Direct screening of living mammalian cell colonies for the identification of DNA repair deficient mutants by a Multiwire Proportional Chamber

Summary Mammalian cell mutants with defective repair of UV-induced DNA damage can be identified by the lack of unscheduled incorporation of radioactive thymidine into DNA after UV irradiation. With a specially designed windowlessMultiWireProportionalChamber (MWPC), colonies with absent or low incorporation of³H-thymidine can be precisely located within a population of normally labelled colonies. The main advantages of MWPC analysis over other more conventional methods for radioactive measurements are: (i), cells are not harmed by the analysis, so that previous replica plating of colonies becomes unnecessary; (ii), measurements are quantitative, hence discrimination between colonies is not subjective; (iii), location of unlabelled, non-repairing colonies is rapid and can be automatized. The method therefore appears suitable for the isolation of mammalian cell mutants with a block in DNA repair.     

Photosynthetic characteristics of diploid honeysuckle (<Emphasis Type="Italic">Lonicera japonica</Emphasis> Thunb.) and its autotetraploid cultivar subjected to elevated ozone exposure

In order to investigate the effect of chromosome doubling on ozone tolerance, we compared the physiological responses of a diploid honeysuckle (Lonicera japonica Thunb.) and its autotetraploid cultivar to elevated ozone (O₃) exposure (70 ng g⁻¹, 7 h d⁻¹ for 31 d). Net photosynthetic rate (P _N) of both cultivars were drastically (P<0.01) impaired by O₃. Although there were significantly positive correlation between P _N and stomatal conductance (g _s) in both cultivars under each treatment, the decreased g _s in O₃ might be the result rather than the cause of decreased P _N as indicated by stable or increasing the ratio of intercellular to ambient CO₂ concentration(C _i/C _a). P _N under saturating CO₂ concentration (P _Nsat) and carboxylation efficiency (CE) significantly decreased under O₃ fumigation, which indicated the Calvin cycle was impaired. O₃ also inhibited the maximum efficiency of photosystem II (PSII) photochemistry in the dark-adapted state (F_v/F_m), actual quantum yield of PSII photochemistry (Φ_PSII), electron transport rate (ETR), photochemical quenching coefficient (qP), non-photochemical quenching (NPQ), the maximum in vivo rate of Rubisco carboxylation (V_cmax) and the maximal photosynthetic electron transport rate (J_max) which demonstrated that the decrease in P _N of the honeysuckle exposed to elevated O₃ was probably not only due to impairment of Calvin cycle but also with respect to the light-harvesting and electron transport processes. Compared to the diploid, the tetraploid had higher relative loss in transpiration rate (E), (g _s), (P _Nsat), V_cmax and J_max. This result indicated that the Calvin cycle and electron transport in tetraploid was damaged more seriously than in diploid. A barely nonsignificant (P=0.086) interaction between O₃ and cultivar on P _N suggested a higher photosynthetic sensitivity of the tetraploid cultivar.     

Biomass Partitioning and Gas Exchange in Dalbergia sissoo seedlings under water stress

