Singlet Oxygen Generated in PS￠ò Particles Promotes the Superoxide Generation

The relationship between superoxide and singlet oxygen in PSⅡ particle of spinach under strong illumination was studied with spin-trap ESR technique. The generation of superoxide was increased under D2 environment which prolonged the half life of singlet oxygen. The generation of superoxide was decreased when histidine existed as a scavenger of singlet oxygen. It is highly possible that the superoxide generated in PSⅡ particle originates from singlet oxygen.     

Preliminary Study on Electron Crystallography of 27 kD LHC-￠ò Complex from Cucumber

Cucumber (Cucumis sativus L.) LHC-Ⅱ complex, which consists of only one subunit (27 kD), was isolated and purified. 2-D crystallization was performed by batch method. The crystal is 0.7 μm×1.0 μm, and diffracts to 30 ?. The projection map of the negatively stained two-dimensional crystal of LHC-Ⅱ complex shows that the crystal has p3 symmetry, lattice constant 15.4 nm×15.4 nm, which is different from the LHC-Ⅱ of spinach (Spinacia oleracea L.) and pea (Pisum satium L.). A continuous tomographic tilt series, containing 12 projections from the two-dimensional crystal was subjected to 3-D reconstruction. The 3-D model represents that LHC-Ⅱ complex consists of 6 monomers. These trimer and dimer interactions build up the six member ring.      

Purification and Spectral Characteristics of Cytochrome b-559 from Oxygen-evolving Photosystem ￠ò Core Complexes of Spinach and Rice

Cytochrome b-559 in photosystem Ⅱ reaction center was purified from spinach (Spinacia oleracea L.) and rice (Oryza sativa L.) by a rapid and simple procedure. Their low temperature fluorescence emission and excitation spectra, ultraviolet fluorescence spectra and absolute absorption spectra were presented. The author's purification methods, which enhanced the yield of pure protein and shorted the time for isolation, have several advantages: 1. use of oxygen-evolving PSⅡ core complexes as the starting material in order to avoid disturbing from other cytochromes; 2. isocratic elution of cytochrome b-559 from a DEAE-Sephacel column for eliminating the impurity and yielding the protein in pure state; 3. a simple column procedure for removal of excess Triton X-100. Purified cytochromes b-559 from these species have similar optical spectra and mobility during gel electrophoresis under native conditions. From the results of novel electrophoresis (Tricine-SDS-PAGE), cytochrome b-559 from both spinach and rice reveal two polypeptide bands (apparent molecular weight 9 kD and 4 kD, respectively). By measuring of 77 K fluorescence spectra, it was shown that for the purified cytochrome b-559 there were two excitation peaks at 439 nm and 413 nm, and two emission peaks at 563 nm and 668 nm. This is the first indication that Cyt b-559 is able to emit fluorescence and also transfer excited electrons to chlorophyll. By the use of ultraviolet fluorescence spectra, it was demonstrated for the first time that the location of Trp residue could be in the hydrophobic transmembrane region of cytochrome b-559.     

Orientation of Pigments in the Isolated Photosystem ￠ò Sub-core Reaction Center CP47/D1/D2/Cyt b-559 Complexes: A Linear Dichrosism Study

Linear dichroism (LD) spectroscopy is an important technique in the study of the orientation and organization of pigments in the photosynthetic membrane complexes in vivo and in vitro. In this work, the orientation of the pigments in the isolated photosystem Ⅱ (PSⅡ) sub-core reaction center complexes was analyzed and characterized by means of low temperature absorption and LD spectroscopy. The preparations containing different amounts of CP47 isolated from spinach (Spinacia oleracea L.) chloroplast were used in order to investigate the orientation of pigments in the PSⅡ sub-core CP47/D1/D2/Cyt b-559 (CP47/D1/D2) complexes. Chlorophyll a (Chl a) absorbing at 680 nm in CP47/D1/D2/Cyt b-559 complex showed an orientation of the Q y transition parallel to the membrane plane. It is proposed that there are two forms of β-carotene (β-Car) in CP47/D1/D2/Cyt b-559 complex, denoted as β-Car (Ⅰ)and β-Car (Ⅱ), with different orientations, β-Car (Ⅰ) at 470 and 505 nm is roughly parallel to the membrane plane, and β-Car (Ⅱ) at 460 and 490 nm seems to be perpendicular orientation. Upon the photoinhibitory experiment β-Car (Ⅱ) was found to be photosensitive and easily photodamaged. It also showed that the positive LD signal observed at 680 nm was quite complicated. This signal is tentatively attributed to P680 and some Chl　a of antenna in CP47 protein based upon our measurements.     

