The buffering capacity of the internal phase of thylakoids and the magnitude of the pH changes inside under flashing light

The buffering capacity inside thylakoids is determined and the magnitude of flash-induced pH changes inside is calibrated in the pH range from 6.4 to 8.1. The work is based on flash-induced absorption changes of neutral red in a chloroplast suspension in which the outer phase is strongly buffered by bovine serum albumin. It is shown that neutral red is bound inside thylakoids. The binding can be described by a simple isotherm with an apparent K_m = 4 μM and saturation at 1 neutral red per 17 chlorophylls. The apparent pK of neutral red is shifted from 6.6 in solution to 7.25 when bound inside. It is demonstrated that neutral red is a clean indicator of pH changes inside, i.e. when properly used it shows no response to other events. Although bound it reports pH changes which occur in the internal osmolar (aqueous) volume of thylakoids. This is obvious from the influence of chemically very different buffers on the magnitude of the absorption changes of neutral red. These act in a manner proportional to their calculated buffering capacity in aqueous solution. The intrinsic buffering capacity of the internal phase is determined with the aid of these buffers, at pH 7.2 it is between 0.8 and 1 mM (at 60 mosM). The absence of large variations in the buffering capacity in the range from pH 6.4 to 8.1 suggests that proteinaceous groups are involved in addition to the lipids which may dominate the buffering capacity at lower pH. The magnitude of the internal pH change is approx. 0.6 (at pH 7.3) under stimulation of both photosystems with a short xenon flash of light.     

99mTc-Labeled HYNIC-DAPI Causes Plasmid DNA Damage with High Efficiency

^99mTc is the standard radionuclide used for nuclear medicine imaging. In addition to gamma irradiation, ^99mTc emits low-energy Auger and conversion electrons that deposit their energy within nanometers of the decay site. To study the potential for DNA damage, direct DNA binding is required. Plasmid DNA enables the investigation of the unprotected interactions between molecules and DNA that result in single-strand breaks (SSBs) or double-strand breaks (DSBs); the resulting DNA fragments can be separated by gel electrophoresis and quantified by fluorescent staining. This study aimed to compare the plasmid DNA damage potential of a ^99mTc-labeled HYNIC-DAPI compound with that of ^99mTc pertechnetate (^99mTcO₄ ⁻). pUC19 plasmid DNA was irradiated for 2 or 24 hours. Direct and radical-induced DNA damage were evaluated in the presence or absence of the radical scavenger DMSO. For both compounds, an increase in applied activity enhanced plasmid DNA damage, which was evidenced by an increase in the open circular and linear DNA fractions and a reduction in the supercoiled DNA fraction. The number of SSBs elicited by ^99mTc-HYNIC-DAPI (1.03) was twice that caused by ^99mTcO₄ ⁻ (0.51), and the number of DSBs increased fivefold in the ^99mTc-HYNIC-DAPI-treated sample compared with the ^99mTcO₄ ⁻ treated sample (0.02 to 0.10). In the presence of DMSO, the numbers of SSBs and DSBs decreased to 0.03 and 0.00, respectively, in the ^99mTcO₄ ^– treated samples, whereas the numbers of SSBs and DSBs were slightly reduced to 0.95 and 0.06, respectively, in the ^99mTc-HYNIC-DAPI-treated samples. These results indicated that ^99mTc-HYNIC-DAPI induced SSBs and DSBs via a direct interaction of the ^99mTc-labeled compound with DNA. In contrast to these results, ^99mTcO₄ ⁻ induced SSBs via radical formation, and DSBs were formed by two nearby SSBs. The biological effectiveness of ^99mTc-HYNIC-DAPI increased by approximately 4-fold in terms of inducing SSBs and by approximately 10-fold in terms of inducing DSBs.     

塔克拉玛干沙漠南缘柽柳和胡杨水势季节变化研究

对塔克拉玛干沙漠南缘的柽柳和胡杨生长周期内的清晨水势和水势日变化的连续野外测定表明,两种植物在整个生长期内均未出现明显的水分亏缺.清晨水势的季节变化幅度不大,正午水势有不同程度的降低;一次性人工灌溉对植物水分状况没有明显影响.采伐利用方式不影响植物的水分状况.地下水是柽柳和胡杨生存与生长的先决条件.维持该区域地下水位的基本稳定是保证该区域柽柳和胡杨恢复重建的重要前提.     

