In vitro production and distribution of flavonoids in Glycyrrhiza uralensis Fisch.

Glycyrrhiza uralensis Fisch. is known as a common Chinese medicinal herb used to harmonize the effects of other ingredients in most Chinese herbal prescriptions. The rapid production of flavonoids in vitro remains unknown in G. uralensis Fisch. To investigate the in vitro adventitious root regeneration and flavonoid accumulation characteristics in G. uralensis for restrictions on collecting wild plants, suspension cultural and freezing microtomy with histochemical assays were carried out. We reported that multiple adventitious roots were initiated from hypocotyls and stems of G. uralensis. Indole-3-butyric acid (IBA) was more conducive than NAA (1-naphthaleneacetic acid) in inducing G. uralensis adventitious roots, but the addition of 6-BA (6-benzylaminopurine) and KT (kinetin) suppressed the formation of adventitious roots. While the concentration of IBA was 1.0 mg L-1, the flavonoid content and yield were the highest at 19.96 mg g-1 and 1.23 mg g-1, respectively. The optimum medium for adventitious root induction was 1/4-strength Murashige and Skoog's medium containing 0.1 mg L-1 IBA. The content of flavonoids in adventitious roots and apicals cultured in vitro was higher than that in suspension callus, reaching 3.87 times the callus flavonoid content. The histochemical localization of flavonoids showed that G. uralensis flavonoids mainly distributed in the epidermal parenchyma cells of the callus outer layers and gradually accumulated in cell wall and cell gaps of the epidermis and endodermis of adventitious roots along with the primary growth of adventitious roots, indicating that there were no flavonoids in the roots at the early stage of adventitious roots formation. The results showed that calli inducing adventitious roots and apicals for 30 days obtained the highest yield of flavonoid, indicating effective production for flavonoids instead of wild culture. AlCl3 ethanol solution was better than NaOH aqueous solution in terms of chromogenic and localization effects. We concluded that the highest yield of flavonoid and effective production for flavonoid instead of wild culture could be obtained from calli inducing adventitious roots and apicals.

The strong competitive role of 2n pollen in several polyploidy hybridizations in Rosa hybrida.

BACKGROUND: 2n pollen play a strong competitive role in hybridization and breeding of multiploids in Rosa hybrida. The ploidy inheritable characteristic of 'Orange Fire' × 'Old Blush' were analyzed. RESULT: The results of the cytological observations indicated that 2n pollen developed from the defeated cytoplasmic division or nuclear division in the meiosis metaphase II of PMC (pollen mother cell) in 'Old Blush'. The natural generation rate of the 2n pollen in 'Old Blush' (2x) was about 1.39 in percentage of all male gametes, whereas the tetraploids in the F1 offspring possessed a high rate, i.e., 44.00%. The temporal and spatial characteristics of 'Old Blush' pollen germination on the stigma and growth in pistil of 'Orange Fire' and 'DEE' were observed, and the results suggested that the germination rate of 2n pollen on the stigma was not superior to that of 1n pollen, but that the proportion of 2n pollen increased to 30.90 and 37.20%, respectively, while it traversed the stigma and entered into style. The callose plug in the 2n pollen tube was significantly thinner than that of 1n pollen tube. And each trait involved in our experiment probably is very important for F1 morphological phenotypes. CONCLUSION: We conclude that 2n pollen are involved in hybridization and have a competitive advantage while it traversed the stigma and entered into style. The callose plug in the 2n pollen tube was may have strongly influenced the competitive process in R. hybrida.

Origin and diversification of leucine-rich repeat receptor-like protein kinase (LRR-RLK) genes in plants.

