Flow cytofluorimetric analysis of anti-LRP4 (LDL receptor-related protein 4) autoantibodies in Italian patients with Myasthenia gravis

Background: Myasthenia gravis (MG) is an autoimmune disease in which 90% of patients have autoanti-bodies against the muscle nicotinic acetylcholine receptor (AChR), while autoantibodies to muscle-specific tyrosine kinase (MuSK) have been detected in half (5%) of the remaining 10%. Recently, the low-density lipoprotein receptor-related protein 4(LRP4), identified as the agrin receptor, has been recognized as a third autoimmune target in a significant portion of the double sero-negative (dSN) myasthenic individuals, with variable frequency depending on different methods and origin countries of the tested population. There is also convincing experimental evidence that anti-LRP4 autoantibodies may cause MG. Methods: The aim of this study was to test the presence and diagnostic significance of anti-LRP4 autoantibodies in an Italian population of 101 myasthenic patients (55 dSN, 23 AChR positive and 23 MuSK positive), 45 healthy blood donors and 40 patients with other neurological diseases as controls. All sera were analyzed by a cell-based antigen assay employing LRP4-transfected HEK293T cells, along with a flow cytofluorimetric detection system. Results: We found a 14.5% (8/55) frequency of positivity in the dSN-MG group and a 13% frequency of co-occurrence (3/23) in both AChR and MuSK positive patients; moreover, we report a younger female prevalence with a mild form of disease in LRP4-positive dSN-MG individuals. Conclusion: Our data confirm LRP4 as a new autoimmune target, supporting the value of including anti-LRP4 antibodies in further studies on Myasthenia gravis

Role of miR-148a in Mitigating Hepatic Ischemia-Reperfusion Injury by Repressing the TLR4 Signaling Pathway via Targeting CaMKIIα in Vivo and in Vitro

Background/Aims: Hepatic ischemia-reperfusion (I/R) injury, which is mainly induced by inflammation and unstable intracellular ions, is a major negative consequence of surgery that compromises hepatic function. However, the exact mechanisms of liver I/R injury have not been determined. Positive crosstalk with the Ca2+/CaMKII pathway is required for complete activation of the TLR4 pathway and inflammation. We previously found that miR-148a, which decreased in abundance with increasing reperfusion time, targeted and repressed the expression of CaMKIIα. In the present study, we examined the role of the miR-148a machinery in I/R-induced Ca2+/CaMKII and TLR4 signaling changes, inflammation, and liver dysfunction in vivo and in vitro. Methods: Liver function was evaluated by serum aminotransferase levels and hematoxylin-eosin (HE) staining. Inflammatory factors were detected by enzyme-linked immunosorbent assay. Gene and protein expression were assessed by RT-PCR and western blot. Small interfering RNA was used to silence target gene expression. HE staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling were used to measure hepatic tissue apoptosis. These assays were performed to identify factors upregulated in hepatic I/R injury and downregulated by miR-148a. Results: We manifested that expression of CaMKIIα and phosphorylation of TAK1 and IRF3 were elevated in hypoxia/reoxygenation (H/R)-treated primary Kupffer cells (KCs) and liver tissue of I/R-treated mice, but these effects were attenuated by treatment with miR-148a mimic and were accompanied by the alleviation of liver dysfunction and hepatocellular apoptosis. Luciferase reporter experiments showed that miR148a suppressed luciferase activity by almost 60%. Moreover, knockdown of CaMKIIα in H/R KCs led to significant deficiencies in p-TAK1, P-IRF3, IL-6, and TNF-α, which was consistent with the effects of miR-148a overexpression. Otherwise, the same trend of activation of TAK1 and IRF3 and inflammatory factors in vitro was observed in the siTAK1 + siIRF3 group compared with the siCaMKIIα group. Conclusion: Taken together, we conclude that miR-148a may mitigate hepatic I/R injury by ameliorating TLR4-mediated inflammation via targeting CaMKIIα in vitro and in vivo

Autoantibodies to Agrin in Myasthenia Gravis Patients

To determine if patients with myasthenia gravis (MG) have antibodies to agrin, a proteoglycan released by motor neurons and is critical for neuromuscular junction (NMJ) formation, we collected serum samples from 93 patients with MG with known status of antibodies to acetylcholine receptor (AChR), muscle specific kinase (MuSK) and lipoprotein-related 4 (LRP4) and samples from control subjects (healthy individuals and individuals with other diseases). Sera were assayed for antibodies to agrin. We found antibodies to agrin in 7 serum samples of MG patients. None of the 25 healthy controls and none of the 55 control neurological patients had agrin antibodies. Two of the four triple negative MG patients (i.e., no detectable AChR, MuSK or LRP4 antibodies, AChR-/MuSK-/LRP4-) had antibodies against agrin. In addition, agrin antibodies were detected in 5 out of 83 AChR+/MuSK-/LRP4- patients but were not found in the 6 patients with MuSK antibodies (AChR-/MuSK+/LRP4-). Sera from MG patients with agrin antibodies were able to recognize recombinant agrin in conditioned media and in transfected HEK293 cells. These sera also inhibited the agrin-induced MuSK phosphorylation and AChR clustering in muscle cells. Together, these observations indicate that agrin is another autoantigen in patients with MG and agrin autoantibodies may be pathogenic through inhibition of agrin/LRP4/MuSK signaling at the NMJ

VPS35 haploinsufficiency increases Alzheimer’s disease neuropathology

The retromer complex component VPS35 prevents activation of the BACE1 and Aβ production and thus plays an essential role in limiting Alzheimer’s disease neuropathology

