Quantification of Silver Nanoparticle Interactions with Yeast Saccharomyces Cerevisiae Studied using Single-Cell ICP-MS

Silver nanoparticles (AgNPs) have been used in many fields due to their anticancer, antimicrobial, and antiviral potential. Single-cell ICP-MS (SC-ICP-MS) is an emerging technology that allows for the rapid characterization and quantification of a metal analyte across a cell population in a single analysis. In this study, a new rapid and sensitive SC-ICP-MS method was developed to quantitatively study the interactions of AgNPs with yeast Saccharomyces cerevisiae. The method can quantify the cell concentration, silver concentration per cell, and profile the nanoparticle distribution in a yeast cell population. AgNP dosing time, concentration, and AgNP size were quantitatively evaluated for their effects on AgNP-yeast cell interactions. The results showed that the initial uptake of AgNPs was rapid and primarily driven by the mass of Ag per cell. The optimal dosing particle concentrations for highest uptake were approximately 1820, 1000, and 300 AgNPs/cell for 10, 20, and 40 nm AgNPs, respectively. Furthermore, this study also validated a washing method for the application to a microorganism for the first time and was used to quantitatively determine the amount of cell surface–adsorbed AgNPs and intracellular AgNPs. These results indicated that the mass (Ag in ag/cell) ratios of intracelluar vs cell surface-adsorbed AgNPs were similar for different AgNP sizes. This high throughput and ultrasensitive SC-ICP-MS method is expected to have many potential applications, such as optimization of methods for green synthesis of AgNPs, nanotoxicity studies, and drug delivery. This is the first quantification study on the interactions of AgNPs and S. cerevisiae using SC-ICP-MS. Graphical abstract: [Figure not available: see fulltext.]

The connection between tricarboxylic acid cycle enzyme mutations and pseudohypoxic signaling in pheochromocytoma and paraganglioma

Pheochromocytomas and paragangliomas (PPGLs) are rare neuroendocrine tumors originating from chromaffin cells, holding significant clinical importance due to their capacity for excessive catecholamine secretion and associated cardiovascular complications. Roughly 80% of cases are associated with genetic mutations. Based on the functionality of these mutated genes, PPGLs can be categorized into distinct molecular clusters: the pseudohypoxia signaling cluster (Cluster-1), the kinase signaling cluster (Cluster-2), and the WNT signaling cluster (Cluster-3). A pivotal factor in the pathogenesis of PPGLs is hypoxia-inducible factor-2α (HIF2α), which becomes upregulated even under normoxic conditions, activating downstream transcriptional processes associated with pseudohypoxia. This adaptation provides tumor cells with a growth advantage and enhances their ability to thrive in adverse microenvironments. Moreover, pseudohypoxia disrupts immune cell communication, leading to the development of an immunosuppressive tumor microenvironment. Within Cluster-1a, metabolic perturbations are particularly pronounced. Mutations in enzymes associated with the tricarboxylic acid (TCA) cycle, such as succinate dehydrogenase (SDHx), fumarate hydratase (FH), isocitrate dehydrogenase (IDH), and malate dehydrogenase type 2 (MDH2), result in the accumulation of critical oncogenic metabolic intermediates. Notable among these intermediates are succinate, fumarate, and 2-hydroxyglutarate (2-HG), which promote activation of the HIFs signaling pathway through various mechanisms, thus inducing pseudohypoxia and facilitating tumorigenesis. SDHx mutations are prevalent in PPGLs, disrupting mitochondrial function and causing succinate accumulation, which competitively inhibits α-ketoglutarate-dependent dioxygenases. Consequently, this leads to global hypermethylation, epigenetic changes, and activation of HIFs. In FH-deficient cells, fumarate accumulation leads to protein succination, impacting cell function. FH mutations also trigger metabolic reprogramming towards glycolysis and lactate synthesis. IDH1/2 mutations generate D-2HG, inhibiting α-ketoglutarate-dependent dioxygenases and stabilizing HIFs. Similarly, MDH2 mutations are associated with HIF stability and pseudohypoxic response. Understanding the intricate relationship between metabolic enzyme mutations in the TCA cycle and pseudohypoxic signaling is crucial for unraveling the pathogenesis of PPGLs and developing targeted therapies. This knowledge enhances our comprehension of the pivotal role of cellular metabolism in PPGLs and holds implications for potential therapeutic advancements

