DSnet: a new dual-branch network for hippocampus subfield segmentation.

The hippocampus is a critical component of the brain and is associated with many neurological disorders. It can be further subdivided into several subfields, and accurate segmentation of these subfields is of great significance for diagnosis and research. However, the structures of hippocampal subfields are irregular and have complex boundaries, and their voxel values are close to surrounding brain tissues, making the segmentation task highly challenging. Currently, many automatic segmentation tools exist for hippocampal subfield segmentation, but they suffer from high time costs and low segmentation accuracy. In this paper, we propose a new dual-branch segmentation network structure (DSnet) based on deep learning for hippocampal subfield segmentation. While traditional convolutional neural network-based methods are effective in capturing hierarchical structures, they struggle to establish long-term dependencies. The DSnet integrates the Transformer architecture and a hybrid attention mechanism, enhancing the network's global perceptual capabilities. Moreover, the dual-branch structure of DSnet leverages the segmentation results of the hippocampal region to facilitate the segmentation of its subfields. We validate the efficacy of our algorithm on the public Kulaga-Yoskovitz dataset. Experimental results indicate that our method is more effective in segmenting hippocampal subfields than conventional single-branch network structures. Compared to the classic 3D U-Net, our proposed DSnet improves the average Dice accuracy of hippocampal subfield segmentation by 0.57%.

A novel machine learning model for efficacy prediction of immunotherapy-chemotherapy in NSCLC based on CT radiomics.

Lung cancer is categorized into two main types: non-small cell lung cancer (NSCLC) and small cell lung cancer. Of these, NSCLC accounts for approximately 85% of all cases and encompasses varieties such as squamous cell carcinoma and adenocarcinoma. For patients with advanced NSCLC that do not have oncogene addiction, the preferred treatment approach is a combination of immunotherapy and chemotherapy. However, the progression-free survival (PFS) typically ranges only from about 6 to 8 months, accompanied by certain adverse events. In order to carry out individualized treatment more effectively, it is urgent to accurately screen patients with PFS for more than 12 months under this treatment regimen. Therefore, this study undertook a retrospective collection of pulmonary CT images from 60 patients diagnosed with NSCLC treated at the First Affiliated Hospital of Wenzhou Medical University. It developed a machine learning model, designated as bSGSRIME-SVM, which integrates the rime optimization algorithm with self-adaptive Gaussian kernel probability search (SGSRIME) and support vector machine (SVM) classifier. Specifically, the model initiates its process by employing the SGSRIME algorithm to identify pivotal image features. Subsequently, it utilizes an SVM classifier to assess these features, aiming to enhance the model's predictive accuracy. Initially, the superior optimization capability and robustness of SGSRIME in IEEE CEC 2017 benchmark functions were validated. Subsequently, employing color moments and gray-level co-occurrence matrix methods, image features were extracted from images of 60 NSCLC patients undergoing immunotherapy combined with chemotherapy. The developed model was then utilized for analysis. The results indicate a significant advantage of the model in predicting the efficacy of immunotherapy combined with chemotherapy for NSCLC, with an accuracy of 92.381% and a specificity of 96.667%. This lays the foundation for more accurate PFS predictions and personalized treatment plans.

CircPRMT5 promotes progression of osteosarcoma by recruiting CNBP to regulate the translation and stability of CDK6 mRNA.

Research has demonstrated that circular RNAs (circRNAs) exert critical functions in the occurrence and progression of numerous malignant tumors. CircPRMT5 was recently reported to be involved in the pathogenesis of cancers. However, the potential role of circPRMT5 in osteosarcoma needs further investigation. In present study, our results suggested that circPRMT5 was highly upregulated in osteosarcoma cells and mainly localizes in the cytoplasm. CircPRMT5 promoted the proliferation, migration and invasion capacities of osteosarcoma cells, and suppressed cell apoptosis. Knockdown of circPRMT5 exerted the opposite effects. Mechanically, circPRMT5 promoted the binding of CNBP to CDK6 mRNA, which enhanced the stability of CDK6 mRNA and facilitated its translation, thereby promoting the progression of osteosarcoma. Knockdown of CDK6 reversed the promoting effect of circPRMT5 on osteosarcoma cells. These findings suggest that circPRMT5 promotes osteosarcoma cell malignant activity by recruiting CNBP to regulate the translation and stability of CDK6 mRNA. Thus, circPRMT5 may represent a promising therapeutic target for osteosarcoma.

