Neurodevelopmental toxicity induced by PM2.5 Exposure and its possible role in Neurodegenerative and mental disorders.

Recent extensive evidence suggests that ambient fine particulate matter (PM2.5, with an aerodynamic diameter ≤2.5 µm) may be neurotoxic to the brain and cause central nervous system damage, contributing to neurodevelopmental disorders, such as autism spectrum disorders, neurodegenerative diseases, such as Alzheimer's disease and Parkinson's disease, and mental disorders, such as schizophrenia, depression, and bipolar disorder. PM2.5 can enter the brain via various pathways, including the blood-brain barrier, olfactory system, and gut-brain axis, leading to adverse effects on the CNS. Studies in humans and animals have revealed that PM2.5-mediated mechanisms, including neuroinflammation, oxidative stress, systemic inflammation, and gut flora dysbiosis, play a crucial role in CNS damage. Additionally, PM2.5 exposure can induce epigenetic alterations, such as hypomethylation of DNA, which may contribute to the pathogenesis of some CNS damage. Through literature analysis, we suggest that promising therapeutic targets for alleviating PM2.5-induced neurological damage include inhibiting microglia overactivation, regulating gut microbiota with antibiotics, and targeting signaling pathways, such as PKA/CREB/BDNF and WNT/ß-catenin. Additionally, several studies have observed an association between PM2.5 exposure and epigenetic changes in neuropsychiatric disorders. This review summarizes and discusses the association between PM2.5 exposure and CNS damage, including the possible mechanisms by which PM2.5 causes neurotoxicity.

Sphingosylphosphorylcholine alleviates hypoxia-caused apoptosis in cardiac myofibroblasts via CaM/p38/STAT3 pathway.

Blockade of hypoxia-caused nonmyocytes apoptosis helps improve survival and mitigate ventricular remodeling and dysfunction during the chronic stage of myocardial infarction. But tools affecting nonmyocyte apoptosis are very rare. Sphingosylphosphorylcholine (SPC), a naturally occurring bioactive sphingolipid in plasma, was proved to protect cardiomyocyte against apoptosis in an ischemic model in our previous study. Here, we showed that SPC also inhibited hypoxia-induced apoptosis in myofibroblasts, an important type of nonmyocytes in the heart. Calmodulin (CaM) is an identified receptor of SPC. We clarified that SPC inhibited myofibroblast apoptosis through CaM as evidenced by decreased cleaved caspase 3, PARP1 and condensed nucleus. Furthermore, the employment of inhibitor and agonist of p38 and STAT3 suggests that SPC inhibits myofibroblast apoptosis by regulating the phosphorylation of p38 and STAT3, and they act as downstream of CaM. The present work may provide new evidence on the regulation of myofibroblasts apoptosis by SPC and a novel target for heart remodeling after hypoxia.

Phytochemical profiles, antioxidant, and antiproliferative activities of red-fleshed apple as affected by in vitro digestion.

The aim of this study was to characterize the phenolic profiles in the extracts and digesta (after in vitro digestion) of different red-fleshed apple fruit parts and to assess the effects of digestion on the in vitro antioxidant capacity and antiproliferative activity. The main polyphenols were identified by UPLC-MS/MS and HPLC. Our results indicate that the digesta had less total phenolics, flavonoids, and anthocyanins, but more free phenolic acids, than the extracts. An analysis of the in vitro antioxidant capacity (including ABTS radical scavenging activity, DPPH radical-scavenging capacity, ferric reducing antioxidant power [FRAP], and cellular antioxidant activity [CAA]) revealed that the digestion decreased the ABTS, DPPH, and FRAP values, but increased the CAA values, relative to the corresponding values for extracts. These results suggest that the digestion improved the effectiveness of the phenolic substances. Moreover, our findings imply that the digestion promoted the antiproliferative activity of red-fleshed apple peels and flesh relative to the extracts. Future in vivo investigations are warranted based on the results of the current study. PRACTICAL APPLICATION: The effects of an in vitro digestion on the phenolic compounds as well as the antioxidative and antiproliferative activities of red-fleshed apple were evaluated. The resulting data may clarify the bioavailability of the polyphenols in red-fleshed apple and enable scientists and consumers to exploit natural polyphenols.

