Dentin erosive wear is reduced by fluoride varnishes containing nanosized sodium trimetaphosphate in vitro.

This study evaluated the effect of fluoride varnishes containing micrometric or nanosized sodium trimetaphosphate (TMP) on dentin erosive wear in vitro. Bovine root dentin blocks were selected by surface hardness and randomly divided into five experimental groups/varnishes (n = 20/group): placebo, 5% sodium fluoride (NaF); 5% NaF+5% micrometric TMP; 5% NaF+2.5% nanosized TMP; and 5% NaF+5% nanosized TMP. Half of the surface of all blocks received a single application of the assigned varnish, with subsequent immersion in artificial saliva for 6 h. Varnishes were then removed and the blocks were immersed in citric acid (90 s, 4×/day, 5 days). After each erosive cycle, ten blocks of each group were immersed in a placebo dentifrice for 15 s (ERO), while the other ten blocks were subjected to abrasion by brushing (ERO+ABR). Dentin erosive wear was assessed by profilometry. Data were submitted to 2-way ANOVA and to the Holm-Sidak test (p<0.05). Dentin erosive wear was significantly higher for ERO+ABR than for ERO for all varnishes. TMP-containing varnishes promoted superior effects against dentin erosive wear compared with 5% NaF alone; and 5% nanosized TMP led to the lowest wear among all varnishes. In conclusion, the addition of TMP to conventional fluoride varnish (i.e., varnish containing only NaF) enhanced its protective effects against bovine root dentin erosion and erosion+abrasion. Additionally, the use of 5% nanosized TMP led to superior effects in comparison to 5% micrometric TMP, both for erosion and erosion+abrasion in vitro.

Gene Expression Reprogramming by Citrate Supplementation Reduces HepG2 Cell Migration and Invasion.

Citrate, which is obtained from oxaloacetate and acetyl-CoA by citrate synthase in mitochondria, plays a key role in both normal and cancer cell metabolism. In this work, we investigated the effect of 10 mM extracellular citrate supplementation on HepG2 cells. Gene expression reprogramming was evaluated by whole transcriptome analysis using gene set enrichment analysis (GSEA). The transcriptomic data were validated through analyzing changes in the mRNA levels of selected genes by qRT-PCR. Citrate-treated cells exhibited the statistically significant dysregulation of 3551 genes; 851 genes were upregulated and 822 genes were downregulated. GSEA identified 40 pathways affected by differentially expressed mRNAs. The most affected biological processes were related to lipid and RNA metabolism. Several genes of the cytochrome P450 family were upregulated in treated cells compared to controls, including the CYP3A5 gene, a tumor suppressor in hepatocellular carcinoma (HCC) that plays an important protective role in HCC metastasis. The citrate-induced dysregulation of cytochromes could both improve the effectiveness of chemotherapeutics used in combination and reduce the aggressiveness of tumors by diminishing cell migration and invasion.

Iron oxide nanoparticles improving multimetal phytoextraction in Helianthus annuus.

