The rs6576457 G > A variant in the MKRN3 gene promoter significantly increases the risk of central precocious puberty and lung cancer in Hubei Chinese population.

Makorin RING finger protein 3 (MKRN3) is a key inhibitor of the hypothalamic-pituitary-gonadal (HPG) axis. The association between MKRN3 gene variants and central precocious puberty (CPP) has been repeatedly examined. In a recent study, MKRN3 has been assigned a role of tumor suppressor in lung carcinogenesis. Therefore, it is hypothesized that MKRN3 may be the link between CPP and lung cancer (LC), and certain MKRN3 gene variants may affect individuals' susceptibility to CPP and LC. The rs12441287, rs6576457 and rs2239669 in the MKRN3 gene were selected as the target variants. Sanger sequencing was applied to genotype them in two sets of case-control cohorts, namely 384 CPP girls and 422 healthy girls, 550 LC patients and 800 healthy controls. The results showed that rs6576457 but not rs12441287 or rs2239669 was significantly associated with the risk of CPP and LC. Their association with CPP risk was further confirmed in the following meta-analysis. Subsequent functional assays revealed that the rs6576457 genotypes were correlated with differentially expressed MKRN3, and the rs6576457 alleles affected the transcription repressor Oct-1 binding affinity to the MKRN3 promoter. Collectively, the MKRN3 gene rs6576457 may participate in the CPP pathology and LC tumorigenesis in the Hubei Chinese population. However, the present findings should be validated in additional investigations with larger samples from different ethnic populations.

Near-Unity Green Luminescent Hybrid Manganese Halides as X-ray Scintillators.

The increasing demands in optoelectronic applications have driven the advancement of organic-inorganic hybrid metal halides (OIMHs), owing to their exceptional optical and scintillation properties. Among them, zero-dimensional (0D) low-toxic manganese-based scintillators have garnered significant interest due to their exceptional optical transparency and elevated photoluminescence quantum yields (PLQYs), making them promising for colorful light-emitting diodes and X-ray imaging applications. In this study, two OIMH single crystals of (Br-PrTPP)2MnBr4 (Br-PrTPP = (3-bromopropyl) triphenylphosphonium) and (Br-BuTPP)2MnBr4 (Br-BuTPP = (4-bromobutyl) triphenylphosphonium) were prepared via a facile saturated crystallization method. Benefiting from the tetrahedrally coordinated [MnBr4]2- polyhedron, both of them exhibited strong green emissions peaked at 517 nm owing to the d-d electron transition of Mn2+ with near-unity PLQYs of 99.33 and 86.85%, respectively. Moreover, benefiting from the high optical transparencies and remarkable luminescence properties, these manganese halides also exhibit excellent radioluminescent performance with the highest light yield of up to 68,000 photons MeV-1, negligible afterglow (0.4 ms), and linear response to X-ray dose rate with the lowest detection limit of 45 nGyair s-1. In X-ray imaging, the flexible film made by the composite of (Br-PrTPP)2MnBr4 and PDMS shows an ultrahigh spatial resolution of 12.78 lp mm-1, which provides a potential visualization tool for X-ray radiography.

A feasibility study of using silkworm larvae as a novel in vivo model to evaluate the biotoxicity of ionic liquids.

Ionic liquids (ILs) have been reported to be a potential water and soil pollutant, whose toxicity has gained much attention in recent years. In this work, silkworm larvae were used as a novel in vivo model to assess the biotoxicity of ILs, which were performed by three steps. The first step was to determine the susceptibility of different silkworm strains to ILs. Data showed that Jingsong×haoyue was the most susceptible one among three silkworm strains (Jingsong×haoyue, P50, and Yi16) for evaluating the biological effects of ILs. The second step was to compare the toxicity of ILs with different structures using the larvae of Jingsong×haoyue. It was found that three representative ILs, 1-octyl-3-methylimidazole chloride ([C8mim]Cl), N-octyl-3-methylpyridine chloride ([C8mpy]Cl), and 1-octyl-3-methylimidazole tetrafluoroborate ([C8mim]BF4), had significant toxic effects on the growth and development of the larvae with 24 h median lethal concentration (24 h-LC50) values of 112.3, 156.3, and 68.9 µg g-1, respectively, indicating that the types of anions and cations had impacts on the toxicity of ILs. The last step was targeted at investigating responses of the larvae to the exposure of ILs. It was observed that remarkable physiological and biochemical responses occurred in different tissues of the larvae. For example, activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) in different tissues increased significantly to form an active protective mechanism for alleviating the toxic effects of ILs. Additionally, an increase of malondialdehyde (MDA) contents was found in the larvae. The data suggested that ILs could induce lipid peroxidation and cellular damage, which may be the main reason for toxicity of ILs to the larvae. Therefore, silkworm larvae could be used as a susceptible and reliable in vivo model to evaluate the toxicity of ILs, and the results are helpful to reveal their toxic mechanism to insects.

