Insufficient KIF15 during porcine oocyte ageing induces HDAC6-based microtubule instability.

During aging, oocytes display cytoskeleton dynamics defects and aneuploidy, leading to embryonic aneuploidy, which in turn causes miscarriages, implantation failures, and birth defects. KIF15 (also known as Hklp2), a member of the kinesin-12 superfamily, is a cytoplasmic motor protein reported to be involved in Golgi and vesicle-related transport during mitosis in somatic cells. However, the regulatory mechanisms of KIF15 during meiosis in porcine oocytes and the connection with postovulatory aging remain unclear. In present study, we found that KIF15 is expressed during porcine oocyte maturation, and its localization is dependent on microtubule dynamics. Furthermore, the level of KIF15 expression decreased in postovulatory aged oocytes. The decrease in KIF15 blocked polar body extrusion, thereby hindering oocyte maturation. We demonstrated that KIF15 defects contributed to abnormal spindle morphologies and chromosome misalignment, possibly due to microtubule instability, as evidenced by microtubule depolymerization after cold treatment. Additionally, our data indicated that KIF15 modulates HDAC6 to affect tubulin acetylation in oocytes. Taken together, these results suggest that KIF15 regulates HDAC6-related microtubule stability for spindle organization in porcine oocytes during meiosis, which may contribute to the decline in maturation competence in aged porcine oocytes.

Biodegradation of various grades of polyethylene microplastics by Tenebrio molitor and Tenebrio obscurus larvae: Effects on their physiology.

Polyethylene (PE) is the most productive plastic product and includes three major polymers including high-density polyethylene (HDPE), linear low-density polyethylene (LLDPE) and low-density polyethylene (LDPE) variation in the PE depends on the branching of the polymer chain and its crystallinity. Tenebrio obscurus and Tenebrio molitor larvae biodegrade PE. We subsequently tested larval physiology, gut microbiome, oxidative stress, and PE degradation capability and degradation products under high-purity HDPE, LLDPE, and LDPE powders (<300 µm) diets for 21 days at 65 ± 5% humidity and 25 ± 0.5 °C. Our results demonstrated the specific PE consumption rates by T. molitor was 8.04-8.73 mg PE ∙ 100 larvae-1⋅day-1 and by T. obscurus was 7.68-9.31 for LDPE, LLDPE and HDPE, respectively. The larvae digested nearly 40% of the ingested three PE and showed similar survival rates and weight changes but their fat content decreased by 30-50% over 21-day period. All the PE-fed groups exhibited adverse effects, such as increased benzoquinone concentrations, intestinal tissue damage and elevated oxidative stress indicators, compared with bran-fed control. In the current study, the digestive tract or gut microbiome exhibited a high level of adaptability to PE exposure, altering the width of the gut microbial ecological niche and community diversity, revealing notable correlations between Tenebrio species and the physical and chemical properties (PCPs) of PE-MPs, with the gut microbiome and molecular weight change due to biodegradation. An ecotoxicological simulation by T.E.S.T. confirmed that PE degradation products were little ecotoxic to Daphnia magna and Rattus norvegicus providing important novel insights for future investigations into the environmentally-friendly approach of insect-mediated biodegradation of persistent plastics.

Microplastics perturb nitrogen removal, microbial community and metabolism mechanism in biofilm system.

Microplastics (MPs) are a significant component of global pollution and cause widespread concern, particularly in wastewater treatment plants. While understanding the impact of MPs on nutrient removal and potential metabolism in biofilm systems is limited. This work investigated the impact of polystyrene (PS) and polyethylene terephthalate (PET) on the performance of biofilm systems. The results revealed that at concentrations of 100 and 1000 µg/L, both PS and PET had almost no effect on the removal of ammonia nitrogen, phosphorus, and chemical oxygen demand, but reduced the removal of total nitrogen by 7.40-16.6%. PS and PET caused cell and membrane damage, as evidenced by increases in reactive oxygen species and lactate dehydrogenase to 136-355% and 144-207% of the control group. Besides, metagenomic analysis demonstrated both PS and PET changed the microbial structure and caused functional differences. Some important genes in nitrite oxidation (e.g. nxrA), denitrification (e.g. narB, nirABD, norB, and nosZ), and electron production process (e.g. mqo, sdh, and mdh) were restrained, meanwhile, species contribution to nitrogen-conversion genes was altered, therefore disturbing nitrogen-conversion metabolism. This work contributes to evaluating the potential risks of biofilm systems exposed to PS and PET, maintaining high nitrogen removal and system stability.

