dataesr/scientific-paragraphs-categorization huggingface dataset:
This may suggest that the substrate binding pocket of the two P450 enzymes could accommodate more than one C 12 molecules and that the binding of one C 12 molecule could facilitate the binding of the other to the same P450 enzyme, particularly at low to medium substrate concentrations (10-80 μM). Notably, the kinetic data of CYP-Sm46Δ29 towards C 14 substrate exhibited apparent and substantial substrate inhibition (Additional file 1: Figure  S9C ). When the substrate concentration was in excess of 60 μM, the rate of 1-tridecene formation declined dramatically, but was not completely abolished even at a high concentration of 300 μM. Rather, it flattened off to a low activity, indicating an incomplete inhibition. Estimation of the kinetic parameters for this substrate was therefore deduced from the truncated dataset using Michaelis-Menten equation after removal of the inhibited rates at high substrate concentrations  [48].
Discussion In this study, we successfully expressed, purified, and biochemically characterized two new CYP152 peroxygenase family members, CYP-Aa162 and CYP-Sm46Δ29, that are capable of decarboxylating straight-chain saturated fatty acids to yield 1-alkenes to different extents. Previously, Chen et al. [49]  compared the in vivo alkene production by expressing a His-tagged CYP-Sm46 (there called OleT SM , GenBank Accession Number: WP_009381667, an identical sequence to EKU50422, 453 amino acids) in S. cerevisiae, and only observed trace amounts of oddnumbered carbon chain alkenes (C 13 , C 15 , and C 17 ) from a 48-h culture of the recombinant yeast cells (total alkene titer 1. 4 μg L -1 , 40 times lower than that of OleT JE 54. 5 μg L -1 ). In that study, a 'full-length' version of CYP-Sm46 was used.
Furthermore, analysis of mRNA sequences of cytochrome P450 family proteins from other Staphylococcus species such as S. agnetis (GenBank: JPRT01000001. 1) and S. pseudintermedius (GenBank: CP002439. 1) also revealed a similar pattern of the strong 5-6 G-C pairs of Shine-Dalgarno complementarity preceding a GUG start codon. Taken together, we reason that CYP-Sm46Δ29 is most likely to be the native functional form of the protein. P420 species was generally thought to be an inactive form of cytochrome P450 enzymes  [52] [53] [54]. Formation of this 'inactive' P420 was suggested to involve the protonation of the native cysteine thiolate to a neutral thiol heme ligand  [44] , the recruitment of a histidine to replace the native cysteinate ligand  [45] , or other unknown reasons. Either way leads to the failure of forming the thiolate-ligated Fe 2+ species or to a distorted/ weakened heme-thiolate bond, thus underlying the P450 → P420 transition  [45].
Molecular cloning The gene sequences encoding CYP-Aa162 from Alicyclobacillus acidocaldarius LAA1 (CYP152A8, GenBank Accession Number: WP_008340313) and CYP-Sm46 from Staphylococcus massiliensis strain S46 (CYP152L2, Gen-Bank Accession Number: EKU50422) were, respectively, codon-optimized and synthesized by Genewiz (Suzhou, China), and cloned into the expression vector pET28b between NdeI/XhoI restriction sites with a hexahistidine tag at the N terminus for expression in E. coli BL21 (DE3). The truncated CYP-Sm46Δ29 gene (WP_039990689) was PCR-amplified from the initial construct of pET28b-CYP-Sm46 using Pfu DNA polymerase and then subcloned into pET28b vector using the same restriction sites as above. The generation of double mutant gene constructs of Aa162/Q85H/V170I and P450 BSβ /Q85H/V170I was achieved by site-directed mutagenesis via overlap extension PCR  [64].
For GC-MS, the gas chromatography equipment was coupled to an Agilent 5975C MSD single quadrupole mass spectrometer operated under electron ionization mode at 70 eV in the scan range of 50-500 m/z. For detection of the hydroxyl fatty acid products, as only authentic standards of α-and β-hydroxy myristic acids were obtained, samples extracted from the C 14 myristic acid reactions were derivatized with an equal volume of N,Obis(trimethylsilyl)trifluoroacetamide (BSTFA) with 1% trimethylchlorosilane at 72 °C for 15 min prior to GC-MS analysis. The GC-MS analysis followed the protocol developed by Rude et al. [13] , except for using the Agilent J&W DB-5MS column (i. d. 0. 25 μm film thickness, 50 m by 0. 25 mm). Peak identity was determined by comparison of retention time and fragmentation pattern with the authentic standard compounds where available and to the National Institute of Standards and Technology, USA mass spectral database.
