Multiscale embedded printing of engineered human tissue and organ equivalents.

Creating tissue and organ equivalents with intricate architectures and multiscale functional feature sizes is the first step toward the reconstruction of transplantable human tissues and organs. Existing embedded ink writing approaches are limited by achievable feature sizes ranging from hundreds of microns to tens of millimeters, which hinders their ability to accurately duplicate structures found in various human tissues and organs. In this study, a multiscale embedded printing (MSEP) strategy is developed, in which a stimuli-responsive yield-stress fluid is applied to facilitate the printing process. A dynamic layer height control method is developed to print the cornea with a smooth surface on the order of microns, which can effectively overcome the layered morphology in conventional extrusion-based three-dimensional bioprinting methods. Since the support bath is sensitive to temperature change, it can be easily removed after printing by tuning the ambient temperature, which facilitates the fabrication of human eyeballs with optic nerves and aortic heart valves with overhanging leaflets on the order of a few millimeters. The thermosensitivity of the support bath also enables the reconstruction of the full-scale human heart on the order of tens of centimeters by on-demand adding support bath materials during printing. The proposed MSEP demonstrates broader printable functional feature sizes ranging from microns to centimeters, providing a viable and reliable technical solution for tissue and organ printing in the future.

Epigenetic Regulation of Leukocyte Inflammatory Mediator Production Dictates Staphylococcus aureus Craniotomy Infection Outcome.

Staphylococcus aureus is a common cause of surgical-site infections, including those arising after craniotomy, which is performed to access the brain for the treatment of tumors, epilepsy, or hemorrhage. Craniotomy infection is characterized by complex spatial and temporal dynamics of leukocyte recruitment and microglial activation. We recently identified unique transcriptional profiles of these immune populations during S. aureus craniotomy infection. Epigenetic processes allow rapid and reversible control over gene transcription; however, little is known about how epigenetic pathways influence immunity to live S. aureus. An epigenetic compound library screen identified bromodomain and extraterminal domain-containing (BET) proteins and histone deacetylases (HDACs) as critical for regulating TNF, IL-6, IL-10, and CCL2 production by primary mouse microglia, macrophages, neutrophils, and granulocytic myeloid-derived suppressor cells in response to live S. aureus. Class I HDACs (c1HDACs) were increased in these cell types in vitro and in vivo during acute disease in a mouse model of S. aureus craniotomy infection. However, substantial reductions in c1HDACs were observed during chronic infection, highlighting temporal regulation and the importance of the tissue microenvironment for dictating c1HDAC expression. Microparticle delivery of HDAC and BET inhibitors in vivo caused widespread decreases in inflammatory mediator production, which significantly increased bacterial burden in the brain, galea, and bone flap. These findings identify histone acetylation as an important mechanism for regulating cytokine and chemokine production across diverse immune cell lineages that is critical for bacterial containment. Accordingly, aberrant epigenetic regulation may be important for promoting S. aureus persistence during craniotomy infection.

Octanal enhances disease resistance in postharvest citrus fruit by the biosynthesis and metabolism of aromatic amino acids.

