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An organic solvent-free method based on limited dosing options (biocatalyst and zinc chloride) for the quick and mild recovery of chlorophyll (Chl) from spinach has been proposed. This tailored, custom-made protocol has been designed to produce stable green natural colorants. The kinetics of pigment extraction turned out to be a very useful tool to identify the proper conditions, in terms of biocatalyst dose (0.10-50 U/g), extraction time (1-48h), and ZnCl2 amount (50-300ppm), both for enhancing the recovery yield and preserving the green color. Considering the extraction kinetics, the recovery yield, and the colorimetric data, the suitable conditions to produce stable green and food-grade colorants are 0.10 U/g of enzyme, 3h, and 150ppm of ZnCl2 at 25°C. The extraction yield of Chl (4863µg/U) was about 51% greater than control, with a higher extraction rate constant (5.43 × 10-4 g/(µg min)). Considering the impact of ZnCl2 amount on Chl, its protective action resulted to be more noticeable toward Chl a: at 150ppm, an increased amount of about 2.5 and 1.5 times was found for Chl a and Chl b, respectively, in comparison to the reference (0ppm ZnCl2). PRACTICAL APPLICATION: This research demonstrates how a suitable kinetic approach helps to provide a tailored protocol, customized for the vegetable matrix, to produce stable green natural colorants from spinach. Lowering enzyme dosage and ZnCl2 amount during the extraction of chlorophyll at low temperature is crucial for its potential use as a colorant in food industry, providing high economic values through saving time and environmental protection.

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The present study successfully synthesized a novel biochar adsorbent (M-L-BC) using litchi seed modified with zinc chloride for PFASs removal in water. M-L-BC greatly enhanced removal of all examined PFASs (>95 %) as compared to the pristine biochar (<40 %). The maximum adsorption capacity was observed for PFOS, reaching 29.6 mg/g. Adsorption kinetics of PFASs followed the pseudo-second-order model (PSO), suggesting the predominance of chemical adsorption. Moreover, characterization and density functional theory (DFT) calculations jointly revealed involvement of surface complexation, electrostatic interactions, hydrogen bonding, and hydrophobic interactions in PFAS adsorption. Robust PFAS removal was demonstrated for M-L-BC across a wide range of pH (3-9), and coexisting ions had limited impact on adsorption of PFASs except PFBA. Furthermore, M-L-BC showed excellent performance in real water samples and retained reusability after five cycles of regeneration. Overall, M-L-BC represents a promising and high-quality adsorbent for efficient and sustainable removal of PFASs from water.

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Plastic food packaging, with its harmful migration of microplastics and nanoplastics into food, presents significant ecological imbalance and human health risks. In this regard, using food and agricultural byproducts as packaging materials reduces environmental and economic concerns and supports their sustainable management. Herein, cellulosic residue from corncob was employed as a renewable source for developing biodegradable packaging films. It was solubilized in ZnCl2 solution, crosslinked with Ca2+ ions, and plasticized with sorbitol to form films and used to improve the shelf-life of raspberries. The optimized film possesses water vapor permeability, tensile strength, and elongation at break of 1.8(4) x10-10 g-1 s-1 Pa-1, 4.7(1) MPa, and 15.4(7)%, respectively. It displays UV-blocking and antioxidant properties and biodegrades within 29 days at 24% soil moisture. It preserves raspberries for 7 and 5 more days at room temperature and refrigeration conditions, respectively, compared to polystyrene film. Overall, more value addition could be envisioned from agricultural residues to minimize post-harvest losses and food waste through biodegradable packaging, which also aids in mitigating plastic perils.

