Effects of Intestinal Microbiota on the Biological Transformation of Arsenic in Zebrafish: Contribution and Mechanism.

Both the gut microbiome and their host participate in arsenic (As) biotransformation, while their exact roles and mechanisms in vivo remain unclear and unquantified. In this study, as3mt-/- zebrafish were treated with tetracycline (TET, 100 mg/L) and arsenite (iAsIII) exposure for 30 days and treated with probiotic Lactobacillus rhamnosus GG (LGG, 1 × 108 cfu/g) and iAsIII exposure for 15 days, respectively. Structural equation modeling analysis revealed that the contribution rates of the intestinal microbiome to the total arsenic (tAs) and inorganic As (iAs) metabolism approached 44.0 and 18.4%, respectively. Compared with wild-type, in as3mt-/- zebrafish, microbial richness and structure were more significantly correlated with tAs and iAs, and more differential microbes and microbial metabolic pathways significantly correlated with arsenic metabolites (P < 0.05). LGG supplement influenced the microbial communities, significantly up-regulated the expressions of genes related to As biotransformation (gss and gst) in the liver, down-regulated the expressions of oxidative stress genes (sod1, sod2, and cat) in the intestine, and increased arsenobetaine concentration (P < 0.05). Therefore, gut microbiome promotes As transformation and relieves As accumulation, playing more active roles under iAs stress when the host lacks key arsenic detoxification enzymes. LGG can promote As biotransformation and relieve oxidative stress under As exposure.

Slide-type waveflex biosensor based on signal enhancement technology for alpha-fetoprotein detection.

The development of signal enhancement technology in optical fiber biosensors is beneficial for the accurate measurement of low-concentration samples. Here, a localized surface plasmon resonance (LSPR)-based fiber biosensor combining a slide-type fiber structure (thus named WaveFlex Biosensor) and low-dimensional materials is proposed for alpha-fetoprotein (AFP) detection. A symmetric transverse offset splicing technology was used to fabricate the multi-mode fiber (MMF-multi-core fiber (MCF)-MMF structure. Furthermore, the MMF on one side was prepared into an S-taper, forming a slide-type fiber structure to generate more energy leakage. The LSPR signal generated by gold nanoparticles (AuNPs) was enhanced by the CeO2 NPs and C3N quantum dots functionalized on the fiber probe. The excellent performance of NPs was conducive to improving the sensitivity of the WaveFlex biosensor and enabling the rapid detection of samples. An AFP antibody was used to identify AFP micro-biomolecules in a specific manner. Based on the combination of the above two methods, the developed fiber probe was applied to detect AFP, and the sensitivity and limit of detection were 32 pm/(ng/mL) and 6.65 ng/mL, respectively. The experimental results demonstrate that the signal-enhanced AFP WaveFlex biosensor has great potential for the rapid and accurate detection of AFP.

Effective splicing technique of different cladding diameter-based optical fibers and performance evaluation.

In this work, the fabrication and sensing performance of fusion structures based on single-mode fiber (SMF) and multimode fiber (MMF) with different cladding diameters are discussed, and the effects of different lengths of MMF and fiber etching on sensing performance are analyzed. First, the transmitted intensity measurement experiment is performed, and the results indicate that the performance of the SMF-MMF-SMF(SMS)-based structure is better for sensing purposes. In addition, the results demonstrate that the performance of etched fiber is better than that of non-etched fiber. The etched fiber structure with lower fiber diameters produces more evanescent waves and is better for sensing purposes. Therefore, the proposed structure has certain development potential as an application of future optical fiber sensors.

Determination of arsenicals in mouse tissues after simulated exposure to arsenic from rice for sixteen weeks and the effects on histopathological features.