This study assessed the phytotoxicity of a mixture of five different trace elements (TEs) frequently found as pollutants in soils: arsenic, cadmium, copper, lead and zinc. On the other hand, the plant response to a magnetite (Fe3O4) nanoparticle amendment on this mixture as well as nanomagnetite remediation potential has been tested. Sunflower (Helianthus annuus) plants were grown for 90 days in soil contaminated with the five mentioned TEs at the limit levels of TEs in soils likely to receive sludge established by French legislation. Depending on the conditions, experimental set-ups were amended or not with 1% dry weight nanomagnetite (NPsMagn), citric acid-coated nanomagnetite (NPsMagn@CA) or micro-sized magnetite (µPs) in order to assess the behavior of nanomagnetites in a TEs-contaminated water-soil-plant system under repeated water-deficiency stress. The mixture of TEs did not induce phytotoxicity as estimated by plant growth, pigment content, maximum quantum yield of photosynthesis, oxidative impact and antioxidant response. Furthermore, both nanomagnetites treatments in a TEs-contaminated soil significantly increased biomass production by 64 % compared to control and antioxidant enzyme activities compared to control and TEs-treated plants. NPsMagn and NPsMagn@CA particularly enhance phytoextraction of Cd and Cu, increasing the amounts of TEs in aerial parts from 1.5 to 4.5 times compared to set-ups without nanomagnetites. Based on Cd, Cu, Pb and Zn contents in soil solutions, both nanomagnetites treatments improved TEs phytoextraction without increasing groundwater contamination. On the contrary, nanomagnetites significantly reduce arsenic uptake by plants and solubilization in dissolved phase. Our results show that modifying surface physicochemical properties of NPsMagn with citric acid coating does not improve their effects compared to bare NPsMagn. NPsMagn and NPsMagn@CA also appear to mitigate the effects of drought stress. This work highlights several positive environmental aspects related to the use of nanomagnetites in phytoremediation.

Dissolution behavior of stone wool fibers in synthetic lung fluids: Impact of iron oxidation state changes induced by heat treatment for binder removal.

Stone wool fiber materials are commonly used for thermal and acoustic insulation, horticulture and filler purposes. Biosolubility of the stone wool fiber (SWF) materials accessed through acellular in vitro dissolution tests can potentially be used in future as an indicator of fiber biopersistence in vivo. To correlate acellular in vitro studies with in vivo and epidemiological investigations, not only a robust dissolution procedure is needed, but fundamental understanding of fiber behavior during sample preparation and dissolution is required. We investigated the influence of heat treatment procedure for binder removal on the SWF iron oxidation state as well as on the SWF dissolution behavior in simulant lung fluids (with and without complexing agents). We used heat treatments at 450 °C for 5 min and 590 °C for 1 h. Both procedures resulted in complete binder removal from the SWF. Changes of iron oxidation state were moderate if binder was removed at 450 °C for 5 min, and there were no substantial changes of SWF's dissolution behavior in all investigated fluids after this heat treatment. In contrast, if binder was removed at 590 °C for 1 h, complete Fe(II) oxidation to Fe(III) was observed and significant increase of dissolution was shown in fluids without complexing agent (citrate). PHREEQC solution speciation modeling showed that in this case, released Fe(III) may form ferrihydrite precipitate in the solution. Precipitation of ferrihydrite solid phase leads to removal of iron cations from the solution, thus shifting reaction towards the dissolution products and increasing total mass loss of fiber samples. This effect is not observed for heat treated fibers if citrate is present in the fluid, because Fe(III) binds with citrate and remains mobile in the solution. Therefore, for developing the most accurate SWF in vitro acellular biosolubility test, SWF heat treatment for binder removal is not recommended in combination with dissolution testing in fluids without citrate as a complexing agent.

Boron deficiency decreased the root activity of Ga-exposed rice seedlings by reducing iron accumulation and increasing Ga in iron plaque.

Gallium (Ga) is an emerging chemical pollutant chiefly associated with high-tech industries. Boron (B) alleviates the negative effects of toxic elements on plant growth. Thereby, the effects of B fertilization on Ga toxicity in rice seedlings was studied to clarify the role of iron plaque in the distribution of Ga, Fe, and B in Ga-treated rice seedlings in the presence or absence of B. Gallium exposure significantly reduced the biomass of rice seedlings. Boron deficiency induced a significant change in the distribution of B in Ga-treated rice seedlings compared with "Ga+B" treatments. Accumulation of Ga in roots, dithionite-citrate-bicarbonate (DCB) extracts, and shoots showed a dose-dependent manner from both +B and -B rice seedlings. Boron nutrition levels affect the distribution of Fe in roots, DCB extracts, and shoots, in which DCB-extractable Fe was significantly decreased from "Ga-B" treatments compared with "Ga+B" treatments. Root activity was significantly decreased in both Ga-exposed rice seedlings; however, B-deficient seedlings showed a severe reduction than +B rice seedlings. These results reveal that Fe plaque might be a temporary sink for B accumulation when plants are grown with proper B, wherein the re-utilization of DCB-extractable B stored in Fe plaque is mandatory for plant growth under B deficiency. Correlation analysis revealed that B deficiency decreased the root activity of Ga-exposed rice seedlings by reducing DCB-extractable Fe and increasing DCB-extractable Ga in Fe plaque. This study enhances our understanding of how B nutritional levels affect Ga toxicity in rice plants.