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This study was aimed to investigate the effects of different dietary zinc sources on the diarrhea rate, intestinal morphology, immune indexes and intestinal microbial composition of weaned piglets. A total of 240 weaned piglets (Duroc × Landrace × Yorkshire), at the age of 21 days, were randomly assigned to five dietary treatments for a four-week feeding trial to determine the effects of different amounts of tetrabasic zinc chloride (TBZC) supplementation on intestinal morphology, intestinal immune indices and intestinal microflora in weaned piglets, compared with the pharmacological dose of ZnO. The dietary treatments included a negative control (CON), (T1) ZnO (ZnO, 1500 mg/kg), (T2) tetrabasic zinc chloride (TBZC, 800 mg/kg), (T3) tetrabasic zinc chloride (TBZC, 1000 mg/kg), and (T4) tetrabasic zinc chloride (TBZC, 1200 mg/kg). Each treatment comprised six replicate pens, with eight pigs (four barrows and four gilts) per pen. Dietary TBZC of 1200 mg/kg improved the duodenum villus height, jejunum villus height and crypt depth of ileum, and increased the ratio of villus height to crypt depth of ileum (p < 0.05). The dietary supplementation of TBZC at a dosage of 1200 mg/kg has the potential to increase the levels of immunoglobulin G (IgG) and immunoglobulin A (IgA) in the duodenal mucosa. Furthermore, it shows a significant increase in the levels of immunoglobulin A (IgA) in the ileum. Compared with CON, TBZC significantly (p < 0.05) decreased pH values of stomach contents. It also increased the number of Firmicutes in intestinal contents. Compared with CON, the abundance of Firmicutes in jejunum contents of other treatments was significantly improved (p < 0.05), while the abundance of Proteobacteria in ileum contents of high-zinc treatments (T2 and T5) was decreased (p < 0.05). In conclusion, dietary TBZC of 1200 mg/kg improved the digestibility of crude protein in weaned piglets, altered the intestinal morphology of piglets, changed the intestinal microflora of piglets, reduced the diarrhea rate, and significantly improved the development of the small intestine of weaned piglets, and its regulation mechanism on intestinal tract needs further study. In summary, TBZC is likely to be an effective substitute source for the pharmacological dose of ZnO to control diarrhea in weaned piglets.

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The inhalation of zinc chloride (ZnCl2) smoke is one of common resources of lung injury, potentially resulting in severe pulmonary complications and even mortality. The influence of ZnCl2 smoke on lysine succinylation (Ksucc) in the lungs remains uncertain. In this study, we used a ZnCl2 smoke inhalation mouse model to perform global proteomic and lysine succinylome analyses. A total of 6781 Ksucc sites were identified in the lungs, with injured lungs demonstrating a reduction to approximately 2000 Ksucc sites, and 91 proteins exhibiting at least five differences in the number of Ksucc sites. Quantitative analysis revealed variations in expression of 384 proteins and 749 Ksucc sites. The analysis of protein-protein interactions was conducted for proteins displaying differential expression and differentially expressed lysine succinylation. Notably, proteins with altered Ksucc exhibited increased connectivity compared with that in differentially expressed proteins. Beyond metabolic pathways, these highly connected proteins were also involved in lung injury-associated pathological reactions, including processes such as focal adhesion, adherens junction, and complement and coagulation cascades. Collectively, our findings contribute to the understanding of the molecular mechanisms underlaying ZnCl2 smoke-induced lung injury with a specific emphasis on lysine succinylation. These findings could pave the way for targeted interventions and therapeutic strategies to mitigate severe pulmonary complications and mortality associated with such injuries in humans.

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Density separation can isolate microplastics from environmental samples containing sediment. Typically, a solution added to sediment causes microplastics with lower densities to float. The solution of choice can influence the recovery of different particles since denser solutions can separate a greater range of microplastics. The equipment and procedural complexity further influence density separation protocols and microplastic recoveries. Zinc chloride (ZnCl2) is frequently used to isolate high-density polymers from environmental samples yet is rarely validated with simple, well-described protocols. A simple overflow method using ZnCl2 to isolate microplastics from sediment samples is described following a 3-step process: (1. Separation) ZnCl2 (1.7 g cm-3) solution is added to a sediment sample, agitated then settled; (2. Overflows) buoyant particles at the surface of the solution are overflowed twice; (3. Filtration) the overflowed solution is filtered. In a validation experiment with polyamide, rubber, polyvinyl chloride and polyethylene terephthalate/polyester, the mean recovery using this overflow method was 96 % ± 0.6 (standard error). This overflow density separation method proposes an accessible and reliable protocol to extract medium and high-density microplastics.•Microplastic separation with concentrated ZnCl2 solution•Simple overflow of buoyant particles•Reliable extraction of microplastics.

