Pseudo-NRBC in the Mindray BC-6800Plus analyzer: A clue for diagnostic anticipation of fungemia. Experimental and preliminary clinical reports.

INTRODUCTION: Candidemia can be a significant cause of death in immunosuppressed or debilitated patients particularly. Abnormalities of the instrumental cytograms of some hematological analyzers, such as Mindray BC-6800Plus, can be related to circulating Candida. We studied the possible diagnostic usefulness of this information. METHODS: A fungal bloodstream infection has been simulated by adding aliquots of Candida albicans, Candida parapsilosis, and Candida glabrata to 75 leftovers and anonymized peripheral blood samples. Cytographic abnormalities like those of experimental samples were used to select patients with possible fungemia. The microscopic review of peripheral blood smears constituted the confirmatory method. RESULTS: In all experimental samples, the various Candida types caused pseudo-NRBC and morphological abnormalities of WNB and DIFF cytograms. Circulating blastospores, free or engulfed by neutrophils, were the microscopic findings in the peripheral blood smears. In the clinical verification, 72 patients were recruited based on the presence of an evocative cluster in the WNB and DIFF cytograms. The microscopic review of 39 out of 72 samples was positive for NRBC. According to blood cultures, light microscopy revealed fungal forms of several Candida or non-Candida types in the remaining 33 samples. Nine of these cases were not yet known to suffer from bloodstream infection. CONCLUSIONS: Although further confirmatory clinical studies are required for these diagnostic abilities, the BC 6800Plus cytographic abnormalities related to fungemia have proven helpful in rapidly monitoring persistent fungemia in already diagnosed patients. In unknown or undiagnosed cases, they could be the trigger point for the subsequent diagnostic-therapeutic pathway.

Probiotics for the prevention of gestational diabetes mellitus: A meta-analysis of randomized controlled trials.

Changes of intestinal microbiota have been shown to be involved in the development of gestational diabetes mellitus (GDM). We performed a meta-analysis to systematically evaluate the potential role of probiotics for the prevention of GDM. Systematic literature search was performed in electronic databases including PubMed, Cochrane library, Embase, Web of Science, Wanfang, and China National Knowledge Infrastructure (CNKI) to obtain relevant randomized controlled studies. A random-effects model was used to pool the results by incorporating the impact of the potential heterogeneity. Meta-regression and subgroup analyses were conducted to evaluate the source of heterogeneity. Fourteen studies involving 3527 pregnant women were included. Results showed that probiotics significantly reduced the incidence of GDM as compared to control (risk ratio [RR]: 0.71, 95% confidence interval [CI]: 0.52-0.96, P = 0.03) with significant heterogeneity (I2 = 73%). The meta-regression showed that body mass index (BMI) of females was positively associated with the RR for the effect of probiotics on GDM (coefficient = 0.084, P = 0.01). The results of subgroup analyses also suggested that probiotics significantly reduced the risk of GDM in women with BMI < 26 kg/m2, but not in those with BMI ≥ 26 kg/m2 (P for subgroup difference = 0.001). In addition, the preventative efficacy of probiotics on GDM was remarkable in women < 30 years, but not in those ≥ 30 years (P for subgroup difference < 0.001). In conclusion, probiotics may be effective in reducing the risk of GDM, particularly for females with lower BMI and younger age.

Modelling the Flow Behaviour of Al Alloy Sheets at Elevated Temperatures Using a Modified Zerilli-Armstrong Model and Phenomenological-Based Constitutive Models.