BACKGROUND: Leucine-rich repeat receptor-like protein kinases (LRR-RLKs) are the largest group of receptor-like kinases in plants and play crucial roles in development and stress responses. The evolutionary relationships among LRR-RLK genes have been investigated in flowering plants; however, no comprehensive studies have been performed for these genes in more ancestral groups. The subfamily classification of LRR-RLK genes in plants, the evolutionary history and driving force for the evolution of each LRR-RLK subfamily remain to be understood. RESULTS: We identified 119 LRR-RLK genes in the Physcomitrella patens moss genome, 67 LRR-RLK genes in the Selaginella moellendorffii lycophyte genome, and no LRR-RLK genes in five green algae genomes. Furthermore, these LRR-RLK sequences, along with previously reported LRR-RLK sequences from Arabidopsis thaliana and Oryza sativa, were subjected to evolutionary analyses. Phylogenetic analyses revealed that plant LRR-RLKs belong to 19 subfamilies, eighteen of which were established in early land plants, and one of which evolved in flowering plants. More importantly, we found that the basic structures of LRR-RLK genes for most subfamilies are established in early land plants and conserved within subfamilies and across different plant lineages, but divergent among subfamilies. In addition, most members of the same subfamily had common protein motif compositions, whereas members of different subfamilies showed variations in protein motif compositions. The unique gene structure and protein motif compositions of each subfamily differentiate the subfamily classifications and, more importantly, provide evidence for functional divergence among LRR-RLK subfamilies. Maximum likelihood analyses showed that some sites within four subfamilies were under positive selection. CONCLUSIONS: Much of the diversity of plant LRR-RLK genes was established in early land plants. Positive selection contributed to the evolution of a few LRR-RLK subfamilies.

Type 5 adenylyl cyclase increases oxidative stress by transcriptional regulation of manganese superoxide dismutase via the SIRT1/FoxO3a pathway.

BACKGROUND: For reasons that remain unclear, whether type 5 adenylyl cyclase (AC5), 1 of 2 major AC isoforms in heart, is protective or deleterious in response to cardiac stress is controversial. To reconcile this controversy we examined the cardiomyopathy induced by chronic isoproterenol in AC5 transgenic (Tg) mice and the signaling mechanisms involved. METHODS AND RESULTS: Chronic isoproterenol increased oxidative stress and induced more severe cardiomyopathy in AC5 Tg, as left ventricular ejection fraction fell 1.9-fold more than wild type, along with greater left ventricular dilation and increased fibrosis, apoptosis, and hypertrophy. Oxidative stress induced by chronic isoproterenol, detected by 8-OhDG was 15% greater, P=0.007, in AC5 Tg hearts, whereas protein expression of manganese superoxide dismutase (MnSOD) was reduced by 38%, indicating that the susceptibility of AC5 Tg to cardiomyopathy may be attributable to decreased MnSOD expression. Consistent with this, susceptibility of the AC5 Tg to cardiomyopathy was suppressed by overexpression of MnSOD, whereas protection afforded by the AC5 knockout (KO) was lost in AC5 KO×MnSOD heterozyous KO mice. Elevation of MnSOD was eliminated by both sirtuin and MEK inhibitors, suggesting both the SIRT1/FoxO3a and MEK/ERK pathway are involved in MnSOD regulation by AC5. CONCLUSIONS: Overexpression of AC5 exacerbates the cardiomyopathy induced by chronic catecholamine stress by altering regulation of SIRT1/FoxO3a, MEK/ERK, and MnSOD, resulting in oxidative stress intolerance, thereby shedding light on new approaches for treatment of heart failure.

Novel mechanisms for caspase inhibition protecting cardiac function with chronic pressure overload.