Molecular response mechanisms of silkworm (Bombyx mori L.) to the toxicity of 1-octyl-3-methylimidazole chloride based on transcriptome analysis of midguts and silk glands.

In a previous study, silkworm larvae were used as a novel model to assess the biotoxicity of ILs, which showed that ILs could cause significant physiological and biochemical changes in midguts and silk glands of the larvae, and result in the death of larvae. In order to investigate the toxicity of 1-octyl-3-methylimidazole chloride ([C8mim]Cl) to the larvae at molecular level, RNA-sequencing technology was used to construct transcriptomic profiles of midguts and silk glands in this work. Results showed that a lot of differentially expressed genes (DEGs) were effectively screened out through bioinformatics software based on the transcriptome data and reference genome. To give more detail, 5118 and 2211 DEGs (926 and 822 DEGs) were obtained in the midguts (silk glands) when the larvae were exposed to [C8mim]Cl for 6 and 12 h, respectively, relative to the controls. In addition, gene ontology (GO) analysis suggested that the DEGs could be divided into three categories (i.e., biological process, cellular component, and molecular function), and were involved in multiple organelle functions and complex biological processes. Kyoto encyclopedia of genes and genomes (KEGG) analysis showed that the DEGs were enriched in a variety of pathways, such as signal transduction, apoptosis, glycolysis, peroxisome, autophagy, hippo signaling pathway, arginine and proline metabolism. Results of quantitative real-time PCR and histopathological observation indicated that molecular mechanism of the larvae against [C8mim]Cl toxicology may be attributed to cell apoptosis regulation via both the mitochondrial pathway and the death receptor-initiated pathway. Thus, these results provided useful data for exploring the toxicity of ILs to insects at molecular level.

Cultivation of Chlorella sorokiniana in a bubble-column bioreactor coupled with cooking cocoon wastewater treatment: effects of initial cell density and aeration rate.

Previous studies have documented that Chlorella sorokiniana could grow well on cooking cocoon wastewater (CCW) with a maximum biomass of 0.49 g/L. In order to further enhance the biomass production and nutrient removals, a bubble-column bioreactor was designed and performed to cultivate C. sorokiniana in CCW, and two main cultivation parameters were investigated in this work. Results showed that a maximum algal biomass, specific growth rate, and biomass productivity of 2.83 g/L, 0.854 d-1, and 476.25 g/L/d, respectively, were achieved when this alga was cultivated in the bioreactor with an initial cell density of 0.8 g/L and an aeration rate of 3.34 L air/L culture/min; meanwhile, removal efficiencies of ammonium, total nitrogen, total phosphorus, and chemical oxygen demand reached 97.96, 85.66, 97.96, and 86.43%, respectively, which were significantly higher than that obtained in our previous studies. Moreover, chemical compositions in the algal cells varied with the changes of cultivation conditions (i.e., initial cell density and aeration rate). Thus, it is concluded that (1) the bubble-column bioreactor was suitable for cultivation of C. sorokiniana coupled with the CCW treatment and (2) initial cell density and aeration rate affected the biomass production, nutrient removals and chemical compositions of this alga.

Using a freshwater green alga Chlorella pyrenoidosa to evaluate the biotoxicity of ionic liquids with different cations and anions.

