Two groups control light-induced schiff base deprotonation and the proton affinity of asp(85) in the Arg(82)His mutant of bacteriorhodopsin

Arg(82) is one of the four buried charged residues in the retinal binding pocket of bacteriorhodopsin (bR). Previous studies show that Arg(82) controls the pK(a)s of Asp(85) and the proton release group and is essential for fast light-induced proton release. To further investigate the role of Arg(82) in light-induced proton pumping, we replaced Arg(82) with histidine and studied the resulting pigment and its photochemical properties. The main pK(a) of the purple-to-blue transition (pK(a) of Asp(85)) is unusually low in R82H: 1.0 versus 2.6 in wild type (WT). At pH 3, the pigment is purple and shows light and dark adaptation, but almost no light-induced Schiff base deprotonation (formation of the M intermediate) is observed. As the pH is increased from 3 to 7 the M yield increases with pK(a) 4.5 to a value approximately 40% of that in the WT. A transition with a similar pK(a) is observed in the pH dependence of the rate constant of dark adaptation, k(da). These data can be explained, assuming that some group deprotonates with pK(a) 4.5, causing an increase in the pK(a) of Asp(85) and thus affecting k(da) and the yield of M. As the pH is increased from 7 to 10.5 there is a further 2.5-fold increase in the yield of M and a decrease in its rise time from 200 &mgr;s to 75 &mgr;s with pK(a) 9. 4. The chromophore absorption band undergoes a 4-nm red shift with a similar pK(a). We assume that at high pH, the proton release group deprotonates in the unphotolyzed pigment, causing a transformation of the pigment into a red-shifted "alkaline" form which has a faster rate of light-induced Schiff base deprotonation. The pH dependence of proton release shows that coupling between Asp(85) and the proton release group is weakened in R82H. The pK(a) of the proton release group in M is 7.2 (versus 5.8 in the WT). At pH < 7, most of the proton release occurs during O --> bR transition with tau approximately 45 ms. This transition is slowed in R82H, indicating that Arg(82) is important for the proton transfer from Asp(85) to the proton release group. A model describing the interaction of Asp(85) with two ionizable residues is proposed to describe the pH dependence of light-induced Schiff base deprotonation and proton release.     

Effect of methylene blue on cardiac output response to exercise in dogs

To determine whether the increase in cardiac output during mild to moderate exercise is related to an increase in the tissue redox potential, we compared the responses of cardiac output, total body oxygen consumption, and arterial blood lactate-to-pyruvate ratio (a measure of NADH/NAD) to treadmill exercise between dogs treated with normal saline and those treated with a hydrogen acceptor, new methylene blue. Normal saline was infused into the left atrium in the first group of dogs at a rate of 0.38 ml/min throughout the treadmill exercise (2.5 mph and 5.0 mph on a 6% incline, each for 20 min). In the second group, methylene blue was administered as a loading dose (4 mg/kg) before exercise, followed by a continuous infusion (0.15 mg X kg-1 X min-1) throughout exercise. A similar infusion of methylene blue was given to a third group of dogs without exercise; it reduced the arterial lactate-to-pyruvate ratio from 6.70 +/- 0.35 to 4.12 +/- 0.27 but had no or little effects on cardiac output, heart rate, arterial pressure, and left ventricular dP/dt and (dP/dt)/P. Treadmill exercise doubled cardiac output and increased total body O2 consumption three- to fourfold in the first two groups but increased arterial blood lactate-to-pyruvate ratio only in group 1 (6.0 +/- 0.54 to 9.97 +/- 0.91). The relationship between cardiac output and total body O2 consumption was unaffected by the simultaneous administration of methylene blue during exercise. Groups 1 and 2 also did not differ in their heart rate, left ventricular dP/dt and (dP/dt)/P, and plasma catecholamine responses to exercise.(ABSTRACT TRUNCATED AT 250 WORDS)     

Complete intra-annual cycle of leaf area index in a Platanus orientalis L. stand

Leaf area index (LAI) analysis of deciduous forest trees is usually restricted to seasonal monitoring involving the assessment of distinct leaf phenological stages within definite time intervals of the year. However, continuous LAI monitoring that includes entire leaf periods is necessary to define the ecophysiological characteristics of deciduous trees. Therefore, this study investigated the intra-annual cycle of the LAI for a Platanus orientalis L. stand in the Bart?n watershed of Turkey. A complete cycle involves three periods: foliation, stable, and defoliation. The foliation period comprises budburst, leaf emergence and flushing sessions, whereas the defoliation period consists of leaf senescence and leaf fall sessions. The stable period is in between these two periods when LAI values are at a climax around maximum. Eight points were determined in the field for the analysis of LAI by a hemispherical photography technique. Over a relatively frequent schedule, photographs were taken almost weekly during the foliation period. Both weekly and approximate monthly photographs were applied during the stable period. Finally, near-monthly photographs were taken for the defoliation period. The foliation period lasted for about 1.5 months from mid-April to May with the mean LAI reaching from 0.16 up to 2.38. Mean LAI was between 2.38 and 2.47 for a stable period over 2 months (June and July). For the defoliation period, mean LAI dropped from 2.42 down to 0.35 over 5 months from August to December. The total foliated period was more than 8 months, which is relatively long for a temperate forest. In addition, correlations between mean LAI and maximum, mean and minimum temperatures were highly significant (P < 0.01) with coefficients (r) of 0.79, 0.90 and 0.93, respectively. By describing the intra-annual LAI pattern, this study fills a gap in the literature on the phenology of Platanus orientalis L.     

Molecular population genetics of the alcohol dehydrogenase locus in the Hawaiian drosophilid D. mimica

Sequence variation among 10 alleles of the alcohol dehydrogenase (Adh) gene of the Hawaiian drosophilid D. mimica was analyzed with reference to the evolutionary history of the Hawaiian subgroup as well as to levels and patterns of polymorphism of the Adh gene in continental drosophilid species. The Adh gene of D. mimica is less polymorphic than that of other drosophilid species, and no replacement substitutions were found. Statistical analyses of the Adh alleles suggested the action of balancing selection and revealed significant linkage disequilibrium among three of the variable sites. The effective population size was estimated to be only slightly smaller than that of continental species and, surprisingly, on the same order of magnitude as the actual size.      

Insecticidal effects of selected biological control agents on the larvae of Plutella xylostella (Lepidoptera: Plutellidae)

To identify a more effective and safe biological control agent against a common cabbage pest, Plutella xylostella (L.) (Lepidoptera: Plutellidae), the insecticidal effects of selected biological agents were evaluated. The highest insecticidal effects determined were 100, 73.5, 45.5, 47 and 55.3% using toxin HD‐1 (isolated from the Harry Dumagae strain of Bacillus thuringiensis), toxin BTS‐1 (isolated from the tenebrionis strain of B. thuringiensis), B. thuringiensis Berliner, B. thuringiensis israelensis and B. thuringiensis kurstaki, respectively.