Biomass, leaf water potential (_l), net photosynthetic rate (P _N), transpiration rate (E), stomatal conductance (g _s), leaf to air temperature difference (T _diff), and instantaneous water use efficiency (WUE) were measured in the seedlings of Dalbergia sissoo Roxb. grown under irrigation of 20 (W₁), 14 (W₂), 10 (W₃), and 8 (W₄) mm. Treatments were maintained by re-irrigation when water content of the soil reached 7.4% in W₁, 5.6% in W₂, 4.3% in W₃, and 3.2% in W₄. Seedlings in a control (W₅) were left without irrigation after maintaining the soil field capacity (10.7%). Seedlings of W₁ had highest biomass that was one tenth in W₅. Biomass allocation was highest in leaf in W₂ and in root in W₄ and W₅ treatments. Difference between predawn leaf water potential (_Pd) and midday (_mid) increased with soil water stress and with vapour pressure deficit (VPD) in April and May slowing down the recovery in plant leaf water status after transpiration loss. P _N, E, and g _s declined and T _diff increased from W₁ to W₅. Their values were highly significant in April and May for the severely stressed seedlings of W₄ and W₅. P _N increased from 08:00 to 10:00 and E increased until 13:00 within the day for most of the seedlings whereas g _s decreased throughout the day from 08:00 to 17:00. P _N and E were highest in March but their values were low in January, February, April, and May. Large variations in physiological variables to air temperature, photosynthetically active radiation, and vapour pressure deficit (VPD) indicated greater sensitivity of the species to environmental factors. WUE increased from W₁ to W₂ but decreased drastically at high water stress particularly during hot summer showing a kind of adaptation in D. sissoo to water stress. However, low biomass and reduced physiological functions at <50% of soil field capacity suggest that this species does not produce significant biomass at severe soil water stress or drought of a prolonged period.     

Reactivity of one-, two-, three- and four-electron reduced forms of α-[P2W18O62] generated by controlled potential electrolysis in water

One, two, three and four electron reduced forms of α-[P₂W₁₈O₆₂]⁶⁻ in aqueous acidic electrolyte media have been selectively generated by bulk electrolysis from a solution that has an initial pH of 3.6. The reactivities of the reduced polyoxometalate anions and identities of products formed in the presence and absence of dioxygen have been assessed via oxygen and hydrogen Clark-type electrodes, a pH electrode and rotating disk electrode voltammetry. [P₂W₁₈O₆₂]⁷⁻ is stable to protons but is slowly oxidized by dioxygen (timescale: hours to days) back to [P₂W₁₈O₆₂]⁶⁻. In contrast, [P₂W₁₈O₆₂]⁸⁻ reacts more rapidly with O₂ and slowly with H⁺, whereas generation of the [P₂W₁₈O₆₂]⁹⁻ and [P₂W₁₈O₆₂]¹⁰⁻ anion is accompanied by a large increase in pH and rapid reaction with O₂ or, in its absence, with H⁺. Consequently, it is concluded that photocatalytic reactions based upon [P₂W₁₈O₆₂]⁶⁻ chemistry are only likely to be of significance if [P₂W₁₈O₆₂]⁹⁻ or more highly reduced species are generated and form part of the catalytic cycle.     

The genetics of colour-patterns in the grasshopper Chorthippus brunneus

Several alleles were found to determine the colour of the dorsal pronotum in Chorthippus brunneus; there was evidence for at least two loci (C and V). Brown (C^B)was the universal recessive and green (C^C) was dominant to all other colours. The white allele (C^W)was codominant with green(C^G)and purple (C^P). Wing-patterns were determined by a separate, probably linked locus (W). A dominant plain wing-pattern (W^P) was associated with colours other than brown. Striped(W^S)and mottled(W^mo) were codominant and a plain recessive allele (W^P) was also found. All three alleles were associated with the brown phenotype. A purple-sided allele (S^Pu) was sometimes obmd with C^pu.. S^Pu was dominant to brown sides (S^B), A series of markings on the dorsal and lateral pronotum (linea intermedia, fascia postocularis, linea media, carina media and zona lateralis) were investigated and found to be controlled at separate loci which may be linked to W. These characters were expressed by dominant alleles. Epistatic effects by modifier loci were shown to have an important effect on the determination of wing phenotype. Allele Wo⁺, for example, suppressed the stripe-wing pattern, linea media, carina media and zona lateralis. It was concluded that colour patterns appear to be under genetic control and that dominant alleles were rare in the wild. Changes in shades of colours were shown to be age-dependent and minor.     