Plant growth promoting bacteria and citric acid promote growth and cadmium phytoremediation in ryegrass.

Based on the growth-promoting effect of plant growth promoting bacteria on plants and the mobilization of Cd by citric acid, an experiment was designed in which the combined treatment of Bacillus megaterium and citric acid promoted ryegrass to repair Cd-contaminated soil. This study aimed to evaluate the effects of different treatments on the antioxidant enzyme activity, photosynthesis intensity, Cd accumulation, and rhizosphere cadmium migration under cadmium contamination conditions. And the soil morphology and structure changes were studied by infrared spectroscopy FourierTransformInfrared(FT-IR) and scanning electron microscope Energy Dispersive Spectrometer(SEM-EDS) before and after different treatments. The results show that the combined treatment of Bacillus megaterium and citric acid significantly improved the oxidative stress defense and plant photosynthesis and increased of rye biomass. rye biomass 1.28 times higher than CK treatment. Joint treatment significantly increased the amount of shoot accumulation of Cd, 2.31 times higher than CK treatment, increased the migration and accumulation of cadmium. FTIR and SEM-EDS also showed that the organic constituents such as O-H, C-O and C-N in soils as a major mechanism for mobilization of the heavy metal Cd. Thus, the combined treatment of Bacillus megaterium and citric acid can promote plant growth, improve the damage to ryegrass caused by single organic acid addition, and improve the plant extraction efficiency, which is a feasible way to repair Cd-contaminated soil through activated extraction system.

The novelty of this study is the combined application of bacteria and chelating agents to ryegrass to improve phytoremediation efficiency. Bacillus giganosus has a good role in promoting the growth of ryegrass. As citrate, a small molecule chelate, can activate heavy metal cadmium and detoxify heavy metals, so it was selected. This study revealed in detail the response of ryegrass to the heavy metal Cd after exogenous addition of Bacillus gigansus and citrate, which is important for the application of cadmium removal by phytoremediation.

[Remediation of Soil Cadmium Contamination by Solanum nigrum L. Enhanced by the Combination of Exogenous Bacteria and Citric Acid].

To determine how to strengthen the Cd-enriched plant Solanum nigrum L. to remediate cadmium(Cd)-contaminated soil, a pot experiment was conducted with five treatments:control treatment(CK), Glomus mosseae(GM), G. mosseae+citric acid(GM+CA), G. mosseae+Bacillus megaterium(GM+BM), and G. mosseae+B. megaterium+citric acid(GM+BM+CA). We measured soil total Cd, available Cd, plant Cd uptake, and microbial community changes and analyzed the effects of exogenous microbial agents and citric acid addition on the remediation effect of Cd contamination by S. nigrum L. The results showed that relative to that of the CK treatment, the root, stem, and leaf biomass of the GM treatment significantly increased by 35.67%, 41.35%, and 65.38%, and the root and stem biomass of the GM+BM+CA treatment significantly increased by 73.38% and 75.38%. The GM+BM+CA treatment significantly increased Cd accumulation in leaves by 226.84%. The GM+BM+CA treatment significantly increased the Cd transport factor from stem to leaves by 52.47%. The GM+BM+CA treatment significantly increased the leaf bioconcentration factor by 120.53%. In addition, the combined restoration also had an impact on the rhizosphere microbial community structure, especially in inducing the relative abundance of some key microbial groups such as Proteobacteria, Actinobacteria, Glomeromycota, and Olpidiomycota to increase by 2.00%-5.77%, 0.76%-9.96%, 2.11%-3.63%, and 0.54%-2.98%, respectively. According to the RDA analysis, Proteobacteria and Actinobacteria were negatively correlated with soil total Cd, whereas Glomeromycota and Olpidiomycota were negatively correlated with soil total Cd. The changes in key microorganisms enhanced the ability of S. nigrum L. to absorb rhizosphere nutrients and resist Cd stress, increased the Cd accumulation ability of S. nigrum L., and effectively reduced the total Cd content in soil. In conclusion, G. mosseae, citric acid, and B. megaterium activated insoluble Cd in the soil by co-inoculation, which contributed to more Cd accumulation by S. nigrum L. and also produced co-remediation with G. mosseae. The enrichment plant-microorganism combined remediation Cd-contaminated soil has good application potential.

