Characterization of nuclear transport pathways of heat shock protein 70 upon ethanol stress in the budding yeast Saccharomyces cerevisiae

The main objective of this thesis is to characterize the mechanisms of nuclear transport of the cytosolic yeast heat shock protein 70 Ssa4p upon ethanol stress and the effect of stress and nutrient availability on transport carriers.Heat shock protein 70 (Hsp70) plays an important role in cell damage recovery and transport of proteins across intracellular membranes. Upon stress, trafficking of the majority of proteins is inhibited with the exception of Hsp70 that accumulates in the nucleus. Using the budding yeast Saccharomyces cerevisiae as model, this thesis specifically demonstrates that Ssa4p transiently and reversibly accumulates in nuclei upon ethanol stress and exports to the cytoplasm during ethanol recovery. The cNLS-mediated classical nuclear import pathway is not required for stress-induced Ssa4p nuclear trafficking, and the N-terminal 236 amino acid residues are sufficient for Ssa4p nuclear accumulation. The importin beta family member Nmd5p forms import complexes with its cargo Ssa4p and translocates cargo into the nucleus. Upon withdrawal of ethanol stress, Ssa4p exports to the cytoplasm by binding to the exporter Msn5p. Transport of Ssa4p in both directions requires the Gsp1p system. Pkc1p and the cell integrity pathway sensors are involved in Ssa4p nuclear import upon ethanol treatment.The thesis also states that ethanol stress regulates Ssa4p nuclear transport by enhancing the import complex Ssa4p/Nmd5p formation and the docking of Nmd5p at the NPC. Stresses regulate Ssa4p nuclear export by regulating exporter Msn5p cellular localization and formation of the export complex Ssa4p/Msn5p. Therefore, the carrier-mediated nuclear trafficking might also be regulated by the transport carrier localization. Furthermore, intracellular localization of yeast nuclear exporters is sensitive to different stresses and regulated by Snf1 kinase and cell nutrient availability.In summary, this thesis defines the nuclear import and export mechanisms for the Hsp70 Ssa4p and their regulation in stressed cells. The novel finding of how the nutritional state and Snf1 kinase regulate exporter localization in general may provide a better understanding of protein transport regulation and the multiple levels cells use to adjust to environmental changes

The immune factors have complex causal regulation effects on inflammatory bowel disease

BackgroundAlthough a correlation between immune cell phenotypes and inflammatory bowel disease (IBD) has been established, a causal relationship remains unestablished.MethodsTo assess causal associations between immune cell phenotypes and IBD and its subtypes, we employed Mendelian randomization (MR) methods and genome-wide association studies (GWAS) summary statistics. The primary outcomes were determined based on the inverse variance weighting (IVW) results, with the assessment of heterogeneity and pleiotropy conducted through Cochrane’s Q-test and MR-Egger. The stability of the MR results was then examined using leave-one-out analysis, and false discovery rate (FDR) correction was applied to evaluate the strength of the causal relationship between exposure and outcome. Furthermore, to identify immunophenotypes strongly associated with IBD, a meta-integration of the effect values of all positive results in both datasets was conducted.ResultsThe analysis of 731 immune cell phenotypes and IBD using MR techniques revealed potential causal associations between 26 phenotypes and IBD. Subsequent meta-integration of the two datasets provided evidence of solid causal associations between 18 immune phenotypes and IBD and its subtypes. Nominal causal associations were also identified in the remaining eight immune phenotypes and IBD and its subtypes.ConclusionOur study confirms causal solid associations between 18 immune phenotypes and IBD, thus guiding future clinical studies

A machine learning-based model for predicting distant metastasis in patients with rectal cancer

BackgroundDistant metastasis from rectal cancer usually results in poorer survival and quality of life, so early identification of patients at high risk of distant metastasis from rectal cancer is essential.MethodThe study used eight machine-learning algorithms to construct a machine-learning model for the risk of distant metastasis from rectal cancer. We developed the models using 23867 patients with rectal cancer from the Surveillance, Epidemiology, and End Results (SEER) database between 2010 and 2017. Meanwhile, 1178 rectal cancer patients from Chinese hospitals were selected to validate the model performance and extrapolation. We tuned the hyperparameters by random search and tenfold cross-validation to construct the machine-learning models. We evaluated the models using the area under the receiver operating characteristic curves (AUC), the area under the precision-recall curve (AUPRC), decision curve analysis, calibration curves, and the precision and accuracy of the internal test set and external validation cohorts. In addition, Shapley’s Additive explanations (SHAP) were used to interpret the machine-learning models. Finally, the best model was applied to develop a web calculator for predicting the risk of distant metastasis in rectal cancer.ResultThe study included 23,867 rectal cancer patients and 2,840 patients with distant metastasis. Multiple logistic regression analysis showed that age, differentiation grade, T-stage, N-stage, preoperative carcinoembryonic antigen (CEA), tumor deposits, perineural invasion, tumor size, radiation, and chemotherapy were-independent risk factors for distant metastasis in rectal cancer. The mean AUC value of the extreme gradient boosting (XGB) model in ten-fold cross-validation in the training set was 0.859. The XGB model performed best in the internal test set and external validation set. The XGB model in the internal test set had an AUC was 0.855, AUPRC was 0.510, accuracy was 0.900, and precision was 0.880. The metric AUC for the external validation set of the XGB model was 0.814, AUPRC was 0.609, accuracy was 0.800, and precision was 0.810. Finally, we constructed a web calculator using the XGB model for distant metastasis of rectal cancer.ConclusionThe study developed and validated an XGB model based on clinicopathological information for predicting the risk of distant metastasis in patients with rectal cancer, which may help physicians make clinical decisions. rectal cancer, distant metastasis, web calculator, machine learning algorithm, external validatio

