Na+ Sensitivity of the KAT2-Like Channel Is a Common Feature of Cucurbits and Depends on the S5-P-S6 Segment.

Inhibition of Shaker K+ channel activity by external Na+ was previously reported in the melon (Cucumis melo L.) inwardly rectifying K+ channel MIRK and was hypothesized to contribute to salt tolerance. In this study, two inward Shaker K+ channels, CsKAT2 from cucumber (Cucumis sativus) and ClKAT2 from watermelon (Citrullus lanatus), were identified and characterized in Xenopus oocytes. Both channels were inwardly rectifying K+ channels with higher permeability to potassium than other monovalent cations and more active when external pH was acidic. Similarly to MIRK, their activity displayed an inhibition by external Na+, thus suggesting a common feature in Cucurbitaceae (Cucumis spp., Citrullus spp.). CsKAT2 and ClKAT2 are highly expressed in guard cells. After 24 h of plant treatment with 100 mM NaCl, the three KAT2-like genes were significantly downregulated in leaves and guard cells. Reciprocal chimeras were obtained between MIRK and Na+-insensitive AtKAT2 cDNAs. The chimera where the MIRK S5-P-S6 segment was replaced by that from AtKAT2 no longer showed Na+ sensitivity, while the inverse chimera gained Na+ sensitivity. These results provide evidence that the molecular basis of the channel blockage by Na+ is located in the S5-P-S6 region. Comparison of the electrostatic property in the S5-P-S6 region in AtKAT2 and MIRK revealed four key amino acid residues potentially governing Na+ sensitivity.

Supportive role of the Na+ transporter CmHKT1;1 from Cucumis melo in transgenic Arabidopsis salt tolerance through improved K+/Na+ balance.

KEY MESSAGE: CmHKT1;1 selectively exports Na+ from plant cells. Upon NaCl stress, its expression increased in a salt-tolerant melon cultivar. Overexpression of CmHKT1;1 increased transgenic Arabidopsis salt tolerance through improved K+/Na+ balance. High-affinity K+ transporters (HKTs) are thought to be involved in reducing Na+ in plant shoots under salt stress and modulating salt tolerance, but their function in a moderately salt-tolerant species of melon (Cucumis melo L.) remains unclear. In this study, a Na+ transporter gene, CmHKT1;1 (GenBank accession number: MK986658), was isolated from melons based on genome data. The transcript of CmHKT1;1 was relatively more abundant in roots than in stems or leaves from melon seedlings. The tobacco transient expression system showed that CmHKT1;1 was plasma-membrane localized. Upon salt stress, CmHKT1;1 expression was more strongly upregulated in a salt-tolerant melon cultivar, 'Bingxuecui' (BXC) compared with a salt-sensitive cultivar, 'Yulu' (YL). Electrophysiological evidence demonstrated that CmHKT1;1 only transported Na+, rather than K+, when expressed in Xenopus laevis oocytes. Overexpression of CmHKT1;1 increased salt sensitivity in Saccharomyces cerevisiae and salt tolerance in Arabidopsis thaliana. Under NaCl treatments, transgenic Arabidopsis plants accumulated significantly lower concentrations of Na+ in shoots than wild type plants and showed a better K+/Na+ balance, leading to better Fv/Fm, root length, biomass, and enhanced plant growth. The CmHKT1;1 gene may serve as a useful candidate for improving crop salt tolerance.

Morphophysiological and molecular evidence supporting the augmentative role of Piriformospora indica in mitigation of salinity in Cucumis melo L.

Salinity is one of the major limiting factors in plant growth and productivity. Cucumis melo L. is a widely cultivated plant, but its productivity is significantly influenced by the level of salinity in soil. Symbiotic colonization of plants with Piriformospora indica has shown a promotion in plants growth and tolerance against biotic stress. In this study, physiological markers such as ion analysis, antioxidant determination, proline content, electrolyte leakage and chlorophyll measurement were assessed in melon cultivar under two concentrations (100 and 200 mM) of NaCl with and without P. indica inoculation. Results showed that the endophytic inoculation consistently upregulated the level of antioxidants, enhanced plants to antagonize salinity stress. The expression level of an RNA editing factor (SLO2) which is known to participate in mitochondria electron transport chain was analyzed, and its full mRNA sequence was obtained by rapid amplification of cDNA ends (RACE). Under salinity stress, the expression level of SLO2 was increased, enhancing the plant's capability to adapt to the stress. However, P. indica inoculation further elevated the expression level of SLO2. These findings suggested that the symbiotic association of fungi could help the plants to tolerate the salinity stress.