Acknowledgements We thank  Prof. Zhiqi Cong  at  Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences  for his valuable discussion and suggestions. We are also grateful to  Dr. Yousong Ding  at  University of Florida  and  Prof. Thomas Pochapsky  at  Rutgers University  for their kind gifts of the plasmids for expression of  P450 BM3  and CamAB, respectively.
Additional file 1 : 1 Figure S1. Protein sequence alignment of CYP-Aa162 from A. acidocaldarius (GenBank Accession Number: WP_008340313), P450 BSβ from Bacillus subtilis str. 168 (GenBank Accession Number: NP_388092), OleT JE from Jeotgalicoccus sp. ATCC 8456 (GenBank Accession Number: ADW41779), CYP-Sm46 (labelled as Sm46 extended) from Staphylococcus massiliensis S46 (GenBank Accession Number: EKU50422), CYP-Sm46Δ29 (labelled as Sm46Δ29) from S. massiliensis (GenBank Accession Number: WP_039990689) and cytochrome P450 enzymes from other Staphylococcus species such as S. agnetis (GenBank Accession Number: KFE42911), S. delphini (GenBank Accession Number: WP_019165531), S. intermedius (GenBank Accession Number: WP_019167377) and S. pseudintermedius HKU10-03 (GenBank Accession Number: ADV05454).
All chemical shifts are quoted in parts per million (ppm) relative to the residual solvent as the internal standard. Data are reported as: multiplicity (ap = apparent, br = broad, s = singlet, d = doublet, t = triplet, q = quartet, qn = quintet, sx = sextet, sp = septet, oct = octet, non = nonet, m = multiplet), integration and coupling constant(s) in Hz. Melting points were obtained on a Büchi Melting Point B-450 apparatus, and high resolution mass spectra were obtained on an Agilent Technologies LCMS (ESI) or (EI) system.
One can therefore suppose that abusers will chose the substance which better fits to modify their behaviour. Overall, present data suggest that mephedrone differs not only from MDMA but also from other cathinones in its pharmacological profile and behavioural and neurotoxic effects  [15]. Although un-frequent, mephedrone was identified in routine forensic casework. As here reported, during the last 2 years, mephedrone was detected in 4 cases, including a fatal intoxication, an alleged drug-facilitated sexual assault, a traffic accident and a fatal drowning and hair was collected to document past exposure to drugs. Table  2  contains the hair results in these 4 expertises. All hair specimens contained mephedrone and out of the 4 samples analysed, only 1 sample did not contain ecstasy. Police investigations confirmed in all cases that the subjects were nightclub and techno festival attendees.
Acknowledgements This work was financially supported by  RAS  within the projects: a) " Valorizzazione di biomasse d'interesse regionale attraverso processi chimici a basso impatto ambientale" (CRP 72-Bando "Capitale Umano ad Alta Qualificazione. Annualità 2015_L. R. 7 agosto 2007, n°7"), b) " Smart Nanostructured Functional Materials: Synthesis and Characterization with Focus on the Specific Interactions between Solid Surfaces and Biomacromolecules " ( RICALTRO_CTC_2017_RAS_MONDUZZI ) and by  Fondazione Banco di Sardegna  with the projects:  RICALTRO_CTC_2017_FBS_MONDUZZI. The " Teach Mob -Teaching Staff Mobility Programme 2016-2017 " ( University of Turin ) is warmly acknowledged by EC and GC. E. C. is grateful to the  Università degli Studi di Cagliari (Italy)  ( Visiting Professor Program   2016-2017 ) for the grant.
, 2004; Dancik et al. , 2013). Although only three chemicals were tested in this comparison, we considered this number sufficient to make a conclusion on which assay to use for further testing, since in addition to data comparisons, multiple practical aspects were evaluated, as mentioned above. Pig skin has been used as a surrogate for human skin due to their structural, physiological and biochemical similarities  (Simon and Maibach, 2000; Herkenne et al. , 2006; Barbero and Frasch, 2009; Jung and Maibach, 2015)  and is accepted by the Scientific Committee on Consumer Safety (SCCS) for use in skin penetration studies  (SCCS, 2010). Therefore, we investigated whether pig skin (obtained as waste from the food industry) could be used as an alternative source of SC for K SC/v and D SC measurements if the human skin was not available in sufficient quantities.