Biomaterial-assisted tumor therapy: A brief review of hydroxyapatite nanoparticles and its composites used in bone tumors therapy

Malignant bone tumors can inflict significant damage to affected bones, leaving patients to contend with issues like residual tumor cells, bone defects, and bacterial infections post-surgery. However, hydroxyapatite nanoparticles (nHAp), the principal inorganic constituent of natural bone, possess numerous advantages such as high biocompatibility, bone conduction ability, and a large surface area. Moreover, nHAp’s nanoscale particle size enables it to impede the growth of various tumor cells via diverse pathways. This article presents a comprehensive review of relevant literature spanning the past 2 decades concerning nHAp and bone tumors. The primary goal is to explore the mechanisms responsible for nHAp’s ability to hinder tumor initiation and progression, as well as to investigate the potential of integrating other drugs and components for bone tumor diagnosis and treatment. Lastly, the article discusses future prospects for the development of hydroxyapatite materials as a promising modality for tumor therapy

SARS-CoV-2 delta (B.1.617.2) spike protein adjuvanted with Alum-3M-052 enhances antibody production and neutralization ability

BackgroundOptimizing adjuvant is one of the critical methods to improve the vaccine. 3M-052, a novel TLR7/8 agonist which was designed for slow dissemination at the injection site, has a potential as adjuvant, but its performance as a vaccine adjuvant for SARS-CoV-2 (B.1.617.2) spike protein has not been studied. The present study aimed to evaluate the effect of Alum-3M-052 as an adjuvant to improve mice serum antibody titers and pseudovirus neutralization efficiency.MethodFemale Balb/c mice were immunized 3 times at day 0, 7 and 21 intramuscularly with SARS-CoV-2 (B.1.617.2) spike protein and adjuvant (Alum or Alum-3M-052). Mice serum was collected weekly since day 7. Antibody titers of mice serum anti-SARS-CoV-2 (B.1.617.2) IgG and IgM were detected by ELISA. Inhibition rates of mice serum blocking SARS-CoV-2 (B.1.617.2) spike protein binding to ACE2 were detected by SARS-CoV-2 (B.1.617.2) Inhibitor Screening Kit. Neutralization efficiencies of mice serum against both SARS-CoV-2 (BA.2.12.1) pseudovirus and SARS-CoV-2 (B.1.617.2) pseudovirus were detected by pseudovirus neutralizing assay.ResultSerum of mice immunized by SARS-CoV-2 (B.1.617.2) spike protein adjuvanted with Alum-3M-052 had highest antibody titers and higher neutralization efficiency against both SARS-CoV-2 (BA.2.12.1) pseudovirus and SARS-CoV-2 (B.1.617.2) pseudovirus. Besides, neutralization efficiency of anti-SARS-CoV-2 (B.1.617.2) spike protein antibody against SARS-CoV-2 (BA.2.12.1) pseudovirus was lower than that of SARS-CoV-2 (B.1.617.2) pseudovirus.ConclusionAlum-3M-052 rapidly increased the titer of anti-SARS-CoV-2 (B.1.617.2) spike protein neutralizing antibodies and enhanced the neutralization ability against pseudoviruses and variants. This study provided evidence for the application of Alum-3M-052 as an adjuvant in COVID-19 vaccines production

High-efficiency polarization multiplexing metalenses

The polarization multiplexing technique is a well-established method that improves the communication capacity of an optical system. In this paper, we designed orthogonal linear and circular polarization multiplexing metalens using a library of rectangle TiO2 nanostructures. The former can independently focus x- and y-linearly polarized incident lights to designed positions with a focusing efficiency of 53.81% and 51.56%, respectively, whereas the latter with two preset focal points can independently control left and right circularly polarized incident lights with a focusing efficiency of 42.45% and 42.46%, respectively. We also show that both metalenses can produce diffraction-limited focal spots for four polarization states with no obvious distortion, which opens up new applications in polarization imaging and polarization detection