Metabolomics analysis of quality components metabolism during the growth process of pepino (Solanum muricatum) fruit.

Pepino (Solanum muricatum), a horticultural crop that has experienced significant growth in the highlands of China over the past two decades, is widely embraced by consumers due to its distinctive taste and nutritional advantages. This study focused on the cultivar 'Qingcanxiang' of pepino grown on the Qinghai-Tibetan Plateau was analyzed using UPLC-QTOF-MS and RNA-seq transcriptome sequencing. Fruit samples were collected at three distinct stages of development, and the results of the metabolomics and transcriptomics were compared and correlated. The study's findings indicate that the 'Qingcanxiang' fruit contained a total of 187 metabolites, comprising 12 distinct categories of compounds, including amino acids and their derivatives, organic acids, sugars and alcohols, phenols and phenolic acids. Of these metabolites, 94 were identified as differential. Significant variations in nutrient composition were observed across the three growth stages of the fruit. Specifically, the stage spanning from the growth to the maturation was identified as the critical stages for nutrient accumulation and flavor development. Transcriptome sequencing analysis revealed a set of highly associated genes between aspartate and quinic acid, namely SIR2, IRAK4, RP-L29, and CCNH. These genes are potentially involved in the regulation of both amino acid and phenolic acid synthesis. Through the application of metabolomics and transcriptomics, this investigation elucidates the alterations in metabolites and the underlying molecular regulatory mechanisms of pepino fruits during three growth stages. The findings furnish a theoretical foundation for the evaluation of nutritional quality and the enhancement of breeding strategies for pepino.

Revealing and Tuning the Photophysics of C=N Containing Photothermal Molecules: Excited State Dynamics Simulations.

Molecular photothermal conversion materials are recently attracting increasing attention for phototherapy applications. Herein we investigate the excitation and de-excitation processes of a photothermal molecule (C1TI) that is among the recently developed class of small-molecule-based photothermal imines with superb photothermal conversion efficiencies (PTCEs) up to 90% and a molecule (M2) that is constructed by replacing the amino group of C1TI with an H atom, via excited-state dynamics simulations based on the time-dependent density functional theory (TD-DFT). The simulations reveal fast (<150 fs of average time) nonradiative decays of the lowest excited singlet (S1) state to a conical intersection (CI) with the ground (S0) state in high yields (C1TI: 93.9% and M2: 87.1%). The fast decays, driven by C=N bond rotation to a perpendicular structural configuration, are found to be barrierless. The slight structural difference between C1TI and M2 leads to drastically different S0-S1 energy surfaces, especially M2 features a relatively much lower CI (0.8 eV in energy) and much more decay energy (1.0 eV) to approach the CI. This work provides insights into the de-excitation mechanisms and the performance tuning of C=N enabled photothermal materials.

The association of serum phthalate metabolites with biomarkers of ovarian reserve in women of childbearing age.

Phthalates (PAEs) are widely used plasticizers drawing increasing concern due to reproductive toxicity. However, studies on serum PAEs metabolites (mPAEs) and their associations with human ovarian function remain very scarce. In this study, from April 2019 to August 2020, a total of 297 women of childbearing age were recruited in Tianjin, China. Eleven mPAEs were analyzed in serum samples and eight mPAEs were detected at frequencies > 65% with median concentrations of 0.43-15.3 ng/mL. In multinomial logistic analysis, an increase in serum mono (2-isobutyl) phthalate (miBP) was associated with decline in antral follicle count (AFC) (OR=1.26, 95% CI: 0.99, 1.61) and 5-mono-(2-ethyl-5-hydroxyhexyl) phthalate (mEHHP) was significantly associated with AFC increase (OR=1.43, 95% CI: 1.06, 1.92), which were aligned with the associations found between mPAEs and AMH through generalized linear regression. In multiple linear regression models, per 10% increase in serum mono (2-ethylhexyl) phthalate (mEHP), mono (2-ethyl-5-oxohexyl) phthalate (mEOHP) (oxo-mEHP), and principal component 1 featured for high concentrations of mono-n-butyl phthalate (mBP), miBP and mEHP were associated with 0.15 (95% CI: -0.29, -0.02), 0.01 (95% CI: -0.01, 0.00) and 0.01 (95% CI: -0.02, 0.00) ln-unit decrease in estradiol (E2) levels, respectively, while mono-[(2-carboxymethyl) hexyl] phthalate (mCMHP) (carboxymethyl-mEHP) was positively associated with 0.05 ln-unit increase of E2 (95% CI: 0.02, 0.08). The observed negative associations between mPAEs and the Anti-Müllerian hormone (AMH) also aligned with the change in AFC. Generalized linear regression also revealed nonlinear associations between mono-ethyl phthalate (mEP), mCMHP and follicle-stimulating hormone (FSH). Overall, serum mEHP and its metabolites were negatively associated with E2. miBP was negatively associated with AFC. The nonlinear associations between mPAEs and FSH, and AMH need further study.