The effect of body size on food consumption, absorption efficiency, respiration, and ammonia excretion by the inland silverside, Menidia beryllina (Cope) (Osteichthyes: Atherinidae)

The inland silverside, Menidia beryllina (Cope), is an annual zooplanktivore that occurs in estuarine and freshwater habitats along the Atlantic and Gulf of Mexico coasts and drainages of the United States. Experiments were conducted at 25 ± 1°C to quantify the relationship between mean dry weight (W_D) and rates of energy gain from food consumption (C), and energy losses as a result of respiration (R) and ammonia excretion (E) during routine activity and feeding by groups of fish. The absorption efficiency of ingested food energy (A) was also quantified. Rates of C, E, and R increased with W_D by factors (b in the equation y = aW_D^b) equal to 0.462, 0.667, and 0.784, respectively. Mean (±SE) rates of energy loss during feeding were 1.6 ± 0.1 (R) and 3.4 ± 0.6 (E) times greater than those for unfed fish. Absorption efficiency was independent of W_D and estimated to be 89% of C. From these measurements, the surplus energy available for growth and activity (G) and growth efficiency (K₁) were estimated. Over the range in sizes of juveniles and adults (5–500 mg W_D), predicted G and K₁ values decreased from 7.42 to 0.20 J mg fish^?1 day^?1 and 63 to 21%, respectively. Measured and predicted bioenergetic parameters are discussed within an ecological context for a northern population of this species.     

Zirconium and hafnium aqua complexes [(H2O)3M(P2W17O61)]: Synthesis, characterization and substitution of water by chiral ligand

The hydroxocomplexes [{(H₂O)M(μ₂-OH)(P₂W₁₇O₆₁)}₂]¹⁴⁻ (M = Zr, Hf) in HCl undergo cleavage of the hydroxo bridges with the formation of monomeric species [(H₂O)₃M(P₂W₁₇O₆₁)]⁶⁻. In the case of Hf single crystals of the composition (Me₂NH₂)_5.5(H)_1.5[(Hf(H₂O)₃)_0.9(WO)_0.1{P₂W₁₇O₆₁}]Cl·9.5H₂O (1), as the result of co-crystallization of [(H₂O)₃Hf(P₂W₁₇O₆₁)]⁶⁻ and [P₂W₁₈O₆₂]⁶⁻ salts, were isolated from these solutions and structurally characterized. Zr gives (Me₂NH₂)₂(H)₄[{(H₂O)₂ZrP₂W₁₇O₆₁}]·8.67H₂O (2), in whose structure chiral polymeric chains {[(H₂O)₂M(P₂W₁₇O₆₁)]}_n⁶ⁿ⁻ are present. Under hydrothermal conditions the water molecules in [(H₂O)₃M(P₂W₁₇O₆₁)]⁶⁻ are replaced by l-malic acid with the formation of stable chiral polyoxoanions, isolated as (NH₂Me₂)₈[M(L-ООССН(ОН)СН₂СОО)P₂W₁₇O₆₁]·7·9H₂O (M = Zr, 3; M = Hf, 4). The structures of 1, 2 and 3 were determined; 3 and 4 were found to be isostructural. The products were also characterized by elemental analysis, thermogravimetry and IR-spectroscopy.     

The oxygen uptake-power regression in cyclists and untrained men: implications for the accumulated oxygen deficit