The potential for citrate to reinforce epigenetic therapy by promoting apoptosis.

Epigenetic drugs induce ATP depletion, promoting a glycolysis-to-oxidative phosphorylation (OXPHOS) shift which sometimes favors tumor growth by promoting necroptosis over apoptosis. To restore effective apoptosis in tumors, we propose that the administration of citrate could inhibit ATP production, activate caspase-8 (a key necroptosis inhibitor), and downregulate key anti-apoptotic proteins (Bcl-xL and MCL1).

Citric acid and hydrogen sulfide cooperate to mitigate chromium stress in tomato plants by modulating the ascorbate-glutathione cycle, chromium sequestration, and subcellular allocation of chromium.

The study aimed to investigate the role of hydrogen sulfide (H2S) in regulating chromium stress (Cr-S) tolerance of tomato plants treated with citric acid (CA). Prior to the Cr treatment, tomato plants were foliar-fed with CA (100 µM) daily for 3 days. Subsequently, the plants were grown for another ten days in a hydroponic system in a 50 µM Cr (VI) solution. Chromium treatment reduced photosynthetic pigments and plant biomass, but boosted the levels of hydrogen peroxide (H2O2) malondialdehyde (MDA), H2S, phytochelatins (PCs), and glutathione (GSH), electrolyte leakage (EL), and antioxidant enzyme activity in tomato plants. However, the foliar spray of CA mitigated the levels of H2O2, MDA, and EL, promoted plant growth and chlorophyll content, enhanced antioxidant enzymes' activities, and increased H2S production in Cr-S-tomato plants. CA also increased the levels of GSH and PCs, potentially reducing the toxicity of Cr through regulated sequestration. Additionally, the application of sodium hydrogen sulfide (NaHS), a donor of H2S, improved CA-induced Cr stress tolerance. The addition of CA promoted Cr accumulation in root cell wall and leaf vacuoles to suppress its toxicity. To assess the involvement of H2S in CA-mediated Cr-S tolerance, 0.1 mM hypotaurine (HT), an H2S scavenger, was provided to the control and Cr-S-plants along with CA and CA + NaHS. HT reduced the beneficial effects of CA by decreasing H2S production in tomato plants. However, the NaHS addition with CA + HT inverted the adverse impacts of HT, indicating that H2S is required for CA-induced Cr-S tolerance in tomato plants.

Mitigating the toxicity of reactive oxygen species induced by cadmium via restoring citrate valve and improving the stability of enzyme structure in rice.

