Concentration In Cells Research Articles

CDK2 and CDK4 targeted liensinine inhibits the growth of bladder cancer T24 cells

Bladder cancer (BCa) is a urinary tumor with limited treatment options and high mortality. Liensinine (LIEN), a natural bisbenzylisoquinoline alkaloid, has shown excellent anti-tumor effects in numerous preclinical studies. However, the anti-BCa effect of LIEN remains unclear. To the best of our knowledge, this is the first study to investigate the molecular mechanism of LIEN in the management of BCa. First, we identified the treatment-related targets of BCa; those that repeatedly occur in more than two databases, including GeneCards, Online Mendelian Inheritance in Man, DisGeNET, Therapeutic Target Database, and Drugbank. The SwissTarget database was used to screen LIEN-related targets, and those with a probability >0 were possible LIEN targets. The prospective targets of LIEN in the treatment of BCa were then determined using a Venn diagram. Second, we discovered that the PI3K/AKT pathway and senescence mediated the anti-BCa action of LIEN by using GO and KEGG enrichment analysis to explore the function of LIEN therapeutic targets. A protein-protein interaction network was created using the String website, and six algorithms of the CytoHubba plug-in were then used in Cytoscape to assess the core targets of LIEN for the therapy of BCa. The outcomes of molecular docking and dynamics simulation demonstrated that CDK2 and CDK4 proteins were the direct targets of LIEN in the management of BCa, among which CDK2 was more stable in binding to LIEN than CDK4. Finally, in vitro experiments showed that LIEN inhibited the activity and proliferation of T24 cells. The expression of p-/AKT, CDK2, and CDK4 proteins progressively decreased, while the expression and fluorescence intensity of the senescence-related protein, γH2AX, gradually increased with increasing LIEN concentration in T24 cells. Therefore, our data suggest that LIEN may promote senescence and inhibit proliferation by inhibiting the CDK2/4 and PI3K/AKT pathways in BCa.

LncRNA XIST inhibition promotes M2 polarization of microglial and aggravates the spinal cord injury via regulating miR-124–3p / IRF1 axis

BackgroundSpinal cord injury (SCI) has a high disability rate and mortality rate. Recently, LncRNA XIST has been found to be involved in the regulation of inflammatory responses. Therefore, we aimed to investigate the role of XIST in the occurrence and development of SCI and the specific regulation mechanism. Methods: 100 ng/mL lipopolysaccharide (LPS) was used to treat mouse microglia BV2 cells. Hitting spinal cord was performed to C57BL/6 mice for establishing SCI model. Real-time reverse transcriptase-polymerase chain reaction (RT-qPCR), Western blot, Immunofluorescence (IF) and Enzyme linked immunosorbent assay (ELISA) experiments were used to explore the function of XIST, miR-124–3p and IRF1 in LPS-induced BV2 cells. RT-qPCR, Nissl staining, IF, Western blot and ELISA experiment were performed to study the function of XIST in SCI mice. Dual-luciferase reporter assay, RNA immunoprecipitation (RIP), RT-qPCR and Western blot assays were utilized to identify the interaction among XIST, miR-124–3p and IRF1. Results: XIST was upregulated in LPS-induced BV2 cells and spinal cord tissues of SCI mice. Overexpression of XIST promoted the M1 microphages polarization and cytokines concentration in LPS-stimulated BV2 cells, aggravated SCI of mice. Downregulated XIST promoted M1-to-M2 conversion of microglial and relieved the injury of SCI mice. Mechanism verification indicated that XIST acted as a molecular sponge of miR-124–3p and regulated IRF1 expression. Increased miR-124–3p or reduced IRF1 inhibited M1 polarization of microglial and decreased the production of inflammatory cytokines in LPS-induced BV2 cells. Increased XIST or decreased miR-124–3p had an opposite of on LPS-induced BV2 cells. Conclusion: Overexpression of XIST enhanced M1 polarization of microglia and promoted the level of inflammatory cytokines through sponging miR-124–3p and regulating IRF1 expression.

