Temperature-sensitive coupling and uncoupling of ATPase-mediated,nonradiative energy dissipation: Similarities between chloroplasts and leaves

The effects of temperature on the dark relaxation kinetics of nonradiative energy dissipation in photosystem II were compared in lettuce (Lactuca sativa L.) chloroplasts and leaves of Aegialitis annulata R. Br. After high levels of violaxanthin de-epoxidation in the light, Aegialitis leaves showed a marked delay in the dark relaxation of nonradiative dissipation, measured as non-photochemical quenching (NPQ) of photosystem II chlorophyll a fluorescence. Aegialitis leaves also maintained a moderately high adenylate energy charge at low temperatures during and after high-light exposure, presumably because of their limited carbon-fixation capacity. Similarly, dark-sustained NPQ could be induced in lettuce chloroplasts after de-epoxidizing violaxanthin and light-activating the ATP synthase. The duration and extent of dark-sustained NPQ were strongly enhanced by low temperatures in both chloroplasts and leaves. Further, the NPQ sustained at low temperatures was rapidly reversed upon warming. In lettuce chloroplasts, low temperatures sharply decreased the ATP-hydrolysis rate while increasing the duration and extent of the resultant trans-thylakoid proton gradient that elicits the NPQ. This was consistent with a higher degree of energy-coupling, presumably due to reduced proton diffusion through the thylakoid membrane at the lower temperatures. The chloroplast adenylate pool was in equilibrium with the adenylate kinase and therefore both ATP and ADP contributed to reverse coupling. The low-temperature-enhanced NPQ quenched the yields of the dark level (F_o) and the maximal (F_m) fluorescence proportionally in both chloroplasts and leaves. The extent of NPQ in the dark was inversely related to the efficiency of photosystem II, and very similar linear relationships were obtained over a wide temperature range in both chloroplasts and leaves. Likewise, the dark-sustained absorbance changes, caused by violaxanthin de-epoxidation (A_508nm) and energy-dependent light scattering (A_536nm) were strikingly similar in chloroplasts and leaves. Therefore, we conclude that the dark-sustained, low-temperature-stimulated NPQ in chloroplasts and leaves is apparently directly dependent on lumen acidification and chloroplastic ATP hydrolysis. In leaves, the ATP required for sustained NPQ is evidently provided by oxidative phosphorylation in the mitochondria. The functional significance of this quenching process and implications for measurements of photo-protection versus photodamage in leaves are discussed.Abbreviations and Symbols A antheraxanthin - Chl chlorophyll - DPS de-epoxidation state of the xanthophyll cycle, ([Z+A]/[V+A+Z]) - F, F steady-state fluorescence in the absence, presence of thylakoid energization - F_o, F_o dark fluorescence level in the absence, presence of thylakoid energization - F_m, F_m maximal fluorescence in absence, presence of thylakoid energization - NPQ nonphotochemical quenching (F_m/F_m)–1 - V violaxanthin - Z zeaxanthin - NRD nonradiative dissipation - PFD photon flux density - [2ATP+ADP] - pH trans-thylakoid proton gradient - S pH-dependent light scattering - _PSII (F_m–F)/F_m, photon yield of PSII photochemistry at the actual reduction state in the light or dark - [ATP+ADP+AMP] We thank Connie Shih for skillful assistance in growing plants and for conducting HPLC analyses. Support from an NSF/USDA/DOE postdoctoral training grant to A.G. is gratefully acknowledged. A.G. also wishes to thank Prof. Govindjee for valuable discussions. C.I.W.-D.P.B. Publication No. 1197.     

Carbon sequestration rates in organic layers of boreal and temperate forest soils — Sweden as a case study

Aim The aim of this work was to estimate C sequestration rates in the organic matter layer in Swedish forests. Location The region encompassed the forested area (23 × 10⁶ ha) of Sweden ranging from about 55° N to 69° N. Methods We used the concept of limit values to estimate recalcitrant litter remains, and combined it with amount of litter fall. Four groups of tree species were identified (pine, spruce, birch and ‘other deciduous species’). Annual actual evapotranspiration (AET) was estimated for 5 × 5 km grids covering Sweden. For each grid, data of forested area and main species composition were available. The annual input of foliar litter into each grid was calculated using empirical relationships between AET and foliar litter fall in the four groups. Litter input was combined with average limit values for decomposition for the four groups of litter, based on empirical data. Finally, C sequestration rate was calculated using a constant factor of the C concentration in the litter decomposed to the limit value, thus forming soil organic matter (SOM). Results We obtained a value of 4.8 × 10⁶ metric tons of C annually sequestered in SOM in soils of mature forests in Sweden, with an average of 180 kg ha^?1 and a range from 40 to 410 kg ha^?1. Norway spruce forests accumulated annually an average of 200 kg C ha^?1. The pine and birch groups had an average of 150 kg ha^?1 and for the group of other deciduous trees, which is limited to south Sweden, the C sequestration was around 400 kg ha^?1. Conclusions There is a clear C sequestration gradient over Sweden with the highest C sequestration in the south‐west, mainly corresponding to the gradient in litter fall. The limit‐value method appears useful for scaling up to a regional level to describe the C sequestration in SOM. A development of the limit value approach in combination with process‐orientated dynamic models may have a predictive value.     