Effects of 2,3',4,4',5-pentachlorobiphenyl exposure during pregnancy on DNA methylation in the testis of offspring in the mouse.

Polychlorinated biphenyls (PCBs) are persistent organic pollutants, and the widespread use of PCBs has had adverse effects on human and animal health. This study experiment explored the effects of 2,3',4,4',5-pentachlorobiphenyl (PCB118) on the mammalian reproductive system. PCB118 was administered to pregnant mice from 7.5 to 12.5 days of gestation; F1 mice were obtained and the reproductive system of F1 male mice was examined. PCB118 damaged the reproductive system in male F1 mice, as evidenced by negative effects on the testicular organ coefficient (testes weight/bodyweight), a decrease in the diameter of seminiferous tubules and a significant reduction in the anogenital distance in 35-day-old F1 mice. In addition, methylation levels of genomic DNA were reduced, with reductions in the expression of the DNA methyltransferases DNMT1, DNMT3A and DNMT3B, as well as that of the epigenetic regulatory factor ubiquitin like with PHD and ring finger domains 1 (Uhrf1). Together, the results of this study provide compelling evidence that exposure of pregnant mice to PCB118 during primordial germ cell migration in the fetus affects the reproductive system of the offspring and decreases global methylation levels in the testis.

Phytochemical profiles, antioxidant, and antiproliferative activities of four red-fleshed apple varieties in China.

Red-fleshed apples are preferred because of their high content of phenolics and antioxidants. In this study, the phenolic characteristics, antioxidant properties, and antihuman cancer cell properties of the four hybrids of Malus sieversii f. niedzwetzkyana (Ledeb.) M. Roem were analyzed. In addition, the antioxidant and anti-proliferation properties of these apples were measured. Compared to "Fuji" apples, the red-fleshed apples were rich in phenolic and flavonoid chemicals, ranging from 1.5- to 2.6-fold and 1.4- to 2.4-fold, respectively. In all antioxidant methods (DPPH radical-scavenging capacity, ABTS radical scavenging activity, ferric reducing antioxidant power, and cell antioxidant capacity), "A38" obtained the highest antioxidant value, whereas "Fuji" got the lowest antioxidant value. The IC50 values ranged from 33.44 ("A38") to 73.36 mg/mL ("Fuji") for MCF-7 and 20.94 ("A38") to 39.39 mg/mL ("Fuji") for MAD-MB-231. The red-fleshed "A38" and "Meihong" exhibited higher antioxidant and antiproliferative activities in vitro because of the higher levels of phenolics, and the higher potential for development and utilization value. PRACTICAL APPLICATION: The phenolic compounds, antioxidant activity, and antiproliferative activity in vitro of four red-fleshed apple cultivars and one white-fleshed apple cultivar were compared in this study. This information should assist to give a reasonable evaluation for scientists to breed new cultivars with high phenolics and to exploit the natural polyphenol.

Fucoxanthin Exerts Cytoprotective Effects against Hydrogen Peroxide-induced Oxidative Damage in L02 Cells.

Several previous studies have demonstrated the excellent antioxidant activity of fucoxanthin against oxidative stress which is closely related to the pathogenesis of liver diseases. The present work was to investigate whether fucoxanthin could protect human hepatic L02 cells against hydrogen peroxide- (H2O2-) induced oxidative damage. Its effects on H2O2-induced cell viability, lactate dehydrogenase (LDH) leakage, intracellular reduced glutathione, and reactive oxygen species (ROS) contents, along with mRNA and protein relative levels of the cytoprotective genes including Nrf2, HO-1, and NQO1, were investigated. The results showed that fucoxanthin could upregulate the mRNA and protein levels of the cytoprotective genes and promote the nuclear translocation of Nrf2, which could be inhibited by the PI3K inhibitor of LY294002. Pretreatment of fucoxanthin resulted in decreased LDH leakage and intracellular ROS content but enhanced intracellular reduced glutathione. Interestingly, pretreatment using fucoxanthin protected against the oxidative damage in a nonconcentration-dependent manner, with fucoxanthin of 5 µM demonstrating the optimal effects. The results suggest that fucoxanthin exerts cytoprotective effects against H2O2-induced oxidative damage in L02 cells, which may be through the PI3K-dependent activation of Nrf2 signaling.

