
Steph. waren gegenüber der 10fachen Basiskonzentration bei direktem Bespritzen der Larven, Kontakt mit einem frisch angetrockneten
sprit zbelag sowie bei peroraler Aufnahme unempfindlich, wie die Frassleistung der Larven, die Fekundit?t der Imagines sowie
die Schlüpfrate der Eier zeigte. Bei praktischen Eins?tzen von Kernpolyeder-Pr?paraten zur Bek?mpfung der Kohleule sind im
Gegensatz zu den üblichen Anwendungen von Phosphors?ureestern keine direkten Nebenwirkungen auf die untersuchten Nutzinsekten
zu erwarten.
Nr. 2 der Serie: Wirkung von Insektenpathogenen auf Entomophagen. Als Nr. 1 gilt:S. Hassan & A. Krieg: über die schonende Wirkung vonBacillus thuringiensis-Pr?paraten auf den ParasitenTrichogramma cacoeciae (Hym.: Trichogrammatidae). — Z. Pfl. Krankh. Pfl. Schutz, 82, 515–521, 1975. 相似文献
Summary The side effect of a water suspension of polyhedral inclusion bodies of nuclnear polyhedrosis virus (NPV) fromMamestra brassicae (L.) in a concentration of 1-, 5-, and 10-fold of the LC100 for the natural host was tested on two beneficial insects. No reduction in the parasitization capacity ofTrichogramma cacoeciae Marchal (Hym.: Trichogrammatidae) was recorded when adults were exposed to a fresh dry film, or fed on contaminated honey-agar. The development of the parasite inSitotroga cerealella Oliv. host eggs was not affected when these were dipped in a suspension of the NPV at different time intervals after parasitization. In additional tests, it was shown that deposit of the NPV suspension had no repellent effect onTrichogramma. The feeding capacity of the predatory larve ofChrysopa carnea Steph., fecundity of the adults as well as viability of the eggs were not affected when the larvae were directly sprayed with the highest tested concentration of the suspension, exposed to fresh dry film or fed on contaminated diet. These experiments indicate that, in contrast to the use of the conventional organophosphorus insecticides, no hazard to the tested beneficial insects should be expected when the preparation of the polyhedral inclusion bodies will be used in the field.
Nr. 2 der Serie: Wirkung von Insektenpathogenen auf Entomophagen. Als Nr. 1 gilt:S. Hassan & A. Krieg: über die schonende Wirkung vonBacillus thuringiensis-Pr?paraten auf den ParasitenTrichogramma cacoeciae (Hym.: Trichogrammatidae). — Z. Pfl. Krankh. Pfl. Schutz, 82, 515–521, 1975. 相似文献
96.
Initiation and culture of callus and cell suspensions of Cinchona ledgeriana and C. succirubra as well as the successful isolation and selection of a high-yielding alkaloid-forming strain derived from the leaf rachis of a C. succirubra plant are described. Results of feeding experiments with L-tryptophan using two different culture procedures are presented and discussed. Maximum alkaloid yields of up to 0.9% (based on dry weight) or 6.35 mg/l have been obtained. 相似文献
97.
Claudine Grégori Claude Besmond Axel Kahn Jean-Claude Dreyfus 《Biochemical and biophysical research communications》1982,104(2):369-375
High molecular weight cellular RNA was isolated from adult and fetal human liver tissue by a procedure of ethanol precipitation in concentrated guanidine-HCl solutions. About 5 mg of RNA were obtained from one gram of liver. RNA was fractionated by sucrose gradient ultracentrifugation. Aldolase B neosynthesized in a reticulocyte lysate cell-free system under the direction of total or fractionated RNA was purified by immunoaffinity microchromatography. Messenger RNA specifying synthesis of aldolase B exhibited a sedimentation coefficient of 16 S both in adult and fetal liver. This enzyme represented 1.3 % of the total neosynthesized proteins in adult liver, 0.1 % in the liver of a 6-month-old fetus and less than 0.01 % in the liver of a 4.5 month-old fetus. 相似文献
98.
To evaluate the hygienic risk involved in using baculoviruses for insect pest control, safety studies are required. Pigs were chosen as representative test animals of commercial and agricultural importance. The tests were aimed at detecting virus propagation, immune reactions, and signs of acute infection (changes in body temperature and hematology profile, swelling of lymph nodes). Four of five animals inoculated with nuclear polyhedrosis virus showed a slight temperature rise at day 2 postinfection. After day 4 postinfection, no differences between infected animals and controls were observed. In the bioassay, virus activity could be recovered from fecal samples; however, no activity was found in organ extracts. The data did not indicate hygienic risks involved in the application of nuclear polyhedrosis virus, especially that from Mamestra brassicae, in biological pest control. 相似文献
99.
Maturation of lactase-phlorizin hydrolase (LPH) (EC 3.2.1.23-62) requires proteolytic processing of precursor (pro-LPH) to mature microvillus membrane enzyme (m-LPH). Subcellular site and function of this processing are unknown. We studied the processing and sorting of human LPH expressed permanently in MDCK cells. LPH was inserted into the apical membrane and small amounts were found basolateral. Of the LPH immunoprecipitated from the apical membrane, 42% was in the mature, i.e. proteoytically processed form; on the basolateral membrane it was 20%. Thus, LPH-processing occurs after sorting and is not necessary for surface expression. 相似文献
100.
Vendela Parrow Eewa Nnberg Jari Heikkil Ulf Hammerling Sven Phlman 《Journal of cellular physiology》1992,152(3):536-544
SH-SY5Y human neuroblastoma cells can be induced to differentiate into a neuronal phenotype by treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA). In other cell systems, TPA treatment frequently leads to down-regulation of protein kinase C (PKC). However, we now report that TPA-treated and non-treated SH-SY5Y cells express PKC-alpha, but not PKC-beta and PKC-gamma, mRNA. Furthermore, only a slight down-regulation of the PKC-alpha protein could be seen during prolonged treatment with 16 nM TPA, the concentration giving optimal differentiation. In contrast, a higher concentration of TPA (1.6 microM) results in a poor neuronal differentiation and a complete down-regulation of PKC-alpha. PKC-alpha was rapidly translocated to the particulate fraction and remained membrane bound for at least 4 days during treatment with 16 nM TPA. In such cells a sustained increased level of the phosphorylated form of a 80,000 Dalton PKC-substrate was found. In addition to this sustained augmented phosphorylation, administration of fresh TPA at day 4 caused a small but reproducible further increased level of phosphorylated substrate. When the PKC activity was measured by the histone phosphorylation assay a substantial fraction of the initial enzyme activity could still be detected after 4 days of TPA treatment. Taken together, the data demonstrate that PKC remains functionally active during TPA induced differentiation of SH-SY5Y cells, which may suggest a continuous role for the enzyme during the differentiation process. 相似文献