The accumulation of arsenic in rice has become a worldwide concern. In this study, dose-dependency in tissues (intestine, liver and kidney) and blood distribution of inorganic arsenicals and their methylated metabolites were investigated in male C57BL/6 mice exposed to four arsenic species (arsenite [iAs]III, arsenate [iAs]V, monomethylarsonate [MMA]V, and dimethylarsinate [DMA]V) at four doses (control [C]: 0 µg/g, simulation [S]: 0.91 µg/g, medium [M]: 9.1 µg/g and high [H]: 30 µg/g) according to the arsenical composition in rice for 8 and 16 weeks. No adverse effects were observed, while body weight gain decreased in group H. Increases in total arsenic concentrations (CtAs) and histopathological changes in the tissues occurred in all of the test groups. CtAs presented a tendency of kidney > intestine > liver > blood and were time-/dose-dependent in the liver and kidney in groups M and H. In the intestine and blood, abundant iAs (23%-28% in blood and 36%-49% in intestine) was detected in groups M and H, and CtAs decreased in group H from the 8th week to the 16th week. PMI decreased in the liver and SMI decreased in the kidney. These results indicate that the three tissues are injured through food arsenic. The intestine can also accumulate food arsenic, and the high arsenic dose will cause a deficiency in the absorbing function of the intestine. Thus, long-term exposure to arsenic-contaminated rice should be taken seriously attention.

Novel Arsenic Markers for Discriminating Wild and Cultivated Cordyceps.

Ophiocordyceps sinensis has been utilized in China and adjacent countries for thousands of years as a rare functional food to promote health and treat diverse chronic diseases. In recent years, adulterants are usually identified in the processed products of wild O. sinensis. However, the effective adulteration examination has to be additionally performed except their routine test, and accordingly is time- and money-consuming. Recently, arsenic determination has become a necessary test for confirming whether the concentrations of inorganic arsenic are over the O. sinensis limit. In this work, the contents of total arsenic and As species in cultivated O. sinensis, Cordyceps militaris, and other edible fungi were determined by ICP-MS and HPLC-ICP-MS. The results suggest that the As speciation exhibits a species-specific behavior, and accompanies the effect of the As background. The proportions of unknown organic As and contents of total As may be considered as sensitive markers for discriminating wild O. sinensis. This result provides a novel clue for discriminating wild and artificially cultivated mushrooms/their products, with emphasis on arsenic markers for authenticating wild O. sinensis.

Determination of Arsenic Species in Ophiocordyceps sinensis from Major Habitats in China by HPLC-ICP-MS and the Edible Hazard Assessment.

This study sought to determine the concentration and distribution of arsenic (As) species in Ophiocordyceps sinensis (O. sinensis), and to assess its edible hazard for long term consumption. The total arsenic concentrations, measured through inductively coupled plasma mass spectrometry (ICP-MS), ranged from 4.00 mg/kg to 5.25 mg/kg. As determined by HPLC-ICP-MS, the most concerning arsenic species­AsB, MMAV, DMAV, AsV, and AsШ­were either not detected (MMAV and DMAV) or were detected as minor As species (AsB: 1.4⁻2.9%; AsV: 1.3⁻3.2%, and AsШ: 4.1⁻6.0%). The major components were a cluster of unknown organic As (uAs) compounds with AsШ, which accounted for 91.7⁻94.0% of the As content. Based on the H2O2 test and the chromatography behavior, it can be inferred that, the uAs might not be toxic organic As. Estimated daily intake (EDI), hazard quotient (HQ), and cancer risk (CR) caused by the total As content; the sum of inorganic As (iAs) and uAs, namely i+uAs; and iAs exposure from long term O. sinensis consumption were calculated and evaluated through equations from the US Environmental Protection Agency and the uncertainties were analyzed by Monte-Carlo Simulation (MCS). EDItotal As and EDIi+uAs are approximately ten times more than EDIiAs; HQtotalAs and HQi+uAs > 1 while HQiAs < 1; and CRtotal As and CRi+uAs > 1 × 10−4 while CRiAs < 1 × 10−4. Thus, if the uAs is non-toxic, there is no particular risk to local consumers and the carcinogenic risk is acceptable for consumption of O. sinensis because the concentration of toxic iAs is very low.

Phenolic compositions and antioxidant activities of grapefruit (Citrus paradisi Macfadyen) varieties cultivated in China.

