Trefoil factor 3 can stimulate Th17 cell response in the development of type 2 diabetes mellitus.

This study aims to evaluate the role of trefoil factor 3 (TFF3) peptides in type 2 diabetes mellitus (T2DM) from an inflammatory perspective. The focus was on exploring how TFF3 affects the function of T cells. TFF3 overexpression model was constructed using lentivirus in Jurkat cell lines. We evaluated the impact of TFF3 on the proliferation, apoptosis, and IL-17A levels of Jurkat cells cultured in high glucose. The T2DM model was induced in TFF3 knockout (KO) mice through streptozotocin combined with high-fat diet. The measurements included glucose tolerance, insulin tolerance, inflammation markers, Th17 cell proportion, and pancreatic pathological changes. The T2DM modeling led to splenomegaly in mice, and increased expression of TFF3 in their spleens. Overexpression of TFF3 increased the proportion of IL-17+ T cells and the levels of Th17-related cytokines in Jurkat cells. There was no difference in body weight and blood glucose levels between wild-type and TFF3 KO mice. However, T2DM mice lacking the TFF3 gene showed improved glucose utilization, ameliorated pancreatic pathology, decreased inflammation levels, and reduced Th17 cell ratio. TFF3 may be involved in the chronic inflammatory immune response in T2DM. Its mechanism may be related to the regulation of the RORγt/IL-17 signaling pathway and its impact on T cell proliferation and apoptosis.

Threonine fuels glioblastoma through YRDC-mediated codon-biased translational reprogramming.

Cancers commonly reprogram translation and metabolism, but little is known about how these two features coordinate in cancer stem cells. Here we show that glioblastoma stem cells (GSCs) display elevated protein translation. To dissect underlying mechanisms, we performed a CRISPR screen and identified YRDC as the top essential transfer RNA (tRNA) modification enzyme in GSCs. YRDC catalyzes the formation of N6-threonylcarbamoyladenosine (t6A) on ANN-decoding tRNA species (A denotes adenosine, and N denotes any nucleotide). Targeting YRDC reduced t6A formation, suppressed global translation and inhibited tumor growth both in vitro and in vivo. Threonine is an essential substrate of YRDC. Threonine accumulated in GSCs, which facilitated t6A formation through YRDC and shifted the proteome to support mitosis-related genes with ANN codon bias. Dietary threonine restriction (TR) reduced tumor t6A formation, slowed xenograft growth and augmented anti-tumor efficacy of chemotherapy and anti-mitotic therapy, providing a molecular basis for a dietary intervention in cancer treatment.

Tuning charge transport by manipulating concentration dependent single-molecule absorption configurations.

Understanding and tuning charge transport in molecular junctions is pivotal for crafting molecular devices with tailored functionalities. Here, we report a novel approach to manipulate the absorption configuration within a 4,4'-bipyridine (4,4'-BPY) molecular junction, utilizing the scanning tunneling microscope break junction technique in a concentration-dependent manner. Single-molecule conductance measurements demonstrate that the molecular junctions exhibit a significant concentration dependence, with a transition from high conductance (HC) to low conductance (LC) states as the concentration decreases. Moreover, we identified an additional conductance state in the molecular junctions besides already known HC and LC states. Flicker noise analysis and theoretical calculations provided valuable insights into the underlying charge transport mechanisms and single-molecule absorption configurations concerning varying concentrations. These findings contribute to a fundamental comprehension of charge transport in concentration-dependent molecular junctions. Furthermore, they offer promising prospects for controlling single-molecule adsorption configurations, thereby paving the way for future molecular devices.

Contrasting fate and binding behavior of Mn and Cu with dissolved organic matter during in situ remediation using multicomponent capping in malodorous black water.

