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The increased use of next‐generation sequencing has led to the detection of pathogenic TP53 variants in the germline setting in patients without a personal or family history consistent with Li–Fraumeni syndrome (LFS). These variants can represent low‐penetrance LFS, mosaic LFS, or clonal hematopoiesis of indeterminate potential. Additionally, TP53 variants of uncertain significance can be detected in patients with a history suspicious for LFS. The interpretation of the significance of these variants can be challenging but is crucial for an accurate diagnosis and appropriate medical management. This retrospective case review provides illustrative examples of the interpretation of challenging TP53 results through multidisciplinary expertise and use of a flowchart. The authors describe eight patients with TP53 variants associated with ambiguous diagnoses and, for each case, describe how the results were interpreted and the medical care that was implemented. This report presents illustrative cases to help guide clinicians to reach definitive diagnoses for patients when confronted with TP53 variants that are inconsistent with the clinical picture and to add to the body of literature regarding interpretation and medical management of TP53 variants discovered on germline testing. [ABSTRACT FROM AUTHOR]

Transcription factor (TF) residence on chromatin translates into quantitative transcriptional or structural outcomes on genome. Commonly used formaldehyde crosslinking fixes TF-DNA interactions cumulatively and compromises the measured occupancy level. Here we mapped the occupancy level of global or individual zinc finger TFs like CTCF and MAZ, in the form of highly resolved footprints, on native chromatin. By incorporating reinforcing perturbation conditions, we established S-score, a quantitative metric to proxy the continuum of CTCF or MAZ retention across different motifs on native chromatin. The native chromatin-retained CTCF sites harbor sequence features within CTCF motifs better explained by S-score than the metrics obtained from other crosslinking or native assays. CTCF retention on native chromatin correlates with local SUMOylation level, and anti-correlates with transcriptional activity. The S-score successfully delineates the otherwise-masked differential stability of chromatin structures mediated by CTCF, or by MAZ independent of CTCF. Overall, our study established a paradigm continuum of TF retention across binding sites on native chromatin, explaining the dynamic genome organization. Synopsis: The measured TF occupancy level is compromised when chromatin is chemically crosslinked. Profiling TF occupancy on native chromatin restores a more unperturbed picture of TF retention and better explains dynamic genome organization. loMNase-seq and N-ChIP identify highly resolved TF footprints on native chromatin. TFs are continuously "stripped off" across binding sites on native chromatin under increasing salt concentration. The S-score derived from such series of N-ChIP experiments delineates TF binding sites with differential stability under stress. The S-score explains a spectrum of stability of TF-mediated chromatin loops under stress. The measured TF occupancy level is compromised when chromatin is chemically crosslinked. Profiling TF occupancy on native chromatin restores a more unperturbed picture of TF retention and better explains dynamic genome organization. [ABSTRACT FROM AUTHOR]

During the machining of aluminum alloys, the adhesion of chips to the tool affects the performance characteristics. Today, different cooling systems are used to eliminate these negativities. In this study, the effects of end milling using HSS and carbide cutting tools of 6061-T6 aluminum alloy on surface roughness, chip thickness ratio and tool wear were examined using different cooling techniques (dry, minimum quantity lubrication (MQL) and nanocutting fluid). Different cutting speeds (180, 200, 220 m/min) and different feed rates (0.05, 0.06, 0.07 mm/rev) were used in the experiments. According to experimental findings, tool wear and surface roughness decreased at low cutting speed and feed rate by using nanocutting fluid with carbide cutting tools. It has been observed that the chip thickness ratio increases with high cutting speeds using nanocutting fluid and decreases with dry machining and high feed rates. The best milling performance of the aluminum alloy was achieved in experiments using carbide cutting tools and nanocutting fluid. [ABSTRACT FROM AUTHOR]

The involvement of the molecular clock in regulating cell physiological processes on a specific time scale is a recognized concept, yet its specific impact on optimizing androgen production in Leydig cells has been unclear. This study aimed to confirm the role of the REVERBA (NR1D1) gene in controlling the transcription of key genes related to Leydig cell steroid production. We investigated daily variations by collecting Leydig cells from rats at various times within a 24‐h period. Chromatin immunoprecipitation study showed a time‐dependent pattern for genes linked to steroid production (Nur77, Star, Cyp11a1, and Cyp17a1), which closely matched the 24‐h REVERBA levels in Leydig cells, peaking between zeitgeber time (ZT) 7–11. To understand the physiological significance of REVERBA's interaction with promoters of steroidogenesis‐related genes, Leydig cells from rats at two different times (ZT7 and ZT16; chosen based on REVERBA expression levels), were treated with either an agonist (GSK4112) or an antagonist (SR8278). The results revealed that the REVERBA agonist stimulated gene transcription, while the antagonist inhibited it, but only when REVERBA was sufficiently present, indicating a reliance on REVERBA's circadian fluctuation. Moreover, this REVERBA‐dependent stimulation had a clear impact on testosterone production in the culture medium, underscoring REVERBA's involvement in the circadian regulation of testosterone. This study indicates that REVERBA, in addition to being a core component of the cellular clock, plays a key role in regulating androgen production in Leydig cells by influencing the transcription of critical steroidogenesis‐related genes. [ABSTRACT FROM AUTHOR]

