Assessing immune hepatotoxicity of troglitazone with a versatile liver-immune-microphysiological-system.

Drug-induced liver injury is a prevalent adverse event associated with pharmaceutical agents. More significantly, there are certain drugs that present severe hepatotoxicity only during the clinical phase, consequently leading to the termination of drug development during clinical trials or the withdrawal from the market after approval. The establishment of an evaluation model that can sensitively manifest such hepatotoxicity has always been a challenging aspect in drug development. In this study, we build a liver-immune-microphysiological-system (LIMPS) to fully demonstrate the liver injury triggered by troglitazone (TGZ), a drug that was withdrawn from the market due to hepatotoxicity. Leveraging the capabilities of organ-on-chip technology allows for the dynamic modulation of cellular immune milieu, as well as the synergistic effects between drugs, hepatocytes and multiple immune cells. Through the LIMPS, we discovered that 1) TGZ can promote neutrophils to adhered hepatocytes, 2) the presence of TGZ enhances the crosstalk between macrophages and neutrophils, 3) the induction of damage in hepatocytes by TGZ at clinically relevant blood concentrations not observed in other in vitro experiments, 4) no hepatotoxicity was observed in LIMPS when exposed to rosiglitazone and pioglitazone, structurally similar analogs of TGZ, even at the higher multiples of blood drug concentration levels. As an immune-mediated liver toxicity assessment method, LIMPS is simple to operate and can be used to test multiple drug candidates to detect whether they will cause severe liver toxicity in clinical settings as early as possible.

Strain Effect on Dielectricity of Elastic Thermoplastic Polyurethanes.

Dielectric elastomers, such as thermoplastic polyurethanes (TPUs), are widely used as the dielectric layer, encapsulation layer, and substrate of flexible and stretchable devices. To construct capacitors and actuators that work stably upon deformation, it has become urgent to investigate the evolution of dielectricity under stress and strain. However, the lack of effective methods for estimating the dielectric constant of elastomers under strain poses a big challenge. This study reports a device for the in situ measurement of the dielectric constant of TPU under strain. It is found that upon stretching TPU to a strain of 400%, its dielectric constant decreases from 8.02 ± 0.01 to 2.88 ± 0.25 (at 1 MHz). In addition, combined Fourier-transform infrared spectroscopy, the X-ray scattering technique, and atomic force microscopy were utilized to characterize the evolution of the microstructure under strain. The investigation under tensile strain reveals a decreased density and average size of polarized hard domains, along with a tendency of the molecular chains to align in parallel with the tensile stress. The evolution of the microstructures results in a reduction in the measured dielectric constant in TPU.

Unveiling the Potential of Bacillus safensis Y246 for Enhanced Suppression of Rhizoctonia solani.

Rhizoctonia solani is a significant pathogen affecting various crops, including tobacco. In this study, a bacterial strain, namely Y246, was isolated from the soil of healthy plants and exhibited high antifungal activity. Based on morphological identification and DNA sequencing, this bacterial strain was identified as Bacillus safensis. The aim of this investigation was to explore the antifungal potential of strain Y246, to test the antifungal stability of Y246 by adjusting different cultivation conditions, and to utilize gas chromatography-mass spectrometry (GC-MS) to predict the volatile compounds related to antifungal activity in Y246. In vitro assays demonstrated that strain Y246 exhibited a high fungal inhibition rate of 76.3%. The fermentation broth and suspension of strain Y246 inhibited the mycelial growth of R. solani by 66.59% and 63.75%, respectively. Interestingly, treatment with volatile compounds derived from the fermentation broth of strain Y246 resulted in abnormal mycelial growth of R. solani. Scanning electron microscopy analysis revealed bent and deformed mycelium structures with a rough surface. Furthermore, the stability of antifungal activity of the fermentation broth of strain Y246 was assessed. Changes in temperature, pH value, and UV irradiation time had minimal impact on the antifungal activity, indicating the stability of the antifungal activity of strain Y246. A GC-MS analysis of the volatile organic compounds (VOCs) produced by strain Y246 identified a total of 34 compounds with inhibitory effects against different fungi. Notably, the strain demonstrated broad-spectrum activity, exhibiting varying degrees of inhibition against seven pathogens (Alternaria alternata, Phomopsis. sp., Gloeosporium musarum, Dwiroopa punicae, Colletotrichum karstii, Botryosphaeria auasmontanum, and Botrytis cinerea). In our extensive experiments, strain Y246 not only exhibited strong inhibition against R. solani but also demonstrated remarkable inhibitory effects on A. alternata-induced tobacco brown spot and kiwifruit black spot, with impressive inhibition rates of 62.96% and 46.23%, respectively. Overall, these findings highlight the significant antifungal activity of B. safensis Y246 against R. solani. In addition, Y246 has an excellent antifungal stability, with an inhibition rate > 30% under different treatments (temperature, pH, UV). The results showed that the VOCs of strain Y246 had a strong inhibitory effect on the colony growth of R. solani, and the volatile substances produced by strain Y246 had an inhibitory effect on R. solani at rate of 70.19%. Based on these results, we can conclude that Y246 inhibits the normal growth of R. solani. These findings can provide valuable insights for developing sustainable agricultural strategies.