Pulmonary toxicology assessment of polyethylene terephthalate nanoplastic particles in vitro.

Nanoplastics are more likely to be suspended in air and pose a risk of respiratory exposure. However, the early health effects of low-dose nanoplastics on the respiratory system, which are expected to reflect the risk of atmospheric nanoplastics, need to be further evaluated. In this study, nanoparticles of polyethylene terephthalate, a representative plastic polymer in air, were prepared by a precipitation method. The toxicity impacts of nano-PET at environmental concentrations on the human lung carcinoma cell A549 cells were evaluated. Although the nano-PET was identified to enter the cells by confocal microscope observation and alkali-assisted thermal depolymerization coupled with LC-MS/MS analysis, the nano-PET exhibited low toxicity on mitochondrial membrane potential levels and cell apoptosis. At low concentrations of 0.10 and 0.98 µg/mL, the nano-PET had a slight promotion effect on cell viability, while an inhibitory effect on cell viability presented at higher nano-PET concentrations of 98.40 and 196.79 µg/mL. The cell survival rate at 98.4 and 196.79 µg/mL of nano-PET are lower than that of the control, and significant oxidative stress in cells caused by the nano-PET exposure at 49.2 µg/mL was observed. A decrease tendency of mitochondrial membrane potential with the increasing nano-PET exposure presents, which is consistent with the change of reactive oxygen species. Furthermore, nano-PET at ≦ 98.4 µg/mL could not increase the sum of apoptotic in the cells, but the late apoptotic cells increased with the increase of the exposure dose. The major mechanism of the toxic effect of nano-PET on cells may be the increase of reactive oxygen species caused by oxidative stress, which in turn induces a decrease in the mitochondrial membrane potential. This study provides information on the toxicity of nano-PET at environmental concentrations in human lung cells, which helps to enrich the risk cognition of nanoplastics in the respiratory system.

Immunotoxicity responses to polystyrene nanoplastics and their related mechanisms in the liver of zebrafish (Danio rerio) larvae.

Nanoplastics in aquatic environments may induce adverse immunotoxicity effects in fish. However, there is insufficient evidence on the visible immunotoxicity endpoints in the larval stages of fish. The liver plays an important role in systemic and local innate immunity in the fish. In this study, the hepatic inflammatory effects of polystyrene (PS) nanoplastic particles (NPs: 100 and 50 nm) and micron PS particles on transgenic zebrafish (Danio rerio) larvae were estimated using fluorescent-labeled neutrophils, macrophages, and liver-type inflammatory binding protein (fabp10a). Particles with smaller size induced higher aggregations of neutrophils and apoptosis of macrophages in the abdomen of the larvae, corresponding to greater hepatic inflammation in the larvae. NPs increased the expression of fabp10a in the larval livers in a dose- and size-dependent manner. PS particles of 50 nm at a concentration of 0.1 mg·L-1 increased the expression of fabp10a in the larval liver by 21.90% (P < 0.05). The plausible mechanisms of these effects depend on their distribution and the generation of reactive oxygen species in the larvae. Metabonomic analysis revealed that the metabolic pathways of catabolic processes, amino acids, and purines were highly promoted by NPs, compared to micron PS particles. NPs also activate steroid hormone biosynthesis in zebrafish larvae, which may lead to the occurrence of immune-related diseases. For the first time, the liver was identified as the target organ for the immunotoxicity effects of NPs in the larval stage of fish.

