Protein instability and functional defects caused by mutations of dihydro-orotate dehydrogenase in Miller syndrome patients

Synopsis Miller syndrome is a recessive inherited disorder characterized by postaxial acrofacial dysostosis. It is caused by dysfunction of the DHODH (dihydroorotate dehydrogenase) gene, which encodes a key enzyme in the pyrimidine de novo biosynthesis pathway and is localized at mitochondria intermembrane space. We investigated the consequence of three missense mutations, G202A, R346W and R135C of DHODH, which were previously identified in patients with Miller syndrome. First, we established HeLa cell lines stably expressing DHODH with Miller syndrome-causative mutations: G202A, R346W and R135C. These three mutant proteins retained the proper mitochondrial localization based on immunohistochemistry and mitochondrial subfractionation studies. The G202A, R346W DHODH proteins showed reduced protein stability. On the other hand, the third one R135C, in which the mutation lies at the ubiquinone-binding site, was stable but possessed no enzymatic activity. In conclusion, the G202A and R346W mutation causes deficient protein stability, and the R135C mutation does not affect stability but impairs the substrate-induced enzymatic activity, suggesting that impairment of DHODH activity is linked to the Miller syndrome phenotype

Spatially restricted drivers and transitional cell populations cooperate with the microenvironment in untreated and chemo-resistant pancreatic cancer

Pancreatic ductal adenocarcinoma is a lethal disease with limited treatment options and poor survival. We studied 83 spatial samples from 31 patients (11 treatment-naïve and 20 treated) using single-cell/nucleus RNA sequencing, bulk-proteogenomics, spatial transcriptomics and cellular imaging. Subpopulations of tumor cells exhibited signatures of proliferation, KRAS signaling, cell stress and epithelial-to-mesenchymal transition. Mapping mutations and copy number events distinguished tumor populations from normal and transitional cells, including acinar-to-ductal metaplasia and pancreatic intraepithelial neoplasia. Pathology-assisted deconvolution of spatial transcriptomic data identified tumor and transitional subpopulations with distinct histological features. We showed coordinated expression of TIGIT in exhausted and regulatory T cells and Nectin in tumor cells. Chemo-resistant samples contain a threefold enrichment of inflammatory cancer-associated fibroblasts that upregulate metallothioneins. Our study reveals a deeper understanding of the intricate substructure of pancreatic ductal adenocarcinoma tumors that could help improve therapy for patients with this disease

Influence of human factors on emergency evacuation efficiency of nuclear power plant accident

Through questionnaire analysis, the cognitive situation and behavioral characteristics of different people around the nuclear power plant for nuclear emergency evacuation were obtained, and the possible behaviors of people during evacuation were predicted by Logistic model. Chi-square test and Logistic regression model were used to make correlation analysis and influence judgment on the evacuation psychology and evacuation behavior of different people in the area, and the optimal allocation algorithm was used to judge the prediction correctness of the model. The results showed that the factors such as whether to purchase nuclear safety-related insurance, the number of nuclear emergency evacuation drills, the understanding of nuclear emergency evacuation, and the presence of non-nuclear emergency protective measures at home had significant effects on the evacuation consciousness and psychology of the population. Gender, knowledge of nuclear emergency evacuation, the number of nuclear emergency evacuation drills participated, whether there are nuclear emergency protection measures at home and whether nuclear safety insurance has been purchased, and other factors have significant influence on crowd evacuation behavior. The simulation shows that people with certain nuclear evacuation experience can make relatively correct evacuation behavior and reduce evacuation time

Application of Quality Control Circle in Promoting the Use of Rubber Dams in the Root Canal Treatment of Primary Teeth

Objective. Study the effect of quality control circle (QCC) in promoting the usage of rubber dams (RD) in root canal treatment of chronic pulpitis in primary teeth. Methods. Set up a quality control group to increase the amount of rubber dams used in the treatment of chronic pulpitis in primary teeth. Monthly monitoring results of the usage amount were counted by the outpatient computer system. Relevant data were collected through questionnaires, and causes of low utilization were analyzed, and the improvement measurements were formulated and implemented. Quality control circle activity was evaluated. Results. Through the quality control circle activity, the consumption of rubber dams in the root canal treatment of chronic pulpitis was significantly improved, children in treatment became more cooperative, and operation time of root canal treatment has also been shortened. Conclusion. The quality control circle activities played a significant role in promoting the use of rubber dams in the root canal treatment of primary teeth, and it can be used as a method to promote new clinical treatment programs

