scRNA+TCR+BCR-seq revealed the proportions and gene expression patterns of dual receptor T and B lymphocytes in NPC and NLH.

While the relationship between single receptor lymphocytes and cancer has been deeply researched, the origin and biological roles of dual receptor lymphocytes in tumor microenvironment (TME) remain largely unknown. And since nasopharyngeal carcinoma (NPC) is a type of cancer closely associated with immune infiltration, studying the TME of NPC holds particular significance. Utilizing single-cell RNA sequencing combined with T cell receptor (TCR) and B cell receptor (BCR) sequencing (scRNA + TCR + BCR-seq), we analyzed data from 7 patients with NPC and 3 patients with nasopharyngeal lymphatic hyperplasia (NLH). In our research, it was firstly found that the presence of dual receptor lymphocytes in both the TME of NPC and the inflammatory environment of NLH. We also confirmed their clonal expansion, suggesting their potential involvement in the immune response. Subsequently, we further discovered the lineage and the pairing characteristics. It was found that the dual receptor lymphocytes in NPC and NLH mainly originate from memory cells, and the predominant pairing type for dual TCR was ß+α1+α2 and dual BCR was heavy+κ+λ. By further analyzing their gene expression, we compared the function of dual receptor cells with single receptor cells in the context of both NPC and NLH. This groundbreaking research has enhanced our comprehension of the features of dual-receptor cells and has contributed to a better understanding of the TME in NPC. By comparing with NLH, it illuminates part of the alterations in the process of malignant transformation in NPC. These findings present the potential to acquire improved diagnostic markers and treatment modalities.

Long non-coding RNA Linc00657 up-regulates Skp2 to promote the progression of cervical cancer through lipid reprogramming and regulation of immune microenvironment.

More and more evidence shows that long non-coding RNA (lncRNA) plays an important role in the biological behavior of many kinds of malignant tumors, but the specific function of lncRNA Linc00657 in cervical cancer is still unknown. The purpose of this study is to explore the effect of Linc00657 on the malignant progression of cervical cancer and its potential mechanism. In two kinds of cervical cancer cell lines and normal cervical epithelial cells, qRT-PCR showed increased expression of Linc00657 in cervical cancer cells. Through MTT, clone formation test, flow cytometry, wound healing test and Transwell test, it has been found that overexpression of Linc00657 could promote the proliferation,migration and invasion of cervical cancer cells，and inhibit apoptosis. Through the StarBase database, it was found that there may be a mutual regulatory relationship between Linc00657 and Skp2, and Skp2 may be the downstream target of Linc00657. QRT-PCR detection confirmed that the expression of Skp2 was increased in cervical cancer cells with overexpression of Linc00657. TIMER2 database found that Skp2 was associated with lipid metabolic enzymes and immune cell infiltration. It was found that Linc00657 knockdown inhibited tumor growth and metastasis and inhibited the expression of Skp2 in vivo. In short, our research shows that Linc00657 has carcinogenic properties in cervical cancer, and LINC00657 promotes the occurrence of cervical cancer by up-regulating the expression of Skp2. We predict that Linc00657/mir30s/Skp2 axis plays a role in the malignant progression of cervical cancer. In addition, Skp2 may participate in cancer immune response and promote lymph node metastasis of cervical cancer through lipid reprogramming. These findings also provide promising targets for the diagnosis and treatment of cervical cancer.

High accuracy calibration method for multi-line structured light three-dimensional scanning measurement system based on grating diffraction.

Multi-line structured light three-dimensional (3D) scanning measurement system enables to obtain the richer 3D profile data of the object simultaneously during one frame, ensuring high accuracy while structured light is deformed for the modulation by the object. Nevertheless, current calibration methods cannot fully take advantage of its high precision. In this paper, a fast and high-accuracy 3D measurement system based on multi-line lasers with a spatially precise structure via integrating a diffraction grating was proposed. This helps achieve precise calibration results of the light planes by introducing spatial constraint relations of the diffractive light, thus improving measurement accuracy. The operating principle and the workflow of the proposed system were described in detail. The measurement accuracy of the developed prototype was verified through contrastive experiments. At a working distance of 400 mm, the results show that the root mean square error (RMSE) of the proposed system is 0.083 mm, which is improved by 37.6% compared to the traditional calibration method of light planes for the ranging system. The system utilizing a grating that facilitates the integration of the device has great application value.