Enhanced anaerobic degradation of azo dyes by biofilms supported by novel functionalized carriers.

Azo dyes are significant organic pollutants known for their adverse effects on humans and aquatic life. In this study, anthraquinone-2-sulfonate (AQS) immobilized on biochar (BC) was employed as a novel carrier in up-flow anaerobic fixed-bed reactors to induce specific biofilm formation and promote the biotransformation efficiency of azo dyes. Novel carrier-packed reactor 1 (R1) and BC-packed reactor 2 (R2) were used to treat red reactive 2 (RR2) under continuous operation for 175 days. The decolorization rates of R1 and R2 were 96-83% and 91-73%, respectively. The physicochemical characteristics and extracellular polymeric substances (EPS) of the biofilm revealed a more stable structure in R1. Furthermore, the microbial community in R1 interacted more closely with each other and contained more keystone genera. Overall, this study provides a feasible method for improving the biotransformation of azo dyes, thus providing support for practical applications in wastewater treatment projects.

Gut Microbiome Associating with Carbon and Nitrogen Metabolism during Biodegradation of Polyethene in Tenebrio larvae with Crop Residues as Co-Diets.

Tenebrio molitor and Tenebrio obscurus (Coleoptera: Tenebrionidae) larvae are two commercial insects that eat plant and crop residues as diets and also biodegrade synthetic plastics polyethylene (PE). We examined biodegradation of low-density PE (LDPE) foam (Mn = 28.9 kDa and Mw = 342.0 kDa) with and without respective co-diets, i.e., wheat brain (WB) or corn flour (CF), corn straw (CS), and rice straw (RS) at 4:1 (w/w), and their gut microbiome and genetic metabolic functional groups at 27.0 ± 0.5 °C after 28 days of incubation. The presence of co-diets enhanced LDPE consumption in both larvae and broad-depolymerized the ingested LDPE. The diet type shaped gut microbial diversity, potential pathways, and metabolic functions. The sequence of effectiveness of co-diets was WB or CF > CS > RS for larval development and LDPE degradation. Co-occurrence networks indicated that the larvae co-fed with LDPE displayed more complex correlations of gut microbiome than the larvae fed with single diets. The primary diet of WB or CF and crop residues CS and RS provided energy and nitrogen source to significantly enhance LDPE biodegradation with synergistic activities of the gut microbiota. For the larvae fed LDPE and LDPE plus co-diets, nitrogen fixation function was stimulated compared to normal diets and associated with LDPE biodegradation.

Impacts of physical-chemical property of polyethylene on depolymerization and biodegradation in yellow and dark mealworms with high purity microplastics.

Yellow and dark mealworms (Tenebrio molitor and Tenebrio obscurus) biodegrade commercial polyethylene (PE) materials at a high rate. We examined the impact of physical and chemical properties on biodegradation using high purity microplastics (MPs). These included high-density polyethylene (HDPE), low-density polyethylene (LDPE), and linear low-density polyethylene (LLDPE), all with different weight average molecular weights (Mw) and different crystallinity degrees in T. molitor and T. obscurus larvae. The biodegradation extent in the two mealworms was similar but strongly depended on the polymer type in sequence, since LDPE > LLDPE> HDPE (with respective Mw of 222.5, 110.5 and 182 kDa). When LDPE MPs with Mw of 0.84, 6.4 and 106.8 kDa and HDPE with Mw of 52, 105 and 132.7 kDa were tested, the PE MPs with lower Mw showed a greater extent of depolymerization. The results of dominance analysis indicated that less branching structure and higher crystallinity degree negatively impacted depolymerization and biodegradation. Py-GC/MS analysis confirmed the breaking of the macromolecule backbone as well as the formation of oxidized functional groups after all the tested PE materials passed through the mealworm intestine. The results demonstrated that molecular weight, PE type, branching, and crystallinity degree significantly affect the biodegradation capability of PE by the mealworms, and possibly by other biological systems as well.

Confirmation of biodegradation of low-density polyethylene in dark- versus yellow- mealworms (larvae of Tenebrio obscurus versus Tenebrio molitor) via. gut microbe-independent depolymerization.

Tenebrio obscurus (Coleoptera: Tenebrionidae) larvae are capable of biodegrading polystyrene (PS) but their capacity for polyethylene (PE) degradation and pattern of depolymerization remains unknown. This study fed the larvae of T. obscurus and Tenebrio molitor, which have PE degrading capacity, two commercial low-density PE (LDPE) foams i.e., PE-1 and PE-2, with respective number-average molecular weights (Mn) of 28.9 and 27.3 kDa and weight-average molecular weights (Mw) of 342.0 and 264.1 kDa, over a 36-day period at ambient temperature. The Mw of residual PE in frass (excrement) of T. obscurus, fed with PE-1 and PE-2, decreased by 45.4 ± 0.4% and 34.8 ± 0.3%, respectively, while the respective decrease in frass of T. molitor was 43.3 ± 0.5% and 31.7 ± 0.5%. Data analysis showed that low molecular weight PE (<5.0 kDa) was rapidly digested while longer chain portions (>10.0 kDa) were broken down or cleaved, indicating a broad depolymerization pattern. Mass balance analysis indicated nearly 40% of ingested LDPE was digested to CO2. Antibiotic suppression of gut microbes in T. molitor and T. obscurus larvae with gentamicin obviously reduced their gut microbes on day 15 but did not stop depolymerization because the Mn, Mw and size- average molecular weight (Mz) decreased. This confirmed that LDPE biodegradation in T. obscurus was independent of gut microbes as observed during previous PS degradation in T. molitor, suggesting that the intestinal digestive system could perform LDPE depolymerization. High-throughput sequencing revealed significant shifts in the gut microbial community during bran-fed and unfed conditions in response to LDPE feeding in both Tenebrio species. The respective predominant gut genera of Spiroplasma sp. and Enterococcus sp. were observed in LDPE-fed T. molitor and T. obscurus larvae.

Biodegradation of polypropylene by yellow mealworms (Tenebrio molitor) and superworms (Zophobas atratus) via gut-microbe-dependent depolymerization.

Polypropylene (PP), a fossil-based polyolefin plastics widely used worldwide, is non-hydrolyzable and resistant to biodegradation as a major source of plastic pollutants in environment. This study focused on feasibility of PP biodegradation in the larvae of two species of darkling beetles (Coleoptera: Tenebrionidae) i.e., yellow mealworms (Tenebrio molitor) and superworms (Zophobas atratus) using PP foam with number-, weight-, and size-average molecular weights (Mn, Mw, and Mz) of 109.8, 356.2, and 765.0 kDa, respectively. The tests were conducted in duplicates with respective larvae (300 T. molitor and 200 Z. atratus each incubator) at 25 °C and 65% humidity for over a 35-day period. The larvae of T. molitor and Z. atratus fed with PP foam as sole diet consumed PP at 1.0 ± 0.4 and 3.1 ± 0.4 mg 100 larvae-1 days-1, respectively; when fed the PP foam plus wheat bran, the consumption rates were enhanced by 68.11% and 39.70%, respectively. Gel permeation chromatography analyses of the frass of T. molitor and Z. atratus larvae fed PP only indicated that Mw was decreased by 20.4 ± 0.8% and 9.0 ± 0.4%; Mn was increased by 12.1 ± 0.4% and 61.5 ± 2.5%; Mz was decreased by 33.8 ± 1.5% and 32.0 ± 1.1%, indicating limited extent depolymerization. Oxidation and biodegradation of PP was confirmed through analysis of the residual PP in frass. Depression of gut microbes with the antibiotic gentamicin inhibited PP depolymerization in both T. molitor and Z. atratus larvae. High throughput 16S rRNA sequencing revealed that Citrobacter sp. and Enterobacter sp. were associated with PP diets in the gut microbiome of Z. atratus larvae while Kluyvera was predominant in the T. molitor larvae. The results indicated that PP can be biodegraded in both T. molitor and Z. atratus larvae via gut microbe-dependent depolymerization with diversified microbiomes.

Simulating a combined lysis-cryptic and biological nitrogen removal system treating domestic wastewater at low C/N ratios using artificial neural network.

In this study, a combined alkaline (ALK) and ultrasonication (ULS) sludge lysis-cryptic pretreatment and anoxic/oxic (AO) system (AO + ALK/ULS) was developed to enhance biological nitrogen removal (BNR) in domestic wastewater with a low carbon/nitrogen (C/N) ratio. A real-time control strategy for the AO + ALK/ULS system was designed to optimize the sludge lysate return ratio (RSLR) under variable sludge concentrations and variations in the influent C/N (⩽ 5). A multi-layered backpropagation artificial neural network (BPANN) model with network topology of 1 input layer, 3 hidden layers, and 1 output layer, using the Levenberg-Marquardt algorithm, was developed and validated. Experimental and predicted data showed significant concurrence, verified with a high regression coefficient (R2 = 0.9513) and accuracy of the BPANN. The BPANN model effectively captured the complex nonlinear relationships between the related input variables and effluent output in the combined lysis-cryptic + BNR system. The model could be used to support the real-time dynamic response and process optimization control to treat low C/N domestic wastewater.

CircAGFG1 sponges miR-203 to promote EMT and metastasis of non-small-cell lung cancer by upregulating ZNF281 expression.

The circRNA circAGFG1 is reported to be important in triple-negative breast cancer progression. However, the mechanism of circAGFG1 in non-small-cell lung cancer (NSCLC) remains unknown. In this study, expression of circAGFG1 was determined by real-time PCR in 20 pairs of NSCLC tissues and adjacent tissues. Next, functional experiments with circAGFG1 were performed in vitro to evaluate the role of circAGFG1 in tumor metastasis and growth. Meanwhile, a dual luciferase reporter assay, RNA pull-down and RNA immunoprecipitation experiments were used to explore the interaction between circAGFG1 and miR-203. Our results revealed that expression levels of circAGFG1 and miR-203 are upregulated in non-small-cell lung cancer tissues. CircAGFG1 enhances NSCLC cell proliferation, invasion, migration and epithelial-mesenchymal transition in vitro. Mechanistic analyses indicated that circAGFG1 acts as a sponge for miR-203 to repress the effect of miR-203 on its target, ZNF281. In conclusion, our study suggests that circAGFG1 promotes NSCLC growth and metastasis though a circAGFG1/miR-203/ZNF281 axis and may represent a novel therapeutic target.

Comparative Analysis of Four Buckwheat Species Based on Morphology and Complete Chloroplast Genome Sequences.

Buckwheat is a nutritional and economically crop belonging to Polygonaceae, Fagopyrum. To better understand the mutation patterns and evolution trend in the chloroplast (cp) genome of buckwheat, and found sufficient number of variable regions to explore the phylogenetic relationships of this genus, two complete cp genomes of buckwheat including Fagopyrum dibotrys (F. dibotrys) and Fagopyrum luojishanense (F. luojishanense) were sequenced, and other two Fagopyrum cp genomes were used for comparative analysis. After morphological analysis, the main difference among these buckwheat were height, leaf shape, seeds and flower type. F. luojishanense was distinguishable from the cultivated species easily. Although the F. dibotrys and two cultivated species has some similarity, they different in habit and component contents. The cp genome of F. dibotrys was 159,320 bp while the F. luojishanense was 159,265 bp. 48 and 61 SSRs were found in F. dibotrys and F. luojishanense respectively. Meanwhile, 10 highly variable regions among these buckwheat species were located precisely. The phylogenetic relationships among four Fagopyrum species based on complete cp genomes was showed. The results suggested that F. dibotrys is more closely related to Fagopyrum tataricum. These data provided valuable genetic information for Fagopyrum species identification, taxonomy, phylogenetic study and molecular breeding.

Plantlet Regeneration of Tartary Buckwheat (Fagopyrum tataricum Gaertn.) in Vitro Tissue Cultures.

Tartary buckwheat is an ancient annual dicotyledonous herb, which is widely distributed around the world, specifically in the high altitude area of southwestern China and in the hill region of Himalayan. The plantlet regeneration of tartary buckwheat via somatic embryogenesis or multiple shoot induction was investigated in two different tartary buckwheats, Yuanzi and Xichang. The regeneration ability of Yuanzi was better than Xichang tartary buckwheat, and the hypocotyls were better than cotyledons as tartary buckwheat plantlet regeneration explants via somatic embryogenesis. The most suitable medium for callus induction was Murashige and Skoog basal medium added 2 mg/L 2, 4- dichlorophenoxyacetic acid and 1 mg/L Kinetin, which could reach up to 98.96% callus induction percentage. The plantlet regeneration percentage from callus of tartary buckwheat could reach up to 55.77%, which induced on 2.0 mg/L Benzyladenine and 1.0 mg/L KT in MS basal medium. In addition, maximum of multiple shoot induction percentage was 69.05%, which was observed in case of Yuanzi tartary buckwheat in MS basal medium with added 3.0 mg/L 6-BA and 1.0 mg/L Thidiazuron. Roots induction of regenerated plants were achieved on 1/2 MS basal medium with added 1mg/L Indole-3-Butytric acid, which has 75% survival after transferred regenerated plants to soil under field conditions.