Effect of substrate on the spin state of cytochrome P-450 in hepatic microsomes

The effect of substrate on the spin state of oxidized cytochrome P-450 in liver microsomes prepared from phenobarbital-pretreated rats has been examined. Formation of the substrate-induced Type I difference spectrum was found to correlate quantitatively with the disappearance of the ferric low-spin esr signal of cytochrome P-450. The dissociation constant of substrate for oxidized cytochrome P-450 obtained by optical methods was found to be the same as that obtained from esr methods provided that the same high microsomal protein concentration was used. However, a decrease in microsomal protein concentration leads to an apparent increase in the affinity of substrate for oxidized cytochrome P-450, indicating a dependence of lipophilic substrate dissociation constants on the membrane concentration.     

Über das Quellungsverhalten von Holz als Substanz mit Ultrafilterstruktur in Abhängigkeit von Grösse und Bau der Quellungsmittelmoleküle

Zusammenfassung An sauber tangential geschnittenen Furnierstreifen einiger Holzarten wird die Quellung in der Tangentialrichtung untersucht. Bei sorgfältig wasserfreiem Arbeiten nehmen in der Reihe der aliphatischen primären Alkohole C₁–C₆ mit zunehmender C-Kette die Quellungsgeschwindigkeit und der Grad der konstanten Endquellung ab und erreichen beim Hexanol praktisch den Nullwert. Iso-Alkohole wirken schwächer quellend als Normalformen. Von den anderen untersuchten Quellungsmitteln erzielt Wasser die stärkste Quellung überhaupt, dann folgen 1,4-Dioxan und Glycerin.Diese Ergebnisse lassen Holz als eine Substanz mit ziemlich starren hydrophilen Poren erscheinen und lassen Rückschlüsse zu auf die plasmatischen Porenstrukturen, die die Permeabilität im Sinne der Ultrafiltertheorie regeln.Unter experimenteller Mitwirkung vonB. Ehrmann Mit 4 Textabbildungen.Herrn Prof. Dr. Dr. h. c.W. Ruhland, meinem hochverehrten Lehrer, zu seinem 75. Geburtstag in Dankbarkeit gewidmet.     

Radiation-induced cytogenetic instability in vivo.

Radiation-induced cytogenetic instability has been well documented in a number of laboratories, and we have hypothesized that such instability is the initiating event in the process leading to radiation-induced cancer. To date most studies of radiation-induced instability have used systems in which cells are rapidly dividing. For this phenomenon to have significance for radiation carcinogenesis, it must be established that instability can be induced in vivo in less rapidly dividing fully differentiated tissues known to be at risk. In the present study, we have examined the kinetics of radiation-induced cytogenetic instability in mammary epithelial cells after irradiation in vivo. Having established that instability could arise in vivo in intact mammary tissue, we subsequently demonstrated a dose-response relationship both in vitro and in vivo and demonstrated a lower frequency of instability after fractionated exposures.     

Molecular breeding for grain yield in barley: an evaluation of QTL effects in a spring barley cross

 We report results from a breeding strategy designed to accumulate favorable QTL alleles for grain yield identified in the SteptoeבMorex’ (SM) barley germplasm. Two map lines (SM73 and SM145) from the original mapping population were selected based on their marker genotype and QTL structure. When crossed, these lines would be expected to produce progeny with most favorable QTL alleles. One hundred doubled haploid (DH) lines from the F₁ hybrid of this cross were genotyped with ten RFLP markers and one morphological marker defining grain yield to monitor QTL segregation. A subset of 24 lines representing various combinations of putatively favorable and unfavorable QTL alleles, together with Steptoe, ‘Morex’, SM73, and SM145, were phenotyped for grain yield in five environments. Multiple regression procedures were used to explore phenotype and genotype relationships. Most target QTLs showed significant effects. However, significance and magnitude of QTL effects and favorable QTL allele phase varied across environments. All target QTLs showed significant QTL-by-environment interaction (QTL×E), and the QTL on chromosome 2 expressed alternative favorable QTL alleles in different environments. Digenic epistatic effects were also detected between some QTL loci. For traits such as grain yield, marker-assisted selection efforts may be better targeted at determining optimum combinations of QTL alleles rather than pyramiding alleles detected in a reference mapping population. Received: 2 June 1998 / Accepted: 17 September 1998     