The common use of peroxides in the remediation of malodorous black water may lead to the activation of heavy metals in sediment when eliminating black and odorous substances. The mechanisms of heavy metal interactions with dissolved organic matter (DOM) in response to in situ capping have not been elucidated, but this information could guide the optimization of capping materials. We developed a capping material consisting of hydrothermally carbonized sediment (HCS), hydrated magnesium carbonate (HMC) and sodium percarbonate (SPC) and used microcosm experiments to investigate the dynamics of Mn and Cu at the sediment-water interface in malodorous black water. The results showed that HCS, HMC and SPC contributed multiple functions of mechanical protection, chemical isolation and oxygen provision to the new caps. HMC promoted the conversion of Mn/Cu into carbonate minerals. The optimal mass proportions were 25 % HCS, 60 % HMC and 15 % SPC based on the mixture design. In situ capping altered the fate and transformation of metals in the sediment-overlying water profile in the short term through Mn immobilization and Cu activation. The complexation of Cu(II) ions was significantly stronger than that of Mn(II) ions. In situ capping had a significant effect on the order of complexation of different fluorescent DOM molecules with Mn(II)/Cu(II) ions: microbial byproducts and fulvic acid-like components were preferentially complexed with Cu(II) ions after capping, while phenolic and humic acid-like components preferentially interacted with Mn(II) ions. Humic-like components bound to Cu were affected the most by capping treatment, whereas protein-like components were relatively weakly affected. Our study provides valuable knowledge on the impact of in situ capping on DOM-metal complexes.

Protective effects of Huang-Qi-Ge-Gen decoction against diabetic liver injury through regulating PI3K/AKT/Nrf2 pathway and metabolic profiling.

ETHNOPHARMACOLOGICAL RELEVANCE: Huang-Qi-Ge-Gen decoction (HGD) is a traditional Chinese medicine prescription that has been used for centuries to treat "Xiaoke" (the name of diabetes mellitus in ancient China). However, the ameliorating effects of HGD on diabetic liver injury (DLI) and its mechanisms are not yet fully understood. AIM OF THE STUDY: To elucidate the ameliorative effect of HGD on DLI and explore its material basis and potential hepatoprotective mechanism. MATERIALS AND METHODS: A diabetic mice model was induced by feeding a high-fat diet and injecting intraperitoneally with streptozotocin (40 mg kg-1) for five days. After the animals were in confirmed diabetic condition, they were given HGD (3 or 12 g kg-1, i. g.) for 14 weeks. The effectiveness of HGD in treating DLI mice was evaluated by monitoring blood glucose and blood lipid levels, liver function, and pathological conditions. Furthermore, UPLC-MS/MS was used to identify the chemical component profile in HGD and absorption components in HGD-treated plasma. Network pharmacology and molecular docking were performed to predict the potential pathway of HGD intervention in DLI. Then, the results of network pharmacology were validated by examining biochemical parameters and using western blotting. Lastly, urine metabolites were analyzed by metabolomics strategy to explore the effect of HGD on the metabolic profile of DLI mice. RESULTS: HGD exerted therapeutic potential against the disorders of glucose metabolism and lipid metabolism, liver dysfunction, liver steatosis, and fibrosis in a DLI model mice induced by HFD/STZ. A total of 108 chemical components in HGD and 18 absorption components in HGD-treated plasma were preliminarily identified. Network pharmacology and molecular docking results of the absorbed components in plasma indicated PI3K/AKT as a potential pathway for HGD to intervene in DLI mice. Further experiments verified that HGD markedly reduced liver oxidative stress in DLI mice by modulating the PI3K/AKT/Nrf2 signaling pathway. Moreover, 19 differential metabolites between normal and DLI mice were detected in urine, and seven metabolites could be significantly modulated back by HGD. CONCLUSIONS: HGD could ameliorate diabetic liver injury by modulating the PI3K/AKT/Nrf2 signaling pathway and urinary metabolic profile.

An improved Back Propagation Neural Network framework and its application in the automatic calibration of Storm Water Management Model for an urban river watershed.

The use of the Storm Water Management Model (SWMM) to simulate flows in urban river watersheds necessitates the proper calibration of the various parameters involved in the process. Back Propagation Neural Network (BPNN) is often used to establish relationship between two sets of multivariate variables, such as parameters and simulation results of SWMM. The aim of this study is to establish an improved BPNN to calibrate SWMM. It was found that when using gauged flow data obtained from the urban river management system as calibration data, only using BPNN was not sufficient. An improved BPNN framework was proposed with integrating Principal Component Analysis (PCA) and Genetic Algorithm (GA) process, abbreviated as PCA-GA-BPNN. It was proved to be effective for calibration. The BPNN combined with GA process made 90 % of the predicted parameters within reasonable range, which was only 8 % using BPNN alone. The PCA process reduced the training time up to 64 %. Using a hydrograph of 196 h, compared with the nondominated sorting genetic algorithm (NSGA), PCA-GA-BPNN training time can be reduced from 18,142 s down to 4.5 s. Nash efficiency coefficients (NSE) of hydrographs fitting was 0.75. Including more rainfall events data in calibration achieved better fitting than including more gauging station data.