Evaluation of Generic Fertiliser as an Alternative Inorganic Nitrogen Source for Ethanolic Glucose Fermentation by Saccharomyces cerevisiae

In the studies and production of bioethanol, the preferred fermenting yeast (Saccharomyces cerevisiae) is usually cultured in liquid broth that contains yeast extract and peptone. However, the use of these laboratory and scientific grade chemicals is costly, making them impractical for mass bioethanol production. Therefore, this study was conducted to evaluate the feasibility of glucose ethanolic fermentation by S. cerevisiae using generic fertiliser formulations to provide inorganic nitrogen, phosphorus, potassium and trace elements (NPK-TE). Fermentation media of different generic fertiliser strength at 0.5X, 1.0X and 2.0X Fertiliser Nitrogen Equivalents (FNE), as compared to the conventional Yeast Extract-Peptone (YEP) medium as control, was used as fermentation broth during the ethanolic fermentation of glucose. Based on the results, S. cerevisiae cultured in YEP broth produced the highest cell concentration for both wet (21.93 g/L) and dry cells (3.87 g/L), with rapid increment observed in the first 72 h of fermentation. By the end of the fermentation period, lactic acid (3.14 g/L) and acetic acid (0.96 g/L) levels were recorded to be the lowest in YEP medium while their concentration (lactic acid, 8.08 g/L) and (acetic acid, 2.67 g/L) were highest in 2.0X FNE fertiliser medium. Results indicated that the best theoretical ethanol yield (TEY) among the fertiliser media was achieved when fermentation was performed in the 0.5X FNE fertiliser medium, with a TEY of 86.18%. TEY yields were 78.68% and 51.54% in broth with 1.0X and 2.0X FNE, respectively. In general, all three fertiliser media supported ethanolic fermentation of glucose, with the 0.5X FNE fertiliser broth showing a yield that is significantly close to the conventional YEP medium, as seen in the statistical analysis. Similarities in other fermentation profiles such as acetic acid, lactic acid, and biomass production, as well as glucose utilisation, between the results from the YEP samples and samples from the fertiliser broths (at 0.5X and 1.0X FNE) have also shown that generic fertiliser has the potential to be used as an alternative medium to replace the conventional YEP to produce ethanol at a lower cost.

Nutritional strategy underlying plant specialization to gypsum soils.

Gypsum soils are amongst the most widespread extreme substrates of the world, occurring in 112 countries. This type of hypercalcic substrate has a suite of extreme physical and chemical properties that make it stressful for plant establishment and growth. Extreme chemical properties include low plant-available nitrogen and phosphorus and high plant-available sulphur and calcium, which impose strong nutritional imbalances on plants. In spite of these edaphic barriers, gypsum soils harbour rich endemic floras that have evolved independently on five continents, with highly specialized species. Plants that only grow on gypsum are considered soil specialists, and they have a foliar elemental composition similar to the elemental availability of gypsum soils, with high calcium, sulphur and magnesium accumulation. However, the physiological and ecological role of the unique foliar elemental composition of gypsum specialists remains poorly understood, and it is unknown whether it provides an ecological advantage over other generalist species on gypsum soils. This article reviews available literature on the impact of gypsum soil features on plant life and the mechanisms underlying plant adaptation to gypsum environments. We conclude with a hypothesis on the potential role of the nutritional strategy underlying plant specialization to gypsum soils: Gypsum specialists primarily use SO42- as a counter anion to tolerate high Ca2+ concentrations in cells and avoid phosphorus depletion, which is one of the most limiting nutrients in gypsum soils.

C(RGDfK) anchored surface manipulated liposome for tumor-targeted tyrosine kinase inhibitor (TKI) delivery to potentiate liver anticancer activity