Introduction: High-risk procedures in interventional cardiology include a wide spectrum of clinical and anatomical scenarios related to a higher periprocedural morbidity and mortality. The prophylactic use of short-term mechanical circulatory support (ST-MCS) may improve both the safety and efficacy of the intervention by leading to more stable procedural hemodynamics. However, the significant costs may limit its use in resource constrained settings. To overcome this limitation, we ideated a modified, low-cost, veno-arterial extracorporeal membrane oxygenator (V-A ECMO) setup. Methods: We conducted an observational prospective study including all patients undergoing a high-risk interventional cardiology procedure at our institution under prophylactic ST-MCS using a modified, low-cost version of V-A ECMO, where some components of the standard V-A ECMO circuit were replaced by supplies used for cardiac surgical cardiopulmonary bypass, achieving a cost reduction of 72%. We assessed in-hospital and mid-term outcomes, including procedural success, post-procedure complications and mortality. Results: Between March 2016 and December 2021, ten patients underwent high-risk IC procedures with prophylactic use of V-A ECMO. Isolated percutaneous intervention (PCI) was performed in six patients, isolated transcatheter aortic valve replacement (TAVR) in two, and a combined procedure (PCI + TAVR) in two. Mean ejection fraction was 34% (range 20–64%). Mean STS PROM was 16.2% (range 9.5–35.8%) and mean EuroScore was 23.7% (range 1.5–60%). The planned intervention was successfully performed in all cases. There were no reports of V-A ECMO malfunction. In nine patients the VA-ECMO was withdrawn immediately after the procedure but one patient required extended - 24 h - support with no significant issues. One patient experienced a periprocedural myocardial infarction and another developed a femoral pseudoaneurysm. In-hospital and 30-day survival were 100%, and 1-year survival was 80%. Conclusions: High-risk procedures in interventional cardiology can be successfully performed under prophylactic ST-MCS using a modified, low-cost V-A ECMO, suitable for limited-resource settings. [ABSTRACT FROM AUTHOR]

In the field of biomedicine, efficiently and non-invasively isolating target cells has always been one of the core challenges. Optical fiber tweezers offer precise and non-invasive manipulation of cells within a medium and can be easily integrated with microfluidic systems. Therefore, this paper investigated the mechanism of cell manipulation using scattering force with optical fiber tweezers. We employed flat-ended single-mode fiber to drive and sort cells and derived the corresponding scattering force formula based on the T-matrix model. A single-mode optical tweezers system for cell sorting was developed, and an optofluidic experimental platform was constructed that effectively integrates the optical system with microfluidic chips. The chip, featuring an expanded cross-channel design, successfully achieved continuous separation of yeast cells (8~10 µm in diameter) and polystyrene microspheres (15~20 µm in diameter), with a sorting efficiency of up to 86% and maintaining viability in approximately 90% of the yeast cells. Compared to other sorting systems, this system does not require labeling and can achieve continuous sorting with cell viability at a lower cost of instrumentation. [ABSTRACT FROM AUTHOR]

We fabricated a microfluidic chip (osteoblast [OB]–osteoclast [OC] chip) that could regulate the mixture amounts of OB and OC supernatants to investigate the effect of different supernatant distributions on osteogenesis or osteoclastogenesis. Computer-aided design was used to produce an OB–OC chip from polydimethylsiloxane. A pressure controller was assembled and different blends of OB and OC supernatants were correctly determined. OB and OC supernatants were placed on the upper panels of the OB–OC chip after differentiation for an in vitro evaluation. We then tested the changes in osteogenesis using MC3T3-E1 cells in the middle chambers. We observed that a 75:25 distribution of OB and OC supernatants was the most potent in osteogenesis. We then primed the osteogenic differentiation of MC3T3-E1 cells using an OB–OC mixed supernatant or an OB supernatant alone (supernatant ratios of 75:25 or 100:0, respectively). These cells were placed on the calvarial defect sites of rats. Microcomputed tomography and histological analyses determined a significantly higher bone formation in the group exposed to the OB–OC supernatant at a ratio of 75:25. In this study, we demonstrate the applicability of an OB–OC chip to evaluate the effect of different supernatant distributions of OB and OC. We observed that the highest bone-forming potential was in MC3T3-E1 cells treated with conditioned media, specifically the OB–OC supernatant at a ratio of 75:25. [ABSTRACT FROM AUTHOR]

Purpose: Cancer cells are characterized as the uncontrolled proliferation, which demands high levels of nucleotides that are building blocks for DNA synthesis and replication. CAD (carbamoyl-phosphate synthetase 2, aspartate transcarbamylase and dihydroorotase) is a trifunctional enzyme that initiates the de novo pyrimidine synthesis, which is normally enhanced in cancer cells to preserve the pyrimidine pool for cell division. Glioma, representing most brain cancer, is highly addicted to nucleotides like pyrimidine to sustain the abnormal growth and proliferation of cells. CAD is previously reported to be dysregulated in glioma, but the underlying mechanism remains unclear. Methods: The expression of CAD and CHIP (carboxyl terminus of Hsc70-interacting protein) protein in normal brain cells and three glioblastoma (GBM) cell lines were measured by immunoblots. Lentiviruses-mediated expression of target proteins or shRNAs were used to specifically overexpress or knock down CAD and CHIP. Cell counting, colony formation, apoptosis and cell cycle assays were used to assess the roles of CAD and CHIP in GBM cell proliferation and survival. Co-immunoprecipitation and ubiquitination assays were used to examine the interaction of CHIP with CAD and the ubiquitination of CAD. The correlation of CAD and CHIP expression with GBM patients' survival was obtained by analyzing the GlioVis database. Results: In this study, we showed that the expression of CAD was upregulated in glioma, which was positively correlated with the tumor grade and survival of glioma patients. Knockdown of CAD robustly inhibited the cell proliferation and colony formation of GBM cells, indicating the essential role of CAD in the pathogenesis of GBM. Mechanistically, we firstly identified that CAD was modified by the K29-linked polyubiquitination, which was mediated by the E3 ubiquitin ligase CHIP. By interacting with and ubiquitinating CAD, CHIP enhanced its proteasomal and lysosomal degradation, which accounted for the anti-proliferative role of CHIP in GBM cells. To sustain the expression of CAD, CHIP is significantly downregulated, which is correlated with the poor prognosis and survival of GBM patients. Notably, the low level of CHIP and high level of CAD overall predict the short survival of GBM patients. Conclusion: Altogether, these results illustrated the essential role of CAD in GBM and revealed a novel therapeutic strategy for CAD-positive and CHIP-negative cancer. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Tiangong University is the property of Journal of Tianjin Polytechnic University and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Chromatin immunoprecipitation (ChIP-seq) is the most common approach to observe global binding of proteins to DNA in vivo. The occupancy of transcription factors (TFs) from ChIP-seq agrees well with an alternative method, chromatin endogenous cleavage (ChEC-seq2). However, ChIP-seq and ChEC-seq2 reveal strikingly different patterns of enrichment of yeast RNA polymerase II (RNAPII). We hypothesized that this reflects distinct populations of RNAPII, some of which are captured by ChIP-seq and some of which are captured by ChEC-seq2. RNAPII association with enhancers and promoters - predicted from biochemical studies - is detected well by ChEC-seq2 but not by ChIP-seq. Enhancer/promoter-bound RNAPII correlates with transcription levels and matches predicted occupancy based on published rates of enhancer recruitment, preinitiation assembly, initiation, elongation, and termination. The occupancy from ChEC-seq2 allowed us to develop a stochastic model for global kinetics of RNAPII transcription which captured both the ChEC-seq2 data and changes upon chemical-genetic perturbations to transcription. Finally, RNAPII ChEC-seq2 and kinetic modeling suggests that a mutation in the Gcn4 transcription factor that blocks interaction with the NPC destabilizes promoter-associated RNAPII without altering its recruitment to the enhancer.