BacHBerry: BACterial Hosts for production of Bioactive phenolics from bERRY fruits

BACterial Hosts for production of Bioactive phenolics from bERRY fruits (BacHBerry) was a 3-year project funded by the Seventh Framework Programme (FP7) of the European Union that ran between November 2013 and October 2016. The overall aim of the project was to establish a sustainable and economically-feasible strategy for the production of novel high-value phenolic compounds isolated from berry fruits using bacterial platforms. The project aimed at covering all stages of the discovery and pre-commercialization process, including berry collection, screening and characterization of their bioactive components, identification and functional characterization of the corresponding biosynthetic pathways, and construction of Gram-positive bacterial cell factories producing phenolic compounds. Further activities included optimization of polyphenol extraction methods from bacterial cultures, scale-up of production by fermentation up to pilot scale, as well as societal and economic analyses of the processes. This review article summarizes some of the key findings obtained throughout the duration of the project

A New Calculation Method of Cutterhead Torque Considering Shield Rolling Angle

The existing cutterhead torque calculation method usually simplifies the characteristics of the shield, which ignores the rolling angle. In this paper, the cross-river shield project of Wuhan Metro Line 8 is taken as the research focus. Firstly, the measured data of the cutterhead torque (CT), the rolling angle and rotation direction were analyzed. Then on this basis, the penetrability, tunneling thrust, and rolling angle were taken as the influential factors to analyze CT sensitivity. Finally, based on the theoretical calculation model, a modified solution of CT was obtained considering the rolling angle. The results show that the rolling angle can be reduced to zero by changing the direction of the cutterhead rotation; the rolling angle has a greater impact on CT than the other two factors as shown through the analysis of the range difference and Statistical Product and Service Solutions (SPSS) method. As the absolute value of the rolling angle increases, CT also increases, and the relationship between them is linear. To a certain extent, the rolling angle of the shield can reflect the difficulty of tunneling and the running status. By monitoring the rolling angle of the shield, the prediction of CT can be more in line with the actual construction conditions

A New Calculation Method of Cutterhead Torque Considering Shield Rolling Angle

The existing cutterhead torque calculation method usually simplifies the characteristics of the shield, which ignores the rolling angle. In this paper, the cross-river shield project of Wuhan Metro Line 8 is taken as the research focus. Firstly, the measured data of the cutterhead torque (CT), the rolling angle and rotation direction were analyzed. Then on this basis, the penetrability, tunneling thrust, and rolling angle were taken as the influential factors to analyze CT sensitivity. Finally, based on the theoretical calculation model, a modified solution of CT was obtained considering the rolling angle. The results show that the rolling angle can be reduced to zero by changing the direction of the cutterhead rotation; the rolling angle has a greater impact on CT than the other two factors as shown through the analysis of the range difference and Statistical Product and Service Solutions (SPSS) method. As the absolute value of the rolling angle increases, CT also increases, and the relationship between them is linear. To a certain extent, the rolling angle of the shield can reflect the difficulty of tunneling and the running status. By monitoring the rolling angle of the shield, the prediction of CT can be more in line with the actual construction conditions

Impact of In-Air Gestures on In-Car Task’s Diver Distraction

As in-vehicle information systems (IVIS) grow increasingly complex, the demand for innovative artificial intelligence-based interaction methods that enhance cybersecurity becomes more crucial. In-air gestures offer a promising solution due to their intuitiveness and individual uniqueness, potentially improving security in human–computer interactions. However, the impact of in-air gestures on driver distraction during in-vehicle tasks and the scarcity of skeleton-based in-air gesture recognition methods in IVIS remain largely unexplored. To address these challenges, we developed a skeleton-based framework specifically tailored for IVIS that recognizes in-air gestures, classifying them as static or dynamic. Our gesture model, tested on the large-scale AUTSL dataset, demonstrates accuracy comparable to state-of-the-art methods and increased efficiency on mobile devices. In comparative experiments between in-air gestures and touch interactions within a driving simulation environment, we established an evaluation system to assess the driver’s attention level during driving. Our findings indicate that in-air gestures provide a more efficient and less distracting interaction solution for IVIS in multi-goal driving environments, significantly improving driving performance by 65%. The proposed framework can serve as a valuable tool for designing future in-air gesture-based interfaces for IVIS, contributing to enhanced cybersecurity

Additional file 1 of Motoneurons innervation determines the distinct gene expressions in multinucleated myofibers

Additional file 1: Figure S1. Difference of nucleus between SR and NSR at NMJs. A. Whole-mount staining of diaphragm muscles from 2-month-old C57BL/6J mice. Note that several nuclei are enriched at R-BTX-labeled synaptic sites (indicated with dotted lines). Up, diaphragm muscles. Down, a single muscle fiber. B. Quantification of number of nucleus in the synaptic region (SR) or the non-synaptic region (NSR) in A. n =16 myofibers. Results were from three mice. C. Representative images of isolated SR and NSR from soleus muscles. Red, R-BTX. D. Immunoblot showing the expression of Tom20 and H3K4me2 in SR and NSR of diaphragm muscles. E. Immunoblot showing the expression of Tom20 and Kdm1a in SR and NSR of diaphragm muscles during development. F. ATP levels in SR and NSR of diaphragm muscles. Unless otherwise specified, three independent experiments were performed; the mean ± SEM is shown. t-test in B, and E, *p < 0.05 and ***p < 0.001