Current anticancer drug research includes tumor-targeted administration as a critical component because it is the best strategy to boost efficacy and decrease toxicity. Low drug concentration in cancer cells, nonspecific distribution, rapid clearance, multiple drug resistance, severe side effects, and other factors contribute to the disappointing results of traditional chemotherapy. As an innovative technique of treatments for hepatocellular carcinoma (HCC) in recent years, nanocarrier-mediated targeted drug delivery systems can overcome the aforesaid limitations via enhanced permeability and retention effect (EPR) and active targeting. Epidermal growth factor receptor (EGFR) inhibitor Gefitinib (Gefi) has dramatic effects on hepatocellular carcinoma. Herein, we developed and assessed an αvβ3 integrin receptor targeted c(RGDfK) surface modified liposomes for better targeting selectivity and therapeutic efficacy of Gefi on HCC cells. The conventional and modified Gefi loaded liposomes, i.e., denoted as Gefi-L and Gefi-c(RGDfK)-L, respectively, were prepared through the ethanol injection method and optimized via Box Behnken design (BBD). The FTIR and 1H NMR spectroscopy verified that the c(RGDfK) pentapeptides had formed an amide bond with the liposome surface. In addition, the particle size, Polydispersity index, zeta potential, encapsulation efficiency, and in-vitro Gefi release of the Gefi-L and Gefi-c(RGDfK)-L were measured and analyzed. As indicated by the MTT assay on HepG2 cells, Gefi-c(RGDfK)-L displayed considerably higher cytotoxicity than Gefi-L or Gefi alone. Throughout the incubation period, HepG2 cells took up significantly more Gefi-c(RGDfK)-L than Gefi-L. According to the in vivo biodistribution analysis, Gefi-c(RGDfK)-L accumulated more strongly at the tumor site than Gefi-L and free Gefi. Furthermore, HCC-bearing rats treated with Gefi-c(RGDfK)-L showed a substantial drop in liver marker enzymes (alanine transaminase, alkaline phosphatase, aspartate transaminase, and total bilirubin levels) compared to the disease control group. Gefi-c(RGDfK)-L suppresses tumour growth more effectively than Gefi-L and free Gefi, according to an in vivo analysis of their anticancer activities. Thus, c(RGDfK)-surface modified liposomes, i.e., Gefi-c(RGDfK)-L may serve as an efficient carrier for the targeted delivery of anticancer drugs.

P-785 Promising Reproductive Outcomes In Patients With Refractory Thin Endometrium After Autologous Bone Marrow Regenerative Cell Therapy (ABM-RCT) – A Case Series

Abstract Study question Does Autologous Bone Marrow Regenerative Cell Therapy (ABM-RCT) improve endometrial thickness and receptivity and restoration of fertility in refractory cases of thin endometrium? Summary answer Selective Autologous Bone Marrow Regenerative Cell Therapy from Seragen (ABM-RCT) improves endometrial thickness, pregnancy, and live birth rate in refractory cases of thin endometrium. What is known already The impaired endometrial function can limit implantation due to insufficient tissue regeneration. Clinical investigation has proved that stem cells existing in the endometrium might originate from bone marrow and provides evidence that bone marrow-derived cells have a non-hematopoietic physiologic contribution to decidual stroma and play a vital role in implantation and pregnancy maintenance and non-hematopoietic BMSCs are able to impact decidual molecular milieu and overcome implantation defects. ABM-RCT is the most commonly used cell therapy and its safety and efficacy are well-established and documented. This forms the basis for exploring and standardizing ABM-RCT in the management of refractory thin endometrium. Study design, size, duration Eleven patients (from age 29 to 43 with refractory thin endometrium resistant to conventional treatment modalities were recruited and obtained informed consent with detailed explanation that the therapy is still experimental and the risk of failure was given to the couple as part of gynec-oncology treatment. Five patients had asherman’s syndrome, two patients had genital tuberculosis, two patients had thin endometrium hypo-responsive/unresponsive to estrogens, with RIF and two patients underwent chemo and radiotherapy Participants/materials, setting, methods BM aspiration was performed under local anesthesia, from the iliac crest using a disposable BM aspiration needle (Jamshidi, 11 G) and collected in heparinized syringes. Progenitor cell enrichment was done by Seragen’s Selective Enrichment Protocol. Peripheral blood was collected to enrich growth factor concentrate. A 2.9mm Hysteroscope was used with an operating channel and egg pickup needle attached to it and cells were implanted in subendometrial zone in all four walls of the uterine cavity. Main results and the role of chance All patients presented to us with refractory thin endometrium in HRT cycle despite giving all possible medications. Hence the decision was taken for Autologous Bone Marrow Cell Concentrate Therapy.Post ABM-RCT, endometrial thickness showed increase 100% (11/11) of more than 8 mm in all cases on day of embryo transfer with average endometrial thickness improvement was 1.8 mm than previous cycles, with uniform triple layer pattern.10 out of 11 (91%) patients conceived after autologous stem cell injection. 9 Out of 10 (90%) patients conceived had delivered healthy babies and 1 patient (9%) had a miscarriage at 12th week of pregnancy. 1 patient did not conceive will be assessed for immunoprofiling and one more rejuvenation cycle. Limitations, reasons for caution Our study outcomes are consistent with several previous studies and the feasibility of treatment at an IVF setup and encouraging results. Well-planned study with more patients is warranted to evaluate safety, effectiveness, and cost of this modality before it becomes integrated in treatment of this frustrating condition during IVF procedures Wider implications of the findings Our results demonstrates selectively enriched ABM-RCT enhances endometrial thickness, tissue remodeling, uneventful gestation, improved pregnancy and live birth rate. Need for Surrogacy in Non-Responsive Thinendometrium May Be Re-Evaluated. Role of hematopoetic stem cell research and knowledge generated is representing reliable strategy in IVF that may be routinely used in clinicalpractice. Trial registration number Not applicable