The regression of oxygen uptake (O₂) on power output and the O₂ demand predicted for suprapeak oxygen uptake (O_2peak) exercise (power output = 432 W) were compared in ten male cyclists [C, mean O_2peak = 67.9 (SD 4.2) ml · kg^–1 · min^–1] and nine active, yet untrained men [UT, mean O_2peak = 54.1 (SD 6.5) ml · kg^–1 · min^–1]. The O₂-power regression was determined using a continuous incremental cycle test (CON4), performed twice, which comprised several 4-min exercise periods progressing in intensity from approximately 40%–85% O_2peak. Minute ventilation (_E), heart rate (HR), respiratory exchange ratio (R), blood lactate concentration ([1a^–]_b) and rectal temperature (T _re) were measured at rest and during CON4. The slope of the O₂-power regression was greater (P 0.05) in C [12.4 (SD 0.7) ml · min^–1. W^–1] compared to UT [11.7 (SD 0.4) ml · min^–1 W^–1]; as a result, the O₂ demand (at 432 W) was also higher (P 0.05) in C [5.97 (SD 0.23) l · min^–1] than UT [5.70 (SD 0.15) 1 · min^–1]. ExerciseR and [la^–]_b were lower (P 0.05) in C .in comparison to UT at all power outputs, whereas _E and HR were relatively lower (P 0.05) in C at power outputs approximating 180 W, 220 W and 270 W. Differences in fat metabolism estimated over the first three power outputs accounted for approximately 19% of the difference in O₂-power slopes between the groups and up to 46% of the difference in O₂ at a given intensity. Although the O₂-power regressions were linear for C [r = 0.997 (SD 0.001)] and UT [r = 0.997 (SD 0.001)], the O₂-power slope was higher at power outputs at or above the lactate threshold (13.2 ml · min^–1 · W^–1 than at lower intensities (11.6 ml · min^–1 · W^–1) in C, an effect which was less profound in UT. As a result, the exclusion of O₂ at the highest power outputs completely abolished the difference in O₂-power slopes between C and UT. Thus, the relatively higher O₂ during incremental exercise in C can be almost entirely attributed to the higher O₂ cost of cycling at higher power outputs. In addition, the presence of non-linear responses in O₂ at higher intensities also confirms the invalidity of describing the O₂ response across a wide range of power outputs using a linear function, and challenges the validity of predicting the O₂ demand of more intense exercise by a linear extrapolation of this same function.     

A Natural Genetic Variant of Granzyme B Confers Lethality to a Common Viral Infection

Many immune response genes are highly polymorphic, consistent with the selective pressure imposed by pathogens over evolutionary time, and the need to balance infection control with the risk of auto-immunity. Epidemiological and genomic studies have identified many genetic variants that confer susceptibility or resistance to pathogenic micro-organisms. While extensive polymorphism has been reported for the granzyme B (GzmB) gene, its relevance to pathogen immunity is unexplored. Here, we describe the biochemical and cytotoxic functions of a common allele of GzmB (GzmB^W) common in wild mouse. While retaining ‘Asp-ase’ activity, GzmB^W has substrate preferences that differ considerably from GzmB^P, which is common to all inbred strains. In vitro, GzmB^W preferentially cleaves recombinant Bid, whereas GzmB^P activates pro-caspases directly. Recombinant GzmB^W and GzmB^P induced equivalent apoptosis of uninfected targets cells when delivered with perforin in vitro. Nonetheless, mice homozygous for GzmB^W were unable to control murine cytomegalovirus (MCMV) infection, and succumbed as a result of excessive liver damage. Although similar numbers of anti-viral CD8 T cells were generated in both mouse strains, GzmB^W-expressing CD8 T cells isolated from infected mice were unable to kill MCMV-infected targets in vitro. Our results suggest that known virally-encoded inhibitors of the intrinsic (mitochondrial) apoptotic pathway account for the increased susceptibility of GzmB^W mice to MCMV. We conclude that different natural variants of GzmB have a profound impact on the immune response to a common and authentic viral pathogen.     

Energy expenditure in relation to activity of lesser mouse deer (Tragulus javanicus)