Lipid and glucose metabolism in senescence

Senescence is an inevitable biological process. Disturbances in glucose and lipid metabolism are essential features of cellular senescence. Given the important roles of these types of metabolism, we review the evidence for how key metabolic enzymes influence senescence and how senescence-related secretory phenotypes, autophagy, apoptosis, insulin signaling pathways, and environmental factors modulate glucose and lipid homeostasis. We also discuss the metabolic alterations in abnormal senescence diseases and anti-cancer therapies that target senescence through metabolic interventions. Our work offers insights for developing pharmacological strategies to combat senescence and cancer

Association between cathepsins and benign prostate diseases: a bidirectional two-sample Mendelian randomization study

ObjectivesThe relationship between cathepsins and prostate cancer (PCa) has been reported. However, there is a lack of research on cathepsins and benign prostate diseases (BPDs). This study investigated the potential genetic link between cathepsins and BPDs through the utilization of Mendelian randomization (MR) analysis to determine if a causal relationship exists.MethodsPublicly accessible summary statistics on BPDs were obtained from FinnGen Biobank. The data comprised 149,363 individuals, with 30,066 cases and 119,297 controls for BPH, and 123,057 individuals, with 3,760 cases and 119,297 controls for prostatitis. The IEU OpenGWAS provided the Genome-wide association data on ten cathepsins. To evaluate the causal relationship between BPDs and cathepsins, five distinct MR analyses were employed, with the primary method being the inverse variance weighted (IVW) approach. Additionally, sensitivity analyses were conducted to examine the horizontal pleiotropy and heterogeneity of the findings.ResultsThe examination of IVW MR findings showed that cathepsin O had a beneficial effect on BPH (IVW OR=0.94, 95% CI 0.89–0.98, P=0.0055), while cathepsin X posed a threat to prostatitis (IVW OR=1.08, 95% CI 1.00–1.16, P=0.047). Through reverse MR analysis, it was revealed that prostatitis had an adverse impact on cathepsin V (IVW OR=0.89, 95% CI 0.80–0.99, P=0.035), while no favorable association was observed between BPH and cathepsins. The results obtained from MR-Egger, weighted median, simple mode, and weighted mode methods were consistent with the findings of the IVW approach. Based on sensitivity analyses, heterogeneity, and horizontal pleiotropy are unlikely to distort the results.ConclusionThis study offers the initial evidence of a genetic causal link between cathepsins and BPDs. Our findings revealed that cathepsin O was beneficial in preventing BPH, whereas cathepsin X posed a potential threat to prostatitis. Additionally, prostatitis negatively affected cathepsin V level. These three cathepsins could be targets of diagnosis and treatment for BPDs, which need further research

Identification and Typing of Human Enterovirus: A Genomic Barcode Approach

Identification and typing of human enterovirus (HEVs) are important to pathogen detection and therapy. Previous phylogeny-based typing methods are mainly based on multiple sequence alignments of specific genes in the HEVs, but the results are not stable with respect to different choices of genes. Here we report a novel method for identification and typing of HEVs based on information derived from their whole genomes. Specifically, we calculate the k-mer based barcode image for each genome, HEV or other human viruses, for a fixed k, 1<k<7, where a genome barcode is defined in terms of the k-mer frequency distribution across the whole genome for all combinations of k-mers. A phylogenetic tree is constructed using a barcode-based distance and a neighbor-joining method among a set of 443 representative non-HEV human viruses and 395 HEV sequences. The tree shows a clear separation of the HEV viruses from all the non-HEV viruses with 100% accuracy and a separation of the HEVs into four distinct clads with 93.4% consistency with a multiple sequence alignment-based phylogeny. Our detailed analyses of the HEVs having different typing results by the two methods indicate that our results are in better agreement with known information about the HEVs

Characterization of Neuraminidases from the Highly Pathogenic Avian H5N1 and 2009 Pandemic H1N1 Influenza A Viruses

