Establishment and optimization of a high-throughput UPLC-MS/MS method for the simultaneous quantitation of cycloicaritin and its valine carbamate prodrug in rat plasma.

An ultra-performance liquid chromatography-tandem mass spectrometry was developed to assay the concentration for the quantification of cycloicaritin and its carbamate prodrug (3-O-L-valyl carbamate prodrug of cycloicaritin) in the plasma of Sprague-Dawley rats. Analytes were separated on an Acquity UPLC BEH C18 (2.1 × 50 mm, 1.7 µm) after liquid-liquid extraction with methyl tert-butyl ether. Acetonitrile and water containing 0.1 % formic acid were the mobile phases of the method. Using electrospray ionization in the positive ion mode, the method was performed with a total run time of 2.60 min. The response of the experiments was linear over the concentration ranges from 1 to 250 ng/mL for cycloicaritin and 1-250 ng/mL for prodrug. The intra- and inter-day precision and accuracy were within the recommended limits of the FDA. The matrix effect that we observed met the criteria. The method was successfully applied to the pharmacokinetics of cycloicaritin and its carbamate prodrug in Sprague-Dawley rats.

Molecular and functional dissection of EARLY-FLOWERING 3 (ELF3) and ELF4 in Arabidopsis.

The circadian clock is an endogenous timekeeper system that generates biological rhythms of approximately 24 h in most organisms. EARLY FLOWERING 3 (ELF3) and ELF4 were initially identified as negative regulators of flowering time in Arabidopsis thaliana. They were then found to play crucial roles in the circadian clock by integrating environmental light and ambient temperature signals and transmitting them to the central oscillator, thereby regulating various downstream cellular and physiological processes. At dusk, ELF3 acts as a scaffold, interacting with ELF4 and the transcription factor LUX ARRHYTHMO (PHYTOCLOCK1) to form an EVENING COMPLEX (EC). This complex represses the transcription of multiple circadian clock-related genes, thus inhibiting hypocotyl elongation and flowering. Subsequent studies have expanded knowledge about the regulatory roles of the EC in thermomorphogenesis and shade responses. In addition, ELF3 and ELF4 also form multiple complexes with other proteins including chromatin remodeling factors, histone deacetylase, and transcription factors, thus enabling the transcriptional repression of diverse targets. In this review, we summarize the recent advances in elucidating the regulatory mechanisms of ELF3 and ELF4 in plants and discuss directions for future research on ELF3 and ELF4.

Neuroimaging-based brain-age prediction of first-episode schizophrenia and the alteration of brain age after early medication.

BACKGROUND: Neuroimaging- and machine-learning-based brain-age prediction of schizophrenia is well established. However, the diagnostic significance and the effect of early medication on first-episode schizophrenia remains unclear. AIMS: To explore whether predicted brain age can be used as a biomarker for schizophrenia diagnosis, and the relationship between clinical characteristics and brain-predicted age difference (PAD), and the effects of early medication on predicted brain age. METHOD: The predicted model was built on 523 diffusion tensor imaging magnetic resonance imaging scans from healthy controls. First, the brain-PAD of 60 patients with first-episode schizophrenia, 60 healthy controls and 21 follow-up patients from the principal data-set and 40 pairs of individuals in the replication data-set were calculated. Next, the brain-PAD between groups were compared and the correlations between brain-PAD and clinical measurements were analysed. RESULTS: The patients showed a significant increase in brain-PAD compared with healthy controls. After early medication, the brain-PAD of patients decreased significantly compared with baseline (P < 0.001). The fractional anisotropy value of 31/33 white matter tract features, which related to the brain-PAD scores, had significantly statistical differences before and after measurements (P < 0.05, false discovery rate corrected). Correlation analysis showed that the age gap was negatively associated with the positive score on the Positive and Negative Syndrome Scale in the principal data-set (r = -0.326, P = 0.014). CONCLUSIONS: The brain age of patients with first-episode schizophrenia may be older than their chronological age. Early medication holds promise for improving the patient's brain ageing. Neuroimaging-based brain-age prediction can provide novel insights into the understanding of schizophrenia.

Arabidopsis NUCLEAR FACTOR Y A8 inhibits the juvenile-to-adult transition by activating transcription of MIR156s.

Vegetative (juvenile-to-adult) and flowering (vegetative-to-reproductive) phase changes are crucial in the life cycle of higher plants. MicroRNA156 (miR156) and its target SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) genes are master regulators that determine vegetative phase changes. The miR156 level gradually declines as a plant ages and its expression is rapidly repressed by sugar. However, the underlying regulatory mechanism of transcriptional regulation of the MIR156 gene remains largely unknown. In this study, we demonstrated that Arabidopsis NUCLEAR FACTOR Y A8 (NF-YA8) binds directly to CCAAT cis-elements in the promoters of multiple MIR156 genes, thus activating their transcription and inhibiting the juvenile-to-adult transition. NF-YA8 was highly expressed in juvenile-stage leaves, and significantly repressed with developmental age and by sugar signals. Our results suggest that NF-YA8 acts as a signaling hub, integrating internal developmental age and sugar signals to regulate the transcription of MIR156s, thus affecting the juvenile-to-adult and flowering transitions.

Arabidopsis ELF4-like proteins EFL1 and EFL3 influence flowering time.

The circadian clock synchronizes internal and external stimuli to ensure numerous biological processes occur at the optimal time. EARLY FLOWERING 4 (ELF4) is a key evening-phased component of the circadian clock and essential for photoperiod-dependent flowering time regulation in Arabidopsis thaliana. There are four homologous ELF4-like (EFL1-EFL4) genes in the Arabidopsis genome but their functions are unknown. Protein sequence alignment and phylogenetic analysis showed that these four EFL proteins contained an evolutionarily conserved domain, DUF1313, of unknown function. To investigate the physical roles of these genes in Arabidopsis, we overexpressed the four homologous EFL genes in the elf4 mutant background. Under both long-day (LD) and short-day (SD) conditions, overexpression of EFL1 not only completely rescued the early flowering phenotype of the elf4 mutant, but also delayed flowering. Overexpression of EFL2, however, failed to rescue this phenotype and overexpression of EFL3 partially rescued the early flowering phenotype. The transcription levels of the key flowering time regulation genes CONSTANS (CO) and FLOWERING LOCUS T (FT) were significantly decreased in the EFL1- and EFL3-overexpressing transgenic lines in a dose-dependent manner, compared with the elf4 mutant. These results suggest that EFL1 and EFL3 are involved in flowering time regulation in Arabidopsis.