Identification of a Lifespan Extending Mutation in the Schizosaccharomyces pombe Cyclin Gene clg1 + by Direct Selection of Long-Lived Mutants

Model organisms such as budding yeast, worms and flies have proven instrumental in the discovery of genetic determinants of aging, and the fission yeast Schizosaccharomyces pombe is a promising new system for these studies. We devised an approach to directly select for long-lived S. pombe mutants from a random DNA insertion library. Each insertion mutation bears a unique sequence tag called a bar code that allows one to determine the proportion of an individual mutant in a culture containing thousands of different mutants. Aging these mutants in culture allowed identification of a long-lived mutant bearing an insertion mutation in the cyclin gene clg1 ⁺. Clg1p, like Pas1p, physically associates with the cyclin-dependent kinase Pef1p. We identified a third Pef1p cyclin, Psl1p, and found that only loss of Clg1p or Pef1p extended lifespan. Genetic and co-immunoprecipitation results indicate that Pef1p controls lifespan through the downstream protein kinase Cek1p. While Pef1p is conserved as Pho85p in Saccharomyces cerevisiae, and as cdk5 in humans, genome-wide searches for lifespan regulators in S. cerevisiae have never identified Pho85p. Thus, the S. pombe system can be used to identify novel, evolutionarily conserved lifespan extending mutations, and our results suggest a potential role for mammalian cdk5 as a lifespan regulator.     

Treadmill desks: A 1‐year prospective trial

Objective: Sedentariness is associated with weight gain and obesity. A treadmill desk is the combination of a standing desk and a treadmill that allow employees to work while walking at low speed. Design and Methods: The hypothesis was that a 1‐year intervention with treadmill desks is associated with an increase in employee daily physical activity (summation of all activity per minute) and a decrease in daily sedentary time (zero activity). Employees (n = 36; 25 women, 11 men) with sedentary jobs (87 ± 27 kg, BMI 29 ± 7 kg/m², n = 10 Lean BMI < 25 kg/m², n = 15 Overweight 25 < BMI < 30 kg/m², n = 11 Obese BMI > 30 kg/m²) volunteered to have their traditional desk replaced with a treadmill desk to promote physical activity for 1 year. Results: Daily physical activity (using accelerometers), work performance, body composition, and blood variables were measured at Baseline and 6 and 12 months after the treadmill desk intervention. Subjects who used the treadmill desk increased daily physical activity from baseline 3,353 ± 1,802 activity units (AU)/day to, at 6 months, 4,460 ± 2,376 AU/day (P < 0.001), and at 12 months, 4,205 ± 2,238 AU/day (P < 0.001). Access to the treadmill desks was associated with significant decreases in daily sedentary time (zero activity) from at baseline 1,020 ± 75 min/day to, at 6 months, 929 ± 84 min/day (P < 0.001), and at 12 months, 978 ± 95 min/day (P < 0.001). For the whole group, weight loss averaged 1.4 ± 3.3 kg (P < 0.05). Weight loss for obese subjects was 2.3 ± 3.5 kg (P < 0.03). Access to the treadmill desks was associated with increased daily physical activity compared to traditional chair‐based desks; their deployment was not associated with altered performance. For the 36 participants, fat mass did not change significantly, however, those who lost weight (n = 22) lost 3.4 ± 5.4 kg (P < 0.001) of fat mass. Weight loss was greatest in people with obesity. Conclusions: Access to treadmill desks may improve the health of office workers without affecting work performance.     