The flow behaviour of AA2060 Al alloy under warm/hot deformation conditions is complicated because of its dependency on strain rates (ε˙), strain (ε), and deformation modes. Thus, it is crucial to reveal and predict the flow behaviours of this alloy at a wide range of temperatures (T) and ε˙ using different constitutive models. Firstly, the isothermal tensile tests were carried out via a Gleeble-3800 thermomechanical simulator at a T range of 100, 200, 300, 400, and 500 °C and ε˙ range of 0.01, 0.1, 1, and 10 s-1 to reveal the warm/hot flow behaviours of AA2060 alloy sheet. Consequently, three phenomenological-based constitutive models (L-MJC, S1-MJC, S2-MJC) and a modified Zerilli-Armstrong (MZA) model representing physically based constitutive models were developed to precisely predict the flow behaviour of AA2060 alloy sheet under a wide range of T and ε˙. The predictability of the developed constitutive models was assessed and compared using various statistical parameters, including the correlation coefficient (R), average absolute relative error (AARE), and root mean square error (RMSE). By comparing the results determined from these models and those obtained from experimentations, and confirmed by R, AARE, and RMSE values, it is concluded that the predicted stresses determined from the S2-MJC model align closely with the experimental stresses, demonstrating a remarkable fit compared to the S1-MJC, L-MJC, and MZA models. This is because of the linking impact between softening, the strain rate, and strain hardening in the S2-MJC model. It is widely known that the dislocation process is affected by softening and strain rates. This is attributed to the interactions that occurred between ε and ε˙ from one side and between ε, ε˙, and T from the other side using an extensive set of constants correlating the constitutive components of dynamic recovery and softening mechanisms.

Serum heme oxygenase-1 level predicts clinical outcome after acute ischemic stroke.

AIMS: The relationship between heme oxygenase-1 (HO-1) and human ischemic stroke outcome remains unclear, which was investigated in this study. METHODS: Acute ischemic stroke patients admitted within 24 h were enrolled. Serum HO-1 levels at baseline were measured via ELISA. Poor 3-month functional outcome was defined as modified Rankin Scale (mRS) score 3-6. Multivariable-adjusted binary logistic regression and restricted cubic spline models were employed to examine association between serum HO-1 and functional outcome. HO-1's additive prognostic utility was assessed by net reclassification index (NRI) and integrated discrimination improvement (IDI). RESULTS: Of 194 eligible patients, 79 (40.7%) developed poor functional outcomes at 3-month follow-up. The highest quartile of serum HO-1 was independently associated with a lower risk of poor functional outcome (adjusted OR 0.13, 95% CI 0.04-0.45; p = 0.001) compared with the lowest HO-1 category. The relationship between higher HO-1 levels and reduced risk of poor functional outcome was linear and dose responsive (p = 0.002 for linearity). Incorporating HO-1 into the analysis with conventional factors significantly improved reclassification for poor functional outcomes (NRI = 41.2%, p = 0.004; IDI = 5.0%, p = 0.004). CONCLUSIONS: Elevated serum HO-1 levels at baseline were independently associated with improved 3-month functional outcomes post-ischemic stroke. Serum HO-1 measurement may enhance outcome prediction beyond conventional clinical factors.

Serum copeptin in women with gestational diabetes mellitus: A meta-analysis.

To address inconsistencies in prior research, this meta-analysis examines the potential link between serum copeptin levels and the risk of developing gestational diabetes mellitus (GDM). Previous studies have reported mixed results regarding the relationship between serum copeptin levels and GDM risk. Our objective was to comprehensively evaluate this association. We systematically reviewed observational studies from Medline, Web of Science, Embase, Wanfang, and Chinese National Knowledge Infrastructure (CNKI) databases up to October 15, 2023, employing a random-effects model to integrate the data while considering heterogeneity. This analysis incorporated 10 studies comprising 625 women with GDM and 1212 healthy pregnant controls. Our findings showed no significant difference in serum copeptin levels between women with GDM and those without (standardized mean difference [SMD] 0.01, 95% confidence interval [CI] -0.22 to 0.24, P = 0.92, I2 = 75%). Univariate meta-analysis indicated a positive correlation between the body mass index (BMI) of the participants and the outcomes (coefficient = 0.11, P = 0.002). Further subgroup analysis demonstrated that women with a mean BMI ≥ 26 kg/m2 and GDM had significantly higher serum copeptin levels compared to their non-GDM counterparts (SMD 0.31, 95% CI 0.05 to 0.57, P = 0.02, I2 = 46%). Conversely, no difference was observed in women with a BMI < 26 kg/m2 (SMD -0.23, 95% CI -0.37 to -0.09, P = 0.002, I2 = 0%, P for subgroup difference = 0.003). Variables such as the country of study, maternal age, timing of blood sampling, copeptin measurement methods, or GDM diagnostic criteria did not significantly affect the results. In summary, the association between serum copeptin levels and GDM risk is influenced by the BMI of pregnant women, indicating that elevated serum copeptin might be linked to GDM in individuals with a BMI ≥ 26 kg/m2.