Deep guided transformer dehazing network.

Single image dehazing has received a lot of concern and achieved great success with the help of deep-learning models. Yet, the performance is limited by the local limitation of convolution. To address such a limitation, we design a novel deep learning dehazing model by combining the transformer and guided filter, which is called as Deep Guided Transformer Dehazing Network. Specially, we address the limitation of convolution via a transformer-based subnetwork, which can capture long dependency. Haze is dependent on the depth, which needs global information to compute the density of haze, and removes haze from the input images correctly. To restore the details of dehazed result, we proposed a CNN sub-network to capture the local information. To overcome the slow speed of the transformer-based subnetwork, we improve the dehazing speed via a guided filter. Extensive experimental results show consistent improvement over the state-of-the-art dehazing on natural haze and simulated haze images.

PPNet: Pyramid pooling based network for polyp segmentation.

Colonoscopy is the gold standard method for investigating the gastrointestinal tract. Localizing the polyps in colonoscopy images plays a vital role when doing a colonoscopy screening, and it is also quite important for the following treatment, e.g., polyp resection. Many deep learning-based methods have been applied for solving the polyp segmentation issue. However, precisely polyp segmentation is still an open issue. Considering the effectiveness of the Pyramid Pooling Transformer (P2T) in modeling long-range dependencies and capturing robust contextual features, as well as the power of pyramid pooling in extracting features, we propose a pyramid pooling based network for polyp segmentation, namely PPNet. We first adopt the P2T as the encoder for extracting more powerful features. Next, a pyramid feature fusion module (PFFM) combining the channel attention scheme is utilized for learning a global contextual feature, in order to guide the information transition in the decoder branch. Aiming to enhance the effectiveness of PPNet on feature extraction during the decoder stage layer by layer, we introduce the memory-keeping pyramid pooling module (MPPM) into each side branch of the encoder, and transmit the corresponding feature to each lower-level side branch. Experimental results conducted on five public colorectal polyp segmentation datasets are given and discussed. Our method performs better compared with several state-of-the-art polyp extraction networks, which demonstrate the effectiveness of the mechanism of pyramid pooling for colorectal polyp segmentation.

An n-valued neutrosophic set method for the assessment of an offshore oil spill risk.

With the development of maritime transportation, oil spill accidents occur frequently. In this paper, a scientific and reasonable assessment of ship oil spills, offshore oil platform oil spills, and subsea pipeline oil spills is carried out, and a risk assessment method of an offshore oil spill based on an n-value neutrosophic set is proposed. First, the oil spill risk evaluation index systems of these three risk sources are constructed, respectively, and the entropy weight method is used to calculate the weight. Second, we establish a risk assessment model under the n-value neutrosophic environment. Furthermore, we use the n-value neutrosophic-weighted arithmetic average (NVNWAA) to calculate the risk levels of the following three risk sources: ship oil spills, offshore oil platform spills, and submarine pipeline spills. Finally, according to the results of the risk assessment, the countermeasures to strengthen the preventive measures of oil spill accidents are put forward.

Psychological Adaptability and Intervention Strategies of College Students' English Learning under the Mixed Foreign Language Teaching Environment Monitoring.