Heat production (HP) of male and female mouse deer during eating, standing and sitting was determined using the open circuit respiration chamber (RC). The time taken for similar activities was also determined in an outdoor enclosure (OD). The animals were fed kangkong (Ipomoea aquatica), sweet potato (Ipomoea batatas) and rabbit pellet ad libitum. Male mouse deer consumed more dry matter (DM), organic matter (OM) and gross energy (GE) than female. The time for each activity of male and female mouse deer kept in RC and OD was similar. The average time spent in RC and OD for both male and female, respectively, for sitting (956 and 896 min/day) was significantly (P<0.01) longer than standing (463 and 520 min/day) and eating (21 and 24 min/day). Heat production for male and female mouse deer, respectively, during eating was the highest (0.44 and 0.43 kJ/kg W^0.75/min) followed by standing (0.37 and 0.33 kJ/kgW^0.75/min) and sitting (0.26 and 0.26 kJ/kg W^0.75/min). The difference in HP per min during standing between male and female was significant (P<0.05). The HP for 08.00–14.00 h and 14.00–20.00 h periods were higher than 20.00–02.00 h and 02.00–08.00 h periods. The overall HP for males during 08.00–14.00 h and 14.00–20.00 h periods were significantly (P<0.05) higher (114.8 and 119.2 kJ/kg W^0.75) than female (107.5 and 110.4 kJ/kg W^0.75), respectively.     

Microelectrode array evaluation of gut pacemaker activity in wild-type and mice

Interstitial cells of Cajal in the myenteric plexus region (ICC-MyP) form a network and generate basal pacemaking electrical activity. This morphological feature leads us to believe that these cells may be essential for the coordinating actions of gastrointestinal (GI) motility. We aim to propose a new method for functional assessment of ICC electrical activity and its network. Field potentials in a 1 mm² region were simultaneously measured using an 8 × 8 microelectrode array (MEA) with a polar distance of 150 μm. The extracellular solution contained nifedipine and tetrodotoxin (TTX) to suppress activities of smooth muscle cells and neurons, respectively. We compared spatial electrical activities between ileal muscle preparations from wild-type (WT) and W/W^v mice. In spatio-temporal analyses, basal electrical activities were well synchronized with a propagation delay in WT, while those in W/W^v were small in amplitude and irregular in occurrence. The power spectrum in WT had a prominent peak corresponding to the frequency of ICC-MyP pacemaker activity, while that of W/W^v lacked it. Consequently, the ratio of the spectral power in 9.4–27.0 cpm was significantly larger in WT than in W/W^v. In conclusion, MEA measurements demonstrated that the network-forming ICC-MyP not only generates but also coordinates basal electrical activities. Disorders of GI motility based on morphological and functional impairments of ICC network with the range of several hundreds of micrometers, could be uncovered in future extensive studies.     

Isolation and characterization of sex chromosome rearrangements generating male muscle dystrophy and female abnormal oogenesis in the silkworm, Bombyx mori

In deletion-mapping of W-specific RAPD (W-RAPD) markers and putative female determinant gene (Fem), we used X-ray irradiation to break the translocation-carrying W chromosome (W ^Ze ). We succeeded in obtaining a fragment of the W ^Ze chromosome designated as Ze ^W, having 3 of 12 W-RAPD markers (W-Bonsai, W-Yukemuri-S, W-Yukemuri-L). Inheritance of the Ze ^W fragment by males indicates that it does not include the Fem gene. On the basis of these results, we determined the relative positions of W-Yukemuri-S and W-Yukemuri-L, and we narrowed down the region where Fem gene is located. In addition to the Ze ^W fragment, the Z chromosome was also broken into a large fragment (Z₁) having the + ^sch (1-21.5) and a small fragment (Z₂) having the + ^od (1-49.6). Moreover, a new chromosomal fragment (Ze ^WZ₂) was generated by a fusion event between the Ze ^W and the Z₂ fragments. We analyzed the genetic behavior of the Z₁ fragment and the Ze ^WZ₂ fragment during male (Z/Z₁ Ze ^WZ₂) and female (Z₁ Ze ^WZ₂/W) meiosis using phenotypic markers. It was observed that the Z₁ fragment and the Z or the W chromosomes separate without fail. On the other hand, non-disjunction between the Ze ^WZ₂ fragment and the Z chromosome and also between the Ze ^WZ₂ fragment and the W chromosome occurred. Furthermore, the females (2A: Z/Ze ^WZ₂/W) and males (2A: Z/Z₁) resulting from non-disjunction between the Ze ^WZ₂ fragment and the W chromosome had phenotypic defects: namely, females exhibited abnormal oogenesis and males were flapless due to abnormal indirect flight muscle structure. These results suggest that Z₂ region of the Z chromosome contains dose-sensitive gene(s), which are involved in oogenesis and indirect flight muscle development.     