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Background Chronic epipharyngitis causes postnasal drip, pharyngeal pain, pharyngeal discomfort, headache, and shoulder stiffness. Additionally, autonomic nervous system symptoms such as dizziness, general fatigue, and sleeplessness may occur. It can also contribute to the development of focal diseases. Although epipharyngeal abrasive therapy (EAT) is effective for chronic epipharyngitis involving the abrasion of the epipharynx with a zinc chloride solution, there is a lack of clear diagnostic criteria, and treatment outcomes are rarely reported. Methodology A classification of the severity of chronic epipharyngitis was attempted in 154 cases based on nasopharyngeal endoscopic findings, with a subsequent examination of treatment outcomes using EAT. Diagnosis of chronic epipharyngitis involved identifying redness, swelling, postnasal drip, and crusting of the epipharyngeal mucosa. Severity classification relied on a four-point scale measuring the degree of redness and swelling, with additional points assigned for the presence of postnasal drip and crusting. This classification also served as a criterion for judging treatment effectiveness. The prevalence and improvement rate of black spots and granular changes were assessed through nasopharyngeal endoscopy with narrow-band imaging. Subjective symptoms were evaluated using before and after treatment questionnaires, employing a four-point scale for symptoms commonly associated with chronic epipharyngitis (headache, postnasal drip, nasal obstruction, pharyngeal discomfort, pharyngeal pain, shoulder stiffness, tinnitus, ear fullness, dizziness, cough, and sputum). A 10-point numerical rating scale (NRS) was used to assess the physical condition. Results Following EAT, the severity of nasopharyngeal endoscopic findings notably improved, with a 76.0% (117/154) improvement rate (remarkable improvement: 19.5% (30), improvement: 56.5% (87)). The improvement rate for the chief complaint reached 85.7% (132/154), demonstrating significant enhancement in the score for each symptom. NRS scores also improved at a rate of 76.0% (117/154). A significant correlation was observed between the improvement in local findings and chief complaints. The prevalence of black spots and granular changes before EAT was 83.8% (129/154) and 64.3% (99/154), exhibiting improvement rates of 65.9% (87/132) and 54.8% (57/104), respectively. Conclusions Nasopharyngeal endoscopy proves valuable for diagnosing and assessing the severity of chronic epipharyngitis, as well as evaluating treatment effectiveness. The findings indicate that EAT is an effective treatment for chronic epipharyngitis, with improvements in local findings correlating with enhancements in the chief complaint. This underscores the importance of employing aggressive EAT in managing patients with chronic epipharyngitis.

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The Expert Panel for Cosmetic Ingredient Safety (Panel) assessed the safety of 27 inorganic and organometallic zinc salts as used in cosmetic formulations; these salts are specifically of the 2+ (II) oxidation state cation of zinc. These ingredients included in this report have various reported functions in cosmetics, including hair conditioning agents, skin conditioning agents, cosmetic astringents, cosmetic biocides, preservatives, oral care agents, buffering agents, bulking agents, chelating agents, and viscosity increasing agents. The Panel reviewed the relevant data for these ingredients, and concluded that these 27 ingredients are safe in cosmetics in the present practices of use and concentration described in this safety assessment when formulated to be non-irritating.

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Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the assessment of the application for the renewal of the authorisation of zinc chloride hydroxide monohydrate as a nutritional feed additive for all animal species (3b609). The applicant reported changes in the production process and requested a change in the specification of the additive. The EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) confirms that the use of the additive under the current authorised conditions of use is safe for the target species and the consumers. The use of the additive in animal nutrition at the proposed use levels for the terrestrial species and land-based aquaculture systems is considered safe for the environment. The FEEDAP Panel cannot conclude on the safety of the additive for marine sediment compartment, when it is used in sea cages. The additive is an eye irritant, but not irritant to the skin and not a skin sensitiser. The exposure through inhalation is likely, but in the absence of data, the Panel is not in a position to conclude on the risk of exposure by inhalation. There was no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

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Highly microporous adsorbents have been under considerable scrutiny for efficient adsorptive storage of H2. Of specific interest are sustainable, chemically activated, microporous carbon adsorbents, especially from renewable and organic precursor materials. In this article, six peat-derived microporous carbon materials were synthesized by chemical activation with ZnCl2. N2 and CO2 gas adsorption data were measured and simultaneously fitted with the 2D-NLDFT-HS model. Thus, based on the obtained results, the use of a low ratio of ZnCl2 for chemical activation of peat-derived carbon yields highly ultramicroporous carbons which are able to adsorb up to 83% of the maximal adsorbed amount of adsorbed H2 already at 1 bar at 77 K. This is accompanied by the high ratio of micropores, 99%, even at high specific surface area of 1260 m2 g-1, exhibited by the peat-derived carbon activated at 973 K using a 1:2 ZnCl2 to peat mass ratio. These results show the potential of using low concentrations of ZnCl2 as an activating agent to synthesize highly ultramicroporous carbon materials with suitable pore characteristics for the efficient low-pressure adsorption of H2.