Performance of the digital cell morphology analyzer MC-100i in a multicenter study in tertiary hospitals in China.

BACKGROUND: This study investigated the performance of the MC-100i, a pre-commercial digital morphology analyzer utilizing a convolutional neural network algorithm, in a multicentric setting involving up to 11 tertiary hospitals in China. METHODS: Blood smears were analyzed by MC-100i, verified by morphologists, and manually differentiated. The classification performance on WBCs and RBCs was evaluated by comparing the classification results using different methods. The PLT and PLT clump counting performance was also assessed. The total assay time including hands-on time was evaluated. RESULTS: The agreements between pre- and post-classification were high for normal WBCs (κ > 0.96) and lower for overall abnormal WBCs (κ = 0.90). The post-classification results correlated well with manual differentials for both normal and abnormal WBCs (r > 0.93), except for basophils (r = 0.8480) and atypical lymphocytes (r = 0.8211). The clinical sensitivity and specificity of each RBC abnormality after verification were above 90 % using microscopy reviews as the reference. The PLTs counted by the MC-100i before and after verification correlated well with those measured by the PLT-O mode (r = 0.98). Moreover, PLT clumps were successfully classified by the analyzer in EDTA-dependent pseudothrombocytopenia blood samples. CONCLUSIONS: The MC-100i is an accurate and reliable digital cell morphology analyzer, offering another intelligent option for hematology laboratories.

A new hematological parameter model for the diagnosis and prognosis of sepsis in emergency department: A single-center retrospective study.

INTRODUCTION: Sepsis, a syndrome of organ dysfunction caused by an unregulated host response to infection. This study aimed to develop a novel sepsis diagnostic model of hematological parameters and evaluate its effectiveness in the early identification and prognosis of sepsis in emergency departments. METHODS: A retrospective study was conducted in Emergency Department. Cell population data parameters related to monocytes and neutrophils were obtained using the Mindary BC-6800 plus hematology analyzer. Receiver operating characteristic (ROC) curve analysis, logistic regression analysis was performed to assess the performance of the parameters and establish a diagnostic and prognostic model of sepsis, which was then verified with a validation cohort. RESULTS: Mon_XW exhibited the best diagnostic performance (area under the ROC curve [AUC] = 0.848, 95% confidence interval [CI]: 0.810-0.885, p < 0.001), followed by Neu_Y and Neu_YW (AUC = 0.777 95% CI: 0.730-0.824, p < 0.001). Logistic regression analysis identified Mon_XW and Neu_Y as independent predictors, which were used to establish a diagnostic model named hematological parameter for sepsis (HPS). HPS demonstrated the best diagnostic performance with an AUC of 0.862 (95% CI: 0.826-0.898, p < 0.001), sensitivity of 70.0%, and specificity of 87.1%, compared to C-reactive protein (CRP) and procalcitonin (PCT). The validation cohort also found that the positive predictive value of HPS was 70.4% and the negative predictive value was 92.2%. CONCLUSION: The developed HPS model showed promising diagnostic efficacy for sepsis in the emergency department, which outperformed CRP and PCT in terms of sensitivity and specificity. By enabling early identification and prognosis of sepsis, that contributes to reducing sepsis-related mortality.

Secondary somatosensory cortex glutamatergic innervation of the thalamus facilitates pain.

ABSTRACT: Although the secondary somatosensory cortex (SII) is known to be involved in pain perception, its role in pain modulation and neuropathic pain is yet unknown. In this study, we found that glutamatergic neurons in deep layers of the SII (SII Glu ) responded to bilateral sensory inputs by changing their firing with most being inhibited by contralateral noxious stimulation. Optical inhibition and activation of unilateral SII Glu reduced and enhanced bilateral nociceptive sensitivity, respectively, without affecting mood status. Tracing experiments revealed that SII Glu sent dense monosynaptic projections to the posterolateral nucleus (VPL) and the posterior nucleus (Po) of the thalamus. Optical inhibition and activation of projection terminals of SII Glu in the unilateral VPL and Po inhibited and facilitated pain on the contralateral side, respectively. After partial sciatic nerve ligation, SII Glu became hyperactive as evidenced by higher frequency of spontaneous firing, but the response patterns to peripheral stimulation remained. Optical inhibition of SII Glu alleviated not only bilateral mechanical allodynia and thermal hyperalgesia but also the negative affect associated with spontaneous pain. Inhibition of SII Glu terminals in the VPL and Po also relieved neuropathic pain. This study revealed that SII Glu and the circuits to the VPL and Po constitute a part of the endogenous pain modulatory network. These corticothalamic circuits became hyperactive after peripheral nerve injury, hence contributes to neuropathic pain. These results justify proper inhibition of SII Glu and associated neural circuits as a potential clinical strategy for neuropathic pain treatment.