The issues with traditional teaching methods in current English education are becoming more and more evident in college classrooms, which significantly impedes the innovative growth of college English education and does not promote the development of college students' all-around English proficiency. This essay focuses on the English learning adaptability and intervention tactics of college students using BFLTM (Blended Foreign Language Teaching Model). This study employs the individual-centered approach to choose college students as its primary subjects and conducts a cluster analysis of the adaptability of various college students' English learning based on the classification indicators. The study makes the following deductions: the general state of students' adaptability to studying English is average (M = 2.17). On the internal dimension, learning easy adaptation (M = 2.165) is average, and cognitive strategy adaptation (M = 2.642) is average, whether the average learning adaptability of English majors is significant (P = 0.001 < 0.05), and it is significant in the dimensions of learning attitude, learning interaction, and learning environment (P < 0.05). In view of the improvement of college students' English learning adaptability under BFLTM, this paper puts forward some strategies to further cultivate students' English autonomous learning ability by creating a suitable classroom atmosphere for autonomous learning, establishing an English autonomous learning center and a virtual community.

Automatic Aesthetics Evaluation of Robotic Dance Poses Based on Hierarchical Processing Network.

Vision plays an important role in the aesthetic cognition of human beings. When creating dance choreography, human dancers, who always observe their own dance poses in a mirror, understand the aesthetics of those poses and aim to improve their dancing performance. In order to develop artificial intelligence, a robot should establish a similar mechanism to imitate the above human dance behaviour. Inspired by this, this paper designs a way for a robot to visually perceive its own dance poses and constructs a novel dataset of dance poses based on real NAO robots. On this basis, this paper proposes a hierarchical processing network-based approach to automatic aesthetics evaluation of robotic dance poses. The hierarchical processing network first extracts the primary visual features by using three parallel CNNs, then uses a synthesis CNN to achieve high-level association and comprehensive processing on the basis of multi-modal feature fusion, and finally makes an automatic aesthetics decision. Notably, the design of this hierarchical processing network is inspired by the research findings in neuroaesthetics. Experimental results show that our approach can achieve a high correct ratio of aesthetic evaluation at 82.3%, which is superior to the existing methods.

Colorectal polyp region extraction using saliency detection network with neutrosophic enhancement.

Colorectal polyp recognition is crucial for early colorectal cancer detection and treatment. Colonoscopy is always employed for colorectal polyp scanning. However, one out of four polyps may be ignored, due to the similarity of polyp and normal tissue. In this paper, we present a novel method called NeutSS-PLP for polyp region extraction in colonoscopy images using a short connected saliency detection network with neutrosophic enhancement. We first utilize the neutrosophic theory to enhance the quality of specular reflections detection in the colonoscopy images. We develop the local and global threshold criteria in the single-valued neutrosophic set (SVNS) domain and define the corresponding T (Truth), I (Indeterminacy), and F (Falsity) functions for each criterion. The well-built neutrosophic images are processed and employed for specular reflection detection and suppressing. Next, we introduce two-level short connections into the saliency detection network, aiming to take advantage of the multi-level and multi-scale features extracted from different stages of the network. Experimental results conducted on two public colorectal polyp datasets achieve 0.877 and 0.9135 mIoU for polyp extraction respectively, and our method performs better compared with several state-of-the-art saliency networks and semantic segmentation networks, which demonstrate the effectiveness of applying the saliency detection mechanism for colorectal polyp region extraction.

TASK-1 regulates mitochondrial function under hypoxia.