The phenolic compounds in different fruit parts including the flavedos, albedos, segment membranes, juice vesicles and seeds of nine grapefruit varieties cultivated in China were determined and their antioxidant capacities were evaluated using three methods. Naringin and neohesperedin were the dominant flavonoids in all grapefruit tested. Fenghongtangmuxun and Jiwei flavedo had the highest contents of naringin (5666.82 µg/g DW) and neohesperedin (1022 µg/g DW), respectively. Gallic acid was the major phenolic acid in all grapefruit tested, and Jiwei juice vesicles had the highest content of gallic acid (343.7 µg/g DW). Fenghongtangmuxun juice vesicles were rich in chlorogenic acid (110.23 µg/g DW), caffeic acid (53.86 µg/g DW) and ferulic acid (23.12 µg/g DW). Overall, the flavedo was rich in flavonoid, while juice vesicle had high amounts of phenolic acid. The Jiwei, Fenghongtangmuxun, Maxu, Huoyan and Hongmaxu grapefruit cultivars contained more phenolics and exhibited higher antioxidant capacities than Shatianyou and Liangpingyou pummelos, and were good sources of natural phytochemical antioxidants.

Long-term arsenic exposure decreases mice body weight and liver lipid droplets.

Arsenic (As) is a widespread global pollutant, and there is significant controversy surrounding its complex relationship with obesity, primarily focused on short-term exposure. Recognizing the prolonged nature of dietary arsenic exposure, this study involved feeding mice with arsenic-contained food for 14 months. The results showed that mice exposed to arsenic developed a non-alcoholic fatty liver condition, characterized by a light-yellow hue on the liver surface and various pathological alterations in the liver cells, including enlarged nuclei, cellular necrosis, inflammatory infiltration, dysfunctional mitochondria, and endoplasmic reticulum disorganization. There were also disruptions in biochemistry indices, with a significant increase in total cholesterol (TC) level and a decrease in high-density lipoprotein (HDL) level. However, some contradictory observations occurred, such as a significant decrease in body weight, triglyceride (TG) level, and the numbers of lipid droplets. Several genes related to lipid metabolism were tested, and a model was used to explain these discrepancies. Besides, examinations of the colon revealed compromised intestinal barrier function and signs of inflammation. Fecal 16S rRNA sequencing and pseudo-targeted metabolomics revealed disruptions in internal homeostasis, such as modules, nodes, connections, and lipid-related KEGG pathways. Fecal targeted metabolomics analyses of short-chain fatty acids (SCFAs) and bile acids (BAs) demonstrated a significant upregulation in three primary bile acids (CA, CDCA, TCDCA), four secondary bile acids (TUDCA, DCA, LCA, GUDCA), and total SCFAs level. Oxidative stress and inflammation were also evident. Additionally, based on correlation analysis and mediation analysis, it was assumed that changes in the microbiota (e.g., Dubosiella) can impact the liver metabolites (e.g., TGs) through alterations in fecal metabolites (e.g., LPCs). These findings provide a theoretical reference for the long-term effect of arsenic exposure on liver lipid metabolism.

WaveFlex biosensor based on S-tapered and waist-expanded technique for detection of glycosylated hemoglobin.

Glycosylated hemoglobin (HbA1c) is considered a new standard for the detection of diabetes mellitus because it is more accurate than regular blood sugar tests and there is no need to take blood on an empty stomach or at a specific time. In this work, we have developed a novel optical fiber biosensor, referred to as the "WaveFlex biosensor," which operates on the principles of localized surface plasmon resonance (LSPR) plasmonic wave. The sensor is fabricated using an innovative S-tapered and waist-expanded technique, enabling it to effectively detect HbA1c. Compared to the HbA1c sensors currently in use, HbA1c optical fiber sensors possess the characteristics of high sensitivity, low cost, and strong anti-interference ability. The gold nanoparticles (AuNPs), cerium oxide (CeO2) nanorods (NRs), and tungsten disulfide (WS2) nanosheets (NSs) are functionalized to improve the effectiveness of the fiber sensor on the probe surface. AuNPs are utilized to generate LSPR by the excitation of evanescent waves to amplify the sensing signal. The CeO2-NRs can have a strong metal-carrier interaction with AuNPs, enhancing the cascade of CeO2-NRs and AuNPs. The WS2-NSs with layered fold structure have a large specific surface area. Therefore, the combination of CeO2-NRs and WS2-NSs is conducive to the binding of antibodies and the addition of sites. The functionalized antibodies on the fiber make the sensor probe capable of specific selection. The developed probe is applied to test the HbA1c solution over concentrations of 0-1000 µg/mL, and the sensitivity and limits of detection of 1.195×10-5 a.u./(µg/mL) and 1.66 µg/mL are obtained, respectively. The sensor probe is also evaluated using assays for reproducibility, reusability, selectivity, and pH. According to the findings, a novel method for detecting blood glucose based on a plasmonic biosensor is proposed.