Myocyte apoptosis is considered a major mechanism in the pathogenesis of heart failure. Accordingly, manipulations that inhibit apoptosis are assumed to preserve cardiac function by maintaining myocyte numbers. We tested this assumption by examining the effects of caspase inhibition (CI) on cardiac structure and function in C57BL/6 mouse with pressure overload model induced by transverse aortic constriction (TAC). CI preserved left ventricular (LV) function following TAC compared with the vehicle. TAC increased apoptosis in non-myocytes more than in myocytes and these increases were blunted more in non-myocytes by CI. Total myocyte number, however, did not differ significantly among control and TAC groups and there was no correlation between myocyte number and apoptosis, but there was a strong correlation between myocyte number and an index of myocyte proliferation, Ki67-positive myocytes. Despite comparable pressure gradients, LV hypertrophy was less in the CI group, likely attributable to decreased wall stress. Since changes in myocyte numbers did not account for protection from TAC, several other CI-mediated mechanisms were identified including: (a) lessening of TAC-induced fibrosis, (b) augmentation of isolated myocyte contractility, and (c) increased angiogenesis and Ki67-positive myocytes, which were due almost entirely to the non-myocyte apoptosis, but not myocyte apoptosis, with CI. CI maintained LV function following TAC not by protecting against myocyte loss, but rather by augmenting myocyte contractile function, myocyte proliferation, and angiogenesis resulting in reduced LV wall stress, hypertrophy, and fibrosis.

Arabidopsis transcription factor ELONGATED HYPOCOTYL5 plays a role in the feedback regulation of phytochrome A signaling.

Phytochrome A (phyA) is the primary photoreceptor responsible for perceiving and mediating various responses to far-red light in Arabidopsis thaliana. FAR-RED ELONGATED HYPOCOTYL1 (FHY1) and its homolog FHY1-LIKE (FHL) are two small plant-specific proteins essential for light-regulated phyA nuclear accumulation and subsequent phyA signaling processes. FHY3 and its homolog FAR-RED IMPAIRED RESPONSE1 (FAR1) are two transposase-derived transcription factors that directly activate FHY1/FHL transcription and thus mediate subsequent phyA nuclear accumulation and responses. Here, we report that ELONGATED HYPOCOTYL5 (HY5), a well-characterized bZIP transcription factor involved in promoting photomorphogenesis, directly binds ACGT-containing elements a few base pairs away from the FHY3/FAR1 binding sites in the FHY1/FHL promoters. We demonstrate that HY5 physically interacts with FHY3/FAR1 through their respective DNA binding domains and negatively regulates FHY3/FAR1-activated FHY1/FHL expression under far-red light. Together, our data show that HY5 plays a role in negative feedback regulation of phyA signaling by attenuating FHY3/FAR1-activated FHY1/FHL expression, providing a mechanism for fine-tuning phyA signaling homeostasis.

Photodissociation dynamics of the thiophenoxy radical at 248, 193, and 157 nm.

The photodissociation dynamics of the thiophenoxy radical (C6H5S) have been investigated using fast beam coincidence translational spectroscopy. Thiophenoxy radicals were produced by photodetachment of the thiophenoxide anion followed by photodissociation at 248 nm (5.0 eV), 193 nm (6.4 eV), and 157 nm (7.9 eV). Experimental results indicate two major competing dissociation channels leading to SH + C6H4 (o-benzyne) and CS + C5H5 (cyclopentadienyl) with a minor contribution of S + C6H5 (phenyl). Photofragment mass distributions and translational energy distributions were measured at each dissociation wavelength. Transition states and minima for each reaction pathway were calculated using density functional theory to facilitate experimental interpretation. The proposed dissociation mechanism involves internal conversion from the initially prepared electronic excited state to the ground electronic state followed by statistical dissociation. Calculations show that SH loss involves a single isomerization step followed by simple bond fission. For both SH and S loss, C-S bond cleavage proceeds without an exit barrier. By contrast, the CS loss pathway entails multiple transition states and minima as it undergoes five membered ring formation and presents a small barrier with respect to products. The calculated reaction pathway is consistent with the experimental translational energy distributions in which the CS loss channel has a broader distribution peaking farther away from zero than the corresponding distributions for SH loss.

Facile fabrication of CoAl-LDH nanosheets for efficient rhodamine B degradation via peroxymonosulfate activation.