To study the precise role of the neuraminidase (NA), and its stalk region in particular, in the assembly, release, and entry of influenza virus, we deleted the 20-aa stalk segment from 2009 pandemic H1N1 NA (09N1) and inserted this segment, now designated 09s60, into the stalk region of a highly pathogenic avian influenza (HPAI) virus H5N1 NA (AH N1). The biological characterization of these wild-type and mutant NAs was analyzed by pseudotyped particles (pseudoparticles) system. Compared with the wild-type AH N1, the wild-type 09N1 exhibited higher NA activity and released more pseudoparticles. Deletion/insertion of the 09s60 segment did not alter this relationship. The infectivity of pseudoparticles harboring NA in combination with the hemagglutinin from HPAI H5N1 (AH H5) was decreased by insertion of 09s60 into AH N1 and was increased by deletion of 09s60 from 09N1. When isolated from the wild-type 2009H1N1 virus, 09N1 existed in the forms (in order of abundance) dimer>>tetramer>monomer, but when isolated from pseudoparticles, 09N1 existed in the forms dimer>monomer>>>tetramer. After deletion of 09s60, 09N1 existed in the forms monomer>>>dimer. AH N1 from pseudoparticles existed in the forms monomer>>dimer, but after insertion of 09s60, it existed in the forms dimer>>monomer. Deletion/insertion of 09s60 did not alter the NA glycosylation pattern of 09N1 or AH N1. The 09N1 was more sensitive than the AH N1 to the NA inhibitor oseltamivir, suggesting that the infectivity-enhancing effect of oseltamivir correlates with robust NA activity

The trans-ancestral genomic architecture of glycemic traits

Glycemic traits are used to diagnose and monitor type 2 diabetes and cardiometabolic health. To date, most genetic studies of glycemic traits have focused on individuals of European ancestry. Here we aggregated genome-wide association studies comprising up to 281,416 individuals without diabetes (30% non-European ancestry) for whom fasting glucose, 2-h glucose after an oral glucose challenge, glycated hemoglobin and fasting insulin data were available. Trans-ancestry and single-ancestry meta-analyses identified 242 loci (99 novel; P < 5 x 10(-8)), 80% of which had no significant evidence of between-ancestry heterogeneity. Analyses restricted to individuals of European ancestry with equivalent sample size would have led to 24 fewer new loci. Compared with single-ancestry analyses, equivalent-sized trans-ancestry fine-mapping reduced the number of estimated variants in 99% credible sets by a median of 37.5%. Genomic-feature, gene-expression and gene-set analyses revealed distinct biological signatures for each trait, highlighting different underlying biological pathways. Our results increase our understanding of diabetes pathophysiology by using trans-ancestry studies for improved power and resolution. A trans-ancestry meta-analysis of GWAS of glycemic traits in up to 281,416 individuals identifies 99 novel loci, of which one quarter was found due to the multi-ancestry approach, which also improves fine-mapping of credible variant sets.Peer reviewe

The relationship between criminology and criminal law: implications for developing Chinese criminology

Abstract During the last four decades, Chinese criminology has grown steadily but modestly. One consensus is the lack of a clear understanding of the relationship between criminology and criminal law. This paper attempts to provide a critical review of the relationship between the two disciplines by first tracing the historical development of criminology in both China and Western countries. It then clarifies the distinctions and overlaps between criminology and criminal law and explains how this relationship has influenced the growth of criminology in China. It concludes by proposing implications for developing criminology in China, particularly discussing potential collaborations that could be forged between criminology and criminal law and how such partnerships can benefit both fields in China

Quantification of Silver Nanoparticle Interactions with Yeast Saccharomyces Cerevisiae Studied using Single-cell ICP-MS

Silver nanoparticles (AgNPs) have been used in many fields due to their anticancer, antimicrobial, and antiviral potential. Single-cell ICP-MS (SC-ICP-MS) is an emerging technology that allows for the rapid characterization and quantification of a metal analyte across a cell population in a single analysis. In this study, a new rapid and sensitive SC-ICP-MS method was developed to quantitatively study the interactions of AgNPs with yeast Saccharomyces cerevisiae. The method can quantify the cell concentration, silver concentration per cell, and profile the nanoparticle distribution in a yeast cell population. AgNP dosing time, concentration, and AgNP size were quantitatively evaluated for their effects on AgNP-yeast cell interactions. The results showed that the initial uptake of AgNPs was rapid and primarily driven by the mass of Ag per cell. The optimal dosing particle concentrations for highest uptake were approximately 1820, 1000, and 300 AgNPs/cell for 10, 20, and 40 nm AgNPs, respectively. Furthermore, this study also validated a washing method for the application to a microorganism for the first time and was used to quantitatively determine the amount of cell surface-adsorbed AgNPs and intracellular AgNPs. These results indicated that the mass (Ag in ag/cell) ratios of intracelluar vs cell surface-adsorbed AgNPs were similar for different AgNP sizes. This high throughput and ultrasensitive SC-ICP-MS method is expected to have many potential applications, such as optimization of methods for green synthesis of AgNPs, nanotoxicity studies, and drug delivery. This is the first quantification study on the interactions of AgNPs and S. cerevisiae using SC-ICP-MS