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Melasma is an acquired chronic condition characterized by hyperchromic patches in photo-exposed areas. The search for new compounds for the treatment of melasma without side effects is constant. In this context, the aim of this study was to investigate the in vitro cytotoxic and antimelanogenic effects of the trace elements Zinc (Zn) and Selenium (Se). In this study, we evaluated the effects of 30 µM hydroquinone, this concentration did not alter mitochondrial function (MTT assay), but increased the percentage of necrotic cells and levels of reactive species. Furthermore, it showed no influence on tyrosinase activity and melanin content. Unlike hydroquinone, exposure for 48 h to 100 µM Zn and 1 and 5 µM Se had no significant influence on the analysis of reactive species, as well as on the percentage of necrotic cells. Still, specifically in relation to 100 µM Zn, it decreased the melanin content. Given the above, the trace elements Zn and Se did not show toxicity at the concentrations tested and Zn showed a promising effect, however, the mechanism needs to be better explored in order to contribute to new and updated research in the fight against melasma with a perspective of therapeutic use.

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The antibiotic ciprofloxacin (CIP) is used to treat a variety of bacterial infections, yet it poses significant health risks to aquatic environments. While adsorption is a promising technique for CIP removal, current adsorption capacities remain limited. In this study, we introduce a diatomite and basic zinc chloride composite (ZnHC-Dt) prepared using a straightforward deposition method, with the ability to achieve highly efficient ciprofloxacin removal. ZnHC-Dt is characterized using field emission scanning electron microscopy (SEM), X-ray diffraction analysis (XRD), Fourier transform infrared spectroscopy (FTIR), and the Brunauer-Emmett-Teller method (BET). We also assess the zeta potential. The optimized ZnHC-Dt adsorbent, achieved at a mass ratio of 0.45 with ZnHC/(ZnHC+Dt), is adopted with a CIP adsorption capacity of 831.96 mg/g at 25 °C, broad pH adaptability (within 3.0-10.0), rapid adsorption rate (reaching equilibrium in 4 h), and stable performance under Na+ ionic strength. The CIP adsorption process follows pseudo-second-order kinetics and aligns well with the Langmuir adsorption model. The high adsorption capacity of ZnHC-Dt can be attributed to electrostatic attraction, hydrogen bonding, surface complexation, and available adsorption sites. During the desorption process, the CIP removal rate retains 65.33% effectiveness after five cycles. The results suggest that ZnHC-Dt holds significant potential for CIP removal in aqueous solutions.

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Most polymeric materials are synthetic and derived from petroleum, hence they accumulate in landfills or the ocean, and recent studies have focused on alternatives to replace them with biodegradable materials from renewable sources. Biodegradable wastes from food and agroindustry, such as spent coffee grounds (SCGs), are annually discarded on a large scale and are rich in organic compounds, such as polysaccharides, that could be used as precursors to produce films. Around 6.5 million tons of SCGs are discarded every year, generating an environmental problem around the world. Therefore, it was the aim of this work to develop films from the SCGs polysaccharide fraction, which is comprised of cellulose, galactomannans and arabinogalactans. Two types of crosslinking were performed: the first forming coordination bonds of calcium ions with polysaccharides; and the second through covalent bonds with 1,4-phenylenediboronic acid (PDBA). The films with Ca2+ ions exhibited a greater barrier to water vapor with a reduction of 44% of water permeability vapor and 26% greater tensile strength than the control film (without crosslinkers). Films crosslinked with PDBA presented 55-81% higher moisture contents, 85-125% greater permeability to water vapor and 67-150% larger elongations at break than the films with Ca2+ ions. Film biodegradability was demonstrated to be affected by the crosslinking density, with the higher the crosslinking density, the longer the time for the film to fully biodegrade. The results are promising and suggest that future research should focus on enhancing the properties of these films to expand the range of possible applications.