Correlation of Serum ß2-MG, HGF, Lp-PLA2 with Carotid Atherosclerosis in Patients with Hypertension Combined with Cerebral Infarction and their Prognostic Value.

Cerebral infarction is characterized by a high morbidity, disability, and fatality rate. This study explored the relationship between serum ß2 microglobulin (ß2-MG), HGF, lipoprotein-associated phospholipase A2 (Lp-PLA2) and carotid atherosclerosis in patients with hypertension combined with cerebral infarction and their prognostic value. A total of 320 patients with cerebral infarction complicated with hypertension who were hospitalized from January 2015 to January 2020 were collected. HGF, Lp-PLA2 and ß2-MG levels were detected. Plaque score (Crouse score) was the patient's cumulative plaque thickness measurements. Additionally, the maximum plaque thickness and the number of plaques were measured.. The correlation was found between high ß2-MG levels and the poor prognosis (HR: 1.29, 95% CI: 1.03-1.52, P = .022). Patients who had elevated levels of HGF were also less likely to have a positive outcome (HR: 1.38, 95% CI: 1.26-1.56, P = .015). High Lp-PLA2 levels were associated with a worse prognosis than low levels (HR: 1.74, 95% CI: 1.29-2.32, P = .015). In conclusion, the levels of ß2-MG, HGF, and Lp-PLA2 in patients with hypertension combined with cerebral infarction were substantially linked with carotid plaques.

[Identification of Peripheral Blood GZMK + CD8 + T Cells As Biomarkers of Alzheimer's Disease Based on Single-Cell Transcriptome].

Objective: Based on single-cell RNA sequencing (scRNA-seq) to explore immune characteristics in the peripheral blood of patients with Alzheimer's disease (AD) as biomarkers. Methods: GSE168522, the scRNA-seq dataset of AD peripheral blood immune cells, was downloaded from the Gene Expression Omnibus (GEO) database and was analyzed in the RAD-Blood web server (http://www.bioinform.cn/RAD-Blood/). The changes in blood cell composition in AD patients were analyzed. The abnormal communications between different types of cells in AD patients were investigated by the CellChat R package. Results: There were two kinds of CD8 + T cells in the blood of AD patients and healthy individuals, one of which highly expressed granzyme K ( GZMK) (false discovery rate [FDR]<0.05), and the other highly expressed GZMA, GZMB, and GZMH (FDR<0.05). In the blood of AD patients, the content of GZMK + CD8 + T cells was increased by 32.9% ( P=5.15E-21), their interactions with other cell types were increased, and they might be associated with AD through the abnormal signal transduction of major histocompatibility complex class Ⅰ (MHC-Ⅰ). Erythrocyte provided the main ligands, that are, human leukocyte antigen (HLA) class Ⅰ molecules, including HLA- A, HLA- B, HLA- C, and HLA- E, for the abnormal MHC-Ⅰ signaling pathway of GZMK + CD8 + T cells. The RESISTIN signaling pathway was specifically enriched in the blood of AD patients. Conclusion: The increased content of peripheral blood GZMK + CD8 + T cells, the increased interaction between GZMK + CD8 + T cells and erythrocytes, and the enhanced RESISTIN pathway are potential blood biomarkers of AD.

Moshen granule ameliorates membranous nephropathy by regulating NF-ƙB/Nrf2 pathways via aryl hydrocarbon receptor signalling.