With the extensive use of ionic liquids (ILs) in various industrial fields, their potential toxicity to aquatic ecosystem has attracted considerable attention. In this work, biotoxicity of ILs with different cations and anions was evaluated by using a freshwater green alga Chlorella pyrenoidosa. Results showed that 1-butyl-3-methylimidazolium chloride ([C4mim]Cl), 1-octyl-3-methylimidazolium chloride ([C8mim]Cl), 1-octyl-3-methylimidazolium nitrate ([C8mim]NO3), 1-octyl-3-methylimidazolium tetrafluoroborate ([C8mim]BF4), and 1-dodecyl-3-methylimidazolium chloride ([C12mim]Cl) had a significant inhibition on the algal growth with EC50 values of 23.48, 4.72, 3.80, 4.44, and 0.10 mg L-1 at the 72 h of exposure, respectively. These data suggested that the toxicity of ILs increased with the increase of side alkyl chain length, while anions had little influences on their toxicity to this alga. Moreover, changes in chlorophyll a content and chlorophyll fluorescence parameters (Fv/Fm and ΦPSII) indicated that the five ILs could damage the photosynthetic system of this alga resulting in the decrease of photosynthetic efficiency. The increased soluble protein content and antioxidase activity could be considered as an active response mechanism of this alga against the exposure of ILs. Content of malondialdehyde (MDA) in this alga increased significantly when it was exposed to ILs, suggesting that reactive oxygen species (ROS) were accumulated in the algal cells, which would cause injury of the algal biofilm and chloroplast. Therefore, results obtained in this work would help to explain the possible underlying toxic mechanisms of ILs to C. pyrenoidosa, and provide a significant theoretical support for assessing the toxicity of ILs to aquatic organisms.

Growth inhibition and oxidative stress induced by 1-octyl-3-methylimidazolium bromide on the marine diatom Skeletonema costatum.

Marine diatom Skeletonema costatum is an important prey in the marine food web and is often used as a standard test organism in ecotoxicological studies. In this study, in vivo experiments were performed to analyze the effects of 1-octyl-3-methylimidazolium bromide ([C8mim]Br) on the growth, photosynthetic activity, and oxidative stress in S. costatum using 96h growth tests with a batch-culture system. The growth of S. costatum was significantly inhibited by [C8mim]Br with 48 and 96h-EC50 of 17.9 and 39.9mgL(-1), respectively. The maximum quantum yield (Fv/Fm) and the light use efficiency (α) were inhibited by [C8mim]Br, which affected the growth of S. costatum. Subsequent biochemical assays in S. costatum revealed that [C8mim]Br induced changes of Chl a content, soluble protein content, and SOD activity, which had significant increases in low [C8mim]Br treatments (≤20mgL(-1)), but decreased in high [C8mim]Br exposures (≥40mgL(-1)). The increase of SOD activity at low concentrations (≤20mgL(-1)) may be considered as an active defense of S. costatum against [C8mim]Br stress by reactive oxygen species (ROS) quenching. In addition, [C8mim]Br increased ROS level and malondialdehyde (MDA) content in S. costatum, suggesting that the physiological effects of [C8mim]Br are resulted from ROS generation.

Digital gene expression analysis in the midgut of 4008 silkworm strain infected with cytoplasmic polyhedrosis virus.

Digital Gene Expression was performed to investigate the midgut transcriptome profile of 4008 silkworm strain orally infected with BmCPV. A total of 4,498,263 and 4,258,240 clean tags were obtained from the control and BmCPV-infected larvae. A total of 752 differentially expressed genes were detected, of which 649 were upregulated and 103 were downregulated. Analysis results of the Kyoto Encyclopedia of Genes and Genomes pathway showed that 334 genes were involved in the ribosome and RNA transport pathways. Moreover, 408 of the 752 differentially expressed genes have a GO category and can be categorized into 41 functional groups according to molecular function, cellular component and biological process. Differentially expressed genes involved in signaling, gene expression, metabolic process, cell death, binding, and catalytic activity changes were detected in the expression profiles. Quantitative real-time PCR was performed to verify the expression of these genes. The upregulated expression levels of Calreticulin, FK506-binding protein, and protein kinase c inhibitor gene probably led to a calcium-dependent apoptosis in the BmCPV-infected cells. The results of this study may serve as a basis for future research not only on the molecular mechanism of BmCPV invasion but also on the anti-BmCPV mechanism of silkworm.