TASK-1, TWIK-related acid-sensitive potassium channel 1, is a member of the two-pore- domain potassium channel family. It is constitutively active at resting potentials and strongly expressed in the heart. However, little is known about the role of TASK-1 channels in hypoxia. A cellular model of hypoxia and reoxygenation from rat heart-derived H9c2 cells or TASK-1 deficient HEK293T cells was employed to explore the role of TASK-1 channels in cytoprotection against hypoxia. The cell viability assay revealed that TASK-1 expression increased the number of viable cells subjected to 2 h of hypoxia followed by 2 h of reoxygenation (H/R). To dissect the protective role of TASK-1 on mitochondrial function, mitochondrial membrane potential (MMP) was assessed by tetramethylrhodamine fluorescence. It was demonstrated that MMP was significantly decreased by H/R, but it was maintained by TASK-1 expression or pretreatment with cyclosporin A, an inhibitor of mitochondrial permeability transition pore (mPTP). The effect of cyclosporin A on MMP was not further altered by TASK-1 expression. Moreover, TASK-1 expression significantly blocked cytochrome c release induced by H/R. While a small fraction of endogenous TASK-1 was found to colocalize with the mitochondrial marker MitoTracker in H9c2 cells, H/R did not alter the extent of colocalization of TASK-1 with MitoTracker. The total TASK-1 protein level was not significantly affected by H/R. In summary, we provided the evidence that TASK-1 channels confer cytoprotection against hypoxia-reoxygenation injury, possibly by their capacity of maintaining the mitochondrial membrane potential via inhibiting MPTP opening.

Kalman Filter for Spatial-Temporal Regularized Correlation Filters.

We consider visual tracking in numerous applications of computer vision and seek to achieve optimal tracking accuracy and robustness based on various evaluation criteria for applications in intelligent monitoring during disaster recovery activities. We propose a novel framework to integrate a Kalman filter (KF) with spatial-temporal regularized correlation filters (STRCF) for visual tracking to overcome the instability problem due to large-scale application variation. To solve the problem of target loss caused by sudden acceleration and steering, we present a stride length control method to limit the maximum amplitude of the output state of the framework, which provides a reasonable constraint based on the laws of motion of objects in real-world scenarios. Moreover, we analyze the attributes influencing the performance of the proposed framework in large-scale experiments. The experimental results illustrate that the proposed framework outperforms STRCF on OTB-2013, OTB-2015 and Temple-Color datasets for some specific attributes and achieves optimal visual tracking for computer vision. Compared with STRCF, our framework achieves AUC gains of 2.8%, 2%, 1.8%, 1.3%, and 2.4% for the background clutter, illumination variation, occlusion, out-of-plane rotation, and out-of-view attributes on the OTB-2015 datasets, respectively. For sporting events, our framework presents much better performance and greater robustness than its competitors.

Risk factors, changes in serum inflammatory factors, and clinical prevention and control measures for puerperal infection.

BACKGROUND: To investigate the risk factors and changes in serum inflammatory factors in puerperal infection, and propose clinical prevention measures. METHODS: A total of 240 subjects with suspected puerperal infection treated in our hospital from January 2017 to December 2017 were collected, among which puerperal infection was definitely diagnosed in 40 cases, and it was excluded in 40 cases. Levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and high-sensitivity C-reactive protein (hs-CRP) were compared between the two groups, and the change trends of IL-6 and hs-CRP were recorded. RESULTS: Levels of IL-6, hs-CRP, and TNF-α in puerperal infection group were higher than those in non-infection group (P < .05). Levels of IL-6 and hs-CRP at enrollment and 1-3 days after enrollment in infection group were higher than those in non-infection group (P < .05). The body mass index >25, placenta previa, placenta accreta, postpartum hemorrhage, premature rupture of membrane, gestational diabetes mellitus, and anemia during pregnancy were relevant and independent risk factors for puerperal infection. Puerperal infection occurred in uterine cavity, vagina, pelvic peritoneum, pelvic tissue, incision, urinary system, etc, and gram-negative (G+) bacteria were dominated in pathogens. CONCLUSION: The inflammatory response of patients with puerperal infection is significantly enhanced.

Functional interaction of the two-pore domain potassium channel TASK-1 and caveolin-3.