Mechanisms of synergistic suppression of ALK-positive lung cancer cell growth by the combination of ALK and SHP2 inhibitors

Lung cancer is a major cause of cancer-related deaths. Alectinib is the first line of treatment for patients with ALK-positive lung cancer, but the survival rate beyond 2–3 years is low. Co-targeting secondary oncogenic drivers such as SHP2 is a potential strategy for improving drug efficacy. This is because SHP2 is expressed ubiquitously, but ALK expression is largely restricted to cancer cells. Thus, the combination of ALK and SHP2 inhibitors may provide a way to restrict synergistic cytotoxicity to cancer cells only, by reducing the dose of SHP2 inhibitors required for anticancer action and minimising SHP2-dependent systemic toxicity. The objective of this study was to investigate whether the combination of a SHP2 inhibitor (SHP099) with alectinib would synergistically suppress the growth of ALK-positive lung cancer cells. Our results demonstrated that the drug combination significantly and synergistically decreased cell viability at relatively low concentrations in ALK-positive H3122 and H2228 cells, due to G1 cell cycle arrest and increased apoptosis because of suppressed downstream RAS/MAPK signalling. The drug combination also induced the expression of mediators of the intrinsic apoptotic pathway, Bim and cleaved caspase-3, and modulated the expression of cell cycle mediators cyclin D1, cyclin B1, and phosphorylated CDK1.

Framework for solving dynamics of Ca2+ ion concentrations in liver cells numerically: Analysis of a non‐negativity‐preserving non‐standard finite‐difference method

In this article, we consider the numerical approximation of dynamics of calcium ion concentrations in liver cells by a non‐linear first‐order system of ordinary differential equations. At first, we summarize some analytical results of the time‐continuous problem formulation which are of importance to numerically solving this system. We continue with the presentation of our numerical framework which contains two ingredients—an optimization approach for determining the unique equilibrium state and, as our main contribution, a non‐standard finite‐difference scheme for numerical approximation of the time‐discretized dynamical system. We prove that our proposed scheme conserves non‐negativity, and we show convergence towards the time‐continuous unique solution. Conclusively, we stress our theoretical findings by numerical examples.

Reduction of EpCAM-Positive Cells from a Cell Salvage Product Is Achieved by Leucocyte Depletion Filters Alone.

Intraoperative cell salvage reduces the need for allogeneic blood transfusion in complex cancer surgery, but concerns about the possibility of it re-infusing cancer cells have hindered its application in oncology. We monitored the presence of cancer cells on patient-salvaged blood by means of flow cytometry; next, we simulated cell salvage, followed by leucodepletion and irradiation on blood contaminated with a known amount of EpCAM-expressing cancer cells, assessing also residual cancer cell proliferation as well as the quality of salvaged red blood cell concentrates (RBCs). We observed a significant reduction of EpCAM-positive cells in both cancer patients and contaminated blood, which was comparable to the negative control after leucodepletion. The washing, leucodepletion and leucodepletion plus irradiation steps of cell salvage were shown to preserve the quality of RBCs in terms of haemolysis, membrane integrity and osmotic resistance. Finally, cancer cells isolated from salvaged blood lose their ability to proliferate. Our results confirm that cell salvage does not concentrate proliferating cancer cells, and that leucodepletion allows for the reduction of residual nucleated cells, making irradiation unnecessary. Our study gathers pieces of evidence on the feasibility of this procedure in complex cancer surgery. Nevertheless, it highlights the necessity of finding a definitive consensus through prospective trials.

3βHSD activity saturates at physiological substrate concentrations in intact cells.