The Effects of Alternative Foods on Life History and Cannibalism of Amblyseius herbicolus (Acari: Phytoseiidae).

The development, survivorship, fecundity, and cannibalism of the predatory phytoseiid mite, Amblyseius herbicolus (Chant), fed six different alternative foods (Oulenziella bakeri, Tyrophagus putrescentiae, Aleuroglyphus ovatus, almond pollen (Prunus armeniaca), apple pollen (Malus pumila), maize pollen (Zea mays)), and natural prey (Tetranychus urticae) were determined in the laboratory. Our findings indicated that A. herbicolus that fed on all six alternative foods could normally complete its developmental and reproductive cycles. The shortest pre-adult developmental duration was observed when A. herbicolus fed on almond pollen (4.91 d) as well as T. urticae (4.90 d), and the longest when it fed on maize pollen (6.24 d). Pre-adult survival rates were higher when the predator fed on almond pollen (0.99), maize pollen (0.96), and O. bakeri (0.93). The highest fecundity was observed when A. herbicolus fed on apple pollen (28.55 eggs/female), almond pollen (26.06 eggs/female), and O. bakeri (26.02 eggs/female) in addition to T. urticae (48.95 eggs/female), and the lowest when it fed on maize pollen (7.84 eggs/female). The highest value of the intrinsic rate of increase (r) was obtained when A. herbicolus fed on O. bakeri (0.202 d-1) in addition to T. urticae (0.210 d-1), followed by almond pollen (0.163 d-1), and the lowest was when it fed on maize pollen (0.064 d-1). Cannibalism of conspecific eggs by adults of A. herbicolus did not occur when O. bakeri and T. urticae were provided. The cannibalism rate of the predatory mite was the lowest when fed on almond pollen, T. putrescentiae, and A. ovatus and the highest on apple pollen. Above all, when fed on O. bakeri and almond pollen, and with no or low cannibalism rate, A. herbicolus had the best development, survivorship, fecundity, and population parameters. Therefore, O. bakeri and almond pollen could be potential alternative foods for mass rearing programs of A. herbicolus or to support its population in the fields.

Effective degradation of 2,4-dihydroxybenzophenone by zero-valent iron powder (Fe0)-activated persulfate in aqueous solution: Kinetic study, product identification and theoretical calculations.