Octanal was found to be able to reduce green mold incidence in citrus fruit by a defense response mechanism. However, the underlying mechanism remains largely unclear. Herein, the metabolomics, RNA-seq and biochemical analyses were integrated to explore the effect of octanal on disease resistance in harvested citrus fruit. Results showed that octanal fumigation at 40 µL L-1 was effective in controlling citrus green mold. Metabolomics analysis showed that octanal mainly led to the accumulation of some plant hormones including methyl jasmonate, abscisic acid, indole-3-butyric acid, indoleacetic acid (IAA), salicylic acid, and gibberellic acid and many phenylpropanoid metabolites including cinnamyl alcohol, hesperidin, dihydrokaempferol, vanillin, quercetin-3-O-malonylglucoside, curcumin, naringin, chrysin, coniferin, calycosin-7-O-ß-D-glucoside, trans-cinnamaldehyde, and 4',5,7-trihydroxy-3,6-dimethoxyflavone. Particularly, IAA and hesperidin were dramatically accumulated in the peel, which might be the contributors to the resistance response. Additionally, transcriptome analysis showed that octanal greatly activated the biosynthesis and metabolism of aromatic amino acids. This was further verified by the accumulation of some metabolites (shikimic acid, tryptophan, tyrosine, phenylalanine, IAA, total phenolics, flavonoids and lignin), increase in some enzyme activities (phenylalanine ammonia-lyase, tyrosine ammonia-lyase, 4-coumarate CoA ligase, cinnamic acid 4-hydroxylase, polyphenol oxidase, and peroxidase), up-regulation of some genes (tryptophan pyruvate aminotransferase, aldehyde dehydrogenase, shikimate kinase and shikimate dehydrogenase) expressions and molecular docking results. Thus, these results indicate that octanal is an efficient strategy for the control of postharvest green mold by triggering the defense response in citrus fruit.

Long-term mortality in patients with end-stage renal disease undergoing hemodialysis and peritoneal dialysis: a propensity score matching retrospective study.

BACKGROUND: Hemodialysis (HD) and peritoneal dialysis (PD) are effective ways to treat end-stage renal disease (ERSD). This study aimed to investigate the differences in survival and the factors that influence it in patients with end-stage renal disease treated with HD or PD. METHODS: We retrospectively analyzed factors related to all-cause death with renal replacement therapy and compared the long-term mortality between HD and PD strategies in patients with ESRD who started HD or PD treatment in our renal HD center between January 1, 2008, and December 1, 2021. RESULTS: Overall, 1,319 patients were included, comprising 690 and 629 patients in the HD and PD groups, respectively, according to the inclusion criteria. After propensity matching, 922 patients remained, with 461 (50%) patients each in the two groups. There were no significant differences in the 1-, 2-, 3-, and 4-year mortality rates between the HD and PD groups (all p > .05). However, the 5- and 10-year mortality rates of the matched patients were 15.8%. 17.6% in the HD group and 21.0%. 27.3% in the PD group, respectively. The 5- and 10-year mortality rates were significantly lower in the HD group (all p < .05) as compared to the PD group. After matching, Kaplan-Meier curve analysis with log-rank test was performed, which showed a significant difference in the survival rates between the two groups (p = .001). Logistic multifactor regression analysis revealed that age, weight, hypertension, serum creatinine, and combined neoplasms influenced the survival rate of patients with ESRD (p < .05). In contrast, age, hypertension, parathyroid hormone (PTH), serum creatinine, and peripheral vascular diseases (PVD) influenced the survival rate of patients in the HD group (p < .05), and age and weight influenced the survival rate of patients in the PD group (p < .05). CONCLUSIONS: This study found that long-term mortality rates were higher in the PD group than that in the HD group, indicating that HD may be superior to PD.

A Dual-network Nerve Adhesive with Enhanced Adhesion Strength Promotes Transected Peripheral Nerve Repair.

Peripheral nerve transection has a high prevalence and results in functional loss of affected limbs. The current clinical treatment using suture anastomosis significantly limits nerve recovery due to severe inflammation, secondary damage, and fibrosis. Fibrin glue, a commercial nerve adhesive as an alternative, avoids secondary damage but suffers from poor adhesion strength. To address their limitations, a highly efficacious nerve adhesive based on dual-crosslinking of dopamine-isothiocyanate modified hyaluronic acid and decellularized nerve matrix is reported in this paper. This dual-network nerve adhesive (DNNA) shows controllable gelation behaviors feasible for surgical applications, robust adhesion strength, and promoted axonal outgrowth in vitro. The in vivo therapeutic efficacy is tested using a rat-based sciatic nerve transection model. The DNNA decreases fibrosis and accelerates axon/myelin debris clearance at 10 days post-surgery, compared to suture and commercial fibrin glue treatments. At 10 weeks post-surgery, the strong adhesion and bioactivity allow DNNA to significantly decrease intraneural inflammation and fibrosis, enhance axon connection and remyelination, aid motor and sensory function recovery, as well as improve muscle contraction, compared to suture and fibrin treatments. Overall, this dual-network hydrogel with robust adhesion provides a rapid and highly efficacious nerve transection treatment to facilitate nerve repair and neuromuscular function recovery.