Effects of ozone exposure on growth and photosynthesis of the seedlings of <Emphasis Type="Italic">Liriodendron chinense</Emphasis> (Hemsl.) Sarg,a native tree species of subtropical China

Little is known about the response of trees to elevated ozone (O₃) in the subtropical region of China, where ambient O₃ concentrations are high enough to damage plants. In this study, pigment content, gas exchange and chlorophyll (Chl) a fluorescence in leaves of Liriodendron chinense (Hemsl.) Sarg seedlings, a deciduous broadleaf tree species native in subtropical regions, were investigated at 15, 40, and 58 days after O₃ fumigation (DAF) at a concentration of 150 mm³ m⁻³ (E-O₃). At the end of experiment, seedlings were harvested for biomass measurement. E-O₃ caused visible injuries on the mature leaves e.g. necrotic patches and accelerated early defoliation. Relative to the charcoal-filtered air (CF) treatment, E-O₃ significantly decreased shoot and root biomass, pigment content, light-saturated net photosynthesis (P _Nsat), stomatal conductance (g _s), maximum rate of carboxylation (V_cmax), photochemical quenching coefficient (q_p) and effective quantum yield of PSII photochemistry (Φ_PSII), and also caused a slight reduction in relative increase of basal diameter. Therefore, L. chinense can be assumed to be an O₃-sensitive tree species, which will be threatened by increasing ambient O₃ concentrations in China.     

Effects of the interaction between ozone and carbon dioxide on gas exchange,ascorbic acid content,and visible leaf symptoms in rice leaves

Tropospheric ozone (O₃) decreases photosynthesis, growth, and yield of crop plants, while elevated carbon dioxide (CO₂) has the opposite effect. The net photosynthetic rate (P _N), dark respiration rate (R _D), and ascorbic acid content of rice leaves were examined under combinations of O₃ (0, 0.1, or 0.3 cm³ m⁻³, expressed as O⁰, O^0.1, O^0.3, respectively) and CO₂ (400 or 800 cm³ m⁻³, expressed as C⁴⁰⁰ or C⁸⁰⁰, respectively). The P _N declined immediately after O₃ fumigation, and was larger under O^0.3 than under O^0.1. When C⁸⁰⁰ was combined with the O₃, P _N was unaffected by O^0.1 and there was an approximately 20 % decrease when the rice leaves were exposed to O^0.3 for 3 h. The depression of stomatal conductance (g _s) observed under O^0.1 was accelerated by C⁸⁰⁰, and that under O^0.3 did not change because the decline under O^0.3 was too large. Excluding the stomatal effect, the mesophyll P _N was suppressed only by O^0.3, but was substantially ameliorated when C⁸⁰⁰ was combined. Ozone fumigation boosted the R _D value, whereas C⁸⁰⁰ suppressed it. An appreciable reduction of ascorbic acid occurred when the leaves were fumigated with O^0.3, but the reduction was partially ameliorated by C⁸⁰⁰. The degree of visible leaf symptoms coincided with the effect of the interaction between O₃ and CO₂ on P _N. The amelioration of O₃ injury by elevated CO₂ was largely attributed to the restriction of O₃ intake by the leaves with stomatal closure, and partly to the maintenance of the scavenge system for reactive oxygen species that entered the leaf mesophyll, as well as the promotion of the P _N.     