As3MT-mediated SAM consumption, which inhibits the methylation of histones and LINE1, is involved in arsenic-induced male reproductive damage.

Studies have demonstrated that arsenic (As) induces male reproductive injury, however, the mechanism remains unknown. The high levels of arsenic (3) methyltransferase (As3MT) promote As-induced male reproductive toxicity. For As-exposed mice, the germ cells in seminiferous tubules and sperm quality were reduced. Exposure to As caused lower S-adenosylmethionine (SAM) and 5-methylcytosine (5 mC) levels, histone and DNA hypomethylation, upregulation of long interspersed element class 1 (LINE1, or L1), defective repair of double-strand breaks (DSBs), and the arrest of meiosis, resulting in apoptosis of germ cells and lower litter size. For GC-2spd (GC-2) cells, As induced apoptosis, which was prevented by adding SAM or by reducing the expression of As3MT. The levels of LINE1, affected by SAM content, were involved in As-induced apoptosis. Furthermore, folic acid (FA) and vitamin B12 (VB12) supplements restored SAM, 5 mC, and LINE1 levels and blocked impairment of spermatogenesis and testes and lower litter size. Exposed to As, mice with As3MT knockdown showed less impairment of spermatogenesis and testes and greater litter size compared to As-exposed wild-type (WT) mice. Thus, the high As3MT levels induced by As consume SAM and block histone and LINE1 DNA methylation, elevating LINE1 expression and evoking impairment of spermatogenesis, which causes male reproductive damage. Overall, we have found a mechanism for As-induced male reproductive damage, which provides biological insights into the alleviation of reproductive injury induced by environmental factors.

A multiplex PCR amplicon sequencing assay to screen genetic hearing loss variants in newborns.

BACKGROUND: Congenital hearing loss is one of the most common birth defects. Early identification and management play a crucial role in improving patients' communication and language acquisition. Previous studies demonstrated that genetic screening complements newborn hearing screening in clinical settings. METHODS: We developed a multiplex PCR amplicon sequencing assay to sequence the full coding region of the GJB2 gene, the most pathogenic variants of the SLC26A4 gene, and hotspot variants in the MT-RNR1 gene. The sensitivity, specificity, and reliability were validated via samples with known genotypes. Finally, a pilot study was performed on 300 anonymous dried blood samples. RESULTS: Of 103 samples with known genotypes, the multiplex PCR amplicon sequencing assay accurately identified all the variants, demonstrating a 100% sensitivity and specificity. The consistency is high in the analysis of the test-retest reliability and internal consistency reliability. In the pilot study, 12.3% (37/300) of the newborns were found to carry at least one pathogenic variant, including 24, 10, and 3 from the GJB2, SLC26A4, and MT-RNR1 gene, respectively. With an allele frequency of 2.2%, the NM_004004.6(GJB2):c.109G>A was the most prevalent variant in the study population. CONCLUSION: The multiplex PCR amplicon sequencing assay is an accurate and reliable test to detect hearing loss variants in the GJB2, SLC26A4, and MT-RNR1 genes. It can be used to screen genetic hearing loss in newborns.

Distribution of arsenic species and pathological characteristics of tissues of the mice fed with arsenic-supplemented food simulating rice.