Streptomyces-based whole-cell biosensors for detecting diverse cell envelope-targeting antibiotics.

Cell envelope-targeting antibiotics are potent therapeutic agents against various bacterial infections. The emergence of multiple antibiotic-resistant strains underscores the significance of identifying potent antimicrobials specifically targeting the cell envelope. However, current drug screening approaches are tedious and lack sufficient specificity and sensitivity, warranting the development of more efficient methods. Genetic circuit-based whole-cell biosensors hold great promise for targeted drug discovery from natural products. Here, we performed comparative transcriptomic analysis of Streptomyces coelicolor M1146 exposed to diverse cell envelope-targeting antibiotics, aiming to identify regulatory elements involved in perceiving and responding to these compounds. Differential gene expression analysis revealed significant activation of VanS/R two-component system in response to the glycopeptide class of cell envelope-acting antibiotics. Therefore, we engineered a pair of VanS/R-based biosensors that exhibit functional complementarity and possess exceptional sensitivity and specificity for glycopeptides detection. Additionally, through promoter screening and characterization, we expanded the biosensor's detection range to include various cell envelope-acting antibiotics beyond glycopeptides. Our genetically engineered biosensor exhibits superior performance, including a dynamic range of up to 887-fold for detecting subtle antibiotic concentration changes in a rapid 2-h response time, enabling high-throughput screening of natural product libraries for antimicrobial agents targeting the bacterial cell envelope.

Panoramic three-dimensional optical digitization system assisted by a bi-mirror.

The digitization of objects' full surfaces finds widespread applications in fields such as virtual reality, art and design, and medical and biological sciences. For the realization of three-dimensional full-surface digitization of objects within complex sceneries, we propose a straightforward, efficient, and robust panoramic three-dimensional optical digitization system. This system contains a laser-based optical three-dimensional measurement system and a bi-mirror. By integrating mirrors into the system, we enable the illumination of the object from all angles using the projected laser beam in a single scanning process. Moreover, the main camera employed in the system can acquire three-dimensional information of the object from several different viewpoints. The rotational scanning method enhances the efficiency and applicability of the three-dimensional scanning process, enabling the acquisition of surface information of large-scale objects. After obtaining the three-dimensional data of the sample from different viewpoints using laser triangulation, mirror reflection transformation was employed to obtain the full-surface three-dimensional data of the object in the global coordinate system. The proposed method has been subjected to precision and validity experiments using samples with different surface characteristics and sizes, resulting in the demonstration of its capability for achieving correct three-dimensional digitization of the entire surface in diverse complex sceneries.

A Novel SiC Trench MOSFET with Self-Aligned N-Type Ion Implantation Technique.

We propose a novel silicon carbide (SiC) self-aligned N-type ion implanted trench MOSFET (NITMOS) device. The maximum electric field in the gate oxide could be effectively reduced to below 3 MV/cm with the introduction of the P-epi layer below the trench. The P-epi layer is partially counter-doped by a self-aligned N-type ion implantation process, resulting in a relatively low specific on-resistance (Ron,sp). The lateral spacing between the trench sidewall and N-implanted region (Wsp) plays a crucial role in determining the performance of the SiC NITMOS device, which is comprehensively studied through the numerical simulation. With the Wsp increasing, the SiC NITMOS device demonstrates a better short-circuit capability owing to the reduced saturation current. The gate-to-drain capacitance (Cgd) and gate-to-drain charge (Qgd) are also investigated. It is observed that both Cgd and Qgd decrease as the Wsp increases, owing to the enhanced screen effect. Compared to the SiC double-trench MOSFET device, the optimal SiC NITMOS device exhibits a 79% reduction in Cgd, a 38% decrease in Qgd, and a 41% reduction in Qgd × Ron,sp. A higher switching speed and a lower switching loss can be achieved using the proposed structure.