Effects of controlled irrigation on water potential,nitrogen uptake and biomass production in Dalbergia sissoo seedlings

Biomass production, pattern of nodulation, nutrient uptake, net photosynthetic rate (P_n), leaf temperature (T_leaf), leaf nitrate reductase (NR) activity and free proline of Dalbergia sissoo seedlings planted in containers with 120 kg soil were studied under different water stress levels to assess the productive potential of the species in dry areas. Seedlings were irrigated at 20 mm (W₁), 14 mm (W₂), 10 mm (W₃), 8 mm (W₄) throughout the experimental period to maintain the respective treatment up to the lowest soil water content of 7.43%, 5.64%, 4.30% and 3.23%, respectively. There was a treatment (W₅) in which seedling were irrigated once to −0.03 MPa and left without re-irrigation. Decreased irrigation level resulted in lowering of leaf water potential (LWP), net photosynthetic rate (P_n), total number of root nodules and nodule dry mass and nitrogen uptake in the seedling. P_n, leaf nitrate reductase (NR) activity and seedling biomass were highest in W₁ indicating a positive relations of NR activity with CO₂ assimilation and biomass production. The decrease in P_n, leaf NR activity and LWP was sharp at W₃ onwards. Monthly changes in the values of P_n, T_leaf and NR activity indicate environmental effect on these physiological variables. Proline was detected only in the seedlings of W₃, W₄ and W₅ treatments after February and was highest in the seedlings of W₅ treatment. The study suggests that severe water deficit adversely affect physiological and biochemical processes that resulted in reduced growth, nutrient uptake and biomass productivity in D. sissoo seedlings. Re-irrigation above W₃ level is recommended for this species.     

Adsorption Characteristics and the Effect of Dissolved Organic Matter on Mercury(II) Adsorption of Various Soils in China

To evaluate the adsorption characteristics and effects of dissolved organic matter (DOM) from wheat straw (DOM_W) and swine manure (DOM_S) on mercury [Hg(II)] adsorption by soils, a series of experiments was conducted on 26 soils with different soil properties.Results showed that the values of K_d (a solid–liquid partition coefficient) of soils varied within a range of 0.768–14.386 L g^?1. K_d values were mainly controlled by soil organic matter (SOM), cation exchange capacity (CEC), soil nitrogen (N), and soil sulfur (S). When DOM_W and DOM_S were added to the soil samples, the amount of Hg(II) adsorbed by the soils decreased significantly. Furthermore, based on the decreased percentage of adsorption of Hg(II) by soil upon adding DOM_W (P_DOMW) or DOM_S (P_DOMS), about 73.08% soils, the values of P_DOMS were smaller than those of P_DOMW. The P_DOMW values were related to SOM, pH, free Fe oxide content, and CEC, whereas P_DOMS values were related to soil free Fe/Al oxide contents. Therefore, we should pay more attention about mercury risk caused by the addition of exogenous organic matter in soils, especially for the soils with low or high pH, less SOM, low CEC, and less free Fe/Al oxide.     

Nifedipine protects against overproduction of superoxide anion by monocytes from patients with systemic sclerosis

We have reported previously that dihydropyridine-type calcium-channel antagonists (DTCCA) such as nifedipine decrease plasma markers of oxidative stress damage in systemic sclerosis (SSc). To clarify the cellular basis of these beneficial effects, we investigated the effects in vivo and in vitro of nifedipine on superoxide anion (O₂ ^•-) production by peripheral blood monocytes. We compared 10 healthy controls with 12 patients with SSc, first after interruption of treatment with DTCCA and second after 2 weeks of treatment with nifedipine (60 mg/day). O₂ ^•- production by monocytes stimulated with phorbol myristate acetate (PMA) was quantified by the cytochrome c reduction method. We also investigated the effects in vitro of DTCCA on O₂ ^•- production and protein phosphorylation in healthy monocytes and on protein kinase C (PKC) activity using recombinant PKC. After DTCCA had been washed out, monocytes from patients with SSc produced more O₂ ^•- than those from controls. Nifedipine treatment considerably decreased O₂ ^•- production by PMA-stimulated monocytes. Treatment of healthy monocytes with nifedipine in vitro inhibited PMA-induced O₂ ^•- production and protein phosphorylation in a dose-dependent manner. Finally, nifedipine strongly inhibited the activity of recombinant PKC in vitro. Thus, the oxidative stress damage observed in SSc is consistent with O₂ ^•- overproduction by primed monocytes. This was decreased by nifedipine treatment both in vivo and in vitro. This beneficial property of nifedipine seems to be mediated by its cellular action and by the inhibition of PKC activity. This supports the hypothesis that this drug could be useful for the treatment of diseases associated with oxidative stress.     