The mechanism of reactive oxygen species (ROS) burst in rice cells induced by cadmium (Cd) stress remains poorly understood. The present study shows that the burst of superoxide anions (O2·-) and hydrogen peroxide (H2O2) in roots and shoots led by Cd stress was attributed to the disturbance of citrate (CA) valve and the damage of antioxidant enzyme structure in the rice seedlings. Cd accumulation in cells altered the molecular structure of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) through attacking glutamate (Glu) and other residues, leading to the significant reduction of their activities in clearing O2·- and decomposing H2O2. Citrate supplementation obviously increased the activity of antioxidant enzymes and decreased ∼20-30% of O2·- and H2O2 contents in roots and shoots. Meanwhile, the synthesis of metabolites/ligands such as CA, α-ketoglutarate (α-KG) and Glu as well as the activities of related enzymes in CA valve were remarkably improved. The activities of antioxidant enzymes were protected by CA through forming stable hydrogen-bonds between CA and antioxidant enzymes, and forming the stable chelates between ligands and Cd. These findings indicate that exogenous CA mitigated the toxicity of ROS under Cd stress by the ways of restoring CA valve function to reduce the production of ROS, and improving the stability of enzyme structure to enhance antioxidant enzymes activity.

New insights into the protective effect of statherin-derived peptide for different acquired enamel pellicle formation times on the native human enamel surfaces.

OBJECTIVES: This study evaluated the protective impact of acquired enamel pellicle (AEP) engineering with statherin-derived peptide (StatpSpS), considering different AEP formation times. DESIGN: A total of 120 native human enamel specimens were divided into 2 main groups: 1) No AEP engineering and 2) AEP engineering with StatpSpS (pretreatment for 1 min; 37 °C, under agitation). Each group was further divided into 4 subgroups: No pellicle, or 1, 60-and-120 min AEP formation times (human saliva; 37 °C). The specimens were then subjected to an erosive challenge (1% citric acid; pH 3.6; 1 min; 25 °C). This procedure was repeated for 5 cycles. Relative surface reflection intensity (%SRI) was measured and scanning electron microscopy (SEM) of the enamel surface was done. RESULTS: All AEP engineering groups protected against initial dental erosion in comparison with No pellicle (p < 0.001), likewise all groups with AEP, independent of engineering or formation times (p 0.001). Furthermore, engineering with StatpSpS even without the presence of AEP protected the enamel when compared to the No engineering/No pellicle group (p < 0.0001). No difference was observed regarding the protection from the different AEP formation times (p > 0.05). Regarding the SEM analysis, in the "No AEP engineering & No AEP" group, a more severe effect of citric acid was observed, with more enamel prism heads and scratches on the surface when compared with the other groups. CONCLUSIONS: AEP provides almost instant protection at formation times even as short as 1 min, protecting the native enamel against erosion. Treatment with StatpSpS by itself provides similar protection as the AEP.

A mixture of organic acids and thymol protects primary chicken intestinal epithelial cells from Clostridium perfringens infection in vitro.

Necrotic enteritis causes economic losses estimated to be up to 6 billion US dollars per year. Clinical and subclinical infections in poultry are also both correlated with decreased growth and feed efficiency. Moreover, in a context of increased antibiotic resistance, feed additives with enhanced antimicrobial properties are a useful and increasingly needed strategy. In this study, the protective effects of a blend of thymol and organic acids against the effects of Clostridium perfringens type A (CP) on chicken intestinal epithelial cells were investigated and compared to bacitracin, a widely used antibiotic in poultry production. Primary chicken intestinal epithelial cells were challenged with CP for a total time of 3 h to assess the beneficial effect of 2 doses of citric acid, dodecanoic acid, and thymol-containing blend, and compare them with bacitracin. During the challenge, different parameters were recorded, such as transepithelial electrical resistance, cell viability, mRNA expression, and reactive oxygen species production. CP induced inflammation with cytokine production and loss of epithelial barrier integrity. It was also able to induce reactive oxygen species production and increase the caspase expression leading to cellular death. The high dose of the blend acted similarly to bacitracin, preventing the disruptive effects of CP and inducing also an increase in zonula occludens-1 mRNA expression. The low dose only partially prevented the disruptive effects of CP but successfully reduced the associated inflammation. This study shows that the usage of thymol combined with 2 organic acids can protect primary chicken intestinal epithelial cells from CP-induced damages creating a valid candidate to substitute or adjuvate the antibiotic treatment against necrotic enteritis.