Conversion of adrenally produced dehydroepiandrosterone (DHEA) to the potent androgen dihydrotestosterone (DHT) is an important mechanism by which prostate cancer reaches castration resistance. At the start of this pathway is a branch point at which DHEA can be converted to Δ4 -androstenedione by the enzyme 3β-hydroxysteroid dehydrogenase (3βHSD) or to Δ5 -androstenediol by 17βHSD. To better understand this process, we studied the kinetics of these reactions in cells. Prostate cancer cells (LNCaP cell line) were incubated with steroids (DHEA and Δ5 -androstenediol) over a range of concentrations and the steroid metabolism reaction products were measured by mass spectrometry or by high-performance liquid chromatography to determine reaction kinetics. To confirm the generalizability of results, experiments were also performed in JEG-3 placental choriocarcinoma cells. The two reactions displayed very different saturation profiles, with only the 3βHSD-catalyzed reaction beginning to saturate within a physiological substrate concentration range. Strikingly, incubating LNCaP cells with low (in the ~10 nM range) concentrations of DHEA resulted in a large majority of the DHEA undergoing 3βHSD-catalyzed conversion to Δ4 -androstenedione, whereas high concentrations of DHEA (in the 100s of nM range) resulted in most of the DHEA undergoing 17βHSD-catalyzed conversion to Δ5 -androstenediol. Contrary to expectations from previous studies that used purified enzyme, cellular metabolism of DHEA by 3βHSD begins to saturate in the physiological concentration range, suggesting that fluctuations in DHEA concentrations could be buffered at the downstream active androgen level.

CNSC-01. GABAERGIC NEURON-TO-GLIOMA SYNAPSES IN DIFFUSE MIDLINE GLIOMAS

Abstract High-grade gliomas include clinically and molecularly distinct subtypes that stratify by anatomical location into diffuse midline gliomas (DMG) such as diffuse intrinsic pontine glioma (DIPG) and hemispheric high-grade gliomas. Neuronal activity drives high-grade glioma progression both through paracrine signaling and direct neuron-to-glioma synapses. Glutamatergic, AMPA receptor-dependent synapses between neurons and malignant glioma cells have been demonstrated in both pediatric and adult high-grade gliomas, but neuron-to-glioma synapses mediated by other neurotransmitters remain largely unexplored. Using whole-cell patch clamp electrophysiology, in vivo optogenetics and patient-derived glioma xenograft models, we have now identified functional, tumor-promoting GABAergic neuron-to-glioma synapses mediated by GABAA receptors in DMGs. GABAergic input has a depolarizing effect on DMG cells due to NKCC1 expression and consequently elevated intracellular chloride concentration in DMG tumor cells. As membrane depolarization increases glioma proliferation, we find that the activity of GABAergic interneurons promotes DMG proliferation in vivo. Increasing GABA signaling with the benzodiazepine lorazepam – a positive allosteric modulator of GABAA receptors commonly administered to children with DMG for nausea or anxiety - increases GABAA receptor conductance and increases glioma proliferation in orthotopic xenograft models of DMG. Conversely, levetiracetam, an anti-epileptic drug that attenuates GABAergic neuron-to-glioma synaptic currents, reduces glioma proliferation in patient-derived DMG xenografts and extends survival of mice bearing DMG xenografts. Concordant with gene expression patterns of GABAA receptor subunit genes across subtypes of glioma, depolarizing GABAergic currents were not found in hemispheric high-grade gliomas. Accordingly, neither lorazepam nor levetiracetam influenced the growth rate of hemispheric high-grade glioma patient-derived xenograft models. Retrospective real-world clinical data are consistent with these conclusions and should be replicated in future prospective clinical studies. Taken together, these findings uncover GABAergic synaptic communication between GABAergic interneurons and DMG cells, underscoring a tumor subtype-specific mechanism of brain cancer neurophysiology with important potential implications for commonly used drugs in this disease context.

Inhibition of USP7 induces p53-independent tumor growth suppression in triple-negative breast cancers by destabilizing FOXM1.