Insertion sequence transposition inactivates CRISPR-Cas immunity.

CRISPR-Cas immunity systems safeguard prokaryotic genomes by inhibiting the invasion of mobile genetic elements. Here, we screened prokaryotic genomic sequences and identified multiple natural transpositions of insertion sequences (ISs) into cas genes, thus inactivating CRISPR-Cas defenses. We then generated an IS-trapping system, using Escherichia coli strains with various ISs and an inducible cas nuclease, to monitor IS insertions into cas genes following the induction of double-strand DNA breakage as a physiological host stress. We identified multiple events mediated by different ISs, especially IS1 and IS10, displaying substantial relaxed target specificity. IS transposition into cas was maintained in the presence of DNA repair machinery, and transposition into other host defense systems was also detected. Our findings highlight the potential of ISs to counter CRISPR activity, thus increasing bacterial susceptibility to foreign DNA invasion.

Surface modification of rare earth Sm-doped WO3 films through polydopamine for enhanced electrochromic energy storage performance.

Electrochromic materials (ECMs) could exhibit reversible color changes upon application of the external electric field, which exhibits huge application prospects in smart windows, energy storage devices, and displays. For the practical application of ECMs, the fast response speed and long cyclic stability are urgent. In this work, the nanoporous Sm-doped WO3 (WSm) films were constructed using hydrothermal technology, then polydopamine (PDA) was modified on the surface of WSm film to obtain the WSm/Px (x = 0.25, 0.5, 1.0, and 2.0) hybrid films. WSm/Px hybrid films displayed high optical contrast and large areal capacitance. In addition, in comparison with WSm film, the WSm/Px hybrid films exhibited faster response speed and better cyclic stability because PDA film enhanced the interface ion transport ability and electrochemical structural stability of the nanoporous WSm film. Notably, the WSm/P1.0 hybrid film displayed the colored/bleached times of 7.4/2.9 s, retained 90.2% of the primitive optical contrast (68.5%) after 5000 electrochromic cycles. Furthermore, the areal capacitance of WSm film could be increased by 224% through the modification of the PDA. Therefore, WSm/Px hybrid films are great prospects for electrochromic energy-saving and storage windows.

Novel Chalcone-Phenazine Hybrids Induced Ferroptosis in U87-MG Cells through Activating Ferritinophagy.

Thirty-seven novel chalcone-phenazine hybrid molecules (C1∼C13 and F1∼F24) with 1,2,3-triazole or ethyl group as linkers were designed and synthesized in this study. Some compounds exhibited selective cytotoxicity against U87-MG cancer cell lines in vitro, in which compound C4 were found to have the best antiproliferative activity. SAR study indicated 1,2,3-triazole group may be crucial for enhancing compounds' cytotoxicity. C4 was verified to induce ferroptosis in U87-MG cells by transcription, lipid peroxidation, lipid ROS assays. Furthermore, C4 was up-regulated LC3-II, degradated FTH1, and then increasing iron resulted in the down-regulation of NCOA4. Together, all above evidences highlighted the potential of compound C4 that triggered ferroptosis by activating ferritinophagy against U87-MG cells.

A thioredoxin reductase 1 inhibitor pyrano [3,2-a] phenazine inhibits A549 cells proliferation and migration through the induction of reactive oxygen species production.

BACKGROUND: Thioredoxin reductase 1 (TrxR1) inhibitor, pyrano [3,2-a] phenazine, named CPUL-1, was synthesized with potential anticancer activity. The aim of the present work was to explore the potential anti-proliferative and anti-metastatic ability of CPUL-1 against A549 cancer cell lines in vitro. METHODS AND RESULTS: First, Cell Counting Kit-8 (CCK8) assay was used to assess cell proliferation. The A549 cell migration was evaluated by wound healing assay and transwell assay. Second, the epithelial-mesenchymal transition (EMT)-related proteins in A549 cells treated with CPUL-1 were analyzed by western blot methods. Then, TrxR1 enzyme activity assay and reactive oxygen species (ROS) assay were conducted to evaluate the effect of CPUL-1 on TrxR1 inhibition and ROS levels. Finally, western blotting was used to explore the mechanism of CPUL-1. The study results revealed that the ability of cell proliferation and migration was decreased under CPUL-1 treatment. CPUL-1 could distinctly restrain the migration and invasion of A549 cells through inhibiting EMT process. The results of TrxR1 enzyme activity assay, ROS assay and western blotting showed that CPUL-1 influenced EMT via inducing ROS-mediated ERK/JNK signaling by inhibiting TrxR1 enzyme activity. CONCLUSIONS: Together, proliferation suppression and anti-metastasis activity of CPUL-1 in A549 cells were demonstrated by all the evidence. Our findings highlight the great potential of phenazine compound CPUL-1 to suppress A549 cells proliferation and metastasis.