Auxin regulates anthocyanin biosynthesis through the Aux/IAA-ARF signaling pathway in apple.

Auxin signaling, which is crucial for normal plant growth and development, mainly depends on ARF-Aux/IAA interactions. However, little is known regarding the regulatory effects of auxin signaling on anthocyanin metabolism in apple (Malus domestica). We investigated the functions of MdARF13, which contains a repression domain and is localized to the nucleus. This protein was observed to interact with the Aux/IAA repressor, MdIAA121, through its C-terminal dimerization domain. Protein degradation experiments proved that MdIAA121 is an unstable protein that is degraded by the 26S proteasome. Additionally, MdIAA121 stability is affected by the application of exogenous auxin. Furthermore, the overexpression of MdIAA121 and MdARF13 in transgenic red-fleshed apple calli weakened the inhibitory effect of MdARF13 on anthocyanin biosynthesis. These results indicate that the degradation of MdIAA121 induced by auxin treatment can release MdARF13, which acts as a negative regulator of the anthocyanin metabolic pathway. Additionally, yeast two-hybrid, bimolecular fluorescence complementation, and pull-down assays confirmed that MdMYB10 interacts with MdARF13. A subsequent electrophoretic mobility shift assay and yeast one-hybrid assay demonstrated that MdARF13 directly binds to the promoter of MdDFR, which is an anthocyanin pathway structural gene. Interestingly, chromatin immunoprecipitation-quantitative real-time PCR results indicated that the overexpression of MdIAA121 clearly inhibits the recruitment of MdARF13 to the MdDFR promoter. Our findings further characterized the mechanism underlying the regulation of anthocyanin biosynthesis via Aux/IAA-ARF signaling.

Effects of PrPC on DF-1 cells' biological processes and RNA-seq-based analysis of differential genes.

To reveal the effects of PrPC on cells' biological processes and on gene expression. We established stable DF-1 (PrPC -knockdown (KD)) cells, and combined with DF-1 (wt) and DF-1 (PrPC -overexpression (OE)) cells that we previously established we studied the effects of chicken PrPC (PrPC ) on DF-1 cells' processes. Then by using high throughput sequencing technology (HTS) and bioinformatics, we analyzed the differentially expressed genes (DEGs) between these cells. The results show that compared with DF-1 (wt) and DF-1 (PrPC -scramble), DF-1 (PrPC -KD) are significantly decreased in adhesion, proliferation, formation rate of colony and cells number of colony, scratch wound healing rate, cells number of invasion and migration, S phase cell populations, but the apoptosis rate and G1 phase cell populations are significantly increased. Conversely, all of these features in DF-1 (PrPC -OE) are opposite. In addition, compared with DF-1 (wt), we found that there are totally 1055 DE genes between DF-1 (PrPC -KD) and DF-1 (PrPC -OE) cells. After Go and pathway enrichment analysis, we know that these DEGs are significantly enriched in cell, cell part, cellular process, and metabolic pathway. In short, we found that PrPC can promote DF-1 cells' processes except apoptosis. Furthermore, PrPC involves in the focal adhesion, cancer, ribosome, metabolic pathways, and so forth, and the overexpression of PrPC can promote the pathway of amoebiasis, but its down-regulation can promote the pathway of serotonergic synapse. However, the details are keeping unknown and that would be our next research.

Association between Polymorphisms in Interleukins 4 and 13 Genes and Chronic Periodontitis in a Han Chinese Population.