Considerable achievements were realized in illuminating underlying pathological mechanisms of patients with idiopathic membranous nephropathy (IMN). Although IMN patients are well diagnosed before they reach renal failure, no currently available drug intervention is effective in halting IMN progression. In this study, we assess Moshen granule (MSG) effect on IMN patients and cationic bovine serum albumin (CBSA)-induced rats. Increasing studies has indicated that activation of aryl hydrocarbon receptor (AHR) was related to oxidative stress and inflammation. We further determine MSG effect on AHR, nuclear factor ƙB (NF-ƙB) and nuclear factor erythroid 2-related factor 2 (Nrf2) in the CBSA-induced rats. MSG markedly reduces proteinuria and improves kidney function in both IMN patients and rats induced by CBSA. MSG markedly inhibits increased mRNA expressions of intrarenal AHR and its four downstream target genes including CYP1A1, CYP1A2, CYP1B1 and COX-2 compared with untreated CBSA-induced rats. This is accompanied by markedly downregulated protein expressions of p-IƙBα and NF-ƙB p65 and its downstream gene products including MCP-1, COX-2, 12-LOX, iNOS, p47phox and p67phox, while markedly preserves protein expressions of Nrf2 and its downstream gene products including catalase, HO-1, GCLM, GCLC, MnSOD and NQO1 in the kidney tissues. These data suggests MSG blunts podocyte damage through inhibiting activation of NF-ƙB/Nrf2 pathway via AHR signaling. This finding may provide a promising therapy for treatment of IMN through oxidative stress and inflammation.

A multiscale analysis of the spatially heterogeneous relationships between non-point source pollution-related processes and their main drivers in Chaohu Lake watershed, China.

A better understanding of the relationships between non-point source (NPS) pollution-related processes and their drivers will help to develop scientific watershed management measures. Although various studies have explored the drivers' impact on NPS pollution-related processes, quantitative knowledge of the properties within these relationships is still needed. This study uses the Integrated Valuation of Ecosystem Services and Trade-offs (InVEST) model to produce three related processes of NPS pollution, quick flow (QF), nitrogen export (NE), and sediment export (SE), in the upstream watershed of Chaohu Lake, China. The spatial distributions of QF, NE, and SE and their responses to multiple natural-socioeconomic drivers at nine spatial scales (1 km2, 10 km2, 20 km2, 30 km2, 50 km2, 75 km2, 100 km2, 200 km2, and town) were compared. The results showed that the spatial scale has little impact on the spatial distributions of NPS pollution-related processes. Across the nine scales, the socioeconomic drivers related to agricultural activities, area proportions of cultivated land (cultivated) and paddy field (paddy), have dominant impacts on NE, while the topographical drivers, the connectivity index (IC) and slope, have dominant impacts on both SE and QF. The magnitudes of single and paired natural-socioeconomic drivers' impacts on NPS pollution-related processes increase logarithmically or linearly with increasing spatial scale, but they tend to reach a stable threshold at a certain coarse scale. Our results emphasized the necessity and importance of embracing spatial scale effects in watershed water environmental management.

Lysine catabolism reprograms tumour immunity through histone crotonylation.

Cancer cells rewire metabolism to favour the generation of specialized metabolites that support tumour growth and reshape the tumour microenvironment1,2. Lysine functions as a biosynthetic molecule, energy source and antioxidant3-5, but little is known about its pathological role in cancer. Here we show that glioblastoma stem cells (GSCs) reprogram lysine catabolism through the upregulation of lysine transporter SLC7A2 and crotonyl-coenzyme A (crotonyl-CoA)-producing enzyme glutaryl-CoA dehydrogenase (GCDH) with downregulation of the crotonyl-CoA hydratase enoyl-CoA hydratase short chain 1 (ECHS1), leading to accumulation of intracellular crotonyl-CoA and histone H4 lysine crotonylation. A reduction in histone lysine crotonylation by either genetic manipulation or lysine restriction impaired tumour growth. In the nucleus, GCDH interacts with the crotonyltransferase CBP to promote histone lysine crotonylation. Loss of histone lysine crotonylation promotes immunogenic cytosolic double-stranded RNA (dsRNA) and dsDNA generation through enhanced H3K27ac, which stimulates the RNA sensor MDA5 and DNA sensor cyclic GMP-AMP synthase (cGAS) to boost type I interferon signalling, leading to compromised GSC tumorigenic potential and elevated CD8+ T cell infiltration. A lysine-restricted diet synergized with MYC inhibition or anti-PD-1 therapy to slow tumour growth. Collectively, GSCs co-opt lysine uptake and degradation to shunt the production of crotonyl-CoA, remodelling the chromatin landscape to evade interferon-induced intrinsic effects on GSC maintenance and extrinsic effects on immune response.