Diet preference of zebrafish (Danio rerio) for bio-based polylactic acid microplastics and induced intestinal damage and microbiota dysbiosis.

The ingestion of petroleum-based microplastics (MPs) by aquatic animals and their toxicological effects are of wide concern. However, the ecological risks of bio-based MPs to aquatic animals remain largely unknown. In the present study, zebrafish (Danio rerio) were exposed to MPs of polylactic acid (PLA), the most widely used bio-based plastic, and polyethylene terephthalate (PET), a high-production volume petroleum-based plastic. PLA MPs were more actively ingested by fish than PET MPs. The abundance of PLA MPs in fish intestines was approximately 170 times greater than that of PET MPs after one day of exposure. The ingestion of PLA MPs caused gastrointestinal damage in zebrafish. In addition, the ingestion of PLA MPs induced specific changes in the diversity of intestinal microbiota and promoted species closely linked with energy metabolism, cellular processes, and fish diseases. This might have been related to the depolymerization of PLA in the digestive tract, which decreased the intestinal pH and changed the carbon source structure. Overall, bio-based MPs may have different ecological effects on aquatic animals than traditional petroleum-based MPs.

Benzotriazole alleviates copper mediated lysosomal membrane damage and antioxidant defense system responses in earthworms (Eisenia fetida).

Benzotriazole (BTR) is a common corrosion inhibitor used to protect copper (Cu) and Cu alloys. To reveal the combined subacute toxicity of BTR and Cu at environmental levels on terrestrial animals, the activity of antioxidative enzymes and the glutathione levels in earthworms (Eisenia fetida) of the single or co-exposure treatments were determined. The activity of both antioxidant enzymes and non-enzymatic antioxidants was affected by BTR in earthworms. Moreover, the analyses of lysosomal neutral red retention time and total antioxidant capacity indicated a detoxification effect of BTR on Cu-induced impairments of the antioxidant defense capacity in earthworms. The apoptotic rate of coelomocytes in earthworms of the co-exposure treatment was lower than that in earthworms treated with Cu only, indicating that BTR alleviates Cu mediated lysosomal membrane damage and antioxidant defense system responses in earthworms.

Maternal exposure to TCDD during gestation advanced sensory-motor development, but induced impairments of spatial learning and memory in adult male rat offspring.

2,3,7,8-Tetrachlorodibenzo-p-Dioxin (TCDD) is an endocrine disrupting chemical (EDC) with high persistency. Even a low amount can pass the placental barrier during gestational exposure. Exposure to TCDD exposure can impair the development of the nervous system in children, leading to impaired learning ability in later-life. But the changes in neurobehavioral developments in infancy and childhood caused by TCDD are unknown. Pregnant Sprague-Dawley rats were given a consecutive daily dose of TCDD (200 or 800 ng/day/kg) or an equivalent volume of vehicle by gavage on gestational days 8-14 (GD 8-14) as the prenatal TCDD exposure model. In the offspring, early neurobehavioral development was assessed at postnatal day 5 (PND5) and eye-opening was monitored from PND10 onwards. Adult male offspring was tested by Morris Water Maze for spatial memory and learning ability evaluation. Hippocampus Nissl's staining and astrocyte GFAP immunohistochemistry were used to evaluate the activity of astrocytes. The results of the behavioral tests showed that gestational TCDD exposure induced premature motor activity and earlier eyes-opening, but lead to serious deficits of spatial memory and learning ability in the adult male offspring. Morphology and number of neurons in the hippocampus CA1 region was not affected, while the activity of astrocytes in the same region was significantly reduced. These data indicate that perinatal TCDD exposure induced premature neurobehavioral development but impaired the spatial learning and memory in adult male rat offspring. The decreased activity of astrocytes in the hippocampus may play a role in these adverse effects.