Layered double hydroxides (LDHs) have been extensively investigated as promising peroxymonosulfate (PMS) activators for the degradation of organic pollutants. However, bulk LDHs synthesized using conventional methods possess a closely stacked layered structure, which seriously blocks active sites and yields low intrinsic activity. In this study, we exfoliated bulk CoAl-LDHs to fabricate CoAl-LDH nanosheets by alkali-etching and Ostwald ripening via a simple hydrothermal process in a KOH solution. The exfoliated LDHs possessed the typical nanosheet structure with more exposed active sites for PMS activation, and hence, boosted the degradation of the pollutants. CoAl-1 exhibited an outstanding catalytic performance as the PMS activator for rhodamine B (RhB) degradation with the apparent rate constant of 0.1687 min-1, which was about 3.63 and 5.02 times higher than that of commercial nano-Co3O4 and bulk CoAl-LDH, respectively. The maximum RhB degradation of 93.1% was achieved at the optimal reaction conditions: catalyst dose 0.1 g L-1, PMS concentration 0.3 mM, pH 7, and temperature 298 K. Further analysis of RhB degradation mechanism illustrated that singlet oxygen (1O2) dominated RhB degradation in the CoAl-1/PMS system, while ˙OH, ˙O2-, and ˙SO4- may mainly serve as the intermediates for the generation of 1O2 and were indirectly involved in the degradation. This study provides a promising strategy for developing two-dimensional LDH nanosheets for wastewater remediation.

Pain Catastrophizing, Kinesiophobia and Exercise Adherence in Patients After Total Knee Arthroplasty: The Mediating Role of Exercise Self-Efficacy.

Purpose: To examine whether exercise self-efficacy mediates the contributions of pain catastrophizing and kinesiophobia to exercise adherence in patients after total knee arthroplasty. Patients and Methods: A cross-sectional study design was conducted. A total 211 post-total knee arthroplasty patients were recruited from three orthopedics units of a tertiary hospital in China. Participants were invited to complete questionnaires on pain catastrophizing, kinesiophobia, exercise self-efficacy, and exercise adherence. Mplus 8.3 software was used to construct mediation models. Results: Pain catastrophizing and kinesiophobia were negatively correlated with exercise adherence (r = -0.509, r = -0.605, p < 0.001 respectively), while exercise self-efficacy were positively associated with exercise adherence (r = 0.799, p < 0.001). The results found exercise self-efficacy mediated the correlations of pain catastrophizing and kinesiophobia with exercise adherence after adjusting for demographic and clinical covariates. Pain catastrophizing indirectly affected patients' exercise adherence through its effect on exercise efficacy (indirect effect: -0.412), while Kinesiophobia is directly associated with exercise adherence and also indirectly through exercise self-efficacy (direct effect: -0.184, indirect effect: -0.415). Conclusion: Patients after total knee arthroplasty who have high levels of psychological distress (pain catastrophizing and kinesiophobia) are vulnerable to be non-adherent to exercise behaviors. Exercise self-efficacy explains the effects of pain catastrophizing and kinesiophobia on exercise adherence and may be a key target for measures to improve exercise behaviors in patients after total knee arthroplasty.

Developmental ablation of Id1 and Id3 genes in the vasculature leads to postnatal cardiac phenotypes.

The Id1 and Id3 genes play major roles during cardiac development, despite their expression being confined to non-myocardial layers (endocardium-endothelium-epicardium). We previously described that Id1Id3 double knockout (dKO) mouse embryos die at mid-gestation from multiple cardiac defects, but early lethality precluded the studies of the roles of Id in the postnatal heart. To elucidate postnatal roles of Id genes, we ablated the Id3 gene and conditionally ablated the Id1 gene in the endothelium to generate conditional KO (cKO) embryos. We observed cardiac phenotypes at birth and at 6 months of age. Half of the Id cKO mice died at birth. Postnatal demise was associated with cardiac enlargement and defects in the ventricular septum, trabeculation and vasculature. Surviving Id cKO mice exhibited fibrotic vasculature, cardiac enlargement and decreased cardiac function. An abnormal vascular response was also observed in the healing of excisional skin wounds of Id cKO mice. Expression patterns of vascular, fibrotic and hypertrophic markers were altered in the Id cKO hearts, but addition of Insulin-Like Growth Factor binding protein-3 (IGFbp3) reversed gene expression profiles of vascular and fibrotic, but not hypertrophic markers. Thus, ablation of Id genes in the vasculature leads to distinct postnatal cardiac phenotypes. These findings provide important insights into the role/s of the endocardial network of the endothelial lineage in the development of cardiac disease, and highlight IGFbp3 as a potential link between Id and its vascular effectors.