Kisspeptin-10 in cryodiluent improves the post-thaw quality of Nili-Ravi buffalo (Bubalus bubalis) bull spermatozoa.

Effects of kisspeptin-10 as antioxidant in cryodiluent were evaluated on post-thaw quality of buffalo spermatozoa. Qualified semen samples from five bulls were pooled, divided into five aliquots and extended in Tris-citric acid cryodiluent containing differential doses of kisspeptin-10 (5, 10, 15, and 20 µmol L-1 and negative control. Extended sperm suspension was cooled to 4°C, packaged in 0.54 ml straws and cryopreserved. At post-thawing, catalase (unit mg-1 ), peroxidase (unit mg-1 ) and reduced glutathione (µmol L-1 ) levels were highest (p < 0.05) with 20 µmol L-1 of kisspeptin-10 as compared to negative control. Moreover, lipid peroxidation (nmol L-1  min-1  mg protein-1 ) level was lowest (p < 0.05) with 20 µmol L-1 of kisspeptin-10. Sperm progressive motility (%), rapid velocity (%) and kinematics were higher (p < 0.05) with 15 and 20 µmol L-1 of kisspeptin-10 as compared to negative control. Supra-vital plasma membrane integrity (%), viable sperm with intact acrosome (%) and DNA integrity (%) were improved (p < 0.05) with all doses of kisspeptin-10 as compared to negative control. It was concluded that the addition of 15 and 20 µmol L-1 kisspeptin-10 in cryodiluent ameliorated the overall frozen-thawed quality parameters of Nili-Ravi buffalo spermatozoa.

Binding of Citrate-Fe3+ to Plastic Culture Dishes, an Artefact Useful as a Simple Technique to Screen for New Iron Chelators.

NaCT mediates citrate uptake in the liver cell line HepG2. When these cells were exposed to iron (Fe3+), citrate uptake/binding as monitored by the association of [14C]-citrate with cells increased. However, there was no change in NaCT expression and function, indicating that NaCT was not responsible for this Fe3+-induced citrate uptake/binding. Interestingly however, the process exhibited substrate selectivity and saturability as if the process was mediated by a transporter. Notwithstanding these features, subsequent studies demonstrated that the iron-induced citrate uptake/binding did not involve citrate entry into cells; instead, the increase was due to the formation of citrate-Fe3+ chelate that adsorbed to the cell surface. Surprisingly, the same phenomenon was observed in culture wells without HepG2 cells, indicating the adsorption of the citrate-Fe3+ chelate to the plastic surface of culture wells. We used this interesting phenomenon as a simple screening technique for new iron chelators with the logic that if another iron chelator is present in the assay system, it would compete with citrate for binding to Fe3+ and prevent the formation and adsorption of citrate-Fe3+ to the culture well. This technique was validated with the known iron chelators deferiprone and deferoxamine, and with the bacterial siderophore 2,3-dihydroxybenzoic acid and the catechol carbidopa.

Bioactive Poly(octanediol-citrate-polyglycol) Accelerates Skin Regeneration through M2 Polarization Immunomodulating and Early Angiogenesis.