Zero-dimensional hybrid tin halides with stable broadband light emissions.

Considering the instability and toxicity of 3D Pb-based perovskite nanocrystals, lead-free low-dimensional organic-inorganic hybrid metal halides have attracted widespread attention as potential substitutes. Herein, two new tin-based 0D halides [H4BAPP]SnBr5·Br and [H4BAPP]SnCl5·Cl·H2O (BAPP = 1,4-bis(3-aminopropyl)piperazine) were synthesized successfully based on [SnX5]3- as an emission center. Typically, [H4BAPP]SnBr5·Br and [H4BAPP]SnCl5·Cl·H2O display broadband yellow and yellow-green light emissions originating from the radiative recombination of self-trapped excitons (STEs). The photoluminescence quantum yields (PLQYs) of the two compounds were calculated to be 19.27% and 2.36%, respectively. Furthermore, the excellent chemical and thermal stability and broadband light emissions reveal their potential application in solid-state white lighting diodes.

Mixotrophic culture of bait microalgae for biomass and nutrients accumulation and their synergistic carbon metabolism.

Microalgae cannot meet the bait demand for aquaculture due to light intensity limitation and other disadvantageous conditions. This research selected 6 mixotrophic microalgae, and the optimal strains and organic carbon were screened. The results showed that Thalassiosira pseudonana and Chlorella sp. are suitable for mixotrophic culture. The maximum cell density of Thalassiosira pseudonana was found to be 1.67 times than that of the photoautotrophic group when glycerol was added. The maximum cell density of Chlorella sp. with acetic acid was 1.69 times than that of the photoautotrophic group. When the concentration of acetic acid was 5.0 g·L-1 and the concentration of KNO3 was 0.2 g·L-1, the maximum biomass of Chlorella sp. could reach 3.54 × 107 cells·mL-1; the maximum biomass of Thalassiosira pseudonana was 5.53 × 106 cells·mL-1 with 10.0 g·L-1 glycerol and 0.2 g·L-1 KNO3. Metabolomic analysis further revealed that mixotrophic bait microalgae could promote the accumulation of lipids and amino acids.

Novel protein of IBP from silkworm, Bombyx mori, involved in cytoplasmic polyhedrosis virus infection.

In the present study, the full-length cDNA of a novel insulin-related peptide-binding protein (named BmIBP2) was identified from silkworm, Bombyx mori, using rapid amplification of cDNA ends. The full-length cDNA of BmIBP2 is 1293 bp, consisting of a 5'-terminal untranslated region (UTR) of 61 bp, and a 3'-UTR of 335 bp with a poly-adenylation signal sequence AATAAA and a poly (A) tail. The BmIBP2 cDNA encodes a polypeptide of 298 amino acids, including an IG domain and an IGc2 domain, with a theoretical isoelectric point of 5.73 and a predicted molecular weight of 33.1 kDa. The BmIBP2 also has a signal peptide of 23 amino acids and a potential N-glycosylation site. The sequence similarity and phylogenic analysis indicated that BmIBP2 belongs to the group of invertebrates IBP and is closer to IGFBP7 than to the other IGFBPs in vertebrates. These findings suggest that BmIBP2 is a putative homolog of vertebrate endocrine factor IGFBP7 and has a functional similarity. By fluorescent quantitative real-time polymerase chain reaction, mRNA transcripts of BmIBP2 were mainly detected in the midgut but were hardly detectable in the hemocytes, vasa mucosa, fat body, silk gland, head, testicle, ovary, and spiracle. After the silkworm larvae were infected by B. mori cytoplasmic polyhedrosis virus (BmCPV), a significant up-regulation in the relative expression level of BmIBP2 was found. All the results suggested that BmIBP2 is a novel protein that plays an important role in the insulin-signal pathway and in the immune response of silkworm to BmCPV infection.

Salinity fluctuation influences the toxicity of 1-octyl-3-methylimidazolium chloride ([C8mim]Cl) to a marine diatom Phaeodactylum tricornutum.