Investigation of SiC Trench MOSFETs' Reliability under Short-Circuit Conditions.

In this paper, the short-circuit robustness of 1200 V silicon carbide (SiC) trench MOSFETs with different gate structures has been investigated. The MOSFETs exhibited different failure modes under different DC bus voltages. For double trench SiC MOSFETs, failure modes are gate failure at lower dc bus voltages and thermal runaway at higher dc bus voltages, while failure modes for asymmetric trench SiC MOSFETs are soft failure and thermal runaway, respectively. The shortcircuit withstanding time (SCWT) of the asymmetric trench MOSFET is higher than that of the double trench MOSFETs. The thermal and mechanical stresses inside the devices during the short-circuit tests have been simulated to probe into the failure mechanisms and reveal the impact of the device structures on the device reliability. Finally, post-failure analysis has been carried out to verify the root causes of the device failure.

Determination of the Protein-Protein Interactions within Acyl Carrier Protein (MmcB)-Dependent Modifications in the Biosynthesis of Mitomycin.

Mitomycin has a unique chemical structure and contains densely assembled functionalities with extraordinary antitumor activity. The previously proposed mitomycin C biosynthetic pathway has caused great attention to decipher the enzymatic mechanisms for assembling the pharmaceutically unprecedented chemical scaffold. Herein, we focused on the determination of acyl carrier protein (ACP)-dependent modification steps and identification of the protein-protein interactions between MmcB (ACP) with the partners in the early-stage biosynthesis of mitomycin C. Based on the initial genetic manipulation consisting of gene disruption and complementation experiments, genes mitE, mmcB, mitB, and mitF were identified as the essential functional genes in the mitomycin C biosynthesis, respectively. Further integration of biochemical analysis elucidated that MitE catalyzed CoA ligation of 3-amino-5-hydroxy-bezonic acid (AHBA), MmcB-tethered AHBA triggered the biosynthesis of mitomycin C, and both MitB and MitF were MmcB-dependent tailoring enzymes involved in the assembly of mitosane. Aiming at understanding the poorly characterized protein-protein interactions, the in vitro pull-down assay was carried out by monitoring MmcB individually with MitB and MitF. The observed results displayed the clear interactions between MmcB and MitB and MitF. The surface plasmon resonance (SPR) biosensor analysis further confirmed the protein-protein interactions of MmcB with MitB and MitF, respectively. Taken together, the current genetic and biochemical analysis will facilitate the investigations of the unusual enzymatic mechanisms for the structurally unique compound assembly and inspire attempts to modify the chemical scaffold of mitomycin family antibiotics.

End-to-end pattern optimization technology for 3D shape measurement.

The nonlinear errors caused by the gamma effect seriously affect the measurement accuracy of phase-measuring profilometry systems. We present a novel robust end-to-end pattern optimization technology for phase-height mapping. This method generates standard sinusoidal patterns on the reference plane on a camera image instead of on a projection image by optimizing the projection image, which can avoid phase errors due to phase conversion from projector to camera. The pixel set mapping is achieved by chessboard calibration; then the projection image is optimized with the multiscale optimization method and the dislocation optimization method based on proportional integral derivative control. The experiments show that the proposed method can effectively avoid the influence of nonlinear effect and achieve high-quality three-dimensional measurement.