H11 kinase/heat shock protein 22 deletion impairs both nuclear and mitochondrial functions of STAT3 and accelerates the transition into heart failure on cardiac overload.

BACKGROUND: Cardiac overload, a major cause of heart failure, induces the expression of the heat shock protein H11 kinase/Hsp22 (Hsp22). METHODS AND RESULTS: To determine the specific function of Hsp22 in that context, a knockout mouse model of Hsp22 deletion was generated. Although comparable to wild-type mice in basal conditions, knockout mice exposed to pressure overload developed less hypertrophy and showed ventricular dilation, impaired contractile function, increased myocyte length and accumulation of interstitial collagen, faster transition into heart failure, and increased mortality. Microarrays revealed that hearts from knockout mice failed to transactivate genes regulated by the transcription factor STAT3. Accordingly, nuclear STAT3 tyrosine phosphorylation was decreased in knockout mice. Silencing and overexpression experiments in isolated neonatal rat cardiomyocytes showed that Hsp22 activates STAT3 via production of interleukin-6 by the transcription factor nuclear factor-κB. In addition to its transcriptional function, STAT3 translocates to the mitochondria where it increases oxidative phosphorylation. Both mitochondrial STAT3 translocation and respiration were also significantly decreased in knockout mice. CONCLUSIONS: This study found that Hsp22 represents a previously undescribed activator of both nuclear and mitochondrial functions of STAT3, and its deletion in the context of pressure overload in vivo accelerates the transition into heart failure and increases mortality.

Cardiomyocyte overexpression of the α1A-adrenergic receptor in the rat phenocopies second but not first window preconditioning.

We examined α(1A)-adrenergic receptor (AR) mediation of preconditioning in a novel α(1A)-AR cardiac transgenic (TG) rat model (α(1A)-TG). Compared with nontransgenic littermates (NTLs), in conscious α(1A)-TG rats, heart rate was reduced, contractility [left ventricle (LV) +dP/dt, ejection fraction, end-systolic elastance] was significantly enhanced, and triple product (LV systolic wall stress × LV +dP/dt × heart rate) was unchanged. However, infarct size (IS)/area at risk (AAR) in response to ischemia-reperfusion (30 min coronary occlusion/3 h reperfusion) was reduced to 35 ± 4.6% in α(1A)-TGs vs. 52 ± 2.2% in NTLs (P < 0.05). Second window preconditioning reduced IS/AAR in NTLs to 29 ± 2.7% but did not afford further protection in α(1A)-TGs. In contrast, with first window preconditioning, IS/AAR was reduced to similar levels in both α(1A)-TGs (12 ± 1.4%) and NTLs (10 ± 1.1%). In untreated α(1A)-TGs, cardioprotection was associated with enhanced myocardial phosphorylated (p)-mitogen/extracellular signal-regulated kinase (MEK), p-extracellular signal-regulated kinase (ERK), and inducible nitric oxide synthase (iNOS) at the protein level, along with a 1.3-fold increase in total nitric oxide synthase activity like in second window preconditioning. Affymetrix microarrays revealed that few genes (4.6% of 3,172 upregulated; 8.8% of 3,498 downregulated) showed directionally similar changes in α(1A)-TGs vs. NTLs subjected to second window preconditioning. Thus, second, but not first, window cardioprotection is evident in α(1A)-TGs in the absence of ischemic preconditioning and is mediated by iNOS activation associated with MEK/ERK phosphorylation. Transcriptionally, however, second window preconditioning is considerably more complex than α(1A)-TG preconditioning, with the alteration of thousands of additional genes affording no further protection than that already available in α(1A)-TG rats.