To reduce uncertainty on D SC , the tape stripping was carried out as quickly as possible. Criteria defined by  Reddy et al. (2002)  for tape stripping time were adhered to. A total of 15 strips were removed and weighed before extraction of the chemical using methanol. Analytical method. The extract solution was directly injected onto an LC/MS-MS system (Shimadzu Nexera LC system with a CTC PAL Autosampler coupled with a mass spectrometer API 3200 (ABSciex, Concord, Ontario, Canada). The analytical system was managed by the software Analyst version 1. 6. The analytical column used was a Kinetex C18 from Phenomenex (Torrance, CA, USA) (50 x 3. 0 mm, dp. 2. 6 μm) and analysis are carried out with gradient elutions with mobile phases of 20 mM of ammonium acetate (A) and acetonitrile (B). The column temperature was fixed at 50°C, the volume of the injection was 10 μl and the flow rate at 0.
D SC ¼ k p *H SC =K SC=v (9) For the third approach, which was used for the reported values from Protocol 2B in Table  2 , Fickˈs 2nd law was used to determine K SC/v and D SC. Specifically, K SC/v *H SC and D SC /H SC 2 were calculated using data from Protocol 2B from the permeation of the chemical through the isolated SC by non-linear regression of the cumulative amounts absorbed per time (Q) (using software JMP Pro 10 (SAS Institute)), according to Eqn (10). These values are reported in Table  2. (Protocol 2B). Q t ð Þ ¼ K SC H SC C v D sc H 2 SC t À 1 6 À 2 π 2 ∑ ∞ n¼1 À1 ð Þ n n 2 e À Dsc H 2 SC n 2 π 2 t ( 10 ) The non-linear regression was performed against the cumulative penetration profile throughout the portion of the plot which showed good linearity and there was good mass balance (90-110%).
• Measurement considerations: For Protocol 1 only, the exposure times may need adjusting according to the chemical. There is some uncertainty of the SC thickness determined by weighing individual tape strips in Protocol 1. In Protocol 2A and B, the thickness of the SC and other layers is afforded by a direct measurement. Unlike Protocol 2B, in Protocol 1, the skin layers require no mounting or structural support. In Protocol 2A for K SC/v , the SC is accurately weighed by removing any added water by drying. This gives a more accurate measurement of the tissue layer mass. • Data handling: For Protocol 1, manual adjustment of curve fitting is sometimes needed. For Protocol 2A, a correction to partially hydrated SC is afforded by equation of  Nitsche et al. (2006). This correction for hydration is not necessary for other tissue layers.
The use of isolated skin layers, though, comes with notable advantages, such as low variability in the data generated for K SC/v and the possibility to apply the same procedures to the other isolated skin layers (unlike Protocol 1). Moreover, two different values are obtained for K SC/v based on different models and assumptions. A comparison of measured parameters from pig and human skin using Protocol 1 support the findings of  Gerstel et al. (2016)  such that the distribution of chemicals was similar in pig and human skin. Indeed, there were only small differences observed in the values generated for the two species, especially for caffeine and resorcinol. A greater difference was observed for 7-EC, which showed some deviation from the theoretical profile. Interestingly, the SC thickness measured on full thickness pig skin using the TEWL protocol was not significantly different from the thickness of human SC.
At the same time, new significantly blue shifted bands appeared at 230e350 nm that is the charac teristic absorption region of non conjugated aromatic and hetero aromatic fragments (Fig. 1 ; Figs. S5eS7, ESI ). To explain this observation, we assumed that dimeric complexes L 2 $Ba 2þ undergo the [2þ2] photocycloaddition reaction of the ethylene double bonds resulting in the formation of cyclobutane derivatives 1be4b, in which conjugation between aromatic rings is disrupted by the cyclobutane ring closure. (Scheme 1). Kinetic studies on the photochemical reactions of the complexes of 1a and 3a with Ba(ClO 4 ) 2 were carried out upon irradiation at 313 nm or 365 nm depending on the position of the absorption maxima of the cor responding species. Analysis of the kinetic data allowed us to determine the values of the quantum yields of the forward and reverse photoisomerisation and photocycloaddition reactions (Table  2 ).