Comparison of Characteristics and Outcomes Between Acute Ischemic Stroke Patients with Different Types of Heart Failure.

Acute ischemic stroke (AIS) can be complicated by heart failure involving preserved ejection fraction (HFpEF) or reduced ejection fraction (HFrEF), and whether or not the prognosis differs between the 2 types of patients remains unclear. We compared the clinical characteristics and outcomes of the 2 types of patients at 3 months after the stroke.We retrospectively analyzed patients who, between 1 January 2018 and 1 January 2021, experienced AIS that was complicated by HFrEF or HFpEF. All patients had been prospectively registered in the Chengdu Stroke Registry. Poor outcome was defined as a modified Rankin Scale (mRS) score of 2-6 at 3 months. Univariate and binary logistic regression was used to assess whether HFpEF was associated with a significantly worse prognosis than HFrEF.Among the final sample of 108 patients (60.2% men; mean age, 73.08 ± 10.82 years), 75 (69.4%) had HFpEF. Compared to HFrEF patients, those with HFpEF were older (P = 0.002), were more likely to have chronic kidney disease (P = 0.033), and were more likely to experience a poor outcome (P = 0.022). After adjustments, HFpEF was associated with significantly greater risk of poor outcome than HFrEF (OR 4.13, 95%CI 1.20-15.79, P = 0.029). However, rates of hemorrhagic transformation or mortality at 3 months after AIS did not differ significantly between the 2 types of heart failure (all P > 0.05).Patients with AIS involving HFpEF experience worse outcomes than those with HFrEF and therefore may require special monitoring and management. Our findings need to be verified in large prospective studies.

Lifetime evaluation of left ventricular structure and function in male ApoE null mice after gamma and space-type radiation exposure.

The space radiation (IR) environment contains high charge and energy (HZE) nuclei emitted from galactic cosmic rays with the ability to overcome current shielding strategies, posing increased IR-induced cardiovascular disease risks for astronauts on prolonged space missions. Little is known about the effect of 5-ion simplified galactic cosmic ray simulation (simGCRsim) exposure on left ventricular (LV) function. Three-month-old, age-matched male Apolipoprotein E (ApoE) null mice were irradiated with 137Cs gamma (γ; 100, 200, and 400 cGy) and simGCRsim (50, 100, 150 cGy all at 500 MeV/nucleon (n)). LV function was assessed using transthoracic echocardiography at early/acute (14 and 28 days) and late/degenerative (365, 440, and 660 days) times post-irradiation. As early as 14 and 28-days post IR, LV systolic function was reduced in both IR groups across all doses. At 14 days post-IR, 150 cGy simGCRsim-IR mice had decreased diastolic wall strain (DWS), suggesting increased myocardial stiffness. This was also observed later in 100 cGy γ-IR mice at 28 days. At later stages, a significant decrease in LV systolic function was observed in the 400 cGy γ-IR mice. Otherwise, there was no difference in the LV systolic function or structure at the remaining time points across the IR groups. We evaluated the expression of genes involved in hemodynamic stress, cardiac remodeling, inflammation, and calcium handling in LVs harvested 28 days post-IR. At 28 days post-IR, there is increased expression of Bnp and Ncx in both IR groups at the lowest doses, suggesting impaired function contributes to hemodynamic stress and altered calcium handling. The expression of Gals3 and ß-Mhc were increased in simGCRsim and γ-IR mice respectively, suggesting there may be IR-specific cardiac remodeling. IR groups were modeled to calculate the Relative Biological Effectiveness (RBE) and Radiation Effects Ratio (RER). No lower threshold was determined using the observed dose-response curves. These findings do not exclude the possibility of the existence of a lower IR threshold or the presence of IR-induced cardiovascular disease (CVD) when combined with additional space travel stressors, e.g., microgravity.