A Facile Strategy for the Fabrication of Cell-laden Porous Alginate Hydrogels Based on Two-phase Aqueous Emulsions.

Porous alginate hydrogels possess many advantages as cell carriers. However, current pore generation methods require either complex or harsh fabrication processes, toxic components, or extra purification steps, limiting the feasibility and affecting the cellular survival and function. In this study, a simple and cell-friendly approach to generate highly porous cell-laden alginate hydrogels based on two-phase aqueous emulsions is reported. The pre-gel solutions, which contain two immiscible aqueous phases of alginate and caseinate, are crosslinked by calcium ions. The porous structure of the hydrogel construct is formed by subsequently removing the caseinate phase from the ion-crosslinked alginate hydrogel. Those porous alginate hydrogels possess heterogeneous pores around 100 µm and interconnected paths. Human white adipose progenitors (WAPs) encapsulated in these hydrogels self-organize into spheroids and show enhanced viability, proliferation, and adipogenic differentiation, compared to non-porous constructs. As a proof of concept, this porous alginate hydrogel platform is employed to prepare core-shell spheres for coculture of WAPs and colon cancer cells, with WAP clusters distributed around cancer cell aggregates, to investigate cellular crosstalk. This efficacious approach is believed to provide a robust and versatile platform for engineering porous-structured alginate hydrogels for applications as cell carriers and in disease modeling.

IL-10 production by granulocytes promotes Staphylococcus aureus craniotomy infection.

BACKGROUND: Treatment of brain tumors, epilepsy, or hemodynamic abnormalities requires a craniotomy to access the brain. Nearly 1 million craniotomies are performed in the US annually, which increase to ~ 14 million worldwide and despite prophylaxis, infectious complications after craniotomy range from 1 to 3%. Approximately half are caused by Staphylococcus aureus (S. aureus), which forms a biofilm on the bone flap that is recalcitrant to antibiotics and immune-mediated clearance. However, the mechanisms responsible for the persistence of craniotomy infection remain largely unknown. The current study examined the role of IL-10 in promoting bacterial survival. METHODS: A mouse model of S. aureus craniotomy infection was used with wild type (WT), IL-10 knockout (KO), and IL-10 conditional KO mice where IL-10 was absent in microglia and monocytes/macrophages (CX3CR1CreIL-10 fl/fl) or neutrophils and granulocytic myeloid-derived suppressor cells (G-MDSCs; Mrp8CreIL-10 fl/fl), the major immune cell populations in the infected brain vs. subcutaneous galea, respectively. Mice were examined at various intervals post-infection to quantify bacterial burden, leukocyte recruitment, and inflammatory mediator production in the brain and galea to assess the role of IL-10 in craniotomy persistence. In addition, the role of G-MDSC-derived IL-10 on neutrophil activity was examined. RESULTS: Granulocytes (neutrophils and G-MDSCs) were the major producers of IL-10 during craniotomy infection. Bacterial burden was significantly reduced in IL-10 KO mice in the brain and galea at day 14 post-infection compared to WT animals, concomitant with increased CD4+ and γδ T cell recruitment and cytokine/chemokine production, indicative of a heightened proinflammatory response. S. aureus burden was reduced in Mrp8CreIL-10 fl/fl but not CX3CR1CreIL-10 fl/fl mice that was reversed following treatment with exogenous IL-10, suggesting that granulocyte-derived IL-10 was important for promoting S. aureus craniotomy infection. This was likely due, in part, to IL-10 production by G-MDSCs that inhibited neutrophil bactericidal activity and TNF production. CONCLUSION: Collectively, these findings reveal a novel role for granulocyte-derived IL-10 in suppressing S. aureus clearance during craniotomy infection, which is one mechanism to account for biofilm persistence.

