A comparison between cisatracurium and rocuronium-induced neuromuscular block on laryngeal electromyography recovery after neostigmine reversal in a porcine model

BackgroundInducing and reversing neuromuscular block is essential to a positive outcome of thyroid surgery, with intraoperative neuromonitoring (IONM) being used to decrease recurrent and superior laryngeal nerve injuries and improve vocal outcome. Neostigmine is a non-specific broad-spectrum and inexpensive reversal agent for neuromuscular blocking agents (NMBAs). The aim of this porcine study was to explore the effect of neostigmine on electromyography (EMG) signal recovery profile following the commonly used NMBAs, cisatracurium and rocuronium. MethodsTwelve piglets were allocated into two groups with six piglets in each group. When stable baseline EMG signals were obtained, a neuromuscular block was induced by intravenous cisatracurium 0.2 mg/kg (group C) or rocuronium 0.6 mg/kg (group R) for each piglet. We compared laryngeal EMG tracing with spontaneous recovery (control) and neostigmine (0.04 mg/kg) reversal for each group. The time course of real-time laryngeal EMG signals was observed for 30 min from NMBA injection. Effects of neostigmine on EMG signal were assessed at 50% EMG recovery and by the maximum neuromuscular block recovery degree from the baseline value. ResultsNeostigmine shortened the recovery time to 50% EMG amplitude in both group C (16.5 [2.5] vs. 29.0 [2.0] min, P<0.01) and group R (16.5[2.5] vs. 26.5 [1.5] min, P<0.05) compared to spontaneous recovery, respectively. Neostigmine reversal also enhanced the maximum degree of EMG amplitude recovery in both group C (83.6 [5.1] vs. 47.2 [6.1] %, P<0.01) and group R (85.6 [18.2]vs. 57.1 [6.3] %, P<0.05) compared to spontaneous recovery, respectively. The reversal effect of neostigmine did not differ significantly between cisatracurium and rocuronium. ConclusionsThis porcine model demonstrated that neostigmine provides an adequate and timely IONM signal suppressed by both cisatracurium and rocuronium. These results can potentially expand the options for precision neuromuscular block management during IONM to improve vocal outcomes in thyroid surgery patients

Upregulation of Pd-L1 by Sars-Cov-2 Promotes Immune Evasion

Patients with severe COVID-19 often suffer from lymphopenia, which is linked to T-cell sequestration, cytokine storm, and mortality. However, it remains largely unknown how severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) induces lymphopenia. Here, we studied the transcriptomic profile and epigenomic alterations involved in cytokine production by SARS-CoV-2-infected cells. We adopted a reverse time-order gene coexpression network approach to analyze time-series RNA-sequencing data, revealing epigenetic modifications at the late stage of viral egress. Furthermore, we identified SARS-CoV-2-activated nuclear factor-κB (NF-κB) and interferon regulatory factor 1 (IRF1) pathways contributing to viral infection and COVID-19 severity through epigenetic analysis of H3K4me3 chromatin immunoprecipitation sequencing. Cross-referencing our transcriptomic and epigenomic data sets revealed that coupling NF-κB and IRF1 pathways mediate programmed death ligand-1 (PD-L1) immunosuppressive programs. Interestingly, we observed higher PD-L1 expression in Omicron-infected cells than SARS-CoV-2 infected cells. Blocking PD-L1 at an early stage of virally-infected AAV-hACE2 mice significantly recovered lymphocyte counts and lowered inflammatory cytokine levels. Our findings indicate that targeting the SARS-CoV-2-mediated NF-κB and IRF1-PD-L1 axis may represent an alternative strategy to reduce COVID-19 severity

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Multiobjective Optimization for Planning the Service Areas of Smart Parcel Locker Facilities in Logistics Last Mile Delivery