Occurrence and impact of carbendazim and hymexazol residues on yeast growth and ochratoxin A contamination during wine production.

BACKGROUND: Grapes are highly vulnerable to infection by carbon black aspergilli, which produce ochratoxin A (OTA), a mycotoxin. Carbendazim and hymexazol are widely applied to control grape diseases. Howerver, fungicides, toxigenic fungi, and OTA can be transferred from grapes to wine causing potential safety issues. The impact of these residues on fungal populations and OTA during vinification are currently unclear. Here we investigated the effects of carbendazim and hymexazol on the viability of Aspergillus carbonarius and OTA contamination during an indoor wine-processing experiment. RESULTS: The population size of A. carbonarius substantially increased at 24 h followed by a significantly decreased at 72 h after destemming and crushing. However, carbendazim and hymexazol notably inhibited the growth of A. carbonarius in must samples. In addition, yeast growth was substantially deleyed by carbendazim, hymexazol, and OTA during the first 3 days in compared with the control. Carbendazim, hymexazol, and OTA residues declined over time, and the processing factors (PFs) for carbendazim and hymexazol throughout vinification were 0.164, 0.074, and 0.185-0.476, respectively. Carbendazim and hymexazol each reduced OTA concentrations. However, there was no significant difference after 48 h. Addition of carbendazim or hymexazol significantly reduced the level of A. carbonarius but had no significant effect on the final concentration of OTA in mature wine. CONCLUSION: The wine-making process can reduce the residues of OTA, carbendazim, and hymexazol in grapes, but it is recommended that grapes chosen to make wine should be free of A. carbonarius contamination. © 2023 Society of Chemical Industry.

A dual-mode fluorescent probe based on carbon dots for detecting solution polarity.

Fluorescent probes always attract huge attention. In particular, carbon dots, due to their unique biocompatibility and variable fluorescence characteristics, can be used in multiple fields and were fulled of expectation by researchers. Since the advent of the dual-mode carbon dots probe, which greatly improved the accuracy of quantitative detection, there are higher hopes for dual-mode carbon dots probes. Herein, we have successfully developed a new dual-mode fluorescent carbon dots probe based on 1,10-phenanthroline (Ph-CDs). Ph-CDs detect the object to be measured based on both down-conversion luminescence and up-conversion luminescence at the same time, different from the reported dual-mode fluorescent probes which are based on the wavelength and intensity changes in down-conversion luminescence. As-prepared Ph-CDs have good linear relationships with the polarity of solvents in down-conversion luminescence (R2 = 0.9909) and up-conversion luminescence (R2 = 0.9374), respectively. Hence, Ph-CDs provide a new in-depth insight into the design of fluorescent probes with dual-mode detection and they can give more accurate, reliable and convenient detection results.

Yishen Huashi granule modulated lipid metabolism in diabetic nephropathy via PI3K/AKT/mTOR signaling pathways.

Aim: Diabetic nephropathy (DN) is the primary cause of end-stage renal disease worldwide. Although etiology for DN is complex and still needs to be fully understood, lipid metabolism disorder is found to play a role in it. Previously, we found Yishen Huashi (YSHS) granule could inhibit diabetic damage and reduce level of microalbuminuria (mALB) in DN animals. To explore its role and mechanism in lipid metabolism under DN settings, this study was designed. Materials and methods: DN rats were induced by streptozotocin (STZ), HepG2 and CaCO2 cells were applied for in vitro study. Hematoxylin-Eosin (HE), periodic acid-Schiff (PAS) staining, and Transmission Electron Microscopy (TEM) were applied for histological observation; 16s Sequencing was used for intestinal microbiota composition analysis; western blotting (WB) and immunofluorescence were carried out for molecular biological study, and enzyme-linked immunosorbent assay (ELISA) was used for lipid determination. Results: YSHS administration significantly reduced levels of total cholesterol (TC), triglyceride (TG), and low-density lipoprotein (LDL-C), while increased level of high-density lipoprotein (HDL-C); meanwhile, histological changes and steatosis of the liver was ameliorated, integrity of the intestinal barrier was enhanced, and dysbacteriosis within intestinal lumen was ameliorated. Mechanism study found that YSHS modulated mitophagy within hepatocytes and inhibited mTOR/AMPK/PI3K/AKT signaling pathway. Conclusion: In conclusion, we found in the present study that YSHS administration could ameliorate lipid metabolism disorder in DN animals, and its modulation on intestinal-liver axis played a significant role in it.