Effects of Force and Inflammatory Factors on the Expressions of Osteogenesis Regulators in Human Periodontal Ligament Cells.

Objective To observe the effects of force signals and inflammatory cytokines on the expressions of functional proteins during the differentiation of periodontal ligament cells(PDLCs) into osteoclasts. Methods The caries-free premolars that needed to be removed for orthodontic treatment were collected,human periodontal ligament cells were cultured in vitro.Human PDLCs were exposed to inflammatory cytokines including interleukin(IL)-1ß,-6,-23,and tumor necrosis factor alpha(TNF-α). Cyclicmechanical tension with a maximum 5% elongation for different durations(0,2,4,8,12,and 24 hours) were applied. Then the expressions of signaling molecules related to osteoclastogenesis(OPG) and receptor activated nuclear factor κB ligand(RANKL) were determined at protein levels by Western blotting. Results Inflammatory cytokines improved the expressions of osteoclastgenesis regulators in hPDLCs,while cyclic-tension force reduced their expressions. However,the combined effect of inflammatory cytokines and cyclic-tension force resulted in high expressions of osteoclastgenesis regulators. Conclusion Inflammatory cytokines can promote the expressions of the osteoclastgenic factors,which can not be offset by cyclic-tension force.

Effects of Periodontal Ligament Cells on Alveolar Bone Metabolism under the Action of Force and Inflammatory Factors and Its Molecular Mechanisms.

Periodontal ligament may have independent response to orthodontic stimulation and thus initiate alveolar bone osteogenesis and osteoclasts. Orthodontic-induced alveolar bone remodeling has no bone loss,while inflammation can lead to alveolar bone loss,suggesting that force signal and inflammatory factors may induce the differentiation of undifferentiated cells in the periodontal ligament via different pathways. The strength of the force and the nature of the force (basal tension and fluid shear force) may affect the differentiation of periodontal ligament cells,and may produce antagonistic or synergistic effect with the inflammatory factors through complex autocrine and paracrine regulation,resulting in local bone reconstruction,which is manifested as bone deposition and bone absorption. Studies have shown that Wnt signaling is an important regulatory pathway for osteoblast differentiation. Inflammatory factors can block the differentiation of stem cells into osteoblasts. The Wnt pathway is closely related to the effects of force and inflammatory factors on the differentiation of periodontal ligament cells.

Mitotherapy for Fatty Liver by Intravenous Administration of Exogenous Mitochondria in Male Mice.

Mitochondrial dysfunction is a major and common mechanism in developing non-alcoholic fatty liver disease (NAFLD). Replacement of dysfunctional mitochondria by functional exogenous mitochondria may attenuate intrahepatic excessive lipid and recover hepatocyte function. However, no data shows that mitochondria can be systemically administrated to animals to date. Here we suggest that mitochondria isolated from hepatoma cells are used as a mitotherapy agent to treat mouse fatty liver induced by high-fat diets. When the mitochondria were intravenously injected into the mice, serum aminotransferase activity and cholesterol level decreased in a dose-dependent manner. Also, the mitotherapy reduced lipid accumulation and oxidation injury of the fatty liver mice, improved energy production, and consequently restored hepatocyte function. The mitotherapy strategy offers a new potential therapeutic approach for treating NAFLD.

Twenty-six circulating antigens and two novel diagnostic candidate molecules identified in the serum of canines with experimental acute toxoplasmosis.