Adaptive Optimization Boosted the Production of Moenomycin A in the Microbial Chassis Streptomyces albus J1074.

Great efforts have been made to improve Streptomyces chassis for efficient production of targeted natural products. Moenomycin family antibiotics, represented by moenomycin (Moe) and nosokomycin, are phosphoglycolipid antibiotics that display extraordinary inhibition against Gram-positive bacteria. Herein, we assembled a completed 34 kb hybrid biosynthetic gene cluster (BGC) of moenomycin A (moe-BGC) based on a 24 kb nosokomycin analogue biosynthetic gene cluster (noso-BGC). The heterologous expression of the hybrid moe-BGC in Streptomyces albus J1074 achieved the production of moenomycin A in the recombinant strain LX01 with a yield of 12.1 ± 2 mg/L. Further strong promoter refactoring to improve the transcriptional levels of all of the functional genes in strain LX02 enhanced the production of moenomycin A by 58%. However, the yield improvement of moenomycin A resulted in a dramatic 38% decrease in the chassis biomass compared with the control strain. To improve the weak physiological tolerance to moenomycin A of the chassis, another copy of the gene salb-PBP2 (P238N&F200D), encoding peptidoglycan biosynthetic protein PBP2, was introduced into the chassis strain, producing strain LX03. Cell growth was restored, and the fermentation titer of moenomycin A was 130% higher than that of LX01. Additionally, the production of moenomycin A in strain LX03 was further elevated by 45% to 40.0 ± 3 mg/L after media optimization. These results suggested that the adaptive optimization strategy of strong promoter refactoring in the BGC plus physiological tolerance in the chassis was an efficient approach for obtaining the desired natural products with high titers.

p-Aminophenylalanine Involved in the Biosynthesis of Antitumor Dnacin B1 for Quinone Moiety Formation.

Actinosynnema species produce diverse natural products with important biological activities, which represent an important resource of antibiotic discovery. Advances in genome sequencing and bioinformatics tools have accelerated the exploration of the biosynthetic gene clusters (BGCs) encoding natural products. Herein, the completed BGCs of dnacin B1 were first discovered in two Actinosynnema pretiosum subsp. auranticum strains DSM 44131T (hereafter abbreviated as strain DSM 44131T) and X47 by comparative genome mining strategy. The BGC for dnacin B1 contains 41 ORFs and spans a 66.9 kb DNA region in strain DSM 44131T. Its involvement in dnacin B1 biosynthesis was identified through the deletion of a 9.7 kb region. Based on the functional gene analysis, we proposed the biosynthetic pathway for dnacin B1. Moreover, p-amino-phenylalanine (PAPA) unit was found to be the dnacin B1 precursor for the quinone moiety formation, and this was confirmed by heterologous expression of dinV, dinE and dinF in Escherichia coli. Furthermore, nine potential PAPA aminotransferases (APAT) from the genome of strain DSM 44131T were explored and expressed. Biochemical evaluation of their amino group transformation ability was carried out with p-amino-phenylpyruvic acid (PAPP) or PAPA as the substrate for the final product formation. Two of those, APAT4 and APAT9, displayed intriguing aminotransferase ability for the formation of PAPA. The proposed dnacin B1 biosynthetic machinery and PAPA biosynthetic investigations not only enriched the knowledge of tetrahydroisoquinoline (THIQ) biosynthesis, but also provided PAPA building blocks to generate their structurally unique homologues.

Clinical evaluation of prophylactic abdominal aortic balloon occlusion in patients with placenta accreta: a systematic review and meta-analysis.