Geochronological results from the Zhela Formation volcanics of the Tethyan Himalaya and their implications for the breakup of eastern Gondwana.

The relationship between the Kerguelen mantle plume and the breakup of eastern Gondwana is still debated. The new Zircon SHRIMP U-Pb dating of 139.9 ± 4.6 Ma, as well as previous ages from the Zhela Formation volcanic rocks in the Tethyan Himalaya, show that the studied Zhela Formation volcanic rocks formed during the Late Jurassic-Early Cretaceous, rather than the Middle Jurassic. The calculated volume of the Comei-Bunbury igneous rocks is ~ 114,250 km3, which is compatible with the large igneous provinces and, consequently, the typical mantle plume models. The new date results, along with existing dates, show that the volcanism attributed to the Kerguelen mantle plume in the Tethyan Himalaya ranges from ca.147 Ma to ca.124 Ma, with two peaks at approximately 141 Ma and 133 Ma. This new finding, together with geochemical and palaeomagnetic data obtained from the Comei-Bunbury igneous rocks, indicate that the Kerguelen mantle plume contributed significantly to the breakup of eastern Gondwana and that eastern Gondwana first disintegrated and dispersed at ca.147 Ma, the Indian plate separated completely from the eastern Gondwana before ca.125 Ma.

The association between hypertensive angiopathy and cerebral amyloid angiopathy in primary intracerebral hemorrhage.

Objective: To determine the association between the burden of cerebral small vessel disease (CSVD) due to hypertensive angiopathy (HA) and cerebral amyloid angiopathy (CAA) on MRI in patients with primary intracerebral hemorrhage (ICH). Methods: Patients with primary ICH admitted to our center from March 2012 to November 2021 were consecutively enrolled. We used multivariate binary and ordinal regression analyses to assess the association between HA-CSVD burden and CAA-CSVD burden. Lobar cerebral microbleeds (CMBs) were categorized into three level of severity: 0-1, 2-4, and ≥ 5 lobar CMBs. A high CAA-CSVD score was defined as a CAA-CSVD score of ≥3. Results: Overall, 222 participants (mean age 59.88 ± 13.56) were included into analysis. Age and ICH etiology differed among different lobar CMB severity and between the presence and absence of high CAA-CSVD score (all p < 0.05). Positive associations between HA-related markers and both lobar CMB severity and high CAA-CSVD score (p < 0.05 for the presence of lacune, deep CMBs ≥5, the presence of WMH, and HA-CSVD score) were observed in univariate analysis. These associations remained significant after adjusting for age, sex, ICH etiology, and potential vascular risk factors. The distribution of CAA-CSVD score was significantly different between patients with and without CMBs ≥5 (adjusted OR 2.351, 95% CI 1.242-4.455, p = 0.009) after correcting for age, sex, ICH etiology, and vascular risk factors. Conclusion: Our study provides evidence of an association between HA-CSVD and CAA-CSVD in patients with primary ICH, which needs to be verified in future studies.

Catalytic Growth of High-Performance Graphite-like Carbon Films on a Nitrided Substrate: Experimental Study and First-Principles Calculations.

This study reports a novel process for the fabrication of high-performance graphite-like carbon (GLC) films on nitrided substrates through successive steps in a plasma nitriding system. Unlike films fabricated via conventional dual treatments (nitriding + film deposition), here-fabricated GLC films were grown on the surface of nitrided steel via a catalytic reaction. A transition zone having a nanocrystalline and amorphous structure was observed at the interface between the nitrided substrate and GLC films, which increased the durability of GLC films because this structure enhanced the adhesion of GLC films on the nitrided substrate, improving resistance to spallation and wear. Experimental study and first-principles calculations showed that the Fe3N phase had a stronger catalytic effect on GLC films than the Fe4N phase, and GLC films grown on the nitrided layer dominated by Fe3N were thicker with stronger adhesion and excellent frictional and wear properties compared with GLC films grown on the nitrided layer dominated by Fe4N because of the beneficial structure formed at the interface. This study reports a simple and inexpensive method to fabricate a dual layer containing high-performance GLC films via a catalytic growth and interface matching mechanism.

Triglyceride-glucose index predicts death in patients with stroke younger than 65.