Increase in water-use efficiency and underlying processes in pine forests across a precipitation gradient in the dry Mediterranean region over the past 30 years

Motivated by persistent predictions of warming and drying in the entire Mediterranean and other regions, we have examined the interactions of intrinsic water-use efficiency (W _i) with environmental conditions in Pinus halepensis. We used 30-year (1974–2003) tree-ring records of basal area increment (BAI) and cellulose ¹³C and ¹⁸O composition, complemented by short-term physiological measurements, from three sites across a precipitation (P) gradient (280–700 mm) in Israel. The results show a clear trend of increasing W _i in both the earlywood (EW) and latewood (LW) that varied in magnitude depending on site and season, with the increase ranging from ca. 5 to 20% over the study period. These W _i trends were better correlated with the increase in atmospheric CO₂ concentration, C _a, than with the local increase in temperature (~0.04°C year⁻¹), whereas age, height and density variations had minor effects on the long-term isotope record. There were no trends in P over time, but W _i from EW and BAI were dependent on the interannual variations in P. From reconstructed C _i values, we demonstrate that contrasting gas-exchange responses at opposing ends of the hydrologic gradient underlie the variation in W _i sensitivity to C _a between sites and seasons. Under the mild water limitations typical of the main seasonal growth period, regulation was directed at increasing C _i/C _a towards a homeostatic set-point observed at the most mesic site, with a decrease in the W _i response to C _i with increasing aridity. With more extreme drought stress, as seen in the late season at the drier sites, the response was W _i driven, and there was an increase in the W _i sensitivity to C _a with aridity and a decreasing sensitivity of C _i to C _a. The apparent C _a-driven increases in W _i can help to identify the adjustments to drying conditions that forest ecosystems can make in the face of predicted atmospheric change.     

High allelic burden of four obesity SNPs is associated with poorer weight loss outcomes following gastric bypass surgery

Genome‐wide association and linkage studies have identified multiple susceptibility loci for obesity. We hypothesized that such loci may affect weight loss outcomes following dietary or surgical weight loss interventions. A total of 1,001 white individuals with extreme obesity (BMI >35 kg/m²) who underwent a preoperative diet/behavioral weight loss intervention and Roux‐en‐Y gastric bypass surgery were genotyped for single‐nucleotide polymorphisms (SNPs) in or near the fat mass and obesity‐associated (FTO), insulin induced gene 2 (INSIG2), melanocortin 4 receptor (MC4R), and proprotein convertase subtilisin/kexin type 1 (PCSK1) obesity genes. Association analysis was performed using recessive and additive models with pre‐ and postoperative weight loss data. An increasing number of obesity SNP alleles or homozygous SNP genotypes was associated with increased BMI (P < 0.0006) and excess body weight (P < 0.0004). No association between the amounts of weight lost from a short‐term dietary intervention and any individual obesity SNP or cumulative number of obesity SNP alleles or homozygous SNP genotypes was observed. Linear mixed regression analysis revealed significant differences in postoperative weight loss trajectories across groups with low, intermediate, and high numbers of obesity SNP alleles or numbers of homozygous SNP genotypes (P < 0.0001). Initial BMI interacted with genotype to influence weight loss with initial BMI <50 kg/m², with evidence of a dosage effect, which was not present in individuals with initial BMI ≥50 kg/m². Differences in metabolic rate, binge eating behavior, and other clinical parameters were not associated with genotype. These data suggest that response to a surgical weight loss intervention is influenced by genetic susceptibility and BMI.