In this work, a marine diatom (Phaeodactylum tricornutum) was exposed to 1-octyl-3-methylimidazolium chloride ([C8mim]Cl) for 96 h at three different salinities (25, 35, and 45 ‰) for investigating their interactive effects. Results showed that values of EC10 and EC50 at 96 h of exposure were 0.29, 1.06, 2.01 µg L-1 and 7.21, 7.71, 7.25 mg L-1 when the salinities were 25, 35, and 45 ‰, respectively, meaning that salinity fluctuation affected the toxicity of [C8mim]Cl to this diatom. Changes in chlorophyll a contents and chlorophyll fluorescence parameters suggested that [C8mim]Cl and salinity fluctuation had a significant interactive effect on the algal photosynthesis. In addition, soluble protein content and activities of antioxidant enzymes in algal cells changed significantly. Increased malondialdehyde contents indicated that the combined stresses could induce excessive production of reactive oxygen species leading to oxidative damage to the algal cells.

Interactive effects of polymethyl methacrylate (PMMA) microplastics and salinity variation on a marine diatom Phaeodactylum tricornutum.

Until now, knowledge about the interactive effects of microplastics and environmental factors on primary producers is quite limited. In this work, a marine diatom (Phaeodactylum tricornutum) was exposed to polymethyl methacrylate (PMMA) microplastics at different salinities (25, 35, and 45‰) for 10 days in order to study their interactive effects. Results showed that growth of P. tricornutum was negatively affected by PMMA microplastics and salinity variation with a minimum EC50 value of 91.75 mg L-1. Photosynthetic activity of P. tricornutum was also inhibited by the two factors, and their interactive effects on chlorophyll fluorescence parameters (Fv/Fm and ΦPSII) were significant. In the algal cells, soluble protein accumulated, activities of two antioxidant enzymes changed, and malondialdehyde (MDA) content increased when this diatom was exposed to the microplastics at different salinities. These data would help to evaluate the risks of microplastics to primary producers under different environmental factors.

Oxidative stress responses caused by dimethyl phthalate (DMP) and diethyl phthalate (DEP) in a marine diatom Phaeodactylum tricornutum.

A marine diatom (Phaeodactylum tricornutum) was exposed to different concentrations of dimethyl phthalate (DMP) and diethyl phthalate (DEP) for 96 h within a batch-culture system to investigate their toxicities. Results showed that P. tricornutum could remove DMP and DEP effectively with removal rates of 0.20-0.30 and 0.14-0.21 mg L-1 h-1, respectively. In addition, DMP and DEP significantly inhibited the photosynthesis and chlorophyll a biosynthesis of P. tricornutum with 96-h EC50 values of 390.5 mg L-1 and 74.0 mg L-1, respectively. Results of reactive oxygen species (ROS) level suggested that the two PAEs could induce excessive ROS production in the diatom. Moreover, activities of antioxidant enzymes (i.e., SOD and POD) in the diatom increased with the increase of DMP and DEP concentrations. The results will help to understand the toxic mechanisms of PAEs, and provide strong evidences for evaluating their ecological risks in the marine environment.

Biomass production and biochemical profiles of a freshwater microalga Chlorella kessleri in mixotrophic culture: Effects of light intensity and photoperiodicity.

In this work, different light conditions (light intensity and photoperiodicity) were set up to investigate the impact of light on growth, chemical compositions and fatty acid profiles of Chlorella kessleri in mixotrophic cultures. Results indicated that C. kessleri could absorb and utilize glucose rapidly when light intensity was ≤ 90 µE m-2 s-1, and a maximum algal biomass of 1.17 g L-1 was obtained in the cultures with 2 g L-1 glucose at a light intensity and light/dark (L/D) cycle of 90 µE m-2 s-1 and 20L:4D, respectively. Additionally, this alga would accumulate a large amount of chlorophyll a (about 30 mg g-1) in the mixotrophic cultures under a low light intensity (≤90 µE m-2 s-1), and the algal chemical compositions changed with light intensity and photoperiodicity. Results of fatty acid profiles suggested that the algal biomass could be used as animal feeds or a good-quality biodiesel feedstock.

