Eman Alefishat

Previous studies on biocatalytic transformations of pinenes by cytochrome P450 (CYP) enzymes reveal the formation of different oxygenated products from a single substrate due to the multistate reactivity of CYP and the many reactive sites in the pinene scaffold. Up until now, the detailed mechanism of these biocatalytic transformations of pinenes have not been reported. Hereby, we report a systematic theoretical study of the plausible hydrogen abstraction and hydroxylation reactions of α- and β-pinenes by CYP using the density functional theory (DFT) method. All DFT calculations in this study were based on B3LYP/LAN computational methodology using the Gaussian09 software. We used the B3LYP functional with corrections for dispersive forces, BSSE, and anharmonicity to study the mechanism and thermodynamic properties of these reactions using a bare model (without CYP) and a pinene-CYP model. According to the potential energy surface and Boltzmann distribution for radical conformers, the major reaction products of CYP-catalyzed hydrogen abstraction from β-pinene are the doublet trans (53.4%) and doublet cis (46.1%) radical conformer at delta site. The formation of doublet cis/trans hydroxylated products released a total Gibbs free energy of about 48 kcal/mol. As for alpha pinene, the most stable radicals were trans-doublet (86.4%) and cis-doublet (13.6%) at epsilon sites, and their hydroxylation products released a total of ~50 kcal/mol Gibbs free energy. Our results highlight the likely C-H abstraction and oxygen rebounding sites accounting for the multi-state of CYP (doublet, quartet, and sextet spin states) and the formation of different conformers due to the presence of cis/trans allylic hydrogen in α-pinene and β-pinene molecules.

<b>Objectives:</b> This study was designed to assess the awareness of women of the risk factors and preventive measures of osteoporosis.<br /> <b>Methods:</b> This is a cross-sectional study that was conducted in Jordan. During the study period, a convenience sample of women were invited to participate via a survey link communicated through social media. Linear regression was used to screen independent variables affecting participants’ awareness of osteoporosis, its risk factors, and preventive measures.<br /> <b>Results:</b> 426 women agreed to participate in this study, with a median age of 38.5 years (IQR=16.0). Results showed that women have a median awareness score of 5.0 out of 9.0 (IQR=4.0) about osteoporosis risk factors, whereas women with higher educational levels (diploma or higher) and those with medical-related degrees have a higher awareness about osteoporosis risk factors compared to others (p≤0.05).<br /> Medical websites were the primary source used to obtain information about osteoporosis by the participating women (n=236, 55.4%). Also, results showed that more than 90% of the women believed that osteoporosis is a chronic silent disease that makes your bones less dense and more fragile (n=418, 98.1%), and that bone strength may increase by taking vitamin D and calcium (n=405, 95.0%).<br /> <b>Conclusion:</b> This study showed that women had an intermediate level of awareness of osteoporosis and prevention measures. The importance of medical websites and social media as sources of information was also demonstrated. More public targeted campaigns are needed to promote awareness of osteoporosis, its risk factors, prevention, and treatment.