The two-pore domain potassium channel TASK-1 is strongly expressed in the heart and has been shown to modulate the resting membrane potential and action potential. However, little is known about the regulation of TASK-1 channels. The present study was designed to determine whether TASK-1 is modulated by caveolin-3, a primary structural protein of cardiac caveolae. Functional studies with the whole-cell voltage clamp technique showed that the expression of caveolin-3 decreased recombinant TASK-1 currents significantly in HEK293T cells, and this effect was prevented by co-expressing the dominant negative mutant caveolin-3 P104L. Immunofluorescence imaging revealed the colocalization of TASK-1 and caveolin-3. Co-immunoprecipitation analysis indicated that caveolin-3 associated with TASK-1. When co-expressed with caveolin-3 P104L, the fluorescence intensity of caveolin-3 on the cell periphery was reduced. This agrees with the functional evidence that caveolin-3 P104L prevented the inhibitory effect of caveolin-3 on TASK-1 currents, possibly via reducing the plasma membrane targeting of caveolin-3. Further, our data from cardiomyocytes suggest that TASK-1 is associated with caveolin-3. In summary, our study indicates that TASK-1 is functionally regulated by caveolin-3, possibly via association with each other on the cell surface. These results point out a novel mechanism in the regulation of TASK-1.

Regulation of protein kinase C-epsilon and its age-dependence.

Protein kinase C (PKC) is an important mediator in the cardioprotection of ischemic preconditioning and has been shown to translocate to mitochondria upon activation. However, little is known about the cellular signaling underlying the translocation of PKC isoforms to mitochondria and its age-dependence. The present study aimed to explore whether adenosine-induced translocation of PKCε to mitochondria is mediated by caveolin-3 and/or adenosine A2B receptor/PI3 kinase mediated signaling, and whether the mitochondrial targeting of PKCε is age-related. Immunofluorescence imaging of isolated mitochondria from cardiomyocytes and H9c2 cells showed that while adenosine-induced increase in mitochondrial PKCε was inhibited by adenosine A1 receptor blocker, pretreatment with adenosine A2B receptor specific inhibitor MRS 1754 or PI3K inhibitor Wortmannin did not significantly reduce adenosine-mediated increase in mitochondrial PKCε. Interestingly, adenosine-induced increase in mitochondrial translocation of PKCε was significantly blocked by suppressing caveolin-3 expression with specific siRNA. When compared to that in young adult rat hearts, the level of mitochondrial PKCε in middle-aged rat hearts was significantly lower at the basal condition and in response to adenosine treatment, along with largely decreased mitochondrial HSP90 and TOM70 protein expression. We demonstrate that adenosine-induced translocation of PKCε to mitochondria is mediated by a caveolin-3-dependent mechanism and this process is age-related, possibly in part, through regulation of HSP90 and TOM70 expression. These results point out a novel mechanism in regulating PKC function in mitochondria.

A Variety of Phase-Transition Behaviors in a Niccolite Series of [NH3 (CH2 )4 NH3 ][M(HCOO)3 ]2.

A niccolite series of [bnH2 (2+) ][M(HCOO)3 ]2 (bnH2 (2+) =1,4-butyldiammonium) shows four kinds of metal-dependent phase transitions, from high temperature para-electric phases to low-temperature ferro-, antiferro-, glass-like, and para-electric phases. The conformational flexibility of bnH2 (2+) and the different size, mass, and bonding character of the metal ion lead to various disorder-order transitions of bnH2 (2+) in the lattice and relevant framework modulations, thus different phase transitions and dielectric responses. The magnetic members display a coexistence or combination of electric and magnetic orderings in the low-temperature region.

Endurance exercise accelerates myocardial tissue oxygenation recovery and reduces ischemia reperfusion injury in mice.