BACKGROUND: The protozoan Toxoplasma gondii is a pathogen that causes severe opportunistic disease in a wide range of hosts. Efficient methods to diagnose acute T. gondii infection are essential for the administration of appropriate treatments and to reduce economic losses. In animals with acute infections, circulating antigens (CAgs) were detected as early as two days post-infection; these CAgs were reliable diagnostic indicators of acute infection. However, only a limited number of CAgs have been identified to date. The objective of this study was to identify a broader spectrum of CAgs and to explore novel diagnostic candidates in serum. METHODS: A canine model of acute toxoplasmiosis was established. For this purpose, six dogs were infected by intraperitoneal inoculation of tachyzoites. The CAgs spectrum in the serum was identified with the immunoprecipitation-shotgun approach. Two CAgs with low homology to other species, coronin protein (TgCOR) and ELMO protein (TgELMO), were heterologously expressed in Escherichia coli. Polyclonal antibodies against these two proteins were prepared, and the presence of these proteins in the serum was verified by Western blotting. The two CAgs were detected and evaluated by indirect ELISA methods. RESULTS: The CAgs levels peaked between two and five days after inoculation, and twenty-six CAgs were identified. Western blotting showed the presence of the two proteins in the serum during acute infection. Based on ELISA tests, the two CAgs were detected during acute infection. CONCLUSIONS: We identified twenty-six CAgs in the serum of canines with experimental acute toxoplasmosis and discovered two novel diagnostic candidates. We also provide new insights into the diagnosis of acute toxoplasmosis.

Effect of Connexin43 on Mechanical Tension-stimulated Osteogenic Transcription Factors of Human Periodontal Ligament Fibroblasts.

OBJECTIVE: To investigate the expression and effect of Connexin43 (Cx43) on tensile tension-stimulated osteogenic transcription factors of human periodontal ligament fibroblasts (hPDLFs). METHODS: After hPDLFs were treated with 5% elongation tension for 1 h, 2 h, 4 h, 8 h, and 24 h, we examined the expressions of Cx43, Osterix, and RUNX2 at the mRNA level. After Cx43 expression was suppressed by siRNA or 18α-GA, the changes The mRNA in hPDLFs of Osterix and RUNX2 were observed. RESULTS: The expressions of Cx43, Osterix, and RUNX2 mRNA in hPDLFs increased in a time-dependent fashion following tensile strain (all P<0.05), with the highest level at 5% elongation for 24 h. After Cx43 expression was blocked by two different methods, the increasing expressions of Osterix and RUNX2 were inhibited. CONCLUSIONS: 5% cyclic tension upregulates Cx43 expression and promotes the expression of Osterix and RUNX2 in a time-dependent manner. Cx43 may be involved in the osteogenic response of hPDLFs to mechanical tension.

Connexin 43 and ERK regulate tension-induced signal transduction in human periodontal ligament fibroblasts.

Periodontal ligament (PDL) fibroblasts play an important role in preserving periodontal homeostasis and transmitting mechanical signals to alveolar bone. Connexin 43 (Cx43), a gap junction protein, is essential for bone homeostasis and regulates bone remodeling. However, the function of Cx43 in human PDL fibroblast-regulated bone remodeling has not yet been elucidated. In this study, human PDL fibroblasts were exposed to cyclic mechanical tension with a maximum 5% elongation for different durations. We then examined the expression of signaling molecules related to osteogenesis and osteoclastogenesis at both the mRNA and protein levels as well as the activity of extracellular signal-regulated kinase (ERK) in human PDL fibroblasts after loading. We found that mechanical tension increased Cx43, which further upregulated osteogenic (e.g., RUNX2, Osterix, and OPG) and down-regulated osteoclastogenic (e.g., RANKL) signaling molecules. Suppressing Cx43 gene (Gja1) by siRNA inhibited the increase in osteogenesis-related molecules but enhanced RANKL expression. Similar to Cx43, activated ERK1/2 was also enhanced by mechanical tension and suppressed by Cx43 siRNA. Inhibition of ERK1/2 signaling using PD98059 reduced the tension-regulated increase in osteogenesis-related molecules but enhanced that of osteoclastogenesis-related ones. These findings suggest that cyclic tension may involve into the osteogenic or osteoclastogenetic differentiation potential of human PDL fibroblasts via the Cx43-ERK1/2 signaling pathway.