Studies on Femtosecond Fluorescence Properties of Light-harvesting Complex in Photosystem ￠ò

Fluorescence properties of light-harvesting complex LHCⅡ） in photosystem Ⅱ isolated from spinach (Spinacia oleracea L.) was investigated with the time-resolved fluorescence spectroscopic technique. The sample was excited by double frequency Ti:Al203 laser with 400 nm wavelength and 120 fs width. Fluorescence signal was detected by Boxcar. Three life-time components （320 fs,4.0 ps and 20.0 ps） were obtained by multi-exponential curve model and nonlinear least-square fitting method. Three fluorescence peaks （652 nm, 672 nm, 691 nm） were obtained by Global analysis and Gauss curve fitting. These components were analyzed according to the structural and fluorescence spectroscopic property of LHCⅡ. The fluorescence ratio of each component to that of all the component was 3.4%, 50% and 46.6% respectively. The possible model of energy transfer in LHCⅡ was discussed.     

Model Compounds of the Mn Cluster in Oxygen-evolving Complex of PS￠ò

Six model compounds have been synthesized and used for probing the structural features of the Mn cluster in oxygen-evolving complex (OEC) of photosystem Ⅱ (PSⅡ). The model compounds contain Mn2(μ-O) 2 and μ-O-μ-carboxylato di-manganese structural units, which offer Mn—Mn, Mn……Mn, and Mn—O(N) structural parameters consistent with the corresponding data of the OEC in PSⅡ, implying that the Mn cluster in OEC may possess similar structural features. Two model compounds containing halide anion have been used for discussing the binding of Cl- to Mn in PSⅡ. It is suggested that in the five S states, ligand exchange would lead to the ligation of chloride to Mn in the S states with Mn of higher valence.     

Stygofauna Abundance and Distribution in the Fissures and Caves of the Nardò (Southern Italy) Fractured Aquifer Subject to Reclaimed Water Injections

The demographic growth in developing countries and the increasing pressure of anthropological activities in industrialized states around the world, are leading to a gradual contamination of the natural habitats of our planet. Although the extent of these effects is unclear, the results can already seen in the quality of natural resources, which are intensely stressed by climate changes (greenhouse effect, nutrients load, water consumption, etc.) and by direct contamination of toxic wastes. This could progressively destroy the variety of faunal species and, indeed, recent warming has caused changes in species distribution and abundance. This paper presents an investigation into the possible effects of climate change and anthropological pressures on the ground water fauna present at the Nardò site (Salento peninsula, Southern Italy). Three ecological categories were examined: stygoxenes, stygophiles and stygobionts. The latter are anophthalmic, without pigment, measure up to 10–12 mm, and live in water which moves throughout fissures and karstic caves of carbonate aquifers. These stygofauna are very sensitive to changes, due to environmental stresses, such as water temperature, dissolved oxygen, water salinity, pH and chemical constituents, in their hypogeous habitat. The stygofauna categories are active organisms which contribute to the biodegradation of organic compounds in wastewater artificially (or naturally) injected in the fractured subsoil. Weak information has been available until now about Salento stygofauna ability to resist water pollution caused by human activities. At the Nardò site 12000 m³/d of 2 ^y effluent from municipal treatment plants have been injected since 1991 in a natural sinkhole. Here, the abundance of the stygofauna, recovered in three wells (Colucci, Brusca and Spundurata cave) at progressive distances from sinkhole, and their distribution have been correlated with ground water constituents. Groundwater quality was monitored on each occasion that stygofauna were collected, during the spring-autumn seasons.     

Comparison Between the Influences of Phosphatidylcholine and Triton X-100 on the Protein Secondary Structures and Oxygen-evolving Activity of Photosystem ￠ò

The structural and functional alterations within the PSⅡ membrane from phosphatidylcholine reconstitution and Triton X-100 (TX-100) treatment were studied by using Fourier transform-infrared (FT-IR) spectroscopic technique and oxygen electrode. Phosphatidylcholine reconstitution showed no significant effect on the protein secondary structures of PSⅡ membrane but an increase of the rate of PSⅡ-mediated oxygen-evolution. The phosphatidylcholine lipids with different length of acyl chains displayed different capabilities to stimulate oxygen-evolution. In contrast, perturbation of the bilayer lipids by TX-100 resulted in obvious changes of the protein secondary structures within the PSⅡ membrane and in the loss of the PSⅡ-mediated oxygen-evolving activity. The results indicate the importance of membrane integrity in maintaining the stability of the photosynthetic membrane proteins.     