The inhibition of inflammation and the promotion of early angiogenesis are paid much attention in skin tissue engineering. Citric acid-based biomaterials are widely used in tissue engineering due to their bioactive structure and biocompatibility, but there are few studies on investigating their role and mechanism in wound repair and skin regeneration. Herein, the potential anti-inflammation mechanism of poly(octanediol-citrate-polyglycol) (POCG) copolymer is reported in regulating skin wound repair. It is found that POCG can modulate macrophages phenotype through downregulating the expression of proinflammatory cytokines (tumor necrosis facor-α (Tnf-α), Interleukin-1ß (IL-1ß), and Interleukin-6 (IL-6) and polarizing macrophages to anti-inflammatory (M2) phenotype. POCG can promote endothelial cell vascularization by increasing the expression of angiogenesis factors (vascular endothelial growth factor (Vegf) and cluster of differentiation 31CD31) mediated by the macrophage polarization. The in vivo study shows that POCG can accelerate skin wound repair through suppressing the acute inflammation and inducing early angiogenesis through the polarization modulation. Furthermore, the POCG polymer has good biocompatibility for both immune cells and tissue cells. This study may provide the important theoretical support on the bioactivity of citrate-based biomaterials and expanding their applications in tissue engineering.

Acid freezes uricolysis in addition to glycolysis: Utility of fluoride-EDTA-citrate tube in the measurement of uric acid for patients administered rasburicase.

BACKGROUND: In samples from patients administered rasburicase, ex vivo uricolysis leads to spuriously low uric acid results. The manufacturer's recommendation of storing the sample in ice-water until analysis, however, does not fully arrest uricolysis. Since uricase activity is affected by pH and metal chelators, we assessed uricolysis inhibition in sodium fluoride-ethylenediaminetetraacetic acid (EDTA)-citrate sample tube (FC Mix tube, Greiner) used primarily for plasma glucose. METHOD: A serum pool was spiked with rasburicase and uric acid measured at 15, 45, 90, 150, 240 and 1080 min in a lithium heparin tube in ice-water, plain tube at room temperature (RT), EDTA tube at RT, FC Mix tube in ice-water, FC Mix tube at RT and FC Mix tube at RT prepared by dissolving FC Mix in serum. RESULTS: The rate of urate decay was lowest in the FC Mix tube independent of temperature, then lithium heparin tube in ice-water, then EDTA tube at RT and highest in the plain tube at RT. Uric acid concentrations in the prepared FC Mix tube at RT and heparin tube in ice-water were, respectively, 98.2% and 93.8% of control values at 90 min, 97.1% and 89.3% of control values at 4 h, and remained higher in the prepared FC Mix tube at all time points. CONCLUSION: NaF-EDTA-citrate mixture largely arrested rasburicase mediated ex vivo uricolysis without the need for sample cooling. We propose that sample tubes containing NaF-EDTA-citrate be used for the measurement of uric acid in patients administered rasburicase.

Citrate Promotes Excessive Lipid Biosynthesis and Senescence in Tumor Cells for Tumor Therapy.

Metabolic disorder is one of the hallmarks of cancers, and reprogramming of metabolism is becoming a novel strategy for cancer treatment. Citrate is a key metabolite and critical metabolic regulator linking glycolysis and lipid metabolism in cellular energy homeostasis. Here it is reported that citrate treatment (both sodium citrate and citric acid) significantly suppresses tumor cell proliferation and growth in various tumor types. Mechanistically, citrate promotes excessive lipid biosynthesis and induces disruption of lipid metabolism in tumor cells, resulting in tumor cell senescence and growth inhibition. Furthermore, ATM-associated DNA damage response cooperates with MAPK and mTOR signaling pathways to control citrate-induced tumor cell growth arrest and senescence. In vivo studies further demonstrate that citrate administration dramatically inhibits tumor growth and progression in a colon cancer xenograft model. Importantly, citrate administration combined with the conventional chemotherapy drugs exhibits synergistic antitumor effects in vivo in the colon cancer models. These results clearly indicate that citrate can reprogram lipid metabolism and cell fate in cancer cells, and targeting citrate can be a promising therapeutic strategy for tumor treatment.

A novel method for explaining the product inhibition mechanisms via molecular docking: inhibition studies for tyrosinase from Agaricus bisporus.