2 ). In all cases, the main photochemical process was the formation of the only one cyclobutane derivative of 1be4b, as revealed by the presence of two doublets in a characteristic region of the cyclo butane ring protons (4. 5e5. 5 ppm), and the only side process was the E Z isomerization of the initial compounds. Notably, irradiation (l 365 nm) of compounds 1ae4a in the absence of barium species resulted in the E Z isomerization only, and no products of photo cycloaddition were detected that is in agreement with the steady state optical spectroscopy data (vide supra). The processes occurring upon addition of barium ions and following photoirradiation were analyzed by NMR in more details for 4 quinoline derivative 2a. The addition of 0. 5 eq. of Ba 2þ to a solution of 2а in CD 3 CN caused two distinctive sets of signals to appear in the 1 Н NMR spectrum (Fig.
After irradiation, the tube was transferred into the NMR spectrometer. To monitor the pathways of the photoreactions, the spectra were recorded at regular time intervals. The concentration of each photoproduct was obtained from the ratio of peak intensities. Absorption spectra were recorded on a Varian Cary 100 double beam spectrophotometer. Preparation and handling of the solu tions were carried out under red light. Photochemical reactions were carried out with a high pressure Hg vapor lamp (120 W) and an immersed Hg photoreactor (125 W). Kinetic studies on the phototransformations of the complexes were performed using a diode array fiber optic spectrometer AvaSpec 2048. Complex for mation of 1ae4a with Mg(ClO 4 ) 2 and Ba(ClO 4 ) 2 was studied by a spectrophotometric titration method in acetonitrile at 20±1 C; the stability constants of the complexes were determined from the photometric data using the SpecFit 32 program.
Acknowledgements This paper was financially supported by the  Ministry of Education and Science of the Russian Federation  on the program to improve competitiveness of  People's Friendship University (RUDN University ) among the world's leading research and education centers in the 2016e2020 and  RFBR 15 03 04695  (optical studies). The NMR measurements were performed with the support of the grant of the  President of Russian Federation  (project  MK   6320. 2016. 3 ).
The well-diffusion method was used to determine the antibacterial efficacy of the T-CE and M + T-CE-mediated AgNPs. Briey, the AgNPs (15 mg mL À1 ) were dissolved in autoclaved water and used immediately. Inoculums were prepared by growing bacterial colonies overnight in nutrient broth. Bacterial lawns were prepared on nutrient agar plates using cotton swabs soaked with each broth culture. The wells made in the agar plates were loaded with 1 mM SNS (positive control), 15 mL of the synthesized AgNPs (15 mg mL À1 ), T-CE and M + T-CE (negative controls) and a commercial antibiotic ampicillin solution (10 mg) (as a standard). The plates were incubated at 37 C for 24 h and the zone of inhibition was measured in mm. The data of each strain were collected from three independent experiments which were repeated thrice.
Regulations usually distinguish between prescription-only (POM) and over-thecounter (OTC) medicines. The former requires medical prescription; the latter are available for SM of common minor or easily treated ailments. However, in the Eastern Mediterranean countries, theoretical prescription medicines can easily be purchased without a prescription, as self-medication (SM) resulting in potential misuse and unnecessary risk for patients. The magnitude of this activity is uncertain. The aim of this article, therefore, is to undertake a comprehensive review to identify the different types of medicines that can easily be purchased as SM in Middle East and recognized as misused. An extensive review of the published literature  was conducted using Pubmed, web of science, Cochrane, and Google Scholar databases, for OTC medicine misuse in the Middle East.
Search strategy Databases, namely, Medline/Pubmed, Web of Science, Cochrane Library, and other sources, were used to identify peer-reviewed papers dealing with the review theme in WHO Eastern Mediterranean countries. Search terms were identified through a pilot review of the literature and were used to identify articles through a systematic, standardized process. The words/strings used for search and inclusion criteria were as follows: using combinations of the following terms: "over the counter", "OTC", "non-prescription", "self prescription medicines", "prescription medicines", "misuse", "abuse", "addiction", "dependency," and "nonmedical use", "irrational use", "inappropriate use", Arab and name of countries belonging to the WHO Eastern Mediterranean Region (EMRO). The search strategy is outlined in figure  1.