Chronic periodontitis (CP) is one of the most common chronic inflammatory diseases and cytokines play a pivotal role in the regulation of immune response. Interleukin-4 (IL-4) and interleukin-13 (IL-13) are anti-inflammatory cytokines and several polymorphisms of them have been proved involved in periodontal disease. This study aimed to evaluate whether three single nucleotide polymorphisms (SNPs), rs2070874 and rs2243248 from IL4 and rs1800925 from IL13, are associated with CP in a Han Chinese population consisting of 440 moderate or severe CP patients and 324 healthy controls. Genomic DNA extracted from buccal epithelial cells of the included participants were genotyped using a matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry method. No significant association between rs2070874 or rs1800925 and CP was found, while the frequencies of rs2243248 and two haplotypes C-G-T and C-T-T showed significant differences between the two groups. The results suggest that the polymorphism rs2243248 and haplotypes C-G-T and C-T-T may be associated with CP susceptibility in the present Han Chinese population.

Exposure to Aroclor-1254 impairs spindle assembly during mouse oocyte maturation.

Polychlorinated biphenyls (PCBs), as typical environmental estrogen disruptors, are a structurally-related group of halogenated aromatic hydrocarbons that are composed of 209 isomers and present as a mixture in the environment. PCBs congener with different numbers and positions of chlorine atoms substituted on the biphenyl moiety. Aroclor-1254 is a mixture of more than 60 PCB congeners. Previous studies have provided the evidence that PCBs have severe negative effects on reproductive functions, but the effects of PCBs on spindle assembly during mouse oocyte maturation in vitro have not been reported. In the present study, female ICR mouse immature oocytes were cultured in M2 medium with 1 and 10 µg mL-1 Aroclor-1254 separately in vitro. The percentage of germinal vesicle breakdown (GVBD) and the first polar body extrusion were recorded. The results showed no significant difference in the percentage of GVBD or the first polar body extrusion between control oocytes and Aroclor-1254-treated oocytes. Further studies showed that the normal localization of γ-tubulin and Aurora-A kinase was interfered and α-tubulin assembling into spindle was affected when mouse oocytes were exposed to Aroclor-1254. The length of spindle from 10 µg mL-1 Aroclor-1254-treated oocytes was longer than that from control oocytes, and the spindle area in the Aroclor-1254-treated groups were decreased. Furthermore, the percentage of DNA damage in cumulus cells revealed an increase after exposed to Aroclor-1254. These results will provide the important reference for the prevention of reproductive disorders caused by PCBs. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1652-1662, 2016.

The association of CSF-1 gene polymorphism with chronic periodontitis in the Han Chinese population.

BACKGROUND: Chronic periodontitis (CP) is a multifactorial complex periodontal disease involving immune response, inflammation, alveolar bone resorption, and attachment loss. Colony stimulating factor-1 (CSF-1) controls the production, differentiation, and function of macrophages and plays a vital role in the innate immune response to the external microbial infections, suggesting the potential role of CSF-1 in the pathogenesis of periodontitis. The objective of this study is to determine the association of single nucleotide polymorphisms (SNPs) rs333967, rs2297706, and rs1058885 with CP in the Han Chinese population. METHODS: Genomic DNA was isolated from buccal epithelial cells obtained from unrelated Chinese participants (440 patients with CP and 324 controls). The SNPs were genotyped by a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry method. RESULTS: Three previously identified SNPs were genotyped in Han Chinese with Shanghai origin, but none of them was statistically significantly associated with CP. However, a T-C-G haplotype in male participants showed an observed P value of 4.52(E-08), with an odds ratio of 0.092. CONCLUSION: None of the individual SNPs among rs333967, rs2297706, and rs1058885 in CSF-1 was found statistically significantly associated with CP in the Han Chinese population with Shanghai origin, whereas a haplotype T-C-G showed an observed statistically significant association with decreased risk of CP susceptibility in males.

[Correlation between single nucleotide polymorphism and periodontitis susceptibility in Shanghai Chinese patients].