The present study aims to investigate the substrate (4-methyl catechol and catechol) specificity and inhibition mechanisms (l-ascorbic acid, citric acid, and l-cysteine) of the tyrosinase enzyme (TYR), which is held responsible for browning in foods and hyperpigmentation in the human skin, through kinetic and molecular docking studies. During the experimental studies, the diphenolase activities of TYR were determined, following which the inhibitory effects of the inhibitors upon the diphenolase activities of TYR. The inhibition types were determined as competitively for l-ascorbic acid and citric acid and noncompetitive for l-cysteine. The kinetic results showed that the substrate specificity was better for catechol while l-cysteine showed the best inhibition profile. As for the in silico studies, they also showed that catechol had a better affinity in line with the experimental results of this study, considering the interactions of the substrates with TYR's active site residues and their distance to CuB metal ion, which is an indicator of diphenolase activity. Besides, the inhibitory mechanisms of the inhibitor molecules were explained by the molecular modeling studies, considering the binding number of the inhibitors with the active site amino acid residues of TYR, the number and length of H bonds, negative binding energy values, and their distance to CuB metal ion. Based on our results, we suggest that the novel method used in this study to explain the inhibitory mechanism of l-cysteine may provide an affordable alternative to the expensive methods available for explaining the inhibitory mechanism of TYR and those of other enzymes. HighlightsThe best affinity for the tyrosinase enzyme occurred with catechol.l-Ascorbic acid, citric acid, l-cysteine inhibited the diphenolic activity of tyrosinase.In silico studies confirmed the best affinity shown by catechol.Product inhibition mechanism of l-cysteine explained by in silico for the first time.Communicated by Ramaswamy H. Sarma.

Citrate-capped gold nanoparticles with a diameter of 14 nm alter the expression of genes associated with stress response, cytoprotection and lipid metabolism in CaCo-2 cells.

Advancements in nanotechnology have provided insight into the unique opportunities for the application of nanomaterials such as gold nanoparticles (AuNPs) in medicine due to their remarkable properties, which includes low toxicity, large surface area, and the ease of synthesis and conjugation to other molecules. Therefore, AuNPs are often preferred for bio-applications. Citrate-capped AuNPs (cAuNPs) have been reported to be non-cytotoxic and are used in numerous studies as drug delivery vehicles to treat various diseases. However, the limitations of bioassays often used to assess the toxicity of AuNPs have been well documented. Herein, we investigate the cytotoxicity of 14 nm cAuNPs in the human colorectal adenocarcinoma (Caco-2) cell line. Treatment conditions (i.e. dose and exposure time) that were established to be non-toxic to Caco-2 cells were used to investigate the effect of cAuNPs on the expression of a Qiagen panel of 86 genes involved in cytotoxicity. Out of 86 studied, 23 genes were differentially expressed. Genes involved in oxidative stress and antioxidant response, endoplasmic reticulum (ER) stress and unfolded protein response, heat shock response, and lipid metabolism were more affected than others. While low concentrations of 14 nm cAuNPs was not cytotoxic and did not cause cell death, cells treated with these nanoparticles experienced ER and oxidative stress, resulting in the activation of cytoprotective cellular processes. Additionally, several genes involved in lipid metabolism were also affected. Therefore, 14 nm cAuNPs can safely be used as drug delivery vehicles at low doses.

Putative Role of Ligands of DNIC in the Physiological Action of the Complex.

In a relatively isolated system of avian embryo, the metabolism of NO, a component of the dinitrosyl iron complexes (DNIC), the main NO donor in most tissues, depends on the ligands that make up the complex. This fact corroborates the earlier hypothesis that these ligands perform a regulatory function in NO metabolism. It is also shown that nitrite injected into the embryo is not oxidized to nitrate like NO in DNIC, but is accumulated outside the amniotic sac. Normally, nitrite is present in an embryo in trace amounts. These facts suggest that NO in the embryo is transferred from the donor molecule to a target in the embryo tissues further transformed with minimum oxidation to nitrite.