Deflecting and routing nematicons via orientation programmable liquid crystal array.

By designing a liquid crystal cell with comb electrode structure, the alignment modulation of nematic liquid crystal in the cell can be realized after the electric field is applied. In different orientation regions, the incident laser beam can deflect at different angles. At the same time, by changing the incident angle of the laser beam, the reflection modulation of the laser beam on the interface of the liquid crystal molecular orientation change can be realized. Based on the above discussion, we then demonstrate the modulation of liquid crystal molecular orientation arrays on nematicon pairs. In different orientation regions of liquid crystal molecules, nematicon pairs can exhibit various combinations of deflections, and these deflection angles are modulable under external fields. Deflection and modulation of nematicon pairs have potential applications in optical routing and optical communication.

Integrating multiple references for single-cell assignment.

Efficient single-cell assignment is essential for single-cell sequencing data analysis. With the explosive growth of single-cell sequencing data, multiple single-cell sequencing data sources are available for the same kind of tissue, which can be integrated to further improve single-cell assignment; however, an efficient integration strategy is still lacking due to the great challenges of data heterogeneity existing in multiple references. To this end, we present mtSC, a flexible single-cell assignment framework that integrates multiple references based on multitask deep metric learning designed specifically for cell type identification within tissues with multiple single-cell sequencing data as references. We evaluated mtSC on a comprehensive set of publicly available benchmark datasets and demonstrated its state-of-the-art effectiveness for integrative single-cell assignment with multiple references.

Integrated transcriptomic-metabolomic analysis reveals that cinnamaldehyde exposure positively regulates the phenylpropanoid pathway in postharvest Satsuma mandarin (Citrus unshiu).

Previously, wax + cinnamaldehyde (WCA) was proven to be able to effectively alleviate fruit decay and induce resistance in harvested Satsuma mandarin (Citrus unshiu). However, the potential molecular mechanism is largely unknown. In the present study, transcriptomics, metabolomics and biochemical analyses were combined to clarify this process. Transcriptomic analysis revealed that the expression of genes involved in secondary metabolites and related to pathogenesis and the phenylpropanoid pathway were significantly influenced by WCA treatment. In addition, metabolite profiling revealed that metabolites in the phenylpropanoid pathway were also predominantly impacted after WCA treatment. Correspondingly, enzymatic activities and gene expression involved in the phenylpropanoid pathway were positively regulated, especially in the first 24 h, resulting in increased levels of total phenolics, flavonoids and other secondary metabolites. Fruit inoculation experiments showed that WCA treatment significantly reduced the development of citrus green mold and sour rot while having no adverse effects on the edible quality of the tested citrus fruit. Our study confirms the potential role of WCA exposure in citrus to induce resistance through the phenylpropanoid pathway.

γ-Cyclodextrin encapsulated thymol for citrus preservation and its possible mechanism against Penicillium digitatum.