PURPOSE: To investigate the correlation between 12 single nucleotide polymorphisms (SNPs) from 7 genes and periodontitis of susceptibility in Shanghai Chinese patients. METHODS: Genotypes of randomly grouped 50 health control subjects and 48 patients with chronic periodontitis were genotyped by MALDI-TOF MS detection system. A questionnaire, medical history, family history, signs, laboratory tests, treatment, follow-up information were collected. Statistical analysis of Chi-square test was performed using SPSS13.0 software package. RESULTS: In the screening of the 12 target SNPs,the GG genotype of rs2891168 correlated significantly with periodontitis, OR=3.071(95%CI:1.018-9.260,P=0.046). CONCLUSION: The GG genotype of rs2891168 may correlate significantly with periodontitis of Shanghai Chinese patients and further studies are needed to verify this finding. Supported by Natural Science Foundation of Shanghai Municipality (Grant No.10ZR1425900) and Research Fund of Bureau of Health of Shanghai Municipality (Grant No.2009202).

Expression changes of neurofilament subunits in the central nervous system of hens treated with tri-ortho-cresyl phosphate (TOCP).

Tri-ortho-cresyl phosphate (TOCP) could induce degeneration of long, large diameter axons within the central and peripheral nervous system of susceptible species including human being and hens, which is referred to as organophosphorus-ester induced delayed neuropathy (OPIDN). The mechanisms involved are not understood. Neuropathologic observations suggested that neurofilament subunits (NFs) could be a main target of TOCP in the peripheral nervous system. Our previous study also showed that NFs in protein levels significantly decreased in sciatic nerves of hens treated with TOCP. In this study, to determine whether the decrement of NFs proteins in sciatic nerves was due to reductions in NF gene expression or protein degradation, hens were treated with a single dose of 750 mg/kg body weight TOCP by gavage, and sacrificed on 21 day post-exposure. Cerebral cortexes and spinal cords were sampled. Transcriptional changes of NFs including high molecular weight neurofilament (NF-H), middle molecular weight neurofilament (NF-M), low molecular weight neurofilament (NF-L), and glyceraldehydes-3-phoaphate dehydrogenase (GAPDH) as inner inference in cerebral cortexes and spinal cords were analyzed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results showed that all of three NFs mRNA in cerebral cortexes down-regulated significantly. However, in spinal cords, there was only NF-M decreased, both of NF-H and NF-L kept unaffected. The protein levels of NFs in pellet and supernatant fractions of cerebral cortexes and spinal cords were also determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. We noticed that all NFs protein declined in pellet of cerebral cortexes, but NF-M reduction was not significant compared with that of control hens. NF-H and NF-M proteins in supernatant of cerebral cortexes exhibited significant increase, while NF-L level showed remarkable decline. In spinal cords, apart from NF-L in pellet were significantly increased, both of NF-H and NF-M in pellet and supernatant, as well as NF-L in supernatant fractions were manifested dramatic reduction compared with the pattern of control. The quantitative analyses revealed that the change magnitude in protein levels was much greater than that in mRNA levels in hens' central nervous system after TOCP administration. These findings suggest that the NFs disturbance in protein levels is closely associated with the decreases in sciatic nerves observed in our previous work after TOCP exposure, rather than that in mRNA levels, and the NFs alterations in protein levels may be one of the responsible factors for the OPIDN.

[Effects of methamidophos on microtubule and microfilament proteins in sciatic nerve of hens].

OBJECTIVE: To investigate the dynamic changes of alpha-tubulin, beta-tubulin and beta-actin in sciatic nerve of hen with organophosphorus ester-induced delayed neuropathy (OPIDN). METHODS: OPIDN was induced in 10-month-old Roman hens by daily subcutaneous administration of 30 mg/kg methamidophos for 15 days. Hens were sacrificed 2, 10, and 23 days respectively after manifesting neuropathy. The sciatic nerves were dissected, homogenized and used for the determination of the alpha-tubulin, beta-tubulin and beta-actin levels by western blotting. RESULTS: The levels of alpha-tubulin in supernatant of sciatic nerves were decreased by 6%, 15% and 25% respectively on day 2, 10 and 23 respectively, while those in pellet remained almost unchanged. beta-tubulin were decreased by 27%, 6%, 19% in pellet and 1%, 21%, 22% in supernatant of sciatic nerves on 2, 10 and 23 days. Beta-actin level in pellet of sciatic nerve increased by 24%, 48% and 17% on day 2, 10 and 23, and little changes were observed in supernatant. CONCLUSION: Methamidophos may induced changes of alpha-tubulin, beta-tubulin and beta-actin levels in sciatic nerve of hen, which may be one of the mechanism of the contribution to the occurrence and development of OPIDN.