FNDC3B (fibronectin type III domain containing 3B) is highly expressed in hepatocellular carcinoma (HCC) and other cancer types, and fusion genes involving FNDC3B have been identified in HCC and leukemia. Growing evidence suggests the significance of FNDC3B in tumorigenesis, particularly in cell migration and tumor metastasis. However, its regulatory mechanisms remain elusive. In this study, we employed bioinformatic, gene regulation, and protein‐DNA interaction screening to investigate the transcription factors (TFs) involved in regulating FNDC3B. Initially, 338 candidate TFs were selected based on previous chromatin immunoprecipitation (ChIP)‐seq experiments available in public domain databases. Through TF knockdown screening and ChIP coupled with Droplet Digital PCR assays, we identified that E2F1 (E2F transcription factor 1) is crucial for the activation of FNDC3B. Overexpression or knockdown of E2F1 significantly impacts the expression of FNDC3B. In conclusion, our study elucidated the mechanistic link between FNDC3B and E2F1. These findings contribute to a better understanding of FNDC3B in tumorigenesis and provide insights into potential therapeutic targets for cancer treatment.

Unfavorable environmental cues severely affect crop productivity resulting in significant economic losses to farmers. In plants, multiple regulatory genes, such as the WRKY transcription factor (TF) family, modulate the expression of defense genes. However, the role of the pineapple WRKY genes is poorly understood. Here, we studied the pineapple WRKY gene, AcWRKY28, by generating AcWRKY28 over-expressing transgenic pineapple plants. Overexpression of AcWRKY28 enhanced the salt stress resistance in transgenic pineapple lines. Comparative transcriptome analysis of transgenic and wild-type pineapple plants showed that “plant-pathogen interaction” pathway genes, including 9 calcium-dependent protein kinases (CPKs), were up-regulated in AcWRKY28 over-expressing plants. Furthermore, chromatin immunoprecipitation and yeast one-hybrid assays revealed AcCPK12, AcCPK3, AcCPK8, AcCPK1, and AcCPK15 as direct targets of AcWRKY28. Consistently, the study of AcCPK12 over-expressing Arabidopsis lines showed that AcCPK12 enhances salt, drought, and disease resistance. This study shows that AcWRKY28 plays a crucial role in promoting salt stress resistance by activating the expression of AcCPK genes.

Spheroids, as three-dimensional (3D) cell aggregates, can be prepared using various methods, including hanging drops, microwells, microfluidics, magnetic manipulation, and bioreactors. However, current spheroid manufacturing techniques face challenges such as complex workflows, the need for specialized personnel, and poor batch reproducibility. In this study, we designed a support-free, 3D-printed microwell chip and developed a compatible low-cell-adhesion process. Through simulation and experimental validation, we rapidly optimized microwell size and the coating process. We successfully formed three types of spheroids—human immortalized epidermal cells (HaCaTs), umbilical cord mesenchymal stem cells (UC-MSCs), and human osteosarcoma cells (MG63s)—on the chip. Fluorescent viability staining confirmed the biocompatibility and reliability of the chip. Finally, drug response experiments were conducted using the chip. Compared to traditional methods, our proposed strategy enables high-throughput production of size-controlled spheroids with excellent shape retention, while enhanced gas exchange during culture improves differentiation marker expression. This platform provides an efficient and cost-effective solution for biosensing applications, such as drug screening, disease modeling, and personalized therapy monitoring. Furthermore, the chip shows significant potential for real-time in vitro monitoring of cellular viability, reaction kinetics, and drug sensitivity, offering valuable advancements in biosensor technology for life sciences and medical applications.