The planning of the location service areas of smart parcel locker facilities became a critical aspect of logistics last mile delivery. In e-commerce, the efficiency of delivering merchandise from retailer warehouses to customers determines the competitiveness of retailers and delivery operators. The considerable increases in e-commerce transactions and safety concerns under the COVID-19 (Coronavirus disease 2019) pandemic made home delivery services even more inefficient than before, which resulted in the considerable increase in social costs. In numerous countries, smart parcel lockers were adopted to increase delivery efficiency, decrease the risk of COVID-19 infection, and reduce the burden on society. This study proposed a multiobjective optimization mathematical model for investigating the planning of the location service areas of smart parcel locker facilities, and then the optimization mathematical model was solved using a combination of the Taguchi method (TA) and nondominant sorting genetic algorithm II (NSGA-II). Finally, this composite approach was applied to a case study in producing favorable solutions for facility location service area planning.</jats:p

Multiobjective Optimization for Planning the Service Areas of Smart Parcel Locker Facilities in Logistics Last Mile Delivery

The planning of the location service areas of smart parcel locker facilities became a critical aspect of logistics last mile delivery. In e-commerce, the efficiency of delivering merchandise from retailer warehouses to customers determines the competitiveness of retailers and delivery operators. The considerable increases in e-commerce transactions and safety concerns under the COVID-19 (Coronavirus disease 2019) pandemic made home delivery services even more inefficient than before, which resulted in the considerable increase in social costs. In numerous countries, smart parcel lockers were adopted to increase delivery efficiency, decrease the risk of COVID-19 infection, and reduce the burden on society. This study proposed a multiobjective optimization mathematical model for investigating the planning of the location service areas of smart parcel locker facilities, and then the optimization mathematical model was solved using a combination of the Taguchi method (TA) and nondominant sorting genetic algorithm II (NSGA-II). Finally, this composite approach was applied to a case study in producing favorable solutions for facility location service area planning

A K-Means Clustering and the Prim’s Minimum Spanning Tree-Based Optimal Picking-List Consolidation and Assignment Methodology for Achieving the Sustainable Warehouse Operations

Rapid industrialization has caused the concentration of greenhouse gases in the atmosphere to increase rapidly, leading to drastic global climate changes and ecological degradation. To establish a sustainable supply chain for consumer electronic products, this study focuses on warehouse operations and develops a K-means clustering and Prim’s minimum spanning tree-based optimal picking-list consolidation and assignment methodology. Compact camera modules are used to demonstrate and verify the effectiveness of this methodology. This methodology can be divided into two parts. First, the K-means clustering method is applied to conduct a picking-list consolidation analysis to create an optimal picking-list consolidation strategy for sustainable warehouse operations. Second, the most similar picking lists in each cluster are connected using Prim’s minimum spanning tree algorithm to generate the connected graph with the minimum spanning tree so as to establish a picking-list assignment strategy for sustainable warehouse operations. In this case study, this to-be model substantially reduced the traveling distance of the electric order-picking trucks within a warehouse and increased the picking efficiency to diminish the carbon emissions toward a sustainable supply chain.</jats:p

Multiobjective Optimization for Planning the Service Areas of Smart Parcel Locker Facilities in Logistics Last Mile Delivery

The planning of the location service areas of smart parcel locker facilities became a critical aspect of logistics last mile delivery. In e-commerce, the efficiency of delivering merchandise from retailer warehouses to customers determines the competitiveness of retailers and delivery operators. The considerable increases in e-commerce transactions and safety concerns under the COVID-19 (Coronavirus disease 2019) pandemic made home delivery services even more inefficient than before, which resulted in the considerable increase in social costs. In numerous countries, smart parcel lockers were adopted to increase delivery efficiency, decrease the risk of COVID-19 infection, and reduce the burden on society. This study proposed a multiobjective optimization mathematical model for investigating the planning of the location service areas of smart parcel locker facilities, and then the optimization mathematical model was solved using a combination of the Taguchi method (TA) and nondominant sorting genetic algorithm II (NSGA-II). Finally, this composite approach was applied to a case study in producing favorable solutions for facility location service area planning