Does Help in Decision-Making in Biology Help in Decision-Making in Human Sciences and Conversely?

A link between biological and human sciences may be established, under the condition that we should admit the existence of reciprocal influences between them. The model for the regulation of agonistic antagonistic couples (MRAAC) is built from the study of biological systems and gives rise to specific types of control. This model can be helpful in decision processes in some human sciences such as management, economical and political strategies. The reason for such an opportunity lies in the fact that MRAAC is a general and phenomenological model able to incorporate the whole of the agonistic antagonistic systems. This type of regulation might be related to the concept of the viability of a system (yet also valid for human science systems) and to a functional and structural pattern which is the basis for agonistic antagonistic networks.     

Calcium signaling in pancreatic β-cells in health and in Type 2 diabetes

Changes in cytosolic free Ca²⁺ concentration ([Ca²⁺]_c) play a crucial role in the control of insulin secretion from the electrically excitable pancreatic β-cell. Secretion is controlled by the finely tuned balance between Ca²⁺ influx (mainly through voltage-dependent Ca²⁺ channels, but also through voltage-independent Ca²⁺ channels like store-operated channels) and efflux pathways. Changes in [Ca²⁺]_c directly affect [Ca²⁺] in various organelles including the endoplasmic reticulum (ER), mitochondria, the Golgi apparatus, secretory granules and lysosomes, as imaged using recombinant targeted probes. Because most of these organelles have specific Ca²⁺ influx and efflux pathways, they mutually influence free [Ca²⁺] in the others. In this article, we review the mechanisms of control of [Ca²⁺] in various compartments and particularly the cytosol, the endoplasmic reticulum ([Ca²⁺]_ER), acidic stores and mitochondrial matrix ([Ca²⁺]_mito), focusing chiefly on the most important physiological stimulus of β-cells, glucose. We also briefly review some alterations of β-cell Ca²⁺ homeostasis in Type 2 diabetes.     

Sex-associated difference in estrogen receptor β expression in N-methyl-N'-nitro-N-nitrosoguanidine-induced gastric cancers in rats

Epidemiologic studies indicate that the incidence of gastric cancer is higher in males than in females. Although the mechanisms mediating this difference are unclear, a role for estrogens has been proposed. We used Western blotting to evaluate the role of estrogen receptor (ER) subtypes ERα and ERβ and proliferating cell nuclear antigen (PCNA) in N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced gastric carcinogenesis in Wistar rats; ERα and ERβ mRNA levels also were analyzed by quantitative real-time RT-PCR analysis. The incidence of gastric cancer was significantly higher in male than female rats. In both sexes, ERα expression was similar in MNNG-treated cancerous and noncancerous tissues and normal gastric tissue. However, ERβ expression in MNNG-treated cancerous and noncancerous tissues was significantly lower in male rats and higher in female rats than that in normal gastric tissue; MNNG-induced cancerous tissue showed the highest ERβ expression. PCNA expression in MNNG-treated cancerous tissues was higher than that in noncancerous tissues, and was higher in male rats than female rats. Western blotting results were consistent with the mRNA changes determined by quantitative real-time RT-PCR. The present study provides evidence of a sex-associated difference in ERβ and PCNA expression in MNNG-induced gastric cancers in Wistar rats.     

Variation in Ambrosia pollen concentration in Southern and Central Poland in 1982–1999

The aim of the study was to investigate theAmbrosia pollen concentrations inselected Polish cities and for Kraków torelate it to some meteorological factors.Sampling was carried out in Kraków in1982–1997 and in Rabka in 1992–1996 with theuse of the gravimetric method. In Zakopane,Kraków, Ostrowiec witokrzyski,Warszawa and Pozna in 1995–1996 both thegravimetric and volumetric methods (Burkardtrap) were employed. In Kraków themonitoring has been performed since 1994 usingthe volumetric method. The results show theragweed pollen presence in August and Septemberwith the tendency to appear more frequently inAugust in some years. In Kraków (1994–1999)Ambrosia pollen was found either in thelast two weeks of August or in the first twoweeks of September which seems to be a regularand repeatable pattern every two years.Seasonal fluctuations of Ambrosia pollenconcentration do not show a clear increasingtendency except at Warszawa and Ostrowiecwitokrzyski in 1996 and at Krakówin 1999. Percentage of Ambrosia pollen inannual sums of total pollen is very low anddoes not exceed 1% except at Ostrowiecwitokrzyski in 1996 (1.2%) and atKraków in 1999 (2%). For Kraków theanalysis of some meteorological factors (Tmax,Tmin, precipitation, wind direction) wasperformed. High temperature and lack of rain orlow precipitation correlate well with ragweedpollen concentrations. During the Ambrosia pollen seasons ESE, E, S, SE, WSW, SWwind directions prevailed which could suggest along-distance transport from Ukraine, the CzechRepublic, Slovakia and also from Hungary, one ofthe most ragweed-polluted countries.     