2,4-Dihydroxybenzophenone (BP-1), a typically known derivative of the benzophenone-type UV filter, has been frequently detected in aqueous environments and poses a potential risk to human health and the entire ecosystem. In this study, an effective advanced oxidation technique using zero-valent iron powder (Fe0)-activated persulfate (PS) was used for the degradation of BP-1. The effects of several experimental parameters, including Fe0 dosages, PS dosages, pH, and common natural water constituents, were systematically investigated. The BP-1 degradation efficiency was enhanced by increasing the Fe0 and PS dosages and decreasing the solution pH. The presence of different concentrations of humic acid (HA) could inhibit BP-1 removal, while the addition of various cations and anions had different effects on the degradation. Moreover, the degradation of BP-1 in five water matrices was also compared, and the removal rates followed the order of ultrapure water > tap water > secondary clarifier effluent > river water > synthetic water. Thirteen oxidation products were identified by liquid chromatography-time-of-flight-mass spectrometry (LC-TOF-MS) analysis, and five possible degradation pathways were proposed. The addition reactions initiated by HO and SO4-, as well as single-electron coupling reactions and ring-closing reactions, were further supported by density functional theory (DFT) calculations. Assessment of toxicity of intermediates of the oxidation of BP-1 suggested decreased toxicity from the parent contaminant. The present work illustrates that BP-1 could be efficiently degraded in the Fe0/PS system, which may provide new insights into the removal of benzophenones in water and wastewater.

Products distribution and contribution of (de)chlorination, hydroxylation and coupling reactions to 2,4-dichlorophenol removal in seven oxidation systems.

We systemically investigated the transformation behavior of 2,4-dichlorophenol (24-DCP) in seven different reaction systems including KMnO4, heat/PS, O3, UV, Fenton, NaClO and K2FeO4 treatment. The results revealed that complete removal of 24-DCP could be reached in minutes, especially for Fe(VI), KMnO4, NaClO, Fenton and O3 system. A total of 41 products were identified by LC-MS, and 10 of them were validated using commercial and self-synthesized standards. Hydroxyl substitution and coupling reactions were commonly observed in the studied systems. Meanwhile, extra routes such as sulfate substitution, (de)chlorination and direct oxidation were also involved for certain oxidation methods. Comparisons showed that a high degree of chlorination (>90%) occurred for NaClO system, while coupling products accounted for ~45% of the removed 24-DCP under PS oxidation. Moreover, low mineralization degree together with high aquatic toxicity was attributed to the occurrence of coupling reaction, which was possibly related to the redox potential of the main oxidative species. Considering the low abundance of coupling products and the gentle reaction condition, UV irradiation is a better option for 24-DCP removal in water and wastewaters. These findings can deepen our understanding on the transformation process of 24-DCP and provide some useful information for the environmental elimination of substituted phenols.

The S. pombe mitochondrial transcriptome.

Mitochondrial gene expression is largely controlled through post-transcriptional processes including mitochondrial RNA (mt-RNA) processing, modification, decay, and quality control. Defective mitochondrial gene expression results in mitochondrial oxidative phosphorylation (OXPHOS) deficiency and has been implicated in human disease. To fully understand mitochondrial transcription and RNA processing, we performed RNA-seq analyses of mt-RNAs from the fission yeast Schizosaccharomyces pombe RNA-seq analyses show that the abundance of mt-RNAs vary greatly. Analysis of data also reveals mt-RNA processing sites including an unusual RNA cleavage event by mitochondrial tRNA (mt-tRNA) 5'-end processing enzyme RNase P. Additionally, this analysis reveals previously unknown mitochondrial transcripts including the rnpB-derived fragment, mitochondrial small RNAs (mitosRNAs) such as mt-tRNA-derived fragments (mt-tRFs) and mt-tRNA halves, and mt-tRNAs marked with 3'-CCACCA/CCACC in S. pombe Finally, RNA-seq reveals that inactivation of trz2 encoding S. pombe mitochondrial tRNA 3'-end processing enzyme globally impairs mt-tRNA 3'-end processing, inhibits mt-mRNA 5'-end processing, and causes accumulation of unprocessed transcripts, demonstrating the feasibility of using RNA-seq to examine the protein known or predicted to be involved in mt-RNA processing in S. pombe Our work uncovers the complexity of a fungal mitochondrial transcriptome and provides a framework for future studies of mitochondrial gene expression using S. pombe as a model system.

The fission yeast Schizosaccharomyces pombe Mtf2 is required for mitochondrial cox1 gene expression.