Prevention of heart failure in mice by an antiviral agent that inhibits type 5 cardiac adenylyl cyclase.

Despite numerous discoveries from genetically engineered mice, relatively few have been translated to the bedside, mainly because it is difficult to translate from genes to drugs. This investigation examines an antiviral drug, which also has an action to selectively inhibit type 5 adenylyl cyclase (AC5), a pharmaceutical correlate of the AC5 knockout (KO) model, which exhibits longevity and stress resistance. Our objective was to examine the extent to which pretreatment with this drug, adenine 9-ß-d-arabinofuranoside (Ara-A), favorably ameliorates the development of heart failure (HF). Ara-A exhibited selective inhibition for AC5 compared with the other major cardiac AC isoform, AC6, i.e., it reduced AC activity significantly in AC5 transgenic (Tg) mice, but not in AC5KO mice and had little effect in either wild-type or AC6Tg mice. Permanent coronary artery occlusion for 3 wk in C57Bl/6 mice increased mortality and induced HF in survivors, as reflected by reduced cardiac function, while increasing cardiac fibrosis. The AC5 inhibitor Ara-A significantly improved all of these end points and also ameliorated chronic isoproterenol-induced cardiomyopathy. As with the AC5KO mice, Ara-A increased mitogen/extracellular signal-regulated kinase (MEK)/extracellular signal-regulated kinase (ERK) phosphorylation. A MEK inhibitor abolished the beneficial effects of the AC5 inhibitor in the HF model, indicating the involvement of the downstream MEK-ERK pathway of AC5. Our data suggest that pharmacological AC5 inhibition may serve as a new therapeutic approach for HF.

Non-destructive Storage Time Prediction of Newhall Navel Oranges Based on the Characteristics of Rind Oil Glands.

How to non-destructively and quickly estimate the storage time of citrus fruit is necessary and urgent for freshness control in the fruit market. As a feasibility study, we present a non-destructive method for storage time prediction of Newhall navel oranges by investigating the characteristics of the rind oil glands in this paper. Through the observation using a digital microscope, the oil glands were divided into three types and the change of their proportions could indicate the rind status as well as the storage time. Images of the rind of the oranges were taken in intervals of 10 days for 40 days, and they were used to train and test the proposed prediction models based on K-Nearest Neighbors (KNN) and deep learning algorithms, respectively. The KNN-based model demonstrated explicit features for storage time prediction based on the gland characteristics and reached a high accuracy of 93.0%, and the deep learning-based model attained an even higher accuracy of 96.0% due to its strong adaptability and robustness. The workflow presented can be readily replicated to develop non-destructive methods to predict the storage time of other types of citrus fruit with similar oil gland characteristics in different storage conditions featuring high efficiency and accuracy.

Plasma-Activated Acidic Electrolyzed Water: A New Food Disinfectant for Bacterial Suspension and Biofilm.

Food-borne diseases are widespread all over the world, and food safety has attracted much attention. This study is the first to use plasma to activate acidic electrolyzed water (AEW) to obtain a new disinfectant for food processing. The germicidal efficacy of plasma-activated acidic electrolyzed water (PA-AEW) on B. subtilis suspension and biofilm was investigated. Furthermore, the synergistic effect of different bactericidal factors was inferred by investigating the physicochemical parameters of PA-AEW and the influencing factors of bactericidal effect. The results demonstrate that PA-AEW is a highly effective and rapid disinfectant. The killing logarithm (KL) value of PA-AEW on B. subtilis suspension could reach 2.33 log10CFU/mL with a sterilization time of 10 s, which is significantly higher than that of AEW (KL = 0.58 log10CFU/mL) and plasma-activated water (PAW) (KL = 0.98 log10CFU/mL) (significant difference, p < 0.01). Moreover, the KL value of the B. subtilis biofilm of PA-AEW was 2.41 log10CFU/mL, better than that of PAW and AEW (significant difference, p < 0.01), indicating that PA-AEW has important application prospects in food processing. The synergistic effect should come from the interaction between reactive chlorine species (RCS) and reactive oxygen and nitrogen species (RONS) in PA-AEW.