Although numerous studies indicate that inhibition of USP7 suppresses tumor growth by activating p53, the precise mechanism by which USP7 contributes to tumor growth through the p53-independent manner is not well understood. p53 is frequently mutated in most triple-negative breast cancers (TNBC), characterized as the very aggressive form of breast cancers with limited treatment options and poor patient outcomes. Here, we found that the oncoprotein Forkhead Box M1 (FOXM1) acts as a potential driver for tumor growth in TNBC and, surprisingly, through a proteomic screen, we identified USP7 as a major regulator of FOXM1 in TNBC cells. USP7 interacts with FOXM1 both in vitro and in vivo. USP7 stabilizes FOXM1 through deubiquitination. Conversely, RNAi-mediated USP7 knockdown in TNBC cells, dramatically reduced the levels of FOXM1. Moreover, based upon the proteolysis targeting chimera (PROTAC) technology, we generated PU7-1 (protein degrader for USP7-1), as a USP7 specific degrader. PU7-1 induces rapid USP7 degradation at low nanomolar concentrations in cells but shows no obvious effect on other USP family proteins. Strikingly, the treatment of TNBC cells with PU7-1 significantly abrogates FOXM1 functions and effectively suppresses cell growth in vitro. By using xenograft mouse models, we found that PU7-1 markedly represses tumor growth in vivo. Notably, ectopic overexpression of FOXM1 can reverse the tumor growth suppressive effects induced by PU7-1, underscored the specific effect on FOXM1 induced by USP7 inactivation. Together, our findings indicate that FOXM1 is a major target of USP7 in modulating tumor growth in a p53-independent manner and reveals the USP7 degrader as a potential therapeutic tool for the treatment of triple-negative breast cancers.

A New Cell Line Derived from the Caudal Fin of the Dwarf Gourami (Trichogaster lalius) and Its Susceptibility to Fish Viruses.

The detection of megalocytiviruses, especially the infectious spleen and kidney necrosis virus (ISKNV), in ornamental fish has increased with the rapid growth of the ornamental fish industry. In this study, dwarf gourami fin (DGF) cells derived from the caudal fin of the dwarf gourami (Trichogaster lalius), which is highly susceptible to red sea bream iridovirus (RSIV) and ISKNV, were established and characterized. The DGF cells were grown at temperatures ranging from 25 °C to 30 °C in Leibovitz's L-15 medium supplemented with 15% fetal bovine serum and were subcultured for more than 100 passages, predominantly with epithelial-like cells. DGF cells had a diploid chromosome number of 2n = 44. Although the initial purpose of this study was to establish a cell line for the causative agents of red sea bream iridoviral disease (RSIV and ISKNV), DGF cells were also susceptible to rhabdoviruses (viral hemorrhagic septicemia virus, hirame rhabdovirus, and spring viraemia of carp virus), exhibiting a significant cytopathic effect characterized by cell rounding and lysis. Additionally, viral replication and virion morphology were confirmed using virus-specific conventional polymerase chain reaction and transmission electron microscopy. Furthermore, both RSIV and ISKNV were replicated at high concentrations in DGF cells compared to other cell lines. Notably, the DGF cells maintained a monolayer during ISKNV infection, indicating the possibility of persistent infection. Thus, DGF can be used for viral diagnosis and may play a critical role in advancing our understanding of ISKNV pathogenesis.

Enhanced phytotoxicity of 4-chloro-3-methyphenol and lindane under sodium and potassium salt stresses

Investigating the uptake of organic pollutants by plants under salt stress is critical for evaluating crop contamination, understanding the mechanism of plant uptake, and implementing phytoremediation. The uptake of a highly phytotoxic contaminant, 4-Chloro-3-Methyphenol (CMP, 45 mg L−1), from solutions by wheat seedling with or without Na+ and K+ was studied to illustrate the synergistic effect of salt on phytotoxicity of CMP, using uptake kinetics, transpiration, Ca2+ leakage and fatty acid saturation as indicators. The influence of Na+ and K+ on the uptake of lindane, a relatively low toxic contaminant, from soil was also explored. Under CMP-Na+ and CMP-K+ exposure, the concentrations of CMP in both root and shoot were lower than those under CMP exposure, as a result of the inhibition of transpiration caused by Na+ and K+ stresses. Low concentration of CMP did not reveal serious toxicity on cell membrane. No apparent difference of MDA generation in root cells was observed, due to the lethal concentration of CMP. The relatively small variation of Ca2+ leakage and fatty acid saturation degree in the root cell under exposure of CMP, CMP-Na+ and CMP-K+, compared to intracellular CMP content, suggested the enhanced phytotoxicity of CMP induced by salt. Higher MDA concentration in shoot cell under CMP-Na+ and CMP-K+ exposure compared with that under CMP exposure again showed the synergetic toxicity of CMP. High Na+ and K+ concentration significantly facilitated the uptake of lindane by wheat seedlings in soils, indicating that it could boost the permeability of cell membrane, thereby increasing the toxicity of linande to wheat seedlings. The short-term effect of low salt concentration on the uptake of lindane was not obvious, but long-term exposure also led to increased uptake. In conclusion, the presence of salt could amplify the phtotoxicity of organic contaminant via several mechanisms.