Mitochondrial gene expression is essential for adenosine triphosphate synthesis via oxidative phosphorylation, which is the universal energy currency of cells. Here, we report the identification and characterization of a homologue of Saccharomyces cerevisiae Mtf2 (also called Nam1) in Schizosaccharomyces pombe. The Δmtf2 mutant with the intron-containing mitochondrial DNA (mtDNA) exhibited impaired growth on a rich medium containing the non-fermentable carbon source glycerol, suggesting that mtf2 is involved in mitochondrial function. mtf2 deletion in a mitochondrial intron-containing background resulted in a barely detectable level of the cox1 mRNA and a reduction in the level of the cob1 mRNA, and severely impaired cox1 translation. In contrast, mtf2 deletion in a mitochondrial intron-less background did not affect the levels of cox1 and cob1 mRNAs. However, Cox1 synthesis could not be restored to the control level in the Δmtf2 mutant with intron-less mtDNA. Our results suggest that unlike its counterpart in S. cerevisiae which plays a general role in synthesis of mtDNA-encoded proteins, S. pombe Mtf2 primarily functions in cox1 translation and the effect of mtf2 deletion on splicing of introns in mtDNA is likely due to a deficiency in the synthesis of intron-encoded maturases.

Persistent spread of the rmtB 16S rRNA methyltransferase gene among Escherichia coli isolates from diseased food-producing animals in China.

A total of 963 non-duplicate Escherichia coli strains isolated from food-producing animals between 2002 and 2012 were screened for the presence of the 16S rRNA methyltransferase genes. Among the positive isolates, resistance determinants to extended spectrum ß-lactamases, plasmid-mediated quinolone resistance genes as well as floR and fosA/A3/C2 were detected using PCR analysis. These isolates were further subjected to antimicrobial susceptibility testing, molecular typing, PCR-based plasmid replicon typing and plasmid analysis. Of the 963 E. coli isolates, 173 (18.0%), 3 (0.3%) and 2 (0.2%) were rmtB-, armA- and rmtE-positive strains, respectively. All the 16S rRNA methyltransferase gene-positive isolates were multidrug resistant and over 90% of them carried one or more type of resistance gene. IncF (especially IncFII) and non-typeable plasmids played the main role in the dissemination of rmtB, followed by the IncN plasmids. Plasmids that harbored rmtB ranged in size from 20kb to 340kb EcoRI-RFLP testing of the 109 rmtB-positive plasmids from different years and different origins suggested that horizontal (among diverse animals) and vertical transfer of IncF, non-typeable and IncN-type plasmids were responsible for the spread of rmtB gene. In summary, our findings highlight that rmtB was the most prevalent 16S rRNA methyltransferase gene, which present persistent spread in food-producing animals in China and a diverse group of plasmids was responsible for rmtB dissemination.

Ex vivo pharmacokinetic and pharmacodynamic analysis of valnemulin against Mycoplasma gallisepticum S6 in Mycoplasma gallisepticum and Escherichia coli co-infected chickens.

Pharmacokinetic and pharmacodynamic (PK/PD) indices against Mycoplasma gallisepticum (MG) S6 were investigated in an ex vivo PK/PD model following oral administration of valnemulin to chickens co-infected with M. gallisepticum and Escherichia coli. The minimum inhibitory concentrations (MICs) for valnemulin against MG S6 in artificial medium and chicken serum were determined. In vitro time-killing curves were established according to a series of multiples of the MIC value in an artificial medium, and ex vivo time-killing curves were established in serum samples obtained from infected chickens at different time points after oral administration with an initial titer of 1 × 10(6) color change units (CCU)/mL MG S6. The sigmoid Emax model was used to provide 24 h area under concentration-time curve/minimum inhibitory concentration ratios (AUC0-24h/MIC) for mycoplasmastasis, mycoplasmacidal activity and mycoplasmal elimination, respectively. The inoculum size and micro or macro methods exhibited little effect on MIC determination of MG, whereas matrix had a large effect. The rapid killing activity observed in in vitro time-killing curves seems to indicate that valnemulin was mycoplasmacidal and concentration dependent against MG. The AUC0-24h/MIC ratio for mycoplasmacidal activity and mycoplasmal elimination was 1321 h and 1960 h, respectively. A dosage regimen of 12.4 mg/kg/day and 18.3 mg/kg/day valnemulin was calculated for mycoplasmacidal activity and mycoplasmal elimination against MG S6, respectively.