Background & aimsThe stimulator of interferon genes (STING)/TANK-binding kinase 1 (TBK1) pathway is vital in mediating innate immune and inflammatory responses during oxidative/endoplasmic reticulum (ER) stress. However, it remains unknown whether macrophage thioredoxin-interacting protein (TXNIP) may regulate TBK1 function and cell death pathways during oxidative/ER stress.MethodsA mouse model of hepatic ischaemia/reperfusion injury (IRI), the primary hepatocytes, and bone marrow-derived macrophages were used in the myeloid-specific TXNIP knockout (TXNIPM-KO) and TXNIP-proficient (TXNIPFL/FL) mice.ResultsThe TXNIPM-KO mice were resistant to ischaemia/reperfusion (IR) stress-induced liver damage with reduced serum alanine aminotransferase (ALT)/aspartate aminotransferase (AST) levels, macrophage/neutrophil infiltration, and pro-inflammatory mediators compared with the TXNIPFL/FL controls. IR stress increased TXNIP, p-STING, and p-TBK1 expression in ischaemic livers. However, TXNIPM-KO inhibited STING, TBK1, interferon regulatory factor 3 (IRF3), and NF-κB activation with interferon-β (IFN-β) expression. Interestingly, TXNIPM-KO augmented nuclear factor (erythroid-derived 2)-like 2 (NRF2) activity, increased antioxidant gene expression, and reduced macrophage reactive oxygen species (ROS) production and hepatic apoptosis/necroptosis in IR-stressed livers. Mechanistically, macrophage TXNIP deficiency promoted cylindromatosis (CYLD), which colocalised and interacted with NADPH oxidase 4 (NOX4) to enhance NRF2 activity by deubiquitinating NOX4. Disruption of macrophage NRF2 or its target gene 2',5' oligoadenylate synthetase-like 1 (OASL1) enhanced Ras GTPase-activating protein-binding protein 1 (G3BP1) and TBK1-mediated inflammatory response. Notably, macrophage OASL1 deficiency induced hepatocyte apoptotic peptidase activating factor 1 (APAF1), cytochrome c, and caspase-9 activation, leading to increased caspase-3-initiated apoptosis and receptor-interacting serine/threonine-protein kinase 3 (RIPK3)-mediated necroptosis.ConclusionsMacrophage TXNIP deficiency enhances CYLD activity and activates the NRF2-OASL1 signalling, controlling IR stress-induced liver injury. The target gene OASL1 regulated by NRF2 is crucial for modulating STING-mediated TBK1 activation and Apaf1/cytochrome c/caspase-9-triggered apoptotic/necroptotic cell death pathway. Our findings underscore a novel role of macrophage TXNIP-mediated CYLD-NRF2-OASL1 axis in stress-induced liver inflammation and cell death, implying the potential therapeutic targets in liver inflammatory diseases.Lay summaryLiver inflammation and injury induced by ischaemia and reperfusion (the absence of blood flow to the liver tissue followed by the resupply of blood) is a significant cause of hepatic dysfunction and failure following liver transplantation, resection, and haemorrhagic shock. Herein, we uncover an underlying mechanism that contributes to liver inflammation and cell death in this setting and could be a therapeutic target in stress-induced liver inflammatory injury.

The thickness of the maximum undistorted chip (agmax) is a crucial component of the machining arguments and significantly affects the quality of the grind. For the purpose of investigating the impact of the agmax on the grinding removal properties of SiO2f/SiO2 composites, grinding experiments of SiO2f/SiO2 composites were conducted. The findings indicate that crack propagation, fiber cracking, matrix breaking, and interface debonding are the primary removal modes of SiO2f/SiO2 composites. Grinding force, surface roughness, and chip size all have a positive correlation with the maximum undeformed chip thickness. Surface brittle fracture brought on by SiO2 fiber microcrack results in little surface roughness. The surface quality degrades when bending fracture and compression fracture take place. Controlling the agmax can enhance the grinding surface quality of SiO2f/SiO2 composites. [ABSTRACT FROM AUTHOR]

Summary: Recursive partitioning of healthy consortia led to the development of the Clonal Hematopoiesis Risk Score (CHRS) for clonal haematopoiesis (CH); however, in the practical setting, most cases of CH are diagnosed after patients present with cytopenias or related symptoms. To address this real‐world population, we characterize the clinical trajectories of 94 patients with CH and distinguish CH harbouring canonical DNMT3A/TET2/ASXL1 mutations alone ('sole DTA') versus all other groups ('non‐sole DTA'). TET2, rather than DNMT3A, was the most prevalent mutation in the real‐world setting. Sole DTA patients did not progress to myeloid neoplasm (MN) in the absence of acquisition of other mutations. Contrastingly, 14 (20.1%) of 67 non‐sole DTA patients progressed to MN. CHRS assessment showed a higher frequency of high‐risk CH in non‐sole DTA (vs. sole DTA) patients and in progressors (vs. non‐progressors). RUNX1 mutation conferred the strongest risk for progression to MN (odds ratio [OR] 10.27, 95% CI 2.00–52.69, p = 0.0053). The mean variant allele frequency across all genes was higher in progressors than in non‐progressors (36.9% ± 4.62% vs. 24.1% ± 1.67%, p = 0.0064). This analysis in the post‐CHRS era underscores the natural history of CH, providing insight into patterns of progression to MN. [ABSTRACT FROM AUTHOR]

Copyright of Food Research & Development is the property of Food Research & Development Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Main conclusion: Six grape centromere-specific markers for cytogenetics were mined by combining genetic and immunological assays, and the possible evolution mechanism of centromeric repeats was analyzed. Centromeric histone proteins are functionally conserved; however, centromeric repetitive DNA sequences may represent considerable diversity in related species. Therefore, studying the characteristics and structure of grape centromere repeat sequences contributes to a deeper understanding of the evolutionary process of grape plants, including their origin and mechanisms of polyploidization. Plant centromeric regions are mainly composed of repetitive sequences, including SatDNA and transposable elements (TE). In this research, the characterization of centromere sequences in the whole genome of grapevine (Vitis vinifera L.) has been conducted. Five centromeric tandem repeat sequences (Vv1, Vv2, Vv5, Vv6, and Vv8) and one long terminal repeat (LTR) sequence Vv24 were isolated. These sequences had different centromeric distributions, which indicates that grape centromeric sequences may undergo rapid evolution. The existence of extrachromosomal circular DNA (eccDNA) and gene expression in CenH3 subdomain region may provide various potential mechanisms for the generation of new centromeric regions. [ABSTRACT FROM AUTHOR]