The exposure and harm of arsenic have attracted wide attention. Rice is an arsenic-rich crop. The purpose of this study was to learn the distribution of arsenic species and the pathological changes in tissues of mice exposed to arsenic-supplemented food simulating rice. Test groups of mice were orally exposed with prepared arsenic feeds supplemented with four arsenic species (arsenite iAsIII, arsenate iAsV, monomethylarsonate MMA, and dimethylarsinate DMA) at three doses (total As concentration: 0.91, 9.1 and 30 µg/g), which simulated the arsenic species ratio in rice. After 112 days, the concentrations of the arsenic species in the spleen, thymus, heart, skin and hair were detected, and histopathology of the spleen, heart and skin was observed. Each arsenic species was detected and their total concentration increased in a dose-dependent manner with a few exceptions. One interesting phenomenon is that ratio of the organic arsenic to inorganic arsenic also increased in a dose-dependent manner. For the other, the order of tissues from high to low arsenic concentration was the same in the medium- and high-dose groups. The histopathological sections of the spleen, heart and skin showed dose-dependent debilitating alterations in tissue architecture. Hyperplasia, hyaline degeneration and sclerosis of fibrous connective tissue occurred in the spleen. Myocardial cell atrophy and interstitial edema occurred in the heart. Hyperpigmentation, hyperkeratosis and atypia of basal cells occurred in the skin. In summary, the long-term intake of high arsenic rice has a health risk. Further studies are needed to assess it.

Arsenic concentrations, diversity and co-occurrence patterns of bacterial and fungal communities in the feces of mice under sub-chronic arsenic exposure through food.

BACKGROUND: Arsenic, a global pollutant and a threshold-free primary carcinogen, can accumulate in rice. Previous studies have focused on arsenic poisoning in drinking water and the effects on gut microbes. The research on arseniasis through food, which involves the bio-transformation of arsenic, and the related changes in gut microbiome is insufficient. METHOD: Mice were exposed from animal feed prepared with four arsenic species (iAsIII, iAsV, MMA, and DMA) at a dose of 30 mg/kg according to the arsenic species proportion in rice for 30 days and 60 days. The levels of total arsenic (tAs) and arsenic species in mice feces and urine samples were determined using ICP-MS and HPLC-ICP-MS, respectively. 16S rRNA and ITS gene sequencing were conducted on microbial DNA extracted from the feces samples. RESULTS: At 30 days and 60 days exposure, the tAs levels excreted from urine were 0.0092 and 0.0093 mg/day, and tAs levels in feces were 0.0441 and 0.0409 mg/day, respectively. We found significant differences in arsenic species distribution in urine and feces (p < 0.05). In urine, the predominant arsenic species were iAsIII (23% and 14%, respectively), DMA (55% and 70%, respectively), and uAs (unknown arsenic, 14% and 10%, respectively). In feces, the proportion of major arsenic species (iAsV, 26% and 21%; iAsIII, 16% and 15%; MMA, 14% and 14%; DMA, 19% and 19%; and uAs, 22% and 29%, respectively) were evenly distributed. Microbiological analysis (MRPP test, α- and ß-diversities) showed that diversity of gut bacteria was significantly related to arsenic exposure through food, but diversity of gut fungi is less affected. Manhattan plot and LEfSe analysis showed that arsenic exposure significantly changes microbial taxa, which might be directly associated with arsenic metabolism and diseases mediated by arsenic exposure, such as Deltaproteobacteria, Polynucleobacter, Saccharomyces, Candida, Amanitaceae, and Fusarium. Network analysis was used to identify the changing hub taxa in feces along with arsenic exposure. Function predicting analysis indicated that arsenic exposure might also significantly increase differential metabolic pathways and would disturb carbohydrates, lipid, and amino acids metabolism of gut bacteria. CONCLUSIONS: The results demonstrate that subchronic arsenic exposure via food significantly changes the gut microbiome, and the toxicity of arsenic in food, especially in staples, should be comprehensively evaluated in terms of the disturbance of microbiome, and feces might be the main pathway through which arsenic from food exposure is excreted and bio-transformed, providing a new insight into the investigation of bio-detoxification for arseniasis.