Exercise training offers cardioprotection against ischemia and reperfusion (I/R) injury. However, few essential signals have been identified to underscore the protection from injury. In the present study, we hypothesized that exercise-induced acceleration of myocardial tissue oxygenation recovery contributes to this protection. C57BL/6 mice (4 weeks old) were trained on treadmills for 45 min/day at a treading rate of 15 m/min for 8 weeks. At the end of 8-week exercise training, mice underwent 30-min left anterior descending coronary artery occlusion followed by 60-min or 24-h reperfusion. Electron paramagnetic resonance oximetry was performed to measure myocardial tissue oxygenation. Western immunoblotting analyses, gene transfection, and myography were examined. The oximetry study demonstrated that exercise markedly shortened myocardial tissue oxygenation recovery time following reperfusion. Exercise training up-regulated Kir6.1 protein expression (a subunit of ATP-sensitive K(+)channel on vascular smooth muscle cells, VSMC sarc-K(ATP)) and protected the heart from I/R injury. In vivo gene transfer of dominant negative Kir6.1AAA prolonged the recovery time and enlarged infarct size. In addition, transfection of Kir6.1AAA increased the stiffness and reduced the relaxation capacity in the vasculature. Together, our study demonstrated that exercise training up-regulated Kir6.1, improved tissue oxygenation recovery, and protected the heart against I/R injury. This exercise-induced cardioprotective mechanism may provide a potential therapeutic intervention targeting VSMC sarc-K(ATP) channels and reperfusion recovery.

MOG1 rescues defective trafficking of Na(v)1.5 mutations in Brugada syndrome and sick sinus syndrome.

BACKGROUND: Loss-of-function mutations in Na(v)1.5 cause sodium channelopathies, including Brugada syndrome, dilated cardiomyopathy, and sick sinus syndrome; however, no effective therapy exists. MOG1 increases plasma membrane (PM) expression of Na(v)1.5 and sodium current (I(Na)) density, thus we hypothesize that MOG1 can serve as a therapeutic target for sodium channelopathies. METHODS AND RESULTS: Knockdown of MOG1 expression using small interfering RNAs reduced Na(v)1.5 PM expression, decreased I(Na) densities by 2-fold in HEK/Na(v)1.5 cells and nearly abolished I(Na) in mouse cardiomyocytes. MOG1 did not affect Na(v)1.5 PM turnover. MOG1 small interfering RNAs caused retention of Na(v)1.5 in endoplasmic reticulum, disrupted the distribution of Na(v)1.5 into caveolin-3-enriched microdomains, and led to redistribution of Na(v)1.5 to noncaveolin-rich domains. MOG1 fully rescued the reduced PM expression and I(Na) densities by Na(v)1.5 trafficking-defective mutation D1275N associated with sick sinus syndrome/dilated cardiomyopathy/atrial arrhythmias. For Brugada syndrome mutation G1743R, MOG1 restored the impaired PM expression of the mutant protein and restored I(Na) in a heterozygous state (mixture of wild type and mutant Na(v)1.5) to a full level of a homozygous wild-type state. CONCLUSIONS: Use of MOG1 to enhance Na(v)1.5 trafficking to PM may be a potential personalized therapeutic approach for some patients with Brugada syndrome, dilated cardiomyopathy, and sick sinus syndrome in the future.

GLUT12 functions as a basal and insulin-independent glucose transporter in the heart.

Glucose uptake from the bloodstream is the rate-limiting step in whole body glucose utilization, and is regulated by a family of membrane proteins called glucose transporters (GLUTs). Although GLUT4 is the predominant isoform in insulin-sensitive tissues, there is recent evidence that GLUT12 could be a novel second insulin-sensitive GLUT. However, its physiological role in the heart is not elucidated and the regulation of insulin-stimulated myocardial GLUT12 translocation is unknown. In addition, the role of GLUT12 has not been investigated in the diabetic myocardium. Thus, we hypothesized that, as for GLUT4, insulin regulates GLUT12 translocation to the myocardial cell surface, which is impaired during diabetes. Active cell surface GLUT (-4 and -12) content was quantified (before and after insulin stimulation) by a biotinylated photolabeled assay in both intact perfused myocardium and isolated cardiac myocytes of healthy and type 1 diabetic rodents. GLUT localization was confirmed by immunofluorescent confocal microscopy, and total GLUT protein expression was measured by Western blotting. Insulin stimulation increased translocation of GLUT-4, but not -12, in the healthy myocardium. Total GLUT4 content of the heart was decreased during diabetes, while there was no difference in total GLUT12. Active cell surface GLUT12 content was increased in the diabetic myocardium, potentially as a compensatory mechanism for the observed downregulation of GLUT4. Collectively, our data suggest that, in contrast to GLUT4, insulin does not mediate GLUT12 translocation, which may function as a basal GLUT located primarily at the cell surface in the myocardium.