Background There is a growing need for percutaneous coronary intervention (PCI) to be performed within the same transcatheter aortic valve implantation (TAVI) procedure. In such cases, cangrelor, a fast-acting intravenous P2Y12-inhibitor with a short offset, is potential clinical utility to minimize bleeding and vascular complications during large-bore arterial access (LBAA) as well as the thrombotic risk associated with concomitant PCI. Case summary We report two cases of TAVI with an indication to concomitant, high-risk PCI. In the first one, cangrelor was started only after LBAA was secured and TAVI completed, just before the initiation of complex PCI. In the second case, due to predicted complex coronary cannulation after TAVI, complex PCI was performed before TAVI and cangrelor started just after LBAA. In both cases, use of cangrelor (vs. pre-treatment with oral P2Y12-i) allowed for a tailored minimization of the risk of bleeding and vascular complications during LBAA while offering full platelet inhibition during a complex/high-risk PCI. Discussion In this case series, we illustrate a possible approach to the use of cangrelor for patients undergoing TAVI and complex/high-risk PCI. In such complex cases, thorough pre-procedural planning might include a cangrelor to minimize vascular, bleeding, and ischaemic complications. [ABSTRACT FROM AUTHOR]

Key Clinical Message: Hematopoietic neoplasms can cause adrenal infarction. In cases of thrombosis occurring at uncommon sites, it is necessary to consider evaluating for the JAK2V617F mutation, even in the absence of notable abnormalities in blood counts. Adrenal infarction, a rare ailment, has been sporadically linked to hematopoietic neoplasms. A 46‐year‐old male encountered left adrenal infarction, which coincided with a progressive rise in platelet counts. Subsequent diagnosis revealed myelodysplastic/myeloproliferative neoplasm‐unclassifiable, featuring a JAK2V617F mutation. Simultaneously, the patient manifested multiple arteriovenous thromboses, necessitating treatment with edoxaban, aspirin, and hydroxyurea. Following thrombosis resolution, he was transferred to a transplantation center. This report delves into the thrombogenicity linked to the JAK2V617F mutation, while also examining documented instances of adrenal infarction in myeloid neoplasms. We should consider evaluating for JAK2V617F mutation even in cases of thrombosis at unusual sites, including adrenal infarction, even if there are no considerable abnormalities in blood counts. [ABSTRACT FROM AUTHOR]

FNDC3B (fibronectin type III domain containing 3B) is highly expressed in hepatocellular carcinoma (HCC) and other cancer types, and fusion genes involving FNDC3B have been identified in HCC and leukemia. Growing evidence suggests the significance of FNDC3B in tumorigenesis, particularly in cell migration and tumor metastasis. However, its regulatory mechanisms remain elusive. In this study, we employed bioinformatic, gene regulation, and protein‐DNA interaction screening to investigate the transcription factors (TFs) involved in regulating FNDC3B. Initially, 338 candidate TFs were selected based on previous chromatin immunoprecipitation (ChIP)‐seq experiments available in public domain databases. Through TF knockdown screening and ChIP coupled with Droplet Digital PCR assays, we identified that E2F1 (E2F transcription factor 1) is crucial for the activation of FNDC3B. Overexpression or knockdown of E2F1 significantly impacts the expression of FNDC3B. In conclusion, our study elucidated the mechanistic link between FNDC3B and E2F1. These findings contribute to a better understanding of FNDC3B in tumorigenesis and provide insights into potential therapeutic targets for cancer treatment. [ABSTRACT FROM AUTHOR]

While machining of high-strength alloy steel materials, poor surface quality and decreased tool life are result by an increase in the cutting zone temperature close to the machining region. To overcome this, surface texture with green manufacturing environment is an alternate path to enhancing machining performance. In the present work, novel Parallel Grooved Textured Tools (PGT) was developed and studied the performance of so developed tools. Further, the Untextured Tool (UT) and PGT performance was compared while machining EN24 alloy steel under Minimum Quantity Lubrication (MQL) condition. It was perceived from result that textured tools reduced the cutting temperature (CT), rake wear (RW), flank wear (FW), surface roughness (Ra) and cutting force (CF) up to 18.96%, 18.79%, 24.31%, 12.69%, and18.96% respectively compared to UT. [ABSTRACT FROM AUTHOR]

Ubiquitination is a cascade reaction involving E1, E2, and E3 enzymes. The orthogonal ubiquitin transfer (OUT) method has been previously established to identify potential substrates of E3 ligases. In this study, we verified the ubiquitination of five substrates mediated by the E3 ligases CHIP and E4B. To further explore the activity of U‐box domains of E3 ligases, two mutants with the U‐box domains interchanged between CHIP and E4B were generated. They exhibited a significantly reduced ubiquitination ability. Additionally, different E3s recruited similar E2 ubiquitin‐conjugating enzymes when ubiquitinating the same substrates, highlighting that U‐box domains determined the E2 recruitment, while the substrate determined the E2 selectivity. This study reveals the influence of substrates and U‐box domains on E2 recruitment, providing a novel perspective on the function of U‐box domains of E3 ligases. [ABSTRACT FROM AUTHOR]