Thermoregulated Expression and Characterization of an NAD(P)H-Dependent 2-Cyclohexen-1-one Reductase in the Plant Pathogenic Bacterium Pseudomonas syringae pv. glycinea

The phytopathogenic bacterium Pseudomonas syringae pv. glycinea PG4180.N9 causes bacterial blight of soybeans and preferably infects its host plant during periods of cold, humid weather conditions. To identify proteins differentially expressed at low temperatures, total cellular protein fractions derived from PG4180.N9 grown at 18 and 28°C were separated by two-dimensional gel electrophoresis. Of several proteins which appeared to be preferentially present at 18°C, a 40-kDa protein with an isoelectric point of approximately 5 revealed significant N-terminal sequence homology to morphinone reductase (MR) of Pseudomonas putida M10. The respective P. syringae gene was isolated from a genomic cosmid library of PG4180, and its nucleotide sequence was determined. It was designated ncr for NAD(P)H-dependent 2-cyclohexen-1-one reductase. Comparison of the 1,083-bp open reading frame with database entries revealed 48% identity and 52% similarity to the MR-encoding morB gene of P. putida M10. The ncr gene was overexpressed in Escherichia coli, and its gene product was used to generate polyclonal antisera. Purified recombinant Ncr protein was enzymatically characterized with NAD(P)H and various morphinone analogs as substrates. So far, only 2-cyclohexen-1-one and 3-penten-2-one were found to be substrates for Ncr. By high-pressure liquid chromatography analysis, flavin mononucleotide could be identified as the noncovalently bound prosthetic group of this enzyme. The distribution of the ncr gene in different Pseudomonas species and various strains of P. syringae was analyzed by PCR and Southern blot hybridization. The results indicated that the ncr gene is widespread among P. syringae pv. glycinea strains but not in other pathovars of P. syringae or in any of the other Pseudomonas strains tested.     

Force-generating capacities and fatigability of the quadriceps femoris in relation to different exercise modes

In this study, we examined whether different exercise modes provoke functional differences in maximal and explosive force-generating capacities and fatigability of the quadriceps femoris (QF). Additionally, the interaction of different functional capacities was studied in competitive athletes. Ten competitive tennis players and 10 endurance athletes participated in the study. Pre-exercise force-generating capacities were determined during maximal voluntary isometric knee extensions (MVC). Fatigability of the QF was studied using sustained isometric contractions with target loads of 20% and 40% of pre-exercise MVC. Postexercise MVCs were conducted 20 seconds, 1 minute, and 3 minutes post task failure. Muscle activation of the QF during the fatiguing exercises and postexercise MVCs was estimated using surface electromyography. Higher explosive force-generating capacities, but no differences in absolute moments, were detected in tennis players compared with endurance athletes. Fatigability of the QF during both fatiguing tasks was approximately the same in both athletic populations. This was indicated by minor group differences in endurance time, postexercise MVC production, and electromyography (EMG)-estimated muscle activation during fatigue. Variability in endurance time was not significantly associated with pre-exercise force-generating capacities in these competitive athletes. In both athletic populations, recovery of MVC was significantly slower after the fatiguing contraction with 20% of MVC compared with that with 40% of MVC. These results may enhance understanding of plasticity of the neuromuscular system and yield interesting information for the optimization of athletic training programs. Explosive strength training might enhance endurance athletes' explosiveness without decreasing muscle fatigue resistance. The exercise profile of competitive tennis is suggested to act as a sufficient trigger to reach high neuromuscular fatigue resistance but may be inadequate to cause significant gains in absolute muscle strength.