The volatility of essential oils greatly limits their industrial applications. Here, we successfully prepared γ-cyclodextrin (γ-CD) inclusion compounds (γ-CDTL) containing thymol (TL) for the control of green mold caused by Penicillium digitatum (P. digitatum) in citrus fruit. In vitro experiment showed that the minimum fungicidal concentration (MFC) of γ-CDTL against the hyphae growth of P. digitatum was 2.0 g/L, and 8 × MFC treatment significantly reduced the occurrence of green mold in citrus fruit and had no adverse effect on fruit quality in vivo test compared to prochloraz. Scanning electron microscopy (SEM), x-ray diffraction (XRD), fourier transform-infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), physical properties and sustained release properties were also performed, results indicated that the hydrogen bonds between TL and γ-CD were the basis for the formation of γ-CDTL. We further investigated the inhibition mechanism of γ-CDTL. SEM and TEM experiments showed that γ-CDTL treatment caused severe damage to the hyphal morphology and cells in 30 min and disrupted the permeability of P. digitatum mycelial cell walls by increasing the chitinase activity, thus accelerating the leakage of intracellular lysates. However, the integrity of the cell membrane was obviously damaged only after 60 min of treatment. In conclusion, we prepared a novel inclusion complex γ-CDTL with obvious antifungal effects and preliminarily elucidated its inclusion mechanism and antifungal mechanism. γ-CDTL might be a potent alternative to chemical fungicides for controlling the postharvest decay of citrus.

The effects of maturation and aging on the rotator cuff tendon-to-bone interface.

Rotator cuff tendon injuries often occur at the tendon-to-bone interface (i.e., enthesis) area, with a high prevalence for the elderly population, but the underlying reason for this phenomenon is still unknown. The objective of this study is to identify the histological, molecular, and biomechanical alterations of the rotator cuff enthesis with maturation and aging in a mouse model. Four different age groups of mice (newborn, young, adult, and old) were studied. Striking variations of the entheses were observed between the newborn and other matured groups, with collagen content, proteoglycan deposition, collagen fiber dispersion was significantly higher in the newborn group. The compositional and histological features of young, adult, and old groups did not show significant differences, except having increased proteoglycan deposition and thinner collagen fibers at the insertion sites in the old group. Nanoindentation testing showed that the old group had a smaller compressive modulus at the insertion site when compared with other groups. However, tensile mechanical testing reported that the old group demonstrated a significantly higher failure stress when compared with the young and adult groups. The proteomics analysis detected dramatic differences in protein content between newborn and young groups but minor changes among young, adult, and old groups. These results demonstrated: (1) the significant alterations of the enthesis composition and structure occur from the newborn to the young time period; (2) the increased risk of rotator cuff tendon injuries in the elderly population is not solely because of old age alone in the rodent model.

The effectiveness of tamsulosin hydrochloride with terazosin combination therapy for chronic prostatitis Type-III b.

Objectives: To study the therapeutic effects of combined tamsulosin hydrochloride and terazosin treatment for patients with chronic prostatitis Type-III b. Methods: This study involved 180 patients with chronic prostatitis Type-III b treated between January 2018 and December 2020 conducted at Nanhua Hospital Affiliated to Nanhua University. Patients were randomly divided into two equal groups: one receiving oral terazosin hydrochloride tablets only (control group), and one orally receiving both tamsulosin hydrochloride sustained-release tablets and terazosin hydrochloride tablets (observation group). Outcome measurements included symptom scoring, inflammatory cytokine levels, as well as white blood cell and lecithin body counts in the prostatic fluid. Results: After 30 days of treatment, the observation group showed greater treatment effectiveness (86.67% vs. 73.33%, P<0.05). QLS, USS, PS, and NIH-CPSI symptom scores were lower in the observation group than the control group (P<0.05). No differences in adverse event distribution and incidence were noted. EPS IL-2 increased more in the observation group, while PGE-2, MIP-1α, and MIP-2 decreased more in the observation group. WBC levels decreased more in the observation group, while lecithin body levels increased more in the observation group. Conclusion: The combination of tamsulosin hydrochloride and terazosin for the treatment of patients with chronic prostatitis Type-III b has a significant effect. This approach reduced patient symptoms, lowered inflammatory biomarkers, and generally improved quality of life. This approach appears to have clinical value worthy of future investigation.

WSe2 Flakelets on N-Doped Graphene for Accelerating Polysulfide Redox and Regulating Li Plating.