Background: The occurrence of somatic mutations in patients with no evidence of hematological disorders is called clonal hematopoiesis (CH). CH, whose subtypes include CH of indeterminate potential and clonal cytopenia of undetermined significance, has been associated with both hematologic cancers and systemic comorbidities. However, CH's effect on patients, especially those with concomitant malignancies, is not fully understood. Methods: We performed a retrospective evaluation of all patients with CH at a tertiary cancer center. Patient characteristics, mutational data, and outcomes were collected and analyzed. Results: Of 78 individuals included, 59 (76%) had a history of cancer and 60 (77%) had moderate to severe comorbidity burdens. DNMT3A, TET2, TP53, and ASXL1 were the most common mutations. For the entire cohort, the 2‐year overall survival rate was 79% (95% CI: 70, 90), while the median survival was not reached. Of 20 observed deaths, most were related to primary malignancies (n = 7, 35%), comorbidities (n = 4, 20%), or myeloid neoplasms (n = 4, 20%). Twelve patients (15%) experienced transformation to a myeloid neoplasm. According to the clonal hematopoiesis risk score, the 3‐year transformation rate was 0% in low‐risk, 15% in intermediate‐risk (p = 0.098), and 28% in high‐risk (p = 0.05) patients. By multivariate analysis, transformation was associated with variant allele frequency ≥0.2 and hemoglobin <10 g/dL. Conclusions: In a population including mostly cancer patients, CH was associated with comorbidities and myeloid transformation in patients with higher mutational burdens and anemia. Nevertheless, such patients were less likely to die of their myeloid neoplasm than of primary malignancy or comorbidities. [ABSTRACT FROM AUTHOR]

Cancer-related somatic genetic alterations are detectable in the blood of individuals without hematologic malignancy, reflecting the outgrowth of a mutated stem/progenitor cell population, a phenomenon termed clonal hematopoiesis (CH). When accompanied by an unexplained cytopenia(s), CH is further refined to clonal cytopenia of undetermined significance (CCUS) whereas, the finding of a mutation/alteration in the setting of a normal complement of blood counts is called clonal hematopoiesis of indeterminate potential (CHIP). CHIP and CCUS are now recognized precursor conditions to myeloid neoplasms. Advances in the understanding of the epidemiology and clonal metrics associated with evolution to a myeloid malignancy has permitted the elaboration of risk stratification tools poised for use in the clinic and initial clinical investigations seeking to disrupt the natural history of high risk CHIP and CCUS. In this review, we focus on CCUS and the current understanding of its classification, risk stratification and potential therapeutic targets

Summary: The widespread usage of next-generation sequencing methods for functional genomics studies requires standardized tools for consistent visualization of the associated data. Here, we present seqNdisplayR, an R package for plotting standard sequencing data coverage within a genomic region of interest in a customizable and reproducible manner. We describe steps for installing software, preparing data files, choosing options, and plotting data. This tool is readily available for users with no prior experience with R through the “Shiny app” interface.For complete details on the use and execution of this protocol, please refer to Lykke-Andersen et al.,1 Gockert et al.,2 and Rouviere et al.3 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

With the development of 5G technology and increasing chip integration, traditional active cooling methods struggle to meet the growing thermal demands of chips. Thermoelectric coolers (TECs) have garnered great attention due to their rapid response, significant cooling differentials, strong compatibility, high stability and controllable device dimensions. In this review, starting from the fundamental principles of thermoelectric cooling and device design, high-performance thermoelectric cooling materials are summarized, and the progress of advanced on-chip TECs is comprehensively reviewed. Finally, the paper outlines the challenges and opportunities in TEC design, performance and applications, laying great emphasis on the critical role of thermoelectric cooling in addressing the evolving thermal management requirements in the era of emerging chip technologies.

The optimum processing conditions for green laver chips were determined using response surface methodology (RSM) to improve taste and reduce off-flavors by applying reaction flavor and air-frying techniques. The optimum composition (w/w) for the chips included 20% green laver, 20% hairtail surimi, and 60% flour. Additional ingredients included distilled water (90 mL) with GDL (3 g), NaHCO₃ (2 g), salt (1 g), sugar (12 g), roasted soybean powder (1.5 g), and reaction flavor solution (RFS, 10 mL). The mixture was kneaded, shaped, dried at 50 °C for 2 h, and air-fried at 195 °C for 80 sec. The resulting green laver chips showed overall acceptance and brittleness values of 7.00 ± 0.74 and 5.89 ± 0.59 N, respectively, with absolute residual errors of 8.43% and 7.07%. The optimum reaction flavor precursors for green laver chips were determined to be threonine (1.0 g%), proline (1.0 g%), glycine (1.4 g%), methionine (0.05 g%), and glucose (2 g%). Flavor analysis revealed that green laver chips with reaction flavor (GLCR) contained 13 alkylpyrazines with corn-like and nutty odors, and 2-acetylpyrrole, which contributed a popcorn-like odor. In contrast, green laver chips without reaction flavor (GLC) predominantly contained straight-chain aldehydes with undesirable odors. The heating process in the air fryer appeared to reduce the aldehyde content and promote pyrazine formation, significantly enhancing GLCR’s flavor.

Chloroplasts are ancient organelles responsible for photosynthesis and various biosynthetic functions essential to most life on Earth. Many of these functions require tightly controlled regulatory processes to maintain homeostasis at the protein level. One such regulatory mechanism is the ubiquitin-proteasome system whose fundamental role is increasingly emerging in chloroplasts. In particular, the role of E3 ubiquitin ligases as determinants in the ubiquitination and degradation of specific intra-chloroplast proteins. Here, we highlight recent advances in understanding the roles of plant E3 ubiquitin ligases SP1, COP1, PUB4, CHIP, and TT3.1 as well as the ubiquitin-dependent segregase CDC48 in chloroplast function.