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The pharmacological activation of the GPR39 receptor has been proposed as a novel strategy for treating seizures; however, this hypothesis has not been verified experimentally. TC-G 1008 is a small molecule agonist increasingly used to study GPR39 receptor function but has not been validated using gene knockout. Our aim was to assess whether TC-G 1008 produces anti-seizure/anti-epileptogenic effects in vivo and whether the effects are mediated by GPR39. To obtain this goal we utilized various animal models of seizures/epileptogenesis and GPR39 knockout mice model. Generally, TC-G 1008 exacerbated behavioral seizures. Furthermore, it increased the mean duration of local field potential recordings in response to pentylenetetrazole (PTZ) in zebrafish larvae. It facilitated the development of epileptogenesis in the PTZ-induced kindling model of epilepsy in mice. We demonstrated that TC-G 1008 aggravated PTZ-epileptogenesis by selectively acting at GPR39. However, a concomitant analysis of the downstream effects on the cyclic-AMP-response element binding protein in the hippocampus of GPR39 knockout mice suggested that the molecule also acts via other targets. Our data argue against GPR39 activation being a viable therapeutic strategy for treating epilepsy and suggest investigating whether TC-G 1008 is a selective agonist of the GPR39 receptor.

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The solubility of zinc-chloride (ZnCl2) in different concentrations of hydrochloric acid have been experimentally studied. Solubility for anhydrous ZnCl2 was found to be the highest in 3-6 M hydrochloric acid solution. Elevating the temperature of the solvent further increased solubility albeit with diminishing returns above 50 °C when the evaporation of hydrochloric acid increases. The best solubility was found to be 26.1 ± 1.17 M in 6 M hydrochloric acid at 50 °C. This information is important for further studies which will be aimed at producing and testing a liquid target for the irradiation of [68Zn]ZnCl2 solution in hydrochloric acid. The testing will involve pressure, irradiation time, acquired activity and other parameters. In the current paper we only describe the experimental results for the solubility measurements for ZnCl2 in different hydrochloric acid concentrations, 68Ga production is not carried out yet.

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The annual production of wastewater from the olive table industry poses a serious problem owing to its high organic matter load, which is highly concentrated in phenolic compounds (PCs) and inorganic materials. This research used adsorption to recover PCs from table olive wastewater (TOWW). Activated carbon was employed as a novel adsorbent. The activated carbon was obtained from olive pomace (OP) and activated using a chemical agent (ZnCl2). Fourier transform infrared spectroscopy analysis (FTIR), Brunauer-Emmett-Teller analysis (BET), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS) were applied to characterize the activated carbon sample. To optimize the biosorption conditions of PCs (adsorbent dose (A), temperature (B), and time (C)), a central composite design (CCD) model was used. An adsorption capacity was 1952.34 mg g-1 for optimal conditions with an activated carbon dose of 0.569 g L-1, a temperature of 39 °C, and a contact time of 239 min. The pseudo-second-order and Langmuir models as kinetic and isothermal mathematical models were proved to be more appropriate for the interpretation of the adsorption phenomenon of PCs. PC recovery was performed in fixed-bed reactors. The results of the adsorption of PCs from TOWW by activated carbon could be an effective process at a low cost.

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Background: Many antimicrobial compounds in mouthwashes can have a negative impact on the oral microbiome. O-cymen-5-ol, a compound derived from a phytochemical, has a targeted mode of action and is being used as an alternative. However, its effect on the native oral microbiome is unknown. Aim: To assess the effect of a mouthwash formulated with o-cymen-5-ol and zinc chloride on the oral microbiome of healthy individuals. Methods: A mouthwash formulated with o-cymen-5-ol and zinc chloride was administered to a cohort of 51 volunteers for 14 days, while another cohort of 49 volunteers received a placebo. The evolution of the oral microbiome in both groups was analysed using a metataxonomic approach. Results: Analysis of the oral microbiome showed that the mouthwash selectively targeted potential oral pathogens while maintaining the integrity of the rest of the microbiome. Specifically, the relative abundance of several potentially pathogenic bacterial taxa, namely Fusobacteriota, Prevotella, Actinomyces, Granulicatella, Abiotrophia, Lautropia, Lachnoanaerobaculum, Eubacterium (nodatum group) and Absconditabacteriales (SR1) decreased, while the growth of Rothia, a nitrate-reducing bacterium beneficial for blood pressure, was stimulated. Conclusions: The use of o-cymen-5-ol and zinc chloride as antimicrobial agents in oral mouthwashes is a valuable alternative to classical antimicrobial agents.