BACKGROUND: Severe obstetric hemorrhage caused by placenta accreta results in significant maternal morbidity and mortality. As a new technology, abdominal aortic balloon occlusion (AABO) is becoming an important treatment for patients with placenta accreta. To evaluate the safety and efficacy of AABO, we conducted a systematic review and meta-analysis of previous studies. METHODS: We used a three-check subset including placenta accreta (placenta previa, percreta, increta, etc.), balloon, and aortic (aortas, aorta, etc.) to form a retrieval formula and searched in MEDLINE, EMBASE, the Cochrane Library, clinicaltrials.gov and Web of Science. All articles regarding placenta previa or placenta accreta and including the use of abdominal aortic balloon occlusion were included in our screening. Two researchers selected articles and extracted data independently. Finally, the Newcastle-Ottawa Quality Assessment Scale was used for quality assessments. RESULTS: We retrieved 776 articles and eventually included 11 clinical studies. Meta-analysis showed that AABO significantly reduced the blood loss volume (MD - 1480 ml, 95% CI -1806 to - 1154 ml, P < 0.001) and blood transfusion volume (MD - 1125 ml, 95% CI -1264 to - 987 ml, P < 0.001). Similarly, obvious reductions in the hysterectomy rate (OR 0.30, 95% CI 0.19 to 0.48, P < 0.001), hospitalization duration (MD - 1.35 days, 95% CI -2.40 to - 0.31 days, P = 0.01), and operative time (MD - 29.23 min, 95% CI -46.04 to - 12.42 min, P < 0.001) were observed in the AABO group. CONCLUSION: The prophylactic use of AABO in patients with placenta accreta is safe and effective.

The Impact of Noncavity-Distorting Intramural Fibroids on the Efficacy of In Vitro Fertilization-Embryo Transfer: An Updated Meta-Analysis.

AIM: To address the impact of noncavity-distorting intramural fibroids on the efficacy of in vitro fertilization-embryo transfer (IVF-ET) outcomes. METHODS: The PubMed, Web of Science, Embase, Cochrane Library, and China National Knowledge Infrastructure were searched systematically. A meta-analysis was performed based on comparative or cohort studies that explored the impact of noncavity-distorting intramural fibroids on the efficacy of IVF-ET treatment. The IVE-ET outcomes of study group (women with noncavity-distorting intramural fibroids) and control group (women without fibroids) were compared, including live birth rate (LBR), clinical pregnancy rate (cPR), implantation rate (IR) , miscarriage rate (MR), and ectopic pregnancy rate (ePR). RESULTS: A total of 28 studies involving 9189 IVF cycles were included. Our meta-analysis showed a significant reduction of LBR in the study group compared to control group (RR = 0.82, 95% CI: 0.73-0.92, and P = 0.005). In addition, it indicated that study group had a significant reduction in cPR (RR = 0.86, 95% CI: 0.80-0.93, P = 0.0001) and IR (RR = 0.90, 95% CI: 0813-1.00, P = 0.04) and have a significantly increase in MR (RR = 1.27, 95% CI: 1.08-1.50, and P = 0.004) compared with control group. CONCLUSIONS: The present evidence suggests that noncavity-distorting intramural fibroids would significantly reduce the IR, cRP, and LBR and significantly increase the MR after IVF treatment, but it would not significantly increase the ePR.

Understanding the Effects of NaCl, NaBr and Their Mixtures on Silver Nanowire Nucleation and Growth in Terms of the Distribution of Electron Traps in Silver Halide Crystals.

In recent years, many research groups have synthesized ultra-thin silver nanowires (AgNWs) with diameters below 30 nm by employing Cl- and Br- simultaneously in the polyol process. However, the yield of AgNWs in this method was low, due to the production of Ag nanoparticles (AgNPs) as an unwanted byproduct, especially in the case of high Br- concentration. Here, we investigated the roles of Cl- and Br- in the preparation of AgNWs and then synthesized high aspect ratio (up to 2100) AgNWs in high yield (>85% AgNWs) using a Cl- and Br- co-mediated method. We found that multiply-twinned particles (MTPs) with different critical sizes were formed and grew into AgNWs, accompanied by a small and large amount of AgNPs for the NaCl and NaBr additives, respectively. For the first time, we propose that the growth of AgNWs of different diameters and yields can be understood based on the electron trap distribution (ETD) of the silver halide crystals. For the case of Cl- and Br- co-additives, a mixed silver halide crystal of AgBr1-xClx was formed, rather than the AgBr/AgCl mixture reported previously. In this type of crystal, the ETD is uniform, which is beneficial for the synthesis of AgNWs with small diameter (30~40 nm) and high aspect ratio. AgNW transparent electrodes were prepared in air by rod coating. A sheet resistance of 48 Ω/sq and transmittance of 95% at 550 nm were obtained without any post-treatment.