Quantification of titanium and zirconium elements in oral mucosa around healthy dental implants: a case–control pilot study

ObjectivesMetallic particles are detected in different sites of the oral cavity, mainly in patients with peri-implantitis lesions. The aim of this pilot study was to analyze the levels of titanium and zirconium elements in the oral mucosa around healthy implants and to investigate the impact of titanium exogenous contamination on the measurements.Materials and MethodsForty-one participants were included in this three-phase study. Two groups of subjects were defined according to presence of titanium or zirconia implants (n: 20) or without any implants nor metallic restorations (n:21). Thirteen patients (n: 5 with zirconia implant; n: 3 with titanium implants; n: 5 control group) took part to the first part designed to optimize and validate the method of detecting titanium (Ti) and zirconium (Zr) elements in the oral mucosa and gingival tissues by the Inductively Coupled Plasma Mass Spectrometry (ICPMS). The second phase compared the levels of Ti and Zr concentrations in patients with implants (n: 12) and without implants (n: 6) who were controlled for their intake of titanium dioxide (TiO2). The last step included ten control subjects without any metallic devices to measure the concentration of Ti and Zr before and after having candies containing TiO2.ResultsIn the first phase, concentrations of Ti and Zr were below the limit of detection (LOD) in most cases, 0.18 μg/L and 0.07 μg/L respectively. In the titanium group, two out of three subjects displayed concentrations above the LOD, 0.21 μg/L and 0.66 μg/L. Zr element was only found in patients with zirconia implants. After controlling the intake of TiO2, all concentrations of Ti and Zr were below the limit of quantification (LOQ). Moreover, in patients with no implants, the Ti concentration in gingiva cells was superior for 75% of the samples after having a TiO2 diet.ConclusionsZirconium was only found in patients with zirconia implants, whereas titanium was detected in all groups even in subjects with no titanium implants. Zirconium and titanium elements were not detected in patients who were controlled for their intake of food and their use of toothpaste irrespective of the presence of implants or not. For 70% of the patients, the titanium detection was directly influenced by the intake of TiO2 contained candies.Clinical relevanceWhen analyzing titanium particles, it is necessary to pay attention to the risk of contamination bias brought by external products. When this parameter was controlled, no titanium particles were detected around clinically healthy implants.

PRMT5 reduces immunotherapy efficacy in triple-negative breast cancer by methylating KEAP1 and inhibiting ferroptosis

BackgroundAs an emerging treatment strategy for triple-negative breast cancer (TNBC), immunotherapy acts in part by inducing ferroptosis. Recent studies have shown that protein arginine methyltransferase 5 (PRMT5) has distinct roles...

Lactate: A critical regulator of cell proliferation via anaphase promoting complex remodeling

Lactate: A critical regulator of cell proliferation via anaphase promoting complex remodeling

Folate concentrations and serum perfluoroalkyl and polyfluoroalkyl substance concentrations in adolescents and adults in the USA (National Health and Nutrition Examination Study 2003–16): an observational study