MicroRNA-21 is a downstream effector of AKT that mediates its antiapoptotic effects via suppression of Fas ligand.

MicroRNA-21 (miR-21) is highly up-regulated during hypertrophic and cancerous cell growth. In contrast, we found that it declines in cardiac myocytes upon exposure to hypoxia. Thus, the objective was to explore its role during hypoxia. We show that miR-21 not only regulates phosphatase and tensin homologue deleted on chromosome 10 (PTEN), but also targets Fas ligand (FasL). During prolonged hypoxia, down-regulation of miR-21 proved necessary and sufficient for enhancing expression of both proteins. We demonstrate here for the first time that miR-21 is positively regulated via an AKT-dependent pathway, which is depressed during prolonged hypoxia. Accordingly, hypoxia-induced down-regulation of miR-21 and up-regulation of FasL and PTEN were reversed by activated AKT and reproduced by a dominant negative mutant, wortmannin, or PTEN. Moreover, the antiapoptotic function of AKT partly required miR-21, which was sufficient for inhibition of caspase-8 activity and mitochondrial damage. In consensus, overexpression of miR-21 in a transgenic mouse heart resulted in suppression of ischemia-induced up-regulation of PTEN and FasL expression, an increase in phospho-AKT, a smaller infarct size, and ameliorated heart failure. Thus, we have identified a unique aspect of the function of AKT by which it inhibits apoptosis through miR-21-dependent suppression of FasL.

Apoptosis in severe, compensated pressure overload predominates in nonmyocytes and is related to the hypertrophy but not function.

It is widely held that myocyte apoptosis in left ventricular hypertrophy (LVH) contributes to left ventricle (LV) dysfunction and heart failure. The main goal of this investigation was to determine if there is a statistical relationship among LV hypertrophy, apoptosis and LV function, and importantly whether the apoptosis occurs in myocytes or nonmyocytes in the heart. We used both rat and canine models of severe LVH induced by chronic thoracic aortic banding with resultant LV-aortic pressure gradients 145-155 mmHg and increases in LV/body weight of 58 and 70%. These models also provided the ability to examine transmural apoptosis in LVH. In both models, the overwhelming majority (88%) of apoptotic cells were nonmyocytes. The regressions for apoptosis vs. LVH were stronger for nonmyocytes than myocytes and also stronger in the subendocardium than the subepicardium. Importantly, LV systolic and diastolic wall stresses were normal, indicating that the apoptosis could not be attributed to LV stretch or heart failure. In addition, there was no relationship between the extent of apoptosis and LV ejection fraction, which actually increased (P < 0.05), in the face of elevated LV systolic pressure, indicating that greater apoptosis did not result in a decrease in LV function. Thus, in response to chronic, severe pressure overload, LVH in the absence of LV dilation, and elevated LV wall stress, apoptosis occurred predominantly in nonmyocytes in the myocardial interstitium, more in the subendocardium than the subepicardium. The extent of apoptosis was linearly related to the amount of LV hypertrophy, but not to LV function.

Decreased sarcolipin protein expression and enhanced sarco(endo)plasmic reticulum Ca2+ uptake in human atrial fibrillation.