Increased Efferent Cardiac Sympathetic Nerve Activity and Defective Intrinsic Heart Rate Regulation in Type 2 Diabetes.

Elevated sympathetic nerve activity (SNA) coupled with dysregulated ß-adrenoceptor (ß-AR) signaling is postulated as a major driving force for cardiac dysfunction in patients with type 2 diabetes; however, cardiac SNA has never been assessed directly in diabetes. Our aim was to measure the sympathetic input to and the ß-AR responsiveness of the heart in the type 2 diabetic heart. In vivo recording of SNA of the left efferent cardiac sympathetic branch of the stellate ganglion in Zucker diabetic fatty rats revealed an elevated resting cardiac SNA and doubled firing rate compared with nondiabetic rats. Ex vivo, in isolated denervated hearts, the intrinsic heart rate was markedly reduced. Contractile and relaxation responses to ß-AR stimulation with dobutamine were compromised in externally paced diabetic hearts, but not in diabetic hearts allowed to regulate their own heart rate. Protein levels of left ventricular ß1-AR and Gs (guanine nucleotide binding protein stimulatory) were reduced, whereas left ventricular and right atrial ß2-AR and Gi (guanine nucleotide binding protein inhibitory regulatory) levels were increased. The elevated resting cardiac SNA in type 2 diabetes, combined with the reduced cardiac ß-AR responsiveness, suggests that the maintenance of normal cardiovascular function requires elevated cardiac sympathetic input to compensate for changes in the intrinsic properties of the diabetic heart.

Impaired relaxation despite upregulated calcium-handling protein atrial myocardium from type 2 diabetic patients with preserved ejection fraction.

BACKGROUND: Diastolic dysfunction is a key factor in the development and pathology of cardiac dysfunction in diabetes, however the exact underlying mechanism remains unknown, especially in humans. We aimed to measure contraction, relaxation, expression of calcium-handling proteins and fibrosis in myocardium of diabetic patients with preserved systolic function. METHODS: Right atrial appendages from patients with type 2 diabetes mellitus (DM, n = 20) and non-diabetic patients (non-DM, n = 36), all with preserved ejection fraction and undergoing coronary artery bypass grafting (CABG), were collected. From appendages, small cardiac muscles, trabeculae, were isolated to measure basal and ß-adrenergic stimulated myocardial function. Expression levels of calcium-handling proteins, sarcoplasmic reticulum Ca2+ ATPase (SERCA2a) and phospholamban (PLB), and of ß1-adrenoreceptors were determined in tissue samples by Western blot. Collagen deposition was determined by picro-sirius red staining. RESULTS: In trabeculae from diabetic samples, contractile function was preserved, but relaxation was prolonged (Tau: 74 ± 13 ms vs. 93 ± 16 ms, non-DM vs. DM, p = 0.03). The expression of SERCA2a was increased in diabetic myocardial tissue (0.75 ± 0.09 vs. 1.23 ± 0.15, non-DM vs. DM, p = 0.007), whereas its endogenous inhibitor PLB was reduced (2.21 ± 0.45 vs. 0.42 ± 0.11, non-DM vs. DM, p = 0.01). Collagen deposition was increased in diabetic samples. Moreover, trabeculae from diabetic patients were unresponsive to ß-adrenergic stimulation, despite no change in ß1-adrenoreceptor expression levels. CONCLUSIONS: Human type 2 diabetic atrial myocardium showed increased fibrosis without systolic dysfunction but with impaired relaxation, especially during ß-adrenergic challenge. Interestingly, changes in calcium-handling protein expression suggests accelerated active calcium re-uptake, thus improved relaxation, indicating a compensatory calcium-handling mechanism in diabetes in an attempt to maintain diastolic function at rest despite impaired relaxation in the diabetic fibrotic atrial myocardium. Our study addresses important aspects of the underlying mechanisms of diabetes-associated diastolic dysfunction, which is crucial to developing new therapeutic treatments.