Background: The triglyceride-glucose index (TGI), a reliable surrogate indicator of insulin resistance (IR), has been proven to be a predictor of the incidence of ischemic stroke. The role of TGI in predicting the outcomes of stroke patients remains controversial. Susceptibility to IR-related diseases varies among patients of different ages. The study aims to evaluate the predictive value of TGI levels on clinical outcomes of patients with ischemic stroke of different ages. Method: This was a retrospective cohort study including patients with ischemic stroke in the Department of Neurology at West China Hospital. TGI was calculated as ln [fasting triglyceride (mg/dL) × fasting glucose (mg/dL)/2]. The patients were subdivided into 3 tertiles according to TGI levels. Multivariate logistic regression analyses were conducted to estimate the association between TGI levels and post-stroke outcomes among the whole patients, younger patients (<65), and older patients (>=65). The outcomes included death and unfavorable functional outcome (modified Rankin scale score 3-6) at 3 and 12 months after stroke. Results: A total of 3,704 patients (men, 65.08%, mean age, 61.44 ± 14.15; women 34.92%, mean age, 65.70 ± 13.69) were enrolled in this study. TGI levels were not associated with 3 month or 12 month death in the whole patients. Patients with higher TGI levels (T2 and T3) had a higher risk of 3 month death than those had lower TGI levels (T1) in the younger group (T2 vs. T1: OR 2.64, 95% CI 1.03-6.79, p = 0.043; T3 vs. T1: OR 2.69, 95% CI 1.00-7.10, p = 0.049) but not in the older group. Additionally, Kaplan-Meier estimate analysis illustrated that the 12 month death risk was significantly higher in the group with the highest TGI among younger patients (p for log-rank test = 0.028) but not among older patients. There was an interactive effect between TGI and age on 3 month death (p for interaction = 0.013) and 12 month death (p for interaction = 0.027). However, TGI was not associated with unfavorable functional outcome at 3 month or 12 month after stroke. Conclusion: Elevated TGI independently predicts death at 3 months and 12 months in patients under 65 with ischemic stroke. Regulating TGI is expected to be an approach to enhance prognosis in young individuals affected by ischemic stroke.

Comparison of the performance of two automatic cell morphology analyzers for peripheral-blood leukocyte morphology analysis: Mindray MC-100i and Sysmex DI-60.

INTRODUCTION: To compare the morphological classification ability of peripheral-blood leukocytes of the automatic cell morphology analyzers MC-100i and DI-60. METHODS: (1) MC-100i and DI-60 were used to analyze leukocytes in 432 venous blood samples collected from three tertiary hospitals across China. The preclassification results were compared with the results reported by senior morphological experts (postclassification results) to evaluate the accuracy, sensitivity, specificity, and consistency of leukocyte preclassification for both instruments. (2) In 200 of the 432 blood samples, morphological experts conducted manual microscopic examination for various types of leukocytes. The correlation between the MC-100i and DI-60 leukocyte postclassification results and the expert microscopist results were analyzed. RESULTS: (1) MC-100i preclassified leukocytes and nucleated red blood cells (RBCs). Compared with the postclassification results, the total leukocyte preclassification accuracy of MC-100i was 97.16%, while that of DI-60 was 87.24%. The sensitivity of MC-100i to abnormal cells (including blasts, promyelocytes, neutrophilic myelocytes, neutrophilic metamyelocytes, reactive lymphocytes, abnormal promyelocytes, plasma cells, abnormal lymphocytes and nucleated RBCs) was 90.24%, which was significantly higher than the 50.72% sensitivity of DI-60. (2) Comparing the postclassification results with manual microscopy, except for reactive lymphocytes and basophils, the MC-100i and DI-60 results had good correlations with various leukocyte types and nucleated RBCs (r > 0.85), and MC-100i was better than DI-60 in the recognition of basophils. CONCLUSION: Both MC-100i and DI-60 have good detection ability for five normal types of leukocytes in peripheral blood. MC-100i has significantly better detection sensitivity for abnormal cells in peripheral blood than DI-60.

Extracellular Vesicle-Encapsulated Adeno-Associated Viruses for Therapeutic Gene Delivery to the Heart.