Literature search results The database search yielded 696 publications for review. Fifty other records were added through other sources like health journals edited in Iran, Pakistan, and Saudi Arabia and previous reviews for reference mining but they were not included. 562 articles were excluded for reasons of duplicated records, review articles, irrelevant topic, time of publication, availability of abstract, outside eastern Mediterranean area, and clinical and/or randomized controlled trial. A secondary search was performed to elaborate the primary concept. A total of 134 full-text articles were assessed of which 49 were excluded as they were related to prescription medications or illicit and alcohol abuse or medication storage and wastage, which did not meet our study objective. Another 21 articles (four out of time frame, two opinion pieces, four letters, five reviews, five articles outside Middle East area, and one animal model) were excluded.
Lack of access to some of the databases in the field and to unpublished research reports was the main limitations in our attempt to shed light on OTC misuse behavior. Uninformative titles and abstracts are also a limitation. However, the main limitation in our analysis is the heterogeneous methodological nature and reporting of the studies. Even finding more studies hidden in obscure repositories would probably not have changed that conclusion. In addition, most studies were consistent and showed similar results on the widespread use of self-medication, so that again missing studies would not change these results.
Experimental Selected experimental procedures and data are presented hereafter. Full details can be found in the ESI. † sec-Butyllithium (1. 3-1. 4 M solution in cyclohexane) was purchased from Sigma-Aldrich or Alfa Aesar and titrated according to a literature method. 21 Diethyl ether, and dichloromethane were purified using a MB SPS-800 solvent purification system (MBRAUN). Other solvents and commercial reagents were used as received, without purification. Petroleum ether refers to the 40-60 °C fraction. The microwave-promoted experiments were run using a CEM Discover Microwave Synthesis System with the temperature and time parameters indicated; the reaction vessels were not flushed with an inert gas. All other reactions were carried out under nitrogen or argon. The temperatures mentioned are the temperatures of the cold baths or the oil baths used.
Acknowledgements We wish to thank  Vincent Jactel  and  Michel Levart  for MS analyses and the following institutions for funding:  École Polytechnique  for grants attributed to P. K. , G. S. and A. B. ; the  French Embassy in Russia  for a 3-month Metchnikov fellowship awarded to O. A. T. and the  Agence Nationale de la Recherche (ANR)  for the grants of C. C. and  R. K. ( ANR-12-BS07-0013  "ACTIMAC"). Finally, Y. S. wishes to thank  Prof. S. Z. Zard  for his continuous encouragement and invaluable support.
Search Strategy A systematic search was conducted in December 2015, by one of the authors in the following databases: Medline [21], SCOPUS  [22] , ISI Web of Knowledge  [23] , ClinicalTrials. gov [24]  and International Clinical Trials Registry Platform  [25]. Search terms in titles and abstracts included a combination with MESH terms: (1) bioactive designation and food rich in flavonol (polyphenol, flavonoid, flavonol, quercetin, kaempferol, galangin, isorhamnetin, jaceidin, kaempferide, morin, myricetin, patuletin, rhamnetin, spinacetin, rutin; spice, caper, saffron, caraway, clove, oregano, onion, shallot, broccoli, spinach, asparagus, bean, chilli pepper, berry, black chokeberry, american cranberry, and lingonberry); (2) type of study and participant (trial, experiment, study, studies, intervention; human, subjects, men, women, patient, volunteer, and participant); and (3) cardiometabolic outcome (flow mediated dilation (FMD), platelet aggregation, blood pressure (BP), total cholesterol (TC), LDL cholesterol, HDL cholesterol, triacylglycerol (TAG), body mass index (BMI), waist circumference (WC), glucose, insulin, insulin resistance (HOMA-IR), glycated hemoglobin (HbA1c) and exercise capacity).
Data Extraction Using a standardized data extraction form, the following data were extracted: (1) publication details-year of publication, name of first author, name and e-mail of corresponding author, and clinical trial registration number (when available); (2) sample characteristics-country, total number of participants, male and female, age mean and age range, ethnicity, health status (healthy, at risk or diagnosed disease), menopausal status, smoking, medication; baseline BMI, diet (assessment method, baseline diet and diet during study), and physical activity level; (3) study characteristics-study design (cross-over or parallel), arms number and description, washout duration, treatment duration, number of participants receiving the test bioactive or food, number of participants receiving the placebo, number of participants completing the study, composition of test and placebo, and dose and mode of administration; (4) information on reported outcomes-type of sample (fasted <8 h, 8-12 h, <12 h, and fed), equipment used (when important to outcome assessment), central measure (mean or median) and dispersion (standard deviation or standard error of the mean) with corresponding units (values before and after test, before and after placebo, or values for test and placebo changes), p value when available, and number of participants with adverse events.