Clinical Profile and Significance of Mucocutaneous Lesions of Primary Sjögren's Syndrome: A Large Cross-sectional Study with 874 Patients.

BACKGROUND: Mucocutaneous lesions are common features of primary Sjögren's syndrome (pSS), but only a few studies have focused on them. To demonstrate the profile of mucocutaneous lesions of pSS and further explore their potential clinical significance, we performed a cross-sectional study on 874 patients. METHODS: Demographic data, clinical manifestations, and laboratory results of 874 pSS patients were collected. Patients were divided into two groups according to the presence of mucocutaneous lesions. Differences in primary symptoms and systemic impairments between the two groups were analyzed. Results of laboratory tests were also compared after excluding those who had taken corticosteroid from both groups. One-year follow-up was done, and occurrences of various new complications were compared. RESULTS: Among the 874 pSS patients, 181 patients had mucocutaneous lesions, accounting for 20.7%. Multiple mucocutaneous manifestations were displayed, and the top four most common types of lesions were purpuric eruptions (39.8%), urticaria (23.8%), Raynaud's phenomenon (14.9%), and angular stomatitis (9.9%). Incidences of pulmonary interstitial fibrosis, pulmonary bullae, leukopenia, and anemia were significantly higher among patients with mucocutaneous lesions (P < 0.05). Increase in IgG and decrease in C4 among patients with mucocutaneous lesions displayed statistical significance after excluding patients from both groups who had taken corticosteroid (P < 0.05). After one-year follow-up, patients with mucocutaneous lesions presented a slightly higher incidence of new complications compared to those without. CONCLUSIONS: Mucocutaneous manifestations of pSS patients were common and diverse. Patients with mucocutaneous manifestations had more systemic damages, higher level of IgG, and lower level of serum C4, suggesting a higher activity of the primary disease.

A comparable study of clinical and optical outcomes after 1.8, 2.0 mm microcoaxial and 3.0 mm coaxial cataract surgery.

AIM: To evaluate the clinical and optical outcomes after clear corneal incision cataract surgery (CICS) with three different incision sizes (1.8, 2.0 and 3.0 mm). METHODS: Eyes of 150 patients with age-related cataract scheduled for coaxial cataract surgery were randomized to three groups: 1.8, 2.0, or 3.0 mm CICS. Intraoperative data and postoperative outcomes including surgically induced astigmatism (SIA), the corneal incision thickness, wavefront aberrations and modulation transfer function (MTF) of cornea were obtained. RESULTS: There were no significant differences among the three groups in demographic characteristics and intraoperative outcome. The 1.8 and 2.0 mm microincisions showed more satisfactory clinical outcomes than the 3.0 mm incision. The 1.8 mm incision showed significantly less SIA than the 2.0 mm incision until postoperative 1mo (P<0.05), but the difference was only 0.14-0.18 D. Combined with less increased incision thickness only at postoperative 1d (P=0.013), the 1.8 mm incision presented better uncorrected distance visual acuity (UCDVA) than the 2.0 mm incision only at 1d postoperatively (P=0.008). For higher-order aberrations and other Zernike coefficients, there were no significant differences between the 1.8 mm group and 2.0 mm group (P>0.05). CONCLUSION: Converting from 3.0 mm CICS to 1.8 or 2.0 mm CICS result in better clinical and optical outcomes. However, when incision is 1.8 mm, the benefits from further reduction in size compared with 2.0 mm are limited. The necessity to reduce the incision size is to be deliberated.

Tris[2,4-dichloro-6-(ethyl-imino-meth-yl)phenolato-κ(2)N,O]cobalt(III).