Interindividual human variation in cisplatinum sensitivity,predictable in an in vitro assay?

Cisplatinum [cis-diamminedichloroplatinum(II)] is one of the most active antitumour agents and its effect is mainly due to the formation of cisplatinum-DNA crosslinks. Formation of cisplatinum-DNA intrastrand crosslinks in nucleated white blood cells of patients was measured, both in (pretreatment) samples exposed in vitro and in cells collected immediately after in vivo exposure to cisplatinum. Large interpatient variations were found. The in vitro results showed a linear correlation with the in vivo data (cc=0.91). The in vitro measurements of crosslinks may allow prediction of response and avoidance of toxicity in individual patients.     

Interindividual human variation in cisplatinum sensitivity, predictable in an in vitro assay?

Cisplatinum [cis-diamminedichloroplatinum(II)] is one of the most active antitumour agents and its effect is mainly due to the formation of cisplatinum-DNA crosslinks. Formation of cisplatinum-DNA intrastrand crosslinks in nucleated white blood cells of patients was measured, both in (pretreatment) samples exposed in vitro and in cells collected immediately after in vivo exposure to cisplatinum. Large interpatient variations were found. The in vitro results showed a linear correlation with the in vivo data (cc = 0.91). The in vitro measurements of crosslinks may allow prediction of response and avoidance of toxicity in individual patients.     

Does variation in morphology correspond with variation in habitat use in freshwater gastropods?

We tested if variations (i.e., breadth) in morphology and habitat use vary predictably among six aquatic gastropod species that were collected across Indiana and Illinois, USA. We predicted that interspecific morphological variation would positively covary with variation in habitat use among species. We used geometric morphometrics (Procrustes technique and relative warp analysis) to quantify morphology and multivariate analyses (PCA) to quantify habitat. Increased morphological breadth did not vary predictably with increased habitat breadth. However, we found that life history traits correspond with patterns in morphological and habitat breadth for these six aquatic gastropods. Pulmonate gastropods (use lungs for respiration) that lack an operculum cover exhibited decreased morphological breadth compared to coenogastropods (use gills for respiration). This pattern may ultimately be a function of behavioral adaptations in freshwater gastropods. Gastropods that are capable of breathing air or using other behavioral modifications such as burrowing to escape predators may not require high morphological breadth. Conversely, selection may favor higher morphological breadth in gastropods with gills that also do not move out of the water column to escape predators.     

Are changes in maximal squat strength during preseason training reflected in changes in sprint performance in rugby league players?

Because previous research has shown a relationship between maximal squat strength and sprint performance, this study aimed to determine if changes in maximal squat strength were reflected in sprint performance. Nineteen professional rugby league players (height = 1.84 ± 0.06 m, body mass [BM] = 96.2 ± 11.11 kg, 1 repetition maximum [1RM] = 170.6 ± 21.4 kg, 1RM/BM = 1.78 ± 0.27) conducted 1RM squat and sprint tests (5, 10, and 20 m) before and immediately after 8 weeks of preseason strength (4-week Mesocycle) and power (4-week Mesocycle) training. Both absolute and relative squat strength values showed significant increases after the training period (pre: 170.6 ± 21.4 kg, post: 200.8 ± 19.0 kg, p < 0.001; 1RM/BM pre: 1.78 ± 0.27 kg·kg(-1), post: 2.05 ± 0.21 kg·kg(-1), p < 0.001; respectively), which was reflected in the significantly faster sprint performances over 5 m (pre: 1.05 ± 0.06 seconds, post: 0.97 ± 0.05 seconds, p < 0.001), 10 m (pre: 1.78 ± 0.07 seconds, post: 1.65 ± 0.08 seconds, p < 0.001), and 20 m (pre: 3.03 ± 0.09 seconds, post: 2.85 ± 0.11 seconds, p < 0.001) posttraining. Whether the improvements in sprint performance came as a direct consequence of increased strength or whether both are a function of the strength and power mesocycles incorporated into the players' preseason training is unclear. It is likely that the increased force production, noted via the increased squat performance, contributed to the improved sprint performances. To increase short sprint performance, athletes should, therefore, consider increasing maximal strength via the back squat.