BACKGROUND: Adeno-associated virus (AAV) has emerged as one of the best tools for cardiac gene delivery due to its cardiotropism, long-term expression, and safety. However, a significant challenge to its successful clinical use is preexisting neutralizing antibodies (NAbs), which bind to free AAVs, prevent efficient gene transduction, and reduce or negate therapeutic effects. Here we describe extracellular vesicle-encapsulated AAVs (EV-AAVs), secreted naturally by AAV-producing cells, as a superior cardiac gene delivery vector that delivers more genes and offers higher NAb resistance. METHODS: We developed a 2-step density-gradient ultracentrifugation method to isolate highly purified EV-AAVs. We compared the gene delivery and therapeutic efficacy of EV-AAVs with an equal titer of free AAVs in the presence of NAbs, both in vitro and in vivo. In addition, we investigated the mechanism of EV-AAV uptake in human left ventricular and human induced pluripotent stem cell-derived cardiomyocytes in vitro and mouse models in vivo using a combination of biochemical techniques, flow cytometry, and immunofluorescence imaging. RESULTS: Using cardiotropic AAV serotypes 6 and 9 and several reporter constructs, we demonstrated that EV-AAVs deliver significantly higher quantities of genes than AAVs in the presence of NAbs, both to human left ventricular and human induced pluripotent stem cell-derived cardiomyocytes in vitro and to mouse hearts in vivo. Intramyocardial delivery of EV-AAV9-sarcoplasmic reticulum calcium ATPase 2a to infarcted hearts in preimmunized mice significantly improved ejection fraction and fractional shortening compared with AAV9-sarcoplasmic reticulum calcium ATPase 2a delivery. These data validated NAb evasion by and therapeutic efficacy of EV-AAV9 vectors. Trafficking studies using human induced pluripotent stem cell-derived cells in vitro and mouse hearts in vivo showed significantly higher expression of EV-AAV6/9-delivered genes in cardiomyocytes compared with noncardiomyocytes, even with comparable cellular uptake. Using cellular subfraction analyses and pH-sensitive dyes, we discovered that EV-AAVs were internalized into acidic endosomal compartments of cardiomyocytes for releasing and acidifying AAVs for their nuclear uptake. CONCLUSIONS: Together, using 5 different in vitro and in vivo model systems, we demonstrate significantly higher potency and therapeutic efficacy of EV-AAV vectors compared with free AAVs in the presence of NAbs. These results establish the potential of EV-AAV vectors as a gene delivery tool to treat heart failure.

Coupling Computational Homogenization with Crystal Plasticity Modelling for Predicting the Warm Deformation Behaviour of AA2060-T8 Al-Li Alloy.

This study aimed to propose a new approach for predicting the warm deformation behaviour of AA2060-T8 sheets by coupling computational homogenization (CH) with crystal plasticity (CP) modeling. Firstly, to reveal the warm deformation behaviour of the AA2060-T8 sheet, isothermal warm tensile testing was accomplished using a Gleeble-3800 thermomechanical simulator at the temperatures and strain rates that varied from 373 to 573 K and 0.001 to 0.1 s-1. Then, a novel crystal plasticity model was proposed for describing the grains' behaviour and reflecting the crystals' actual deformation mechanism under warm forming conditions. Afterward, to clarify the in-grain deformation and link the mechanical behaviour of AA2060-T8 with its microstructural state, RVE elements were created to represent the microstructure of AA2060-T8, where several finite elements discretized every grain. A remarkable accordance was observed between the predicted results and their experimental counterparts for all testing conditions. This signifies that coupling CH with CP modelling can successfully determine the warm deformation behaviour of AA2060-T8 (polycrystalline metals) under different working conditions.

Effects of La Addition on Microstructure Evolution and Thermal Stability of Cu-2.35Ni-0.59Si Sheet.

A Cu-2.35Ni-0.69Si alloy with low La content was designed in order to study the role of La addition on microstructure evolution and comprehensive properties. The results indicate that the La element demonstrates a superior ability to combine with Ni and Si elements, via the formation of La-rich primary phases. Owing to existing La-rich primary phases, restricted grain growth was observed, due to the pinning effect during solid solution treatment. It was found that the activation energy of the Ni2Si phase precipitation decreased with the addition of La. Interestingly, the aggregation and distribution of the Ni2Si phase, around the La-rich phase, was observed during the aging process, owing to the attraction of Ni and Si atoms by the La-rich phase during the solid solution. Moreover, the mechanical and conductivity properties of aged alloy sheets suggest that the addition of the La element showed a slight reducing effect on the hardness and electrical conductivity. The decrease in hardness was due to the weakened dispersion and strengthening effect of the Ni2Si phase, while the decrease in electrical conductivity was due to the enhanced scattering of electrons by grain boundaries, caused by grain refinement. More notably, excellent thermal stabilities, including better softening resistance ability and microstructural stability, were detected for the low-La-alloyed Cu-Ni-Si sheet, owing to the delayed recrystallization and restricted grain growth caused by the La-rich phases.

Predicting the emergence of malignant brain oedema in acute ischaemic stroke: a prospective multicentre study with development and validation of predictive modelling.

Background: We aimed to develop and validate a prognostic model for predicting malignant brain oedema in patients with acute ischaemic stroke in a real-world setting of practice. Methods: A prospective multicentre study enrolled adult patients with acute ischaemic stroke with brain CT < 24 h of onset of symptoms admitted to nine tertiary-level hospitals in China between September 2017 and December 2019. Malignant brain oedema was defined as any patient who had decompressive craniectomy, discharge in coma, or in-hospital death attributed to symptomatic brain swelling. The derivation cohort was a consecutive cohort of patients from one centre and the validation cohort was non-consecutive patients from the other centres. Multivariable logistic regression was used to define independent predictors from baseline clinical characteristics, imaging features, complications, and management. A web-based nomogram and a risk score were developed based on the final model. Model performance was assessed for discrimination and calibration in both derivation and validation cohorts. The study is registered, NCT03222024. Findings: Based on the derivation cohort (n = 1627), the model was developed with seven variables including large infarct (adjusted odds ratio [OR] 40.90, 95% CI 20.20-82.80), National Institutes of Health Stroke Scale (NIHSS) score (OR 1.09, 1.06-1.12), thrombolysis (OR 2.11, 1.18-3.78), endovascular treatment (OR 2.87, 1.47-5.59), pneumonia (OR 2.47, 1.53-3.97), brain atrophy (OR 0.57, 0.37-0.86), and recanalisation (OR 0.36, 0.17-0.75). The classification threshold of a predicted probability ≥0.14 showed good discrimination and calibration in both derivation cohort (area under the receiver-operating curve [AUC] 0.90, 0.87-0.92; sensitivity 0.95, 0.92-0.98) and validation cohort (n = 556, AUC 0.88, 0.82-0.95; sensitivity 0.84, 0.73-0.95). The risk score based on this model had a total point that ranged from -1 to 20, with an optimal score of ≥10 showing good discrimination and calibration in both derivation (AUC 0.89, 0.87-0.92; sensitivity 0.95, 0.92-0.98) and validation (AUC 0.88, 0.82-0.95; sensitivity 0.84, 0.73-0.95) cohorts. Interpretation: The INTEP-AR model (i.e. large Infarct, NIHSS score, Thrombolysis, Endovascular treatment, Pneumonia, brain Atrophy, and Recanalisation) incorporating multiple clinical and radiological characteristics has shown good prognostic value for predicting malignant brain oedema after acute ischaemic stroke. Funding: National Natural Science Foundation of China; Science and Technology Department of Sichuan Province; West China Hospital.

Serendipita indica chitinase protects rice from the blast and bakanae diseases.

Serendipita indica, a multifunctional and useful endophyte fungus, has been intensively investigated in promoting plant growth and resistance towards biotic and abiotic stress. Multiple chitinases from microorganisms or plants have been identified to have a high antifungal activity as a biological control. However, chitinase of S. indica still needs to be characterized. We functionally characterized a chitinase (SiChi) in S. indica. The result showed that the purified SiChi protein confers high chitinase activity; importantly, SiChi inhibits the conidial germination of Magnaporthe oryzae and Fusarium moniliforme. After the successful colonization of rice roots by S. indica, both the rice blast disease and bakanae disease were significantly reduced. Interestingly, the purified SiChi could promptly induce rice disease resistance towards M. oryzae and F. moniliforme pathogens when sprayed on rice leaves. Like S. indica, SiChi could upregulate rice pathogen-resistant proteins and defense enzymes. In conclusion, chitinase of S. indica has direct antifungal activity and indirect induced resistance activity, implying an efficient and economic strategy for rice disease control by applying S. indica and SiChi.