The practical application of lithium-sulfur batteries is still limited by the lithium polysulfides (LiPSs) shuttling effect on the S cathode and uncontrollable Li-dendrite growth on the Li anode. Herein, elaborately designed WSe2 flakelets immobilized on N-doped graphene (WSe2 /NG) with abundant active sites are employed to be a dual-functional host for satisfying both the S cathode and Li anode synchronously. On the S cathode, the WSe2 /NG with a strong interaction towards LiPSs can act as a redox accelerator to promote the bidirectional conversion of LiPSs. On the Li anode, the WSe2 /NG with excellent lithiophilic features can regulate the uniform Li plating/stripping to mitigate the growth of Li dendrite. Taking advantage of these merits, the assembled Li-S full batteries exhibit remarkable rate performance and stable cycling stability even at a higher sulfur loading of 10.5 mg cm-2 with a negative to positive electrode capacity (N/P) ratio of 1.4 : 1.

Macrophage depletion in stellate ganglia alleviates cardiac sympathetic overactivation and ventricular arrhythmogenesis by attenuating neuroinflammation in heart failure.

Cardiac sympathetic overactivation is involved in arrhythmogenesis in patients with chronic heart failure (CHF). Inflammatory infiltration in the stellate ganglion (SG) is a critical factor for cardiac sympathoexcitation in patients with ventricular arrhythmias. This study aims to investigate if macrophage depletion in SGs decreases cardiac sympathetic overactivation and ventricular arrhythmogenesis in CHF. Surgical ligation of the coronary artery was used for induction of CHF. Clodronate liposomes were microinjected into bilateral SGs of CHF rats for macrophage depletion. Using cytokine array, immunofluorescence staining, and Western blot analysis, we found that macrophage expansion and expression of TNFα and IL-1ß in SGs were markedly increased in CHF rats. Flow cytometry data confirmed that the percentage of macrophages in SGs was higher in CHF rats than that in sham rats. Clodronate liposomes significantly reduced CHF-elevated proinflammatory cytokine levels and macrophage expansion in SGs. Clodronate liposomes also reduced CHF-increased N-type Ca2+ currents and excitability of cardiac sympathetic postganglionic neurons and inhibited CHF-enhanced cardiac sympathetic nerve activity. ECG data from 24-h, continuous telemetry recording in conscious rats demonstrated that clodronate liposomes not only restored CHF-induced heterogeneity of ventricular electrical activities, but also decreased the incidence and duration of ventricular tachycardia/fibrillation in CHF. Macrophage depletion with clodronate liposomes attenuated CHF-induced cardiac sympathetic overactivation and ventricular arrhythmias through reduction of macrophage expansion and neuroinflammation in SGs.

Association between the variability of cerebral oxygen saturation during cardiopulmonary bypass and delayed postoperative neurocognitive recovery in cardiac valve surgical patients: A pilot study.

AIMS OF THE STUDY: The association between regional cerebral oxygen saturation (rSO2) and postoperative cognitive decline is controversial. In this study, we investigated the association between the real variability of regional cerebral oxygen saturation during cardiopulmonary bypass (CPB) and postoperative delayed neurocognitive recovery in patients undergoing heart valve surgery. METHODS USED TO CONDUCT THE STUDY: A total of 71 patients who underwent cardiac valve surgery were enrolled in this study. Patients were assessed for cognitive function using the Mini-Mental State Examination (MMSE) and the Montreal Cognitive Assessment Scale (MOCA) on the day before surgery and the 7th day after surgery. The real variability of regional cerebral oxygen saturation (rSO2), real variability of the brain bispectral index of EEG (BIS), real variability of mean arterial pressure (MAP) and body temperature were monitored during CPB. Patients were divided into two groups according to neural cognitive function scores to explore the relationship between postoperative delayed neurocognitive recovery and the real variability of cerebral oxygen saturation, BIS, MAP, and body temperature during CPB. RESULTS OF THE STUDY: Twenty-seven patients were diagnosed with postoperative delayed neurocognitive recovery. The occurrence of postoperative delayed neurocognitive recovery after surgery was closely related to the large variability of rSO2 during the rewarming phase of CPB (P < .05). Logistic analysis showed that preoperative arrhythmia, a lower level of serum albumin after surgery and greater rSO2 variability during the rewarming phase were risk factors for postoperative delayed neurocognitive recovery (P < .05). In this study, there was no correlation between postoperative delayed neurocognitive recovery and BIS, MAP or body temperature variability (P > .05). CONCLUSIONS DRAWN FROM THE STUDY AND CLINICAL IMPLICATIONS: The real variability of rSO2 during the CPB rewarming phase was related to postoperative delayed neurocognitive recovery in patients who underwent cardiac surgery.

Cinnamaldehyde promotes the defense response in postharvest citrus fruit inoculated with Penicillium digitatum and Geotrichum citri-aurantii.

Induced resistance in harvested fruit and vegetables is a superior strategy to reduce postharvest decay. In the present study, Cinnamaldehyde (CA) was applied to investigate for its induced resistance against Penicillium digitatum and Geotrichum citri-aurantii. The results showed that 5250 mg CA/L wax was effective concentration in inducing the resistance of citrus fruit to green mold and sour rot. Wax+ CA (WCA) reduced significantly green mold and sour rot incidences at different exposure times, with 24 h being the optimal exposure time. The host reactions under infection with different pathogens were similar. During initial exposure, treatment with 5250 mg CA/L wax enhanced significantly the activities of phenylalanine ammonia-lyase (PAL), peroxidase (POD), polyphenol oxidase (PPO), ß-1, 3-glucanase (GLU) and chitinase (CHT) in the presence of direct contact with the pathogen. Simultaneously, WCA induced an increase in total phenolic, flavanone and dihydroflavonol, flavone and flavonol, and lignin contents. Thus, our results suggest that treatment using 5250 mg CA/L wax can be applied early to control diseases by provoking response reactions in citrus fruit.

Inhibition of Pyk2 and Src activity improves Cx43 gap junction intercellular communication.

Identification of proteins that interact with Cx43 has been instrumental in the understanding of gap junction (GJ) regulation. An in vitro phosphorylation screen identified that Protein tyrosine kinase 2 beta (Pyk2) phosphorylated purified Cx43CT and this led us to characterize the impact of this phosphorylation on Cx43 function. Mass spectrometry identified Pyk2 phosphorylates Cx43 residues Y247, Y265, Y267, and Y313. Western blot and immunofluorescence staining using HeLaCx43 cells, HEK 293 T cells, and neonatal rat ventricular myocytes (NRVMs) revealed Pyk2 can be activated by Src and active Pyk2 interacts with Cx43 at the plasma membrane. Overexpression of Pyk2 increases Cx43 phosphorylation and knock-down of Pyk2 decreases Cx43 phosphorylation, without affecting the level of active Src. In HeLaCx43 cells treated with PMA to activate Pyk2, a decrease in Cx43 GJ intercellular communication (GJIC) was observed when assayed by dye transfer. Moreover, PMA activation of Pyk2 could be inhibited by the small molecule PF4618433. This partially restored GJIC, and when paired with a Src inhibitor, returned GJIC to the no PMA control-level. The ability of Pyk2 and Src inhibitors to restore Cx43 function in the presence of PMA was also observed in NRVMs. Additionally, an animal model of myocardial infarction induced heart failure showed a higher level of active Pyk2 activity and increased interaction with Cx43 in ventricular myocytes. Src inhibitors have been used to reverse Cx43 remodeling and improve heart function after myocardial infarction; however, they alone could not fully restore proper Cx43 function. Our data suggest that Pyk2 may need to be inhibited, in addition to Src, to further (if not completely) reverse Cx43 remodeling and improve intercellular communication.

TLR2 and caspase-1 signaling are critical for bacterial containment but not clearance during craniotomy-associated biofilm infection.

BACKGROUND: A craniotomy is required to access the brain for tumor resection or epilepsy treatment, and despite precautionary measures, infectious complications occur at a frequency of 1-3%. Approximately half of craniotomy infections are caused by Staphylococcus aureus (S. aureus) that forms a biofilm on the bone flap, which is recalcitrant to antibiotics. Our prior work in a mouse model of S. aureus craniotomy infection revealed a critical role for myeloid differentiation factor 88 (MyD88) in bacterial containment and pro-inflammatory mediator production. Since numerous receptors utilize MyD88 as a signaling adaptor, the current study examined the importance of Toll-like receptor 2 (TLR2) and TLR9 based on their ability sense S. aureus ligands, namely lipoproteins and CpG DNA motifs, respectively. We also examined the role of caspase-1 based on its known association with TLR signaling to promote IL-1ß release. METHODS: A mouse model of craniotomy-associated biofilm infection was used to investigate the role of TLR2, TLR9, and caspase-1 in disease progression. Wild type (WT), TLR2 knockout (KO), TLR9 KO, and caspase-1 KO mice were examined at various intervals post-infection to quantify bacterial burden, leukocyte recruitment, and inflammatory mediator production in the galea, brain, and bone flap. In addition, the role of TLR2-dependent signaling during microglial/macrophage crosstalk with myeloid-derived suppressor cells (MDSCs) was examined. RESULTS: TLR2, but not TLR9, was important for preventing S. aureus outgrowth during craniotomy infection, as revealed by the elevated bacterial burden in the brain, galea, and bone flap of TLR2 KO mice concomitant with global reductions in pro-inflammatory mediator production compared to WT animals. Co-culture of MDSCs with microglia or macrophages, to model interactions in the brain vs. galea, respectively, also revealed a critical role for TLR2 in triggering pro-inflammatory mediator production. Similar to TLR2, caspase-1 KO animals also displayed increased S. aureus titers coincident with reduced pro-inflammatory mediator release, suggestive of pathway cooperativity. Treatment of caspase-1 KO mice with IL-1ß microparticles significantly reduced S. aureus burden in the brain and galea compared to empty microparticles, confirming the critical role of IL-1ß in limiting S. aureus outgrowth during craniotomy infection. CONCLUSIONS: These results demonstrate the existence of an initial anti-bacterial response that depends on both TLR2 and caspase-1 in controlling S. aureus growth; however, neither pathway is effective at clearing infection in the WT setting, since craniotomy infection persists when both molecules are present.

TiO2 -Based Heterostructures with Different Mechanism: A General Synergistic Effect toward High-Performance Sodium Storage.

The general synergistic effect of TiO2 -based heterostructures has been discovered to improve the sodium storage of anodes, involving conversion, alloying, and insertion mechanism materials. Herein, metal sulfides (MS2 , M = Sn2+ , Co2+ , Mo2+ ), metallic Sb and Sn, as well as, carbon nanotubes (CNTs) are chosen as the model examples from the three kinds. The electrochemical testing demonstrates a better performance of heterostructrues involving TiO2 than the pristine anode components. The introduction of TiO2 into the MS2 and Sb or Sn systems induces a built-in electric field as the charge transfer force at the heterojunctions, greatly reducing the ion transfer resistance and promoting interfacial electron transfer. In the CNT/TiO2 structure, the chemical growth of TiO2 nanoparticles on the outer surface of CNTs makes the interface more compact than the physical blending case, offering better improvement of electrochemistry. The synergy should work via the growth of heterostructures, relying on the interface effects, which always plays the promotion role through the formation of driving force or grain boundaries and/or condense phase interface to facilitate charge transfer at the interface during the storage process. Therefore, the construction of reasonable heterostructures can endow materials with intriguing electrochemical performance based on the synergistic effect.