Impacts of four ionic liquids exposure on a marine diatom Phaeodactylum tricornutum at physiological and biochemical levels.

With the increasing applications in various industrial areas, toxicity of ionic liquids (ILs) has gained much attention in recent years. In this work, 1-octyl-3-methylimidazolium nitrate ([C8mim]NO3), 1-octyl-3-methylimidazolium chloride ([C8mim]Cl), N-octyl-3-metylpyridinium chloride ([C8mpy]Cl) and N-octyl-3-metylpyridinium bromide ([C8mpy]Br) were used to investigate the impacts of different types of cations and anions on their toxicity to a marine diatom Phaeodactylum tricornutum. Results showed that the four ILs had poor degradability under the culture conditions used in this study, and significantly inhibited the growth of this diatom with 96-h EC50 values of 24.0, 33.6, 16.1 and 14.4mgL-1 for [C8mim]NO3, [C8mim]Cl, [C8mpy]Cl and [C8mpy]Br, respectively. The data indicated that types of cations and anions would affect the toxicity of ILs to this diatom. Additionally, photosynthesis of this diatom was inhibited because significant decrease of chlorophyll a content and damage of PSII reaction centers were observed when it was exposed to the four ILs. Remarkable physiological and biochemical responses occurred in the cells of this diatom to alleviate the damaging effects of the four ILs. For example, content of soluble protein and activities of superoxide dismutase (SOD) and catalase (CAT) in the cells of this diatom increased significantly to form a protective mechanism against the toxicity of ILs, although they decreased in the cultures with high concentrations of ILs (≥20mgL-1). Thus, these results would not only provide strong evidences for evaluating the ecological risks and toxicity of ILs to marine ecosystem, but also help for understanding their toxic mechanisms to marine diatoms.

Feasibility of Growing Chlorella sorokiniana on Cooking Cocoon Wastewater for Biomass Production and Nutrient Removal.

The feasibility of microalgae cultivation using cooking cocoon wastewater (CCW) collected from a silk production factory was investigated in this work. Results showed that Chlorella sorokiniana grew well on the CCW whether it was autoclaved or not. After 7-day cultivation, the biomass increased by 1.57, 2.78, 3.33, and 3.14 times, and by 3.65, 4.03, 3.27, and 2.82 times when this alga was cultivated in the raw CCW (R-CCW) and autoclaved CCW (A-CCW) at the initial dry cell densities of 0.01, 0.04, 0.08, and 0.16 g/L, respectively. The algal photosynthetic growth was not affected when this alga grew on the R-CCW at an initial dry cell density of ≥ 0.04 g/L, while it was significantly inhibited when the initial dry cell density was 0.01 g/L. Additionally, this alga could remove nutrients rapidly from the CCW, and the removal efficiency increased with the increase of initial dry cell density. Thus, it was concluded that the CCW could be used as a good-quality medium for the algal growth, which is worthy of further study and promotion.

Identification of archaeon-producing hyperthermophilic alpha-amylase and characterization of the alpha-amylase.

The extremely thermophilic anaerobic archaeon strain, HJ21, was isolated from a deep-sea hydrothermal vent, could produce hyperthermophilic alpha-amylase, and later was identified as Thermococcus from morphological, biochemical, and physiological characteristics and the 16S ribosomal RNA gene sequence. The extracellular thermostable alpha-amylase produced by strain HJ21 exhibited maximal activity at pH 5.0. The enzyme was stable in a broad pH range from pH 5.0 to 9.0. The optimal temperature of alpha-amylase was observed at 95 degrees C. The half-life of the enzyme was 5 h at 90 degrees C. Over 40% and 30% of the enzyme activity remained after incubation at 100 degrees C for 2 and 3 h, respectively. The enzyme did not require Ca(2+) for thermostability. This alpha-amylase gene was cloned, and its nucleotide sequence displayed an open reading frame of 1,374 bp, which encodes a protein of 457 amino acids. Analysis of the deduced amino acid sequence revealed that four homologous regions common in amylases were conserved in the HJ21 alpha-amylase. The molecular weight of the mature enzyme was calculated to be 51.4 kDa, which correlated well with the size of the purified enzyme as shown by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis.