The 16α-Hydroxylation of Progesterone by Cytochrome P450 107X1 from Streptomyces avermitilis.

Cytochrome P450 enzymes (CYPs or P450s) are ubiquitous heme-dependent enzymes that catalyze the monooxygenation of non-activated C-H bonds to modify the structure of the substrate. In this study, we heterologously expressed CYP107X1 from Streptomyces avermitilis and conducted in vitro substrate screening using the alternative redox partners putidaredoxin and putidaredoxin reductase. CYP107X1 catalyzed the 16α-hydroxylation of progesterone with regio- and stereoselectivity. The spectroscopic analyses showed that CYP107X1 bound progesterone with a relatively high Kd value of 65.3±38.9 µM. The Km and kcat values for progesterone were estimated to be 47.7±12.0 µM and 0.30 min-1 , respectively. Furthermore, a crystal structure was obtained of CYP107X1 bound with glycerol from the buffer solution. Interestingly, a conserved threonine was replaced with asparagine in CYP107X1, indicating that it may adopt an unnatural proton transfer process and play a crucial role in its catalytic activity.

Effects of a 12-hour neuromuscular electrical stimulation treatment program on the recovery of upper extremity function in sub-acute stroke patients: a randomized controlled pilot trial.

[Purpose] This study investigated the effects of a 12-hour neuromuscular electrical stimulation program in the evening hours on upper extremity function in sub-acute stroke patients. [Subjects and Methods] Forty-five subjects were randomized to one of three groups: 12-hour neuromuscular electrical stimulation group (n=15), which received 12 hours of neuromuscular electrical stimulation and conventional rehabilitation for the affected upper extremity; neuromuscular electrical stimulation group (n=15), which received 30 min of neuromuscular electrical stimulation and conventional rehabilitation; and control group (n=15), which received conventional rehabilitation only. The Fugl-Meyer assessment, Action Research Arm Test, and modified Ashworth scale were used to evaluate the effects before and after intervention, and 4 weeks later. [Results] The improvement in the distal (wrist-hand) components of the Fugl-Meyer assessment and Action Research Arm Test in the 12-hour neuromuscular electrical stimulation group was more significant than that in the neuromuscular electrical stimulation group. No significant difference was found between the two groups in the proximal component (shoulder-elbow) of the Fugl-Meyer assessment. [Conclusion] The 12-hour neuromuscular electrical stimulation group achieved better improvement in upper extremity motor function, especially in the wrist-hand function. This alternative therapeutic approach is easily applicable and can be used in stroke patients during rest or sleep.

Diethyl 4,4'-(3,6-dioxaoctane-1,8-diyl-dioxy)dibenzoate.

The title compound, C(24)H(30)O(8), was obtained by reaction of ethyl 4-hy-droxy-benzoate with 1,2-dichloro-ethane. The mol-ecule occupies a crystallographic inversion center, with its central ethyl-ene bridge in an anti conformation. The other ethyl-ene bridge has a gauche conformation, with the corresponding O-C-C-O torsion angle being 74.2 (1)°. The benzene rings are almost coplanar with the adjacent eth-oxy-carbonyl groups, with an r.m.s. deviation of 0.078 Å.

miR-744-5p inhibits cellular proliferation and invasion via targeting ARF1 in epithelial ovarian cancer.

miR-744-5p has been demonstrated to play an important role in cancer progression. However, the functions of miR-744-5p in epithelial ovarian cancer (EOC) are not well clarified. In this study, our aim was to investigate the role of miR-744-5p and its underlying molecular mechanism in cell progression of EOC. EOC clinical tissues and matched adjacent ovarian epithelial tissues were collected from 18 patients. Tissues and cell lines were analyzed by qPCR or Western blot to investigate the expression of miR-744-5p and ARF1 in EOC. Cell proliferative capacity was assessed by CCK8 and colony formation assays. Wound healing and transwell assays were performed to evaluate cell migration and invasion. The potential binding relation between miR-744-5p and IRF1 was demonstrated by dual luciferase report assay. The results showed that expression of miR-744-5p was low in EOC clinical tissues and cells. Overexpression of miR-744-5p inhibited proliferation, migration, and invasion of EOC cells. Further mechanistic study identified that ARF1 is a target of miR-744-5p, which is negatively correlated with the expression of miR-744-5p, and overexpression of ARF1 could reverse the inhibition of miR-744-5p on the proliferation, migration, and invasion of EOC cells. Taken together, our results indicated that miR-744-5p attenuated EOC progression via targeting IRF1, providing potential guidance for the clinical treatment of ovarian cancer.

[Estimation of winter wheat leaf nitrogen accumulation based on coupling ground- and space-remotely sensed information].

By coupling the SPOT-5 multi-spectral RS images, ground-spectrum, and field measured data of different winter wheat ecological zones, a pure pixel spectrum extraction method was developed based on spectral response function and pixel unmixed, and the quantitative relationships between leaf nitrogen accumulation (LNA) and simulated, measured, and pure pixel spectra were analyzed. The estimation accuracy for LNA was in the sequence of simulated pixel spectra > pure pixel spectra > measured pixel spectra. However, the LNA monitoring model based on simulated pixel spectra couldn't be extrapolated directly to spatial level. The results of model verification also indicated that the monitoring model based on pure pixel spectra performed well in two different wheat ecological zones. Therefore, the pure pixel spectrum extraction method could be applied to other varied and remotely sensed data with different spatial and spectral resolutions by making use of the merits of ground- and space- remote sensing simultaneously, which provided a technological basis for estimating winter wheat nitrogen status in regional scale.