The asymmetric unit of the title compound, [Co(C(9)H(8)Cl(2)NO)(3)], contains three independent mol-ecules. In each mol-ecule, the Co(III) ion is coordinated by an O atom and an N atom from three bidentate 2,4-dichloro-6-(ethyl-imino-meth-yl)phenolate ligands in a slightly distorted octa-hedral environment. In the crystal, a weak C-H⋯Cl hydrogen bond is observed.

[Resveratrol attenuates oxidant-induced mitochondrial damage in embryonic rat cardiomyocytes via inactivating GSK-3ß].

OBJECTIVE: To investigate the underlying mechanism of the protective effects of resveratrol on oxidant-induced mitochondrial damage in embryonic rat cardiomyocytes. METHODS: H9c2 cells, a permanent cell line derived from embryonic rat cardiac tissue, and then randomly divided into control group [PBS, cells exposed to H2O2 (600 µmol/L) for 20 min to induce mitochondrial oxidant damage], resveratrol group (0.01, 0.1, 1, 5, 10 and 20 µmol/L for 20 min at 20 min before exposing to H2O2), resveratrol plus inhibitor group (1 µmol/L KT5823 for 10 min at 10 min before 5 µmol/L resveratrol treatment) and inhibitor group (1 µmol/L KT5823 for 10 min). Mitochondrial membrane potential (ΔΨm) was measured by staining cells with tetramethylrhodamine ethyl ester (TMRE) and the mitochondrial permeability transition pore (mPTP) opening was evaluated by measuring the decrease of TMRE fluorescence intensity. Immunofluorescence assay was used to observe GSK-3ß phosphorylation. The phosphorylation of GSK-3ß and VASP were determined by Western blot. To detect intracellular NO, cells were loaded with DAF-FM DA (specific fluorescent dye of NO) and imaged with confocal microscopy. RESULTS: Compared to the control group, resveratrol (0.01-5 µmol/L) attenuated H2O2-induced mitochondrial damage reflected by attenuating the H2O2-induced TMRE fluorescence intensity decrease in a dose-dependent manner and the efficacy of 10 and 20 µmol/L resveratrol was significantly lower than that of 5 µmol/L resveratrol. Resveratrol also significantly upregulated the protein expression of VASP and increased GSK-3ß Ser(9) phosphorylation, which could lead the inactivation of GSK-3ß. These effects of resveratrol could be significantly abolished by protein kinase G inhibitor KT5823, while KT5823 alone did not affect GSK-3ß and VASP phosphorylation. Confocal microscopy showed that DAF-FM (specific NO indicator) was similar between resveratrol and control group, suggesting that resveratrol did not produce NO. CONCLUSIONS: Resveratrol could attenuate oxidant-induced mitochondrial damage in embryonic rat cardiomyocytes by inactivating GSK-3ß via cGMP/PKG signaling pathway independent of NO-related mechanism.

trans-Bis[4-amino-N-(pyrimidin-2-yl-κN)benzene-sulfonamidato-κN]bis(N,N-dimethyl-formamide-κO)cobalt(II).

The title complex, [Co(C(10)H(9)N(4)O(2)S)(2)(C(3)H(7)NO)(2)], lies across an inversion center. The Co(II) atom is coordinated in a slightly distorted octa-hedral geometry by four N atoms from two bidentate 4-amino-N-(pyrimidin-2-yl)benzene-sulfonamidate (sulfadiazine) anions and two O atoms from two dimethyl-formamide (DMF) ligands. The dihedral angle between the benzene and pyrimidine rings is 82.37 (13)°. A three-dimensional network is generated by N-H⋯O hydrogen bonds between the amino groups and of the sulfonamidate O atoms of neighbouring mol-ecules. The DMF ligand is disordered over two sets of sites in a 0.559 (4):0.441 (4) ratio.

Bis[4-bromo-2-(ethyl-imino-meth-yl)phenolato-κN,O]nickel(II).