2,5-Hexanedione induced reduction in protein content and mRNA expression of neurofilament in rat cerebral cortex.

Exposure chronically to n-hexane produces central-peripheral axonopathy mediated by 2,5-hexanedione (HD). Studies have shown neurofilament (NF) subunit proteins are decreased substantially in cerebral cortices, optic axons, spinal cords, and sciatic nerves from HD-exposed rats. To deeply investigate the alterations in NF contents in HD neuropathy, the relative levels of NF-L, NF-M, and NF-H in rat cerebral cortex were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. HD was administrated to Wistar rats by intraperitoneal injection at dosage of 200 or 400mg/kg. Rats were sacrificed after 6 weeks of treatment, and cerebral cortices were dissected, homogenized, and used for the determination of NF subunit proteins. The results, except for supernatant NF-L and NF-M that could not be assayed, showed HD intoxication resulted in significant decreases by 32-67% (P<0.01) in NF subunits in both of the pellet and supernatant fractions of cerebral cortex homogenate. As an initial investigation to determine how such changes in NF proteins might occur, the gene expression of NF-L, NF-M, and NF-H subunit mRNA was quantified using reverse transcription-polymerase chain reaction (RT-PCR). Statistical analysis revealed that HD exposure caused a significant reduction in the expression of NF-L and NF-H gene (P<0.05 or P<0.01), while the levels of NF-M mRNA kept unaffected (P>0.05). These suggest that the observed reduction in NF gene expression might be related to diminished levels of subunit proteins, while the actual contribution might be uncertain. The functional significance of the reduced protein contents and the regulation of gene expression remain to be determined.

Tri-ortho-cresyl phosphate (TOCP) decreases the levels of cytoskeletal proteins in hen sciatic nerve.

Tri-ortho-cresyl phosphate (TOCP) is an organophosphorus ester. It is capable of producing organophosphorus ester induced delayed neurotoxicity (OPIDN) in human being and sensitive animals, which is characterized by ataxia that progresses to paralysis after 1-3 weeks following exposure to some organophosphorus ester. In present study, 18 adult hens were divided randomly into three groups, i.e. two experimental groups and control group (n = 6 each group). All hens were 10 months old and weighted 1.5-2.0 kg. The hens in two experimental groups were treated with TOCP by gavage at single dosages of 375 and 750 mg/kg respectively. TOCP was dissolved in corn oil and administered at 0.65 ml/kg. Six control hens received an equivalent volume of corn oil by gavage. All hens were sacrificed after 21 days of treatment and the sciatic nerves were dissected, homogenized and used for the determination of cytoskeletal proteins by western blotting. The levels of neurofilament (NF) subunits were decreased both in supernatant and pellet of sciatic nerves, and the most noticeable decrease in levels of NF subunits protein was observed in neurofilament medium (NF-M). Compared to the control hens, neurofilament heavy (NF-H) level decreased by 36 and 38% (P < 0.01) in the pellet and by 27 and 26% (P < 0.05) in the supernatant of sciatic nerves of hens treated with 375 and 750 mg/kg TOCP respectively. The reduction of NF-M were 36 and 68% (P < 0.01) in pellet, 50 and 67% (P < 0.01) in supernatant at 375 and 750 mg/kg dosage respectively. The neurofilament light (NF-L) lessened slightly, but the relative percentage of integrated optical density (IOD) was no significant alteration when compared to the control hens. There were significant decreases in levels of alpha-tubulin, beta-tubulin in pellet and alpha-tubulin, beta-tubulin, beta-actin in supernatant of sciatic nerves in TOCP-treated hens. Thus, the decreases of cytoskeletal proteins suggested the possible involvement of them in delayed neurotoxicity.