Comparison of the efficacy of different biodegradable membranes in guided bone/tissue regeneration: a systematic review and network meta-analysis.

Guided bone/tissue regeneration (GBR/GTR) is commonly used in dental treatment. The desired bone/tissue regeneration is achieved by placing a barrier membrane over the defect to avoid the downward growth of faster-growing connective and epithelial tissue into the defect. This review aimed to evaluate osteogenic properties, degradation characteristics, and postoperative complications of eight biodegradable membranes in animal experiments, including non-crosslinked collagen membrane (NCCM), crosslinked collagen membrane (CCM), silk membrane (SM), polylactic-co-glycolic acid, polylactic acid, polyethylene glycol hydrogel, polycaprolactone (PCL), and magnesium alloys. Seven electronic databases (PubMed, Embase, Web of Science, Cochrane Library, Science Direct, Wiley, Scopus and Google Scholar) were screened. Study selection, data extraction and quality assessment were made in duplicate. The SYRCLE assessment tool, CERQual (Confidence in the Evidence from Reviews of Qualitative Research) tool and GRADE tool were used to grade the risk of bias and level of evidence. A total of 2512 articles were found in the electronic database. Finally, 94 articles were selected, of which 53 were meta-analyzed. Surface under the cumulative ranking curve showed the best results for new bone formation in the magnesium barrier membrane group, followed by SM, PCL, NCCM, and CCM. Qualitative analysis showed good biocompatibility for natural polymer membranes and a longer degradation time for synthetic polymer membranes. In addition, 34 studies all showed high bias risks, while other studies had unclear bias risks. Natural polymer membranes were more effective for bone regeneration and magnesium alloys were proved to be promising barrier materials that warrant future research.

The Protective Effect of the Asystasia chelonoides Extracts on Hypertensive Nephropathy Rats.

BACKGROUND: Hypertensive nephropathy (HN) is one kind of kidney disorders caused by long-term uncontrolled hypertension, usually resulting in severe kidney damage, including inflammation and oxidative stress, no matter in cells or tissues, from patients with nephropathy. In recent years, nephropathy accompanied by hypertension is becoming one of the main causes for kidney replacement therapy, but few effective treatments have been reported for HN treatment. Asystasia chelonoides (AC) is a kind of plant with the effects of anti-inflammation, lowering blood pressure, and anti-oxidative stress. Still, the therapeutic effect of AC in HN rats is not clear. METHODS: To establish HN model by feeding high sugar and high fat diet spontaneously hypertensive rats. Blood measurement, HE staining, PAS staining and biochemical analysis and were used to assess the therapeutic effects of AC extracts and western blotting analyzed the underlying mechanisms of AC extracts treatment in the HN rat model. RESULTS: AC extracts could significantly lower systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean blood pressure (MBP) in HN rats; and reduce the expression of total protein (TP), blood urea nitrogen (BUN), microalbuminuria (MALB), creatinine (Cr), total cholesterol (TC), triglyceride (TG), and low-density lipoprotein-cholesterol (LDL-C) concentrations, and also could down-regulate expression of IL-6, MDA and AGEs, up-regulate the expression of SOD in HN rats; HE staining and PAS staining demonstrated that AC extracts could alleviate the histopathological changes in HN rats; western blotting demonstrated that AC extracts could up-regulate the expression of PPARγ and down-regulate the expression of TGFß1 and NF-кB in HN rats. CONCLUSION: The finding of the article demonstrated that AC extracts had the better therapeutic effect for HN, and provided the pharmacological evidences for AC extracts treatment for HN.