A new series of chiral metal formate frameworks of [HONH3][M(II)(HCOO)3] (M = Mn, Co, Ni, Zn, and Mg): synthesis, structures, and properties.

We report the synthesis, crystal structures, IR, and thermal, dielectric, and magnetic properties of a new series of ammonium metal formate frameworks of [HONH(3)][M(II)(HCOO)(3)] for M = Mn, Co, Ni, Zn, and Mg. They are isostructural and crystallize in the nonpolar chiral orthorhombic space group P2(1)2(1)2(1), a = 7.8121(2)-7.6225(2) Å, b = 7.9612(3)-7.7385(2) Å, c = 13.1728(7)-12.7280(4) Å, and V = 819.27(6)-754.95(4) Å(3). The structures possess anionic metal formate frameworks of 4(9)·6(6) topology, in which the octahedral metal centers are connected by the anti-anti formate ligands and the hydroxylammonium is located orderly in the channels, forming strong O/N-H···O(formate) hydrogen bonds with the framework. HONH(3)(+) with only two non-H atoms favors the formation of the dense chiral 4(9)·6(6) frameworks, instead of the less dense 4(12)·6(3) perovskite frameworks for other monoammoniums of two to four non-H atoms because of its small size and its ability to form strong hydrogen bonding. However, the larger size of HONH(3)(+) compared to NH(4)(+) resulted in simple dielectric properties and no phase transitions. The three magnetic members (Mn, Co, and Ni) display antiferromagnetic long-range ordering of spin canting, at Néel temperatures of 8.8 K (Mn), 10.9 K (Co), and 30.5 K (Ni), respectively, and small spontaneous magnetizations for the Mn and Ni members but large magnetization for the Co member. Thermal and IR spectroscopic properties are also reported.

Molecular mechanism underlying adenosine receptor-mediated mitochondrial targeting of protein kinase C.

Activation of protein kinase C (PKC) via adenosine receptors is known to be involved in the cardioprotection of ischemic preconditioning (IPC). Specifically, activation of PKCε is critical for cardioprotection. There is ample evidence that PKCε resides in cardiac mitochondria. However, the signals that promote translocation of PKCε are largely unknown. The present study was designed to determine whether and how adenosine receptor activation induces translocation of PKCε to mitochondria. Freshly isolated adult rat cardiac myocytes and rat heart-derived H9c2 were used in the study. Immunofluorescence imaging of isolated mitochondria showed that PKCε but not PKCδ was localized in mitochondria and this mitochondrial localization of PKCε was significantly increased by adenosine treatment. The adenosine-induced increase in PKCε-positive mitochondria was largely prevented not only by PKC inhibitor chelerythrine, but also by the HSP90 inhibitor geldanamycin and by siRNA targeting HSP90. Immunoblot analysis from percoll-purified mitochondria further demonstrated that adenosine mediated a significant increase in mitochondrial PKCε but not PKCδ. This effect was blocked by inhibiting PKC activity with chelerythrine and bisindolylmaleimide. Furthermore, co-immunoprecipitation data showed that PKCε but not PKCδ was associated with TOM70 and HSP90, and this association was enhanced by adenosine treatment. Moreover, adenosine-induced association of PKCε with TOM70 was reduced by suppressing HSP90 expression with siRNA. In conclusion, we demonstrate that adenosine induces HSP90-dependent translocation of PKCε to mitochondria, possibly through mitochondrial import machinery TOM70. These results point out a novel mechanism in regulating PKC in mitochondria and suggest an important implication in ischemic preconditioning or postconditioning.