A new approach was used in drilling woven glass fiber reinforced polymer (GFRP) composites to clamping the specimen by a controllable force at drill entry and exit sides. The influence of cutting and support conditions on thrust force, push-out and peel-up delaminations, and burr factor was investigated. A series of ASTM tests were implemented to determine tensile, in-plane shear, Mode I and Mode II fracture toughness properties, which were used to predict the critical thrust force (CTF) at delamination initiation. The delamination and burr areas were accurately measured via inexpensive, easy, and accurate image processing technique. Result from drilling processes revealed that the supported specimens are approximately free from burr accompanied with insignificant peeling within the drill (10 mm)/support hole (12 mm) tolerance. By using support plates, the thrust force was increased up to 8.3% compared to the unsupported conditions. On the other hand, the delaminations were decreased by about 65.5%. The delaminations were dominated by Mode I fracture toughness, which is lower than that of Mode II by 41%. The predicted CTF using concentrated load models are slightly lower than the measured values, which interpret the occurrence of delamination damage on the cutting zone.

Background: Red blood cell distribution width (RDW) is calculated in every blood count test and reflects variability in erythrocyte size. High levels mirror dysregulated erythrocyte homeostasis and have been associated with clonal hematopoiesis as well as higher mortality in several conditions.We aimed to determine the impact of preprocedural RDW levels on functional outcomes after transcatheter aortic valve implantation (TAVI). Methods: In this single-center retrospective study, we analyzed 176 consecutive patients receiving TAVI between 2017 and 2021. RDW upper limit of normal was

Microfluidic chips are driven by microfluidic pumps and controlled by microfluidic valves in microfluidic systems to achieve accurate manipulation of microfluidics. With the continuous improvement of microfluidic chip integration, liquid channels have become more complex and have smaller volumes. How to achieve fluid drive and control at the micro scale has become a key focus in chip design. The paper elaborates on the current research status, key technologies, and development trends of micro pumps and micro valves both domestically and internationally, aiming to provide reference and guidance for the development of related industries and technological research.

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Simple Summary: The honeybee subspecies Apis mellifera jemenitica is highly adapted to extreme summer temperatures. This adaptation can be explained by many anatomical, behavioral and molecular components. Basically, many genes are upregulated/silenced in response to stressors. Histone post-translational modifications (PTMs) represent an important gene regulation mechanism in insects. Here, we investigated how the methylation/demethylation of histone (H3) lysine (K4 and K27) impacts the profusion of lethal(2)-essential-for-life-like (l(2)efl), histone methyltransferases (HMTs) and Polycomb genes in A. m. jemenitica and A. m. carnica under thermal stress. The results revealed significant alterations in histone methylation states leading to a higher profusion of l(2)efl and harmonized changes in HMTs and Polycomb genes in response to heat stress with a substantial variation between both honeybee subspecies. Apparently, histone PTMs can diminish the impact of thermal stress and increase the fitness of both honeybee subspecies. In conclusion, the methylation/demethylation of H3K4 and H3K27 is a key epigenetic mechanism in regulating l(2)efl, histone methyltransferases (HMTs) (trx) and Polycomb genes in A. mellifera exposed to heat stress. Histone post-translational modifications (PTMs) represent a key mechanism in the thermal adaptation of the honeybee Apis mellifera. In this study, a chromatin immunoprecipitation assay and qPCR were employed to explore the changes in the methylation states of H3K4m2, H3K4m3, H3K27m2 and H3K27m3 associated with l2efl (ID: 72474, 724405, 724488), histone methyltransferases (HMTs) ((trx) and PR-set7) and Polycomb (Pc) and (Su(z)12) genes in A. m. jemenitica (tolerant subspecies) and A. m. carnica (susceptible subspecies) in response to heat treatment (42 °C for 1 h). The results revealed significant enrichment fold changes in the methylation/demethylation of most H3K4 and H3K27 marks at all targeted genes. These changes increased the profusion of l2efl (ID: 72474, 724405, 724488), histone methyltransferases (HMTs) (trx) and Polycomb (Pc) and Su(z)12 and decreased the profusion of HMT (PR-set7) in both honeybee subspecies. The changes in the methylation enrichment folds of histone methyltransferases (HMTs) ((trx), PR-set) and Polycomb (Pc), Su(z)12 genes demonstrate the well-harmonized coordination of epigenetic gene regulation in response to heat treatment. Compared to the control, the changes in the methylation enrichment folds of H3K4m3 at Polycomb Su(z)12 were about 30× and 100× higher in treated A. m. jemenitica and A.m. carnica, respectively. Similarly, changes in the methylation/demethylation enrichment folds of HMT (trx) and Polycomb (Pc) and Su(z)12 were 2–3× higher in A. m. carnica than in A. m. jemenitica after treatment (42 °C). It is evident that post-translational chromatin modification in both honeybee subspecies can diminish heat stress impact by (I) increasing the transcriptional provision of l2efl associated with survival and (II) increasing the silencing of genes associated with general cellular activities. [ABSTRACT FROM AUTHOR]