Analysis of a rare progeria variant of Barrier-to-autointegration factor in Drosophila connects centromere function to tissue homeostasis.

Barrier-to-autointegration factor (BAF/BANF) is a nuclear lamina protein essential for nuclear integrity, chromatin structure, and genome stability. Whereas complete loss of BAF causes lethality in multiple organisms, the A12T missense mutation of the BANF1 gene in humans causes a premature aging syndrome, called Néstor-Guillermo Progeria Syndrome (NGPS). Here, we report the first in vivo animal investigation of progeroid BAF, using CRISPR editing to introduce the NGPS mutation into the endogenous Drosophila baf gene. Progeroid BAF adults are born at expected frequencies, demonstrating that this BAF variant retains some function. However, tissue homeostasis is affected, supported by studies of the ovary, a tissue that depends upon BAF for stem cell survival and continuous oocyte production. We find that progeroid BAF causes defects in germline stem cell mitosis that delay anaphase progression and compromise chromosome segregation. We link these defects to decreased recruitment of centromeric proteins of the kinetochore, indicating dysfunction of cenBAF, a localized pool of dephosphorylated BAF produced by Protein Phosphatase PP4. We show that DNA damage increases in progenitor germ cells, which causes germ cell death due to activation of the DNA damage transducer kinase Chk2. Mitotic defects appear widespread, as aberrant chromosome segregation and increased apoptosis occur in another tissue. Together, these data highlight the importance of BAF in establishing centromeric structures critical for mitosis. Further, these studies link defects in cenBAF function to activation of a checkpoint that depletes progenitor reserves critical for tissue homeostasis, aligning with phenotypes of NGPS patients.

Reduction in Five Harmful Substances in Fried Potato Chips by Pre-Soaking Treatment with Different Tea Extracts.

Thermally processed food always contains various types of harmful substances. Control of their levels in food is important for human health. This work used the extracts from green tea dust, old green tea, yellow tea, white tea, oolong tea, and black tea to simultaneously mitigate diverse harmful substances in fried potato chips. The six tea extracts (30 g/L) all showed considerable inhibitory effects on the formation of 5-hydroxymethylfurfural (reduced by 19.8%-53.2%), glyoxal (26.9%-36.6%), and methylglyoxal (16.1%-75.1%). Green tea and black tea extracts exhibited better inhibitory abilities than the other three teas and were further investigated for other harmful compounds by various concentration treatments. Finally, pre-soaking of fresh potato slices in 50 g/L extracts of green tea dust displayed, overall, the most promising inhibitory capacity of HMF (decreased by 73.3%), glyoxal (20.3%), methylglyoxal (69.7%), acrylamide (21.8%), and fluorescent AGEs (42.9%) in fried potato chips, while it exhibited the least impact on the color and texture. The high level of catechins in green tea dust may contribute most to its outstanding inhibitory effect, whereas the distinguished inhibitory effect of black tea extract was speculated to be attributable to the high levels of theaflavins and amino acids in the fully fermented tea. This study indicated that green tea dust, a predominant waste of the tea industry, had great potential to be exploited to improve food quality and safety.

Immunohistochemical Analysis of Nuclear Lamina Structures in the Drosophila Ovary Using CRISPR-Tagged Genes.

The Drosophila ovary represents an outstanding model for investigating tissue homeostasis. Females continuously produce oocytes throughout their lifetime. However, as females age, fecundity declines, in part, due to changes in ovarian niche function and germline stem cell (GSC) homeostasis. Understanding the dynamics of GSC maintenance will provide needed insights into how coordinated tissue homeostasis is lost during aging. Critical regulators of GSC maintenance are proteins that reside in the nuclear lamina (NL), including the NL proteins emerin and Barrier-to-Autointegration Factor (BAF). Continued investigation of how emerin, BAF, and other NL proteins contribute to GSC function depends upon the availability of antibodies for NL proteins, a limiting resource. In this chapter, we discuss strategies for using clustered regularly interspaced short palindromic repeats (CRISPR) genomic editing to produce endogenously tagged NL genes to circumvent this obstacle, using the generation of the gfp-baf allele as an example. We describe strategies for validation of tagged alleles. Finally, we outline methods for immunohistochemical analysis of resulting tagged-NL proteins.