Sarcolipin (SLN), a key regulator of cardiac sarco(endo)plasmic reticulum (SR) Ca(2+) ATPase, is predominantly expressed in atria and mediates ß-adrenergic responses. Studies have shown that SLN mRNA expression is decreased in human chronic atrial fibrillation (AF) and in aortic banded mouse atria; however, SLN protein expression in human atrial pathology and its role in atrial SR Ca(2+) uptake are not yet elucidated. In the present study, we determined the expression of major SR Ca(2+) handling proteins in atria of human AF patients and in human and in a mouse model of heart failure (HF). We found that the expression of SR Ca(2+) uptake and Ca(2+) release channel proteins are significantly decreased in atria but not in the ventricles of pressure-overload induced HF in mice. In human AF and HF, the expression of SLN protein was significantly decreased; whereas the expressions of other major SR Ca(2+) handling proteins were not altered. Further, we found that the SR Ca(2+) uptake was significantly increased in human AF. The selective downregulation of SLN and enhanced SR Ca(2+) uptake in human AF suggest that SLN downregulation could play an important role in abnormal intracellular Ca(2+) cycling in atrial pathology.

Ion-pair dissociation dynamics of H2S in the photon energy range 15.26-15.55 eV.

The H(+) velocity map images from the ion-pair dissociation of H(2)S + hν → SH(-)(X(1)Σ(+), υ = 0, 1) + H(+) have been measured at the excitation energies 15.259, 15.395, and 15.547 eV, respectively. The experimental results show that most of the available energies are transformed into the translational energies. The angular distributions of the fragments SH(-)(X(1)Σ(+), υ = 0) indicate that the dissociation occurs via pure parallel transition with limiting anisotropy parameter of +2. Because the ion-pair dissociation usually occurs via the predissociation of Rydberg states, this suggests that the ion cores of the excited Rydberg states have linear geometries. The geometries and electronic structures of the linear H(2)S(+) have been calculated employing the quantum chemistry calculation method at the CASPT2/avqz level. The electronic structures for the ion-pair states have been calculated at the CASSCF/avtz level, which indicates that the equilibrium geometries of the ion-pair states have bent geometries.

Distinct roles of GSK-3alpha and GSK-3beta phosphorylation in the heart under pressure overload.

Glycogen synthase kinase-3 (GSK-3) is a master regulator of growth and death in cardiac myocytes. GSK-3 is inactivated by hypertrophic stimuli through phosphorylation-dependent and -independent mechanisms. Inactivation of GSK-3 removes the negative constraint of GSK-3 on hypertrophy, thereby stimulating cardiac hypertrophy. N-terminal phosphorylation of the GSK-3 isoforms GSK-3alpha and GSK-3beta by upstream kinases (e.g., Akt) is a major mechanism of GSK-3 inhibition. Nonetheless, its role in mediating cardiac hypertrophy and failure remains to be established. Here we evaluated the role of Serine(S)21 and S9 phosphorylation of GSK-3alpha and GSK-3beta in the regulation of cardiac hypertrophy and function during pressure overload (PO), using GSK-3alpha S21A knock-in (alphaKI) and GSK-3beta S9A knock-in (betaKI) mice. Although inhibition of S9 phosphorylation during PO in the betaKI mice attenuated hypertrophy and heart failure (HF), inhibition of S21 phosphorylation in the alphaKI mice unexpectedly promoted hypertrophy and HF. Inhibition of S21 phosphorylation in GSK-3alpha, but not of S9 phosphorylation in GSK-3beta, caused phosphorylation and down-regulation of G1-cyclins, due to preferential localization of GSK-3alpha in the nucleus, and suppressed E2F and markers of cell proliferation, including phosphorylated histone H3, under PO, thereby contributing to decreases in the total number of myocytes in the heart. Restoration of the E2F activity by injection of adenovirus harboring cyclin D1 with a nuclear localization signal attenuated HF under PO in the alphaKI mice. Collectively, our results reveal that whereas S9 phosphorylation of GSK-3beta mediates pathological hypertrophy, S21 phosphorylation of GSK-3alpha plays a compensatory role during PO, in part by alleviating the negative constraint on the cell cycle machinery in cardiac myocytes.