In the title complex, [Ni(C(9)H(9)BrNO)(2)], the Ni(II) ion lies on an inversion centre and is coordinated in a slightly distorted square-planar geometry by two N atoms and two O atoms from two symmetry-related bidentate 4-bromo-2-(ethyl-imino-meth-yl)phenolate ligands. The complex forms a one-dimensional chain in the crystal structure through short C-H⋯Br contacts (H⋯Br = 3.009 Å).

From nanoplates to microtubes and microrods: a surfactant-free rolling mechanism for facile fabrication and morphology evolution of Ag2S films.

By a simple and facile wet-chemistry technique without any surfactant, various shapes of Ag(2)S crystals--including leaflike pentagonal nanoplates, crinkly nanoscrolls, hexagonal prismlike microtubes, and microrods--were fabricated in situ on a large-area silver-foil surface separately. Detailed experiments revealed that the Ag(2)S nanoplates were formed just by immersing the silver foil in a sulfur/ethanol solution at room temperature and atmospheric pressure, and they subsequently rolled into nanoscrolls and further grew into microtubes and microrods under solvothermal conditions. Inspired by the natural curling of a piece of foliage, we proposed a surfactant-free rolling mechanism to interpret the observed morphological evolution from lamellar to tubular structures. Based on these simple, practical, and green chemical synthetic routes, we can easily synthesize lamellar, scrolled, tubular, and clubbed Ag(2)S crystals by simply adjusting the reaction temperature, pressure, and time. It is very interesting to note that the current rolling process is quite different from the previous reported rolling mechanism that highly depends on the surfactants; we revealed that the lamellar Ag(2)S could be rolled into tubular structures without using any surfactant or other chemical additives, just like the natural rolling process of a piece of foliage. Therefore, this morphology-controlled synthetic route of Ag(2)S crystals may provide new insight into the synthesis of metal sulfide semiconducting micro-/nanocrystals with desired morphologies for further industrial applications. The optical properties of the pentagonal Ag(2)S nanoplates/film were also investigated by UV/Vis and photoluminescence (PL) techniques, which showed large blue-shift of the corresponding UV/Vis and PL spectra.

A K+ channel from salt-tolerant melon inhibited by Na+.

• The possible roles of K(+) channels in plant adaptation to high Na(+) conditions have not been extensively analyzed. Here, we characterize an inward Shaker K(+) channel, MIRK (melon inward rectifying K(+) channel), cloned in a salt-tolerant melon (Cucumis melo) cultivar, and show that this channel displays an unusual sensitivity to Na(+) . • MIRK expression localization was analyzed by reverse-transcription PCR (RT-PCR). MIRK functional analyses were performed in yeast (growth tests) and Xenopus oocytes (voltage-clamp). MIRK-type activity was revealed in guard cells using the patch-clamp technique. • MIRK is an inwardly rectifying Shaker channel belonging to the 'KAT' subgroup and expressed in melon leaves (especially in guard cells and vasculature), stems, flowers and fruits. Besides having similar features to its close homologs, MIRK displays a unique property: inhibition of K(+) transport by external Na(+) . In Xenopus oocytes, external Na(+) affected both inward and outward MIRK currents in a voltage-independent manner, suggesting a blocking site in the channel external mouth. • The degree of MIRK inhibition by Na(+) , which is dependent on the Na(+) /K(+) concentration ratio, is predicted to have an impact on the control of K(+) transport in planta upon salt stress. Expressed in guard cells, MIRK might control Na(+) arrival to the shoots via regulation of stomatal aperture by Na(+) .

Extranuclear estrogen receptors mediate the neuroprotective effects of estrogen in the rat hippocampus.