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This study evaluated the characteristics of zinc chloride modified vitex doniana seed activated carbon (VDZnCl2) for the removal of methylene blue. VDZnCl2 was characterized for textural properties, surface morphology and surface chemistry. Batch adsorption of methylene blue by VDZnCl2 was evaluated for the effects of concentration, contact time, adsorbent dosage, and solution pH. The surface area increased from 14 to 933 m2/g with porous texture to facilitate adsorption. The SEM micrograph showed varieties of pores with widened cavities. The FTIR spectra showed the characteristics of O-H and C=C groups commonly found in carbonaceous materials. The maximum methylene blue adsorption was recorded as 238 mg/g at concentration range of 1-800 mg/L and VDZnCl2 dosage of 50 mg. Sips isotherm fitted well with the equilibrium data, suggesting that the adsorption by VDZnCl2 was a physical process onto its heterogeneous surface, while the applicability of pseudo-first-order kinetics implies that external diffusion was the rate controlling mechanism. The performance put up by VDZnCl2 suggested that it is a potential adsorbent substitute for dye wastewater treatment.

Vitex doniana commonly known as black plum is a deciduous flowering tree that grows around West Africa. Over 80% of the fruit is seed, and often discarded. To date, this seed has received marginal recognition as activated carbon feedstock for the removal of dyes and organic pollutants. However, there is no known study on the synthesis of zinc chloride (ZnCl₂) modified activated carbon from vitex doniana seed and its adsorption of methylene blue from wastewater. Therefore, this study was a first-time attempt of the synthesis of ZnCl₂ modified activated carbon adsorbent from vitex doniana and its removal of methylene blue from wastewater. This work systematically encouraged indigenous adsorbent synthesis for wastewater treatment and contribution to sustainable development goals.

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The G-protein coupled receptor 39 (GPR39) is gaining increasing attention as a target for future drugs, yet there are gaps in the understanding of its pharmacology. Zinc is an endogenous agonist or an allosteric modulator, while TC-G 1008 is a synthetic, small molecule agonist. Zinc is also a positive allosteric modulator for the activity of TC-G 1008 at GPR39. Activation of GPR39 by TC-G 1008 facilitated the development of epileptogenesis in the pentylenetetrazole (PTZ)-induced kindling model of epilepsy. Congruently, TC-G 1008 decreased the seizure threshold in the maximal electroshock seizure threshold (MEST) test. Here, we investigated the effects of TC-G 1008 under the condition of zinc deficiency. Mice were fed a zinc-adequate diet (ZnA, 50 mg Zn/kg) or a zinc-deficient diet (ZnD, 3 mg Zn/kg) for 4 weeks. Following 4 weeks of dietary zinc restriction, TC-G 1008 was administered as a single dose and the MEST test was performed. Additional groups of mice began the PTZ-kindling model during which TC-G 1008 was administered repeatedly and the diet was continued. TC-G 1008 administered acutely decreased the seizure threshold in the MEST test in mice fed the ZnD diet but not in mice fed the ZnA diet. TC-G 1008 administered chronically increased the maximal seizure severity and the percentage of fully kindled mice in those fed the ZnA diet, but not in mice fed the ZnD diet. Our data showed that the amount of zinc in a diet is a factor contributing to the effects of TC-G 1008 in vivo.

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Tetracycline antibiotics (TCs) and heavy metals are commonly used in livestock and poultry farming, leading to their coexistence in the aquatic environment. This coexistence causes combined toxicity to aquatic organisms. Here, zebrafish embryos were exposed to chlortetracycline (CTC), oxytetracycline (OTC), zinc chloride (ZnCl2), and their combinations for 120 h to evaluate their adverse effects on the growth, antioxidant system, immune system, and endocrine system during the early stage of life. OTC/ZnCl2 combined exposure significantly reduced the body weight, whereas the TCs/ZnCl2 combination significantly increased the heart rate of zebrafish larvae, suggesting growth impairment induced by TCs and ZnCl2. Further, combined groups showed more prominent toxicity to the antioxidant system than single groups, as revealed by related levels of enzyme activity and gene expression. In addition, the levels of most pro-inflammatory genes were downregulated, and those of NF-κB-related genes were upregulated in all treatment groups, indicating an immunosuppressive response and the potential role of NF-κB signaling, while the combined treatment was not more toxic than TCs or ZnCl2 alone. Similarly, hormone and endocrine related gene levels were determined. Although both single and combined exposures caused certain endocrine-disrupting effects, the combined exposure did not result in higher toxicity than a single exposure. Our findings showed that a mixture of TCs and ZnCl2 might exert greater toxic effects as compared to a single compound on some systems, providing fundamental data on the toxic effects of single and combined TC and ZnCl2 exposure on aquatic organisms, although studies are needed to explore the underlying mechanisms.

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