The association between serum leptin and post stroke depression: results from a cohort study.

BACKGROUND: Depression is a frequent mood disorder that affects around a third of stroke patients and has been associated with poorer outcomes. Our aim was to determine whether there was a relationship between inflammatory markers (leptin) and post-stroke depression (PSD). METHODS: One hundred and ninety-one ischemic stroke patients admitted to the hospital within the first 24 hours after stroke onset were consecutively recruited and followed up for 3 months. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum levels of leptin at admission. Based on the symptoms, diagnoses of depression were made in accordance with DSM-IV criteria for post-stroke depression at 3 month. RESULTS: Forty-four patients (23.0%) were diagnosed as having major depression at 3 month. Patients with depression showed higher serum leptin levels at 3 month after stroke (32.2 [IQR, 20.8-57.7] v. 9.9 [IQR, 4.6-13.1]ng/ml, respectively; P = 0.000). Serum levels of leptin ≥20 ng/ml were independently associated with PSD [odds ratio (OR) 20.23, 95% confidence interval (CI) 9.11-51.26, P =  0.000], after adjusting for possible confounders. CONCLUSIONS: Serum leptin levels elevated at admission were found to be associated with PSD and may provide a new proposal for the treatment of PSD.

Proteomic analysis of the effect of diclazuril on second-generation merozoites of Eimeria tenella.

Diclazuril has long been used as an effective benzeneacetonitrile anticoccidial for the control of Eimeria tenella that causes coccidiosis. However, the molecular mechanism underlying the anticoccidial effects of diclazuril remains elusive. In this study, a proteomic analysis of the effect of diclazuril on second-generation merozoites of E. tenella was performed. Using two-dimensional gel electrophoresis and real-time quantitative polymerase chain reaction (RT-PCR), 13 target proteins were found to be significantly affected by diclazuril treatment, with 11 of these proteins being identified as annotated proteins from E. tenella or other Apicomplexa parasites. These proteins contribute to various functions, including metabolism, protein synthesis, and host cell invasion. Using RT-PCR, we identified the potential pattern of transcriptional regulation induced by diclazuril, and we suggest some promising targets for the intervention of E. tenella infection.

A novel chemiluminescent ELISA for detecting furaltadone metabolite, 3-amino-5-morpholinomethyl-2-oxazolidone (AMOZ) in fish, egg, honey and shrimp samples.

In this study, an indirect competitive enzyme-linked immunosorbent assay with chemiluminescent (CLELISA) detection for 3-amino-5-morpholinomethyl-2-oxazolidone (AMOZ) was developed. A monoclonal antibody (MAb) against AMOZ was prepared through immunizing BALB/c mice with 4-carboxybenzaldehyle derivatized AMOZ (CPAMOZ), conjugated with bovine serum albumin (BSA) as antigen. The effects of the substrates luminol, p-iodophenol and urea peroxide on the performance of the assay were studied and optimized. In addition, the specificity of the MAb, estimated as the cross-reactivity values with 4-nitrobenzaldehyde derivatized AMOZ (NPAMOZ), CPAMOZ and AMOZ, was 100%, 27.45% and 0.18%, respectively. The sensitivity of the developed CLELISA was estimated as 50% inhibitory concentration (IC50) value (0.14µg/l) with a linear working range between 0.03 and 64µg/l, and a limit of detection of 0.01µg/l. The CLELISA described in this study was 5-fold more sensitive than the indirect competitive ELISA previously developed in our laboratory. Finally, this new CLELISA was compared with a commercial kit to detect NPAMOZ in spiked fish, shrimp, honey and egg samples. The recovery values from four spiked fish, shrimp, honey and egg samples with different concentrations of NPAMOZ in CLELISA were 92.1-107.7%. Thus, the immunoassay method described here has a broad detection range and high sensitivity and is a valid and cost-effective means for high throughput monitoring of residual AMOZ levels in fish, shrimps, honey and eggs with potential applications in other animal tissues.