BACKGROUND: 17beta-estradiol (E2) has been implicated to exert neuroprotective effects in the brain following cerebral ischemia. Classically, E2 is thought to exert its effects via genomic signaling mediated by interaction with nuclear estrogen receptors. However, the role and contribution of extranuclear estrogen receptors (ER) is unclear and was the subject of the current study. METHODOLOGY/PRINCIPAL FINDINGS: To accomplish this goal, we employed two E2 conjugates (E2 dendrimer, EDC, and E2-BSA) that can interact with extranuclear ER and exert rapid nongenomic signaling, but lack the ability to interact with nuclear ER due to their inability to enter the nucleus. EDC or E2-BSA (10 microM) was injected icv 60 min prior to global cerebral ischemia (GCI). FITC-tagged EDC or E2-BSA revealed high uptake in the hippocampal CA1 region after icv injection, with a membrane (extranuclear) localization pattern in cells. Both EDC and E2-BSA exerted robust neuroprotection in the CA1 against GCI, and the effect was blocked by the ER antagonist, ICI182,780. EDC and E2-BSA both rapidly enhanced activation of the prosurvival kinases, ERK and Akt, while attenuating activation of the proapoptotic kinase, JNK following GCI, effects that were blocked by ICI182,780. Administration of an MEK or PI3K inhibitor blocked the neuroprotective effects of EDC and E2-BSA. Further studies showed that EDC increased p-CREB and BDNF in the CA1 region in an ERK- and Akt-dependent manner, and that cognitive outcome after GCI was preserved by EDC in an ER-dependent manner. CONCLUSIONS/SIGNIFICANCE: In conclusion, the current study demonstrates that activation of extranuclear ER results in induction of ERK-Akt-CREB-BDNF signaling in the hippocampal CA1 region, which significantly reduces ischemic neuronal injury and preserves cognitive function following GCI. The study adds to a growing literature that suggests that extranuclear ER can have important actions in the brain.

Isolation and characterization of a GS2 gene in melon (Cucumis melo L.) and its expression patterns under the fertilization of different forms of N.

We isolated a novel glutamine synthetase (GS, EC 6.3.1.2) gene M-GS2 (accession: AY773090) by the RACE approach from melon. The full-length cDNA of M-GS2 is 1807 bp and contains a 1296 bp open reading frame (ORF) encoding 432 amino acids. The deduced protein contains conserved structural domains among plant GS2 proteins and shares extensive sequence homology with GS2 enzymes from other higher plants. M-GS2 expresses with specificity in leaf, and identification of a chloroplast transit peptide (cTP) in M-GS2 suggests that it localizes to the chloroplast. As shown by real-time quantitative PCR, distinct forms of nitrogen (N) found in fertilizers transcriptionally regulated M-GS2 differently. Ammonium and nitrate feeding only significantly regulated M-GS2 transcripts in leaf; starving (0.75 mM) or moderate (3.75 mM) N levels dramatically increased M-GS2 transcripts for 1 day, decreasing to a constant low level after 2-3 days, while sufficient N level (7.5 mM) had a minor effect throughout 3 days compared to controls. Glutamate feeding, however, not only significantly regulated M-GS2 transcripts in leaf (decreased initially then increased to higher levels than controls), but also in root, where it was up-regulated continuously. Our results suggested that M-GS2 is the first GS gene cloned and characterized in melon and melon responds to the variations in N fertilization by differentially expressing M-GS2.

Protective effect of magnolol against hydrogen peroxide-induced oxidative stress in human lens epithelial cells.

Oxidative stress plays a significant role in the progression of cataract. We aimed to investigate the protective effect of magnolol, a compound extracted from the Chinese herb Magnolia officinalis, against oxidative stress in human lens epithelial (HLE) cells as well as the possible molecular mechanism involved. In this study, magnolol was observed to protect against H2O2-induced cytotoxicity in HLE B-3 cells. Magnolol inhibited the generation of reactive oxygen species (ROS), loss of mitochondrial membrane potential (Delta psi m) and release of cytochrome c from mitochondria caused by H2O2 into cytosol in HLE B-3 cells. Magnolol also inhibited H2O2-induced expressions of caspase-9 and caspase-3 and reduction of Bcl-2/Bax ratio. Moreover, magnolol attenuated the deactivation of ERK/MAPK (extracellular signal-regulated kinase/mitogen activated protein kinase) and the enhanced activation of p38, JNK (c-Jun N-terminal kinase) induced by H2O2. Magnolol could be useful in protecting against oxidative stress in HLE cells, suggesting a potential protective effect against cataractogenesis effect against cataractogenesis.