Catalyst-Free Decarboxylative Trifluoromethylation/Perfluoroalkylation of Benzoic Acid Derivatives in Water–Acetonitrile

This study presents a mild and practical method for the catalyst-free decarboxylative trifluoromethylation/perfluoroalkylation of benzoic acid derivatives by using inexpensive NaSO₂CF₃ as a CF₃-source and solid Na₂S₂O₈ as an initiator. The environment-friendly transformation is performed at 30 °C in water–acetonitrile, and it can be easily scaled up to the gram level with a good yield

A Microimage-Processing-Based Technique for Detecting Qualitative and Quantitative Characteristics of Plant Cells

When plants encounter external environmental stimuli, they can adapt to environmental changes through a complex network of metabolism–gene expression–metabolism within the plant cell. In this process, changes in the characteristics of plant cells are a phenotype that is responsive and directly linked to this network. Accurate identification of large numbers of plant cells and quantitative analysis of their cellular characteristics is a much-needed experiment for in-depth analysis of plant metabolism and gene expression. This study aimed to develop an automated, accurate, high-throughput quantitative analysis method, ACFVA, for single-plant-cell identification. ACFVA can quantitatively address a variety of biological questions for a large number of plant cells automatically, including standard assays (for example, cell localization, count, and size) and complex morphological assays (for example, different fluorescence in cells). Using ACFVA, phenomics studies can be carried out at the plant cellular level and then combined with ever-changing sequencing technologies to address plant molecular biology and synthetic biology from another direction

Immune responses to SARS-CoV-2 infection in Humans and ACE2 humanized mice

The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) represents a major public health threat worldwide. Insight into protective and pathogenic aspects of SARS-CoV-2 immune responses is critical to work out effective therapeutics and develop vaccines for controlling the disease. Here, we review the present literature describing the innate and adaptive immune responses including innate immune cells, cytokine responses, antibody responses and T cell responses against SARS-CoV-2 in human infection, as well as in AEC2-humanized mouse infection. We also summarize the now known and unknown about the role of the SARS-CoV-2 immune responses. By better understanding the mechanisms that drive the immune responses, we can tailor treatment strategies at specific disease stages and improve our response to this worldwide public health threat

Dietary Quality and Relationships with Metabolic Dysfunction-Associated Fatty Liver Disease (MAFLD) among United States Adults, Results from NHANES 2017–2018

Metabolic dysfunction-associated fatty liver disease (MAFLD) is a new definition for the evidence of hepatic steatosis and metabolic dysfunctions. The specific role of the dietary factors in the development and progress of the disease are not well illuminated. Thus, we conducted this study on the associations between dietary quality assessed by five dietary quality indexes (Dietary Inflammatory Index, DII; Mediterranean diet, MED; Dietary Approach to Stop Hypertension, DASH; Alternate Healthy Eating Index diet, AHEI; Healthy Eating Indices, HEI) and MAFLD phenotypes. This study was extracted from the latest NHANES 2017–2018 wave. Demographic information, health status, lifestyles, and dietary habits were reported in the questionnaire. Multivariate logistic regression and multivariate ordinal logistic regression methods were applied to explore the associations between dietary quality indexes and MAFLD or MAFLD with liver fibrosis. The weighted prevalence of Non-MAFLD, MAFLD without fibrosis, and MAFLD with fibrosis were 47.05%, 36.67%, and 16.28%, respectively, at the cutoff value of a median Controlled Attenuation Parameter (CAP) 248 dB/m and a median Liver Stiffness Measurement (LSM) 6.3 kPa. When the diagnostic cutoff values of CAP changed to 285 dB/m, the weighted prevalence of Non-MAFLD, MAFLD without liver fibrosis, and MAFLD with fibrosis turned to 64.62%, 22.08%, and 13.30%, respectively. All five dietary quality indexes, including DII, HEI-2015, AHEI, DASH, and MED, were all significantly associated with MAFLD phenotypes. DII was positively associated with MAFLD phenotypes, while other four dietary quality indexes, including HEI-2015, AHEI, DASH, and MED, were significantly associated with lower risk of MAFLD phenotypes. MAFLD is becoming a threatening public health concern among adult Americans and dietary quality is markedly associated with MAFLD phenotypes