Duck hunter activity,success, and satisfaction on public hunting areas

Because hunting disturbance can influence local distribution of ducks and their availability to hunters, managers often limit access to hunting areas to improve hunting success and satisfaction. Few studies have quantified the effectiveness of public area access restrictions on duck hunter activity, harvest, or hunters' satisfaction with their hunting experience. We used a cross-over design over 6 consecutive hunting seasons (2008–2009 through 2013–2014) on State Wildlife Areas (SWAs) in northeastern Colorado, USA, to compare the effects of restricted hunting access regulations and regulations without these restrictions on duck hunter activity, harvest success, harvest levels, and satisfaction. We also considered effects of SWA types, duck abundance, temperature, precipitation, use of equipment by duck hunting parties, and, for hunter satisfaction, hunting success, hunting parties' satisfaction with ducks seen, habitat conditions, crowding from other hunters, and SWA regulations. The number of days when duck hunters had access to restricted properties was about half that on unrestricted properties, and unrestricted properties were used by about twice as many duck hunting parties, but the mean number of hunting parties per available hunting day and mean party size were similar under the 2 types of regulations. Most (56%) duck hunting parties did not bag any ducks; hunting success (harvest of ≥1 duck by a hunting party) was best explained by a model that included a regulation type × hunting season interaction, a SWA type × month interaction, hunter density the previous day, an index of hunter investment (number of decoys used and whether dogs and calls were used), and temperature. Successful hunting parties harvested 1.92 ± 1.60 (SD) ducks/hunter/day (range = 0.1–7.0); the best model predicting the number of ducks harvested per hunter in successful parties included a regulation type × hunting season interaction, a SWA type × month interaction, hunter density the previous day, an index of hunter investment, temperature, and precipitation. Overall satisfaction of duck hunting parties with a day's hunt averaged 3.62 ± 1.20 based on a rank scale of 1 (very unsatisfied) to 5 (very satisfied), and was best explained by a model that included hunting party success; hunter investment; temperature; and satisfaction with duck numbers, habitat conditions, hunting regulations, and crowding. While greatly reducing days available for hunting, access restrictions were associated with improved chances of hunting parties successfully harvesting ≥1 duck in 5 of 6 hunting seasons, and substantially greater numbers of ducks harvested by hunters in successful parties in 1 of 6 hunting seasons. Restrictions did not have a strong direct effect on hunting parties' satisfaction with a day's hunt. Uncontrolled factors, including weather and use of equipment by hunters, had important influences on hunter success, harvest, and satisfaction. Managers should carefully assess hunting activity, hunter expectations and desires, and hunting area characteristics when considering access restrictions on public hunting areas.     

Non‐destructive dry matter estimation of Alhagi sparsifolia vegetation in a desert oasis of Northwest China

Question: Can above‐ground biomass of naturally growing Alhagi sparsifolia shrubs be estimated non‐destructively? Location: Qira oasis (37° 01′N, 80° 48′E, 1365 ma.s.l.) at the southern fringe of the Taklamakan desert, Xinjiang, NW China. Methods: Two methods were compared to estimate above‐ground biomass (AGB) of Alhagi. At first shrub AGB was estimated by manual ground measurements (called ‘allometric approach’) of length, width and height of 50 individuals. Subsequently regression equations were established between calculated shrub canopy volume and shrub AGB (r²= 0.96). These equations were used to calculate AGB from manual ground measurements in 20 sample plots within the Alhagi field. Secondly, kite‐based colour aerial photography coupled with the use of a Geographic Information System (called ‘GIS approach’) was tested. First and second order polynomial regressions between AGB data of the 50 individual shrubs and their respective canopy area allowed to automatically calculate the AGB of all remaining shrubs covered by the photograph (r²= 0.92 to 0.96). The use of non‐linear AGB regression equations required an automatised separation of shrubs growing solitary or in clumps. Separation criteria were the size and shape of shrub canopies. Results: The allometric approach was more reliable but also more time‐consuming than the GIS‐based approach. The latter led to an overestimation of Alhagi dry matter in densely vegetated areas. However, this systematic error decreased with increasing size of the surveyed area. Future research in this field should focus on improvements of AGB estimates in areas of high shrub density.     

多枝柽柳气体交换特性研究

在美国柽柳（Tamarix L.）被列为臭名昭著的十大我来杂草之一,而在中国,它则作为一种濒危物种生存状况堪忧。对多枝柽柳（Tamarix ramosissima L bd.）气体交换进行研究,探讨其蒸汽压差（VPD）、叶水势、气孔导度（g）和水分利用效率（WUE）之间的联系,结果表明,柽柳之所以能在沙漠生存又能在美国河岸地带疯狂扩展,主要原因可能是：①柽柳属地下水湿生植物,发达的根系能直达地下水。②柽柳适应性生境很广,作为一种阳性树种,柽柳非常耐旱,能够在很低的叶水势（φ）条件下（-4.59Mpa）进行光合作用,而较低的光补偿点和暗呼吸速率又反映出其耐阴湿的特点。③灵敏的气孔适应性调整：其g对VPD的灵敏性随着干湿季其体内的水分状况变化而变化,调整蒸腾速率,从而影响其WUE;gs与φ成显著正相关（P≤0.05）;gs与WUE显著相关（P≤0.05）,但只有当gs处于正在下降的时候,光合作用和蒸腾作用两个交换过程受轻微限制时,可得到水消耗和CO2吸收的最优协调,WUE达到最大值。这些适应性调整可能是柽柳能在美国河岸地带疯狂扩展的主要原因。实验结果表明,柽柳完全能够适应荒漠生长环境,因而导致柽柳在我国处于濒危状况的根本原因不是传统认为的干旱的自然环境,而主要是人为因素造成地下水位下降以及过度砍伐所致。