Orthostatic Hypotension and Elevated Resting Heart Rate Predict Low-Energy Fractures in the Population: The Malm? Preventive Project

BackgroundAutonomic disorders of the cardiovascular system, such as orthostatic hypotension and elevated resting heart rate, predict mortality and cardiovascular events in the population. Low-energy-fractures constitute a substantial clinical problem that may represent an additional risk related to such autonomic dysfunction.AimsTo test the association between orthostatic hypotension, resting heart rate and incidence of low-energy-fractures in the general population.ConclusionOrthostatic blood pressure decline and elevated resting heart rate independently predict low-energy fractures in a middle-aged population. These two measures of subclinical cardiovascular dysautonomia may herald increased risks many years in advance, even if symptoms may not be detectable. Although the effect sizes are moderate, the easily accessible clinical parameters of orthostatic blood pressure response and resting heart rate deserve consideration as new risk predictors to yield more accurate decisions on primary prevention of low-energy fractures.     

Reemergence and second brood in the bark beetle Ips typographus

The knowledge about reemergence of parent spruce bark beetles Ips typographus . their dispersal, and production of a second brood is reviewed. A majority of the beetles reemerge after their first brood. The process is mainly determined by temperature but high breeding density decreases the average time spent in the tree. The difference between males and females in reemergence seems to be small. In the field, a positive relationship between residence time and fat content at reemergence seems to exist, whereas the opposite tendency is found under laboratory conditions. In the laboratory, the survival of beetles reemerging late is on the average lower than that for early reemerging beetles. The time of reemergence and size of the first brood do not, however, seem to influence the production of a second brood. A large proportion of the reemerging beetles are able to establish a new brood in the laboratory, but calculations based on the number of attacked trees and estimated reemergence in forests in South Norway suggest that only about one third of the beetles actually produce a second brood. In northern Europe, successful colonization of new trees by the reemerging beetles seems to require a period of several warm days during the main reemergence period.     

Protein gene product 9.5-immunoreactive nerve fibres and cells in human skin

Summary Sections of human skin were processed according to the indirect immunofluorescence technique with a rabbit antiserum against human protein gene product 9.5 (PGP 9.5). Immunoreactivity was detected in intraepidermal and dermal nerve fibres and cells. The intraepidermal nerves were varicose or smooth with different diameters, running as single processes or branched, straight or bent, projecting in various directions and terminating in the stratum basale, spinosum or granulosum. The density of the intraepidermal nerves varied between the different skin areas investigated. PGP 9.5-containing axons of the lower dermis were found in large bundles. They separated into smaller axon bundles within the upper dermis, entering this portion of the skin perpendicular to the surface. Then they branched into fibres mainly arranged parallel to the epidermal-dermal junctional zone. However, the fibres en route to the epidermis traversed the upper dermis more or less perpendicularly. Furthermore, immunoreactive dermal nerve fibres were found in the Meissner corpuscles, the arrector pili muscles, hair follicles, around the eccrine and apocrine sweat glands and around certain blood vessels. Such fibres were also observed around most subcutaneous blood vessels, sometimes heavily innervating these structures. Numerous weakly-to-strongly PGP 9.5-immunoreactive cells were found both in the epidermis and in the dermis.     

Gas chromatography with nitrogen-selective detection of metoprolol from human plasma after reaction with phosgene

A method for the determination of therapeutic levels of metoprolol in human plasma is presented. Metoprolol and the internal standard are extracted from the buffered plasma sample to an organic phase containing 4 x 10^?3M phosgene. After 10 min the organic phase is taken to dryness. The residue is dissolved in ethyl acetate and the formed oxazolidine derivatives are analyzed by gas chromatography with nitrogen-selective detection. With packed columns, rectilinear standard curves through the origin were obtained down to 80 nmoles/l of plasma. The precision of the method at 200 nmoles/l was 1.5% (n = 8). The sensitivity of the method was improved by using capillary column gas chromatography. Linear standard curves were obtained down to 10 nmoles/1 of metoprolol in plasma. The precision of the method at the 50 nmoles/1 level was 2.2% (n = 7). With this simple and straightforward method using extractive derivatization 30 samples can be handled in a day.     

The effect of factors released from the tumor-transformed cells on DNA synthesis, mitosis, and cellular enlargement in 3T3 fibroblasts

Quiescent serum-starved 3T3 cells can be stimulated to initiate DNA synthesis after addition of conditioned media from spontaneously tumor-transformed 3T3 cells (3T6-cells) or from SV-40-transformed 3T3 cells (SV-3T3 cells). The conditioned media were found to stimulate both the chromosome cycle (i.e., DNA synthesis and cell division) and the growth cycle (i.e., cellular enlargement). Furthermore, addition of conditioned media to quiescent 3T3 cells increased the activity of HMG CoA reductase--an enzyme previously proposed to exercise some control on cell proliferation in 3T3 cells (Larsson and Zetterberg: J. Cell. Physiol. 129:99-102, 1986. The increased activity of HMG CoA reductase after treatment with tumor cell conditioned media was correlated to the stimulatory effects on DNA synthesis. By treating 3T3 cells stimulated to resume proliferation by addition of conditioned media with mevinolin (a competitive inhibitor of HMG CoA reductase) the activity of HMG CoA reductase as well as the DNA synthesis and cell division were efficiently inhibited. In contrast, HMG CoA activity was not coupled to the cellular enlargement. Therefore, it is proposed that one set of factors present in tumor cell conditioned media preferentially stimulates the chromosome cycle by increasing the HMG-CoA reductase activity, whereas another set of factors is responsible for growth in cell size. Both types of factors are required for balanced growth.