Perfluoroalkyl and polyfluoroalkyl substances (PFAS) are a family of highly fluorinated aliphatic compounds, which are widely used in commercial applications, including food packaging, textiles, and non-stick cookware. Folate might counteract the effects of environmental chemical exposures. We aimed to explore the relationship between blood folate biomarker concentrations and PFAS concentrations. This observational study pooled cross-sectional data from the National Health and Nutrition Examination Survey (NHANES) 2003 to 2016 cycles. NHANES is a population-based national survey that measures the health and nutritional status of the US general population every 2 years by means of questionnaires, physical examination, and biospecimen collection. Folate concentrations in red blood cells and in serum, and perfluorooctanoic acid (PFOA), perfluorooctanesulfonic acid (PFOS), perfluorononanoic acid (PFNA), and perfluorohexane sulfonic acid (PFHxS) concentrations in serum were examined. We used multivariable regression models to assess the percentage change in serum PFAS concentrations in relation to changes in folate biomarker concentrations. We additionally used models with restricted cubic splines to investigate the shape of these associations. This study included 2802 adolescents and 9159 adults who had complete data on PFAS concentrations, folate biomarkers, and covariates, were not pregnant, and had never had a cancer diagnosis at the time of the survey. The mean age was 15·4 years (SD 2·3) for adolescents and 45·5 years (17·5) for adults. The proportion of male participants was slightly higher in adolescents (1508 [54%] of 2802 participants) than in adults (3940 [49%] of 9159 participants). We found negative associations between red blood cell folate concentrations and serum concentrations of PFOS (percentage change for a 2·7 fold-increase in folate level -24·36%, 95% CI -33·21 to -14·34) and PFNA (-13·00%, -21·87 to -3·12) in adolescents, and PFOA (-12·45%, -17·28 to -7·35), PFOS (-25·30%, -29·67 to -20·65), PFNA (-21·65%, -26·19 to -16·82), and PFHxS (-11·70%, -17·32 to 5·70) in adults. Associations for serum folate concentrations and PFAS were in line with those found for red blood cell folate levels, although the magnitude of the effects was lower. Restricted cubic spline models suggested linearity of the observed associations, particularly for associations in adults. In this large-scale, nationally representative study, we found consistent inverse associations for most examined serum PFAS compounds in relation to folate concentrations measured in either red blood cells or serum among both adolescents and adults. These findings are supported by mechanistic in-vitro studies that show the potential of PFAS to compete with folate for several transporters implicated in PFAS toxicokinetics. If confirmed in experimental settings, these findings could have important implications for interventions to reduce the accumulated PFAS body burden and mitigate the related adverse health effects. United States National Institute of Environmental Health Sciences.

Comparing two extracellular additives to facilitate extended storage of red blood cells in a supercooled state.

Background: Adenosine triphosphate (ATP) levels guide many aspects of the red blood cell (RBC) hypothermic storage lesions. As a result, efforts to improve the quality of hypothermic-stored red cell concentrates (RCCs) have largely centered around designing storage solutions to promote ATP retention. Considering reduced temperatures alone would diminish metabolism, and thereby enhance ATP retention, we evaluated: (a) whether the quality of stored blood is improved at -4°C relative to conventional 4°C storage, and (b) whether the addition of trehalose and PEG400 can enhance these improvements. Study Design and Methods: Ten CPD/SAGM leukoreduced RCCs were pooled, split, and resuspended in a next-generation storage solution (i.e., PAG3M) supplemented with 0-165mM of trehalose or 0-165mM of PEG400. In a separate subset of samples, mannitol was removed at equimolar concentrations to achieve a fixed osmolarity between the additive and non-additive groups. All samples were stored at both 4°C and -4°C under a layer of paraffin oil to prevent ice formation. Results: PEG400 reduced hemolysis and increased deformability in -4°C-stored samples when used at a concentration of 110mM. Reduced temperatures did indeed enhance ATP retention; however, in the absence of an additive, the characteristic storage-dependent decline in deformability and increase in hemolysis was exacerbated. The addition of trehalose enhanced this decline in deformability and hemolysis at -4°C; although, this was marginally alleviated by the osmolarity-adjustments. In contrast, outcomes with PEG400 were worsened by these osmolarity adjustments, but at no concentration, in the absence of these adjustments, was damage greater than the control. Discussion: Supercooled temperatures can allow for improved ATP retention; however, this does not translate into improved storage success. Additional work is necessary to further elucidate the mechanism of injury that progresses at these temperatures such that storage solutions can be designed which allow RBCs to benefit from this diminished rate of metabolic deterioration. The present study suggests that PEG400 could be an ideal component in these solutions.

C-Methylated Spermidine Derivatives: Convenient Syntheses and Antizyme-Related Effects.

The biogenic polyamines, spermidine (Spd) and spermine (Spm), are present at millimolar concentrations in all eukaryotic cells, where they participate in the regulation of vitally important cellular functions. Polyamine analogs and derivatives are a traditional and important instrument for the investigation of the cellular functions of polyamines, enzymes of their metabolism, and the regulation of the biosynthesis of antizyme-a key downregulator of polyamine homeostasis. Here, we describe convenient gram-scale syntheses of a set of C-methylated analogs of Spd. The biochemical properties of these compounds and the possibility for the regulation of their activity by moving a methyl group along the polyamine backbone and by changing the stereochemistry of the chiral center(s) are discussed.