Background: Cell concentration in body fluid is an important factor for clinical diagnosis. The traditional method involves clinicians manually counting cells under microscopes, which is labor-intensive. Automated cell concentration estimation can be achieved using flow cytometers; however, their high cost limits accessibility. Microfluidic systems, although cheaper than flow cytometers, still require high-speed cameras and syringe pumps to drive the flow and ensure video quality. In this paper, we present SmartFlow, a low-cost solution for cell concentration estimation using smartphone-based computer vision on 3D-printed, pump-free microfluidic platforms. Objective: The objective was to design and fabricate microfluidic chips, coupled with clinical utilities, for cell counting and concentration analysis. We answered the following research questions (RQs): RQ1, Can gravity drive the flow within the microfluidic chips, eliminating the need for external pumps? RQ2, How does the microfluidic chip design impact video quality for cell analysis? RQ3, Can smartphone-captured videos be used to estimate cell count and concentration in microfluidic chips? Methods: To answer the 3 RQs, 2 experiments were conducted. In the cell flow velocity experiment, diluted sheep blood flowed through the microfluidic chips with and without a bottleneck design to answer RQ1 and RQ2, respectively. In the cell concentration analysis experiment, sheep blood diluted into 13 concentrations flowed through the microfluidic chips while videos were recorded by smartphones for the concentration measurement. Results: In the cell flow velocity experiment, we designed and fabricated 2 versions of microfluidic chips. The ANOVA test (Straight: F 6, 99=6144.45, P <.001; Bottleneck: F 6, 99=3475.78, P <.001) showed the height difference had a significant impact on the cell velocity, which implied gravity could drive the flow. The video sharpness analysis demonstrated that video quality followed an exponential decay with increasing height differences (video quality=100 e–k×Height) and a bottleneck design could effectively preserve video quality (Straight: R2=0.95, k=4.33; Bottleneck: R2=0.91, k=0.59). Samples from the 13 cell concentrations were used for cell counting and cell concentration estimation analysis. The accuracy of cell counting (n=35, 60-second samples, R2=0.96, mean absolute error=1.10, mean squared error=2.24, root mean squared error=1.50) and cell concentration regression (n=39, 150-second samples, R2=0.99, mean absolute error=0.24, mean squared error=0.11, root mean squared error=0.33 on a logarithmic scale, mean average percentage error=0.25) were evaluated using 5-fold cross-validation by comparing the algorithmic estimation to ground truth. Conclusions: In conclusion, we demonstrated the importance of the flow velocity in a microfluidic system, and we proposed SmartFlow, a low-cost system for computer vision–based cellular analysis. The proposed system could count the cells and estimate cell concentrations in the samples. [ABSTRACT FROM AUTHOR]

Dry cutting is an effective method to realize the concept of green cutting today. However, in the process of cutting bearing steel, the high temperatures and high pressures produced by the cutting tool and chip under dry friction seriously affect the machining performance of the tool. Therefore, a bionic microstructure tool based on bionics is proposed to improve the cutting performance and reduce friction by changing the size parameters of the microstructure. On the basis of finite element simulation and cutting tests, the cutting force, surface roughness, and chip shape are used to evaluate the cutting performance. It is found that composite bionic micro-textured tools have a significantly reduced cutting force compared with non-micro-textured tools; composite bionic micro-textured tools lead to a reduction in surface roughness of 10–25%; and composite bionic micro-textured tools are more prone to enhancing the curling and breaking of chips. In addition, with the increase in the microstructure area occupancy, the cutting performance of the tool was also significantly improved. Moreover, it was found that the cutting performance of the tool was improved when the area occupancy of the micro-texture on the front face of the tool was increased. [ABSTRACT FROM AUTHOR]

The effect of cutting edge radius (rß) on the chip formation mechanism during the finish turning of Ti6Al4V is studied. An increase in rß decreases the cutting to thrust force ratio and produces lower chip thickness due to the increase in plowing zone depth. Larger radius tools produce thinner loosely curled chips whereas tightly curled thicker chips are formed for the unprepared edge tool (36 µm). As rß increases, the chip flow angle (ß) increases and free-flowing chips with reduced up-curl but increased side and lateral curl and consequently larger resultant chip curl radius are produced. The chip serration height to overall chip thickness ratio increases whereas chip core thickness reduces owing to the increased plowing action as rß increases. Chip segmentation analysis reveals increased serration height, thermal instability, and shear localisation in the shear zone due to the increased ploughing effect beyond ~60 µm edge radius. [ABSTRACT FROM AUTHOR]

Post‐translational modifications of Tau are emerging as key players in determining the onset and progression of different tauopathies such as Alzheimer's disease, and are recognized to mediate the structural diversity of the disease‐specific Tau amyloids. Here we show that the E3 ligase CHIP catalyzes the site‐specific ubiquitination of Tau filaments both in vitro and in cellular models, proving that also Tau amyloid aggregates are direct substrate of PTMs. Transmission electron microscopy and mass spectrometry analysis on ubiquitin‐modified Tau amyloids revealed that the conformation of the filaments restricts CHIP‐mediated ubiquitination to specific positions of the repeat domain, while only minor alterations in the structure of the fibril core were inferred using seeding experiments in vitro and in a cell‐based tauopathy model. Overexpression of CHIP significantly increased the ubiquitination of exogenous PHF, proving that the ligase can interact and modify Tau aggregates also in a complex cellular environment. [ABSTRACT FROM AUTHOR]

Controlling stem cell behavior at the material interface is crucial for the development of novel technologies in stem cell biology and regenerative medicine. The composition and presentation of bio-factors on a surface strongly influence the activity of stem cells. Herein, we designed an electroactive surface that mimics the initial process of trabecular bone formation, by immobilizing chondrocyte-derived plasma membrane nanofragments (PMNFs) on its surface for rapid mineralization within 2 days. Moreover, the electroactive surface was based on the conducting polymer polypyrrole (PPy), which enabled dynamic control of the presentation of PMNFs on the surface via electrochemical redox switching, further resulting in the formation of bone minerals with different morphologies. Furthermore, bone minerals with contrasting surface morphologies had differential effects on the differentiation of human bone marrow-derived stem cells (hBMSCs) cultured on the surface. Together, this electroactive surface showed multifunctional characteristics, not only allowing dynamic control of PMNF presentation but also promoting the formation of bone minerals with different morphologies within 2 days. This electroactive substrate could be valuable for more precise control of stem cell growth and differentiation, and further development of more suitable microenvironments containing bone apatite for housing a bone marrow stem cell niche, such as biochips/bone-on-chips. [ABSTRACT FROM AUTHOR]