diff --git "a/README.md" "b/README.md" --- "a/README.md" +++ "b/README.md" @@ -89,267 +89,201 @@ a corpus of pre-annotated text for other tasks (e.g. figure caption to graphic, ### Data Instances ```json { - "accession_id": "PMC8515580", - "pmid": "34661004", + "accession_id": "PMC3222825", + "pmid": "22038649", "introduction": [ "Introduction", - "

Medicinal plants are\nknown to possess several primary and secondary\nmetabolites. The secondary metabolites in particular are involved\nin many biological activities. They are also effective against managing\noxidative stress and hence responsible to fight against various diseases\ncaused by stress. ##REF## , ##REF## The diverse chemical structures\nof such secondary metabolites have significant contributions toward\nnew drug development processes. It has been observed in the past several\ndecades that natural resources provide an enhanced biological activity,\nwhile the demands for the development of similar synthetic lead compound\nare limited. ##UREF## Antioxidants have the capacity\nto scavenge the free radicals, which are generated by metabolism and\nto some extent by various diseases. ##REF## The\nendogenous antioxidant system itself may be able to fight against\nthe deleterious effects produced by oxidative stress. This includes\nthe enzymes such as catalase, superoxide dismutase, glutathione,and\nso on that can scavenge free radicals and protect biological systems\nto some extent. The endogenous antioxidant mechanism reinforcement\nor external antioxidant supplements may help overcome the situation. ##REF##

", - "

Genus Clerodendrum belonging to\nfamily Verbenaceae or Lamiaceae has more than 500 species. This genus\nis distributed in herbs and small trees. Clerodendrum species are reported to possess anti-inflammatory, anti-diabetic,\nanti-cancer, anti-malarial properties, and so on. They comprise various\nclasses of constituents like flavonoids, phenolics, terpenes, steroids,\nvolatile constituents, and so on. ##UREF## Natural sources are more widely promoted and recommended\nfor they display minimal side effects, as compared to synthetic agents. ##UREF## Drug discovery from natural resources is highly\nchallenging as the processes like authentication, screening, isolation,\nstructural elucidation, and so on. require expertise\nand well-experienced support. On the other hand, there has always\nbeen a huge demand on these natural sources for their excellent safety\nparameters. ##UREF## Clerodendrum\npaniculatum (C. paniculatum) leaves, roots, and so on. are reported for their\nanti-oxidant, anti-inflammatory, hepatoprotective, and anti-diabetic\nactivities and so on. However, the flower counterpart of this plant\nhas not been investigated for any of the medicinal activities including\ntheir hepatoprotective roles in animal models. Herein, we have evaluated\nthe hepatoprotective role of the alcoholic extract of the flower part\nof C. paniculatum against CCl4-induced hepatotoxicity in female Wistar Albino rats and justified\nits pharmacological implications toward its use as traditional medicine.

" + "

RNA is increasingly viewed not only as an intermediary between DNA and protein, but also as a key regulator of gene expression and catalysis (Boisvert et al. ##REF## ; Korostelev et al. ##REF## ; Steitz ##REF## ; Breaker ##REF## ; Wahl et al. ##REF## ; Newman and Nagai ##REF## ). Methods to obtain very high-resolution structures and site specific dynamics of RNA molecules are therefore critical. Nuclear magnetic resonance (NMR) spectroscopy has emerged as an effective method for solving RNA 3D structures and for probing their motions. However, RNA molecules that are bigger than 40 nucleotides are difficult to characterize using NMR spectroscopy because of severe chemical shift overlap and rapid signal decay. Selective labeling techniques, first introduced to extend the utility of NMR spectroscopy for protein analysis, are increasingly being applied to RNA as well (Dayie ##REF## ; Lu et al. ##REF## ).

", + "

Of the three primary methods for incorporating selective isotopic labels into nucleic acids, biomass production using E. coli variants (Johnson et al. ##REF## ; Johnson and Hoogstraten ##REF## ; Dayie and Thakur ##REF## ; Thakur et al. ##REF## , ##REF## ) appears to be more general and cost-effective than chemical synthesis (Milecki ##UREF## ) or de novo biosynthesis (Schultheisz et al. ##REF## ; Schultheisz et al. ##REF## ). Selective introduction of 13C isotopes into the ribose ring using chemical synthesis entails difficult regio- and stereo-selective synthesis with low yields (Milecki ##UREF## ). The de novo biosynthetic method requires a large number of enzymes of which only a few are commercially available (Schultheisz et al. ##REF## ; Schultheisz et al. ##REF## ). Additionally, the de novo method requires prohibitively expensive precursors that are limited in the labeling pattern that can be achieved with the biomass methods.

", + "

Unfortunately a biomass method based on E. coli metabolism, until recently, suffered from the disadvantage of isotopic scrambling in both the ribose and nucleobase atoms. This scrambling is caused by the admixture of the metabolic flux from both the oxidative and non-oxidative pentose phosphate pathways (noPPP) and a very efficient TCA cycle (Johnson et al. ##REF## ; Johnson and Hoogstraten ##REF## ). Hoffman and Holland earlier showed that wild-type E. coli grown using selectively labeled acetate enables the incorporation of 13C-labels at either the protonated or non-protonated carbon sites in the RNA ribose and nucleobases. LeMaster and coworkers also showed that a modified bacterial strain, in which the TCA cycle enzymes succinate dehydrogenase and malate dehydrogenase were disabled (DL323), could be grown on selectively labeled glycerol to provide alternative site labeling that removed the 13C-13C coupling in protein NMR spectra (LeMaster and Richards ##REF## ; LeMaster and Kushlan ##UREF## ). Hoogstraten and coworkers then showed that the DL323 strain grown on selectively labeled glycerol also enabled site selective labeling within the ribose ring (Johnson et al. ##REF## ; Johnson and Hoogstraten ##REF## ). Earlier, Pardi and colleagues introduced labeling using 13C-formate in a background of 12C-glucose which site specifically labels purine C8 position exclusively; results were not presented for purine C2 position (Latham et al. ##REF## ). Building on these previous ideas, we showed that an isolated two spin system could be achieved for both base and ribose depending on the strain and complementation of the growth media with formate (Dayie and Thakur ##REF## ; Thakur et al. ##REF## , ##REF## ). Even under these newer conditions, the purine C2 positions were only labeled to ~26%, indicating a general limitation of the formate supplementation methodology (Dayie and Thakur ##REF## ; Thakur et al. ##REF## , ##REF## ).

", + "

However until now, most biomass production of nucleotides had focused on using symmetric carbon sources such as glycerol, glucose, and acetate. For instance if glucose is the carbon source, then at the onset of the TCA cycle this input carbon source is already diluted to 50% by glycolysis and similarly the purine precursor 3-phosphoglycerate (3PG) is only 50% labeled. Similarly if [1, 3-13C]-glycerol is the carbon source, selectivity in the ribose and base regions are again reduced because of the symmetric nature of glycerol such that the purine C5 and all the ribose C1\u2032 have substantial residual coupling that can render relaxation measurements inaccurate (Johnson et al. ##REF## ; Dayie and Thakur ##REF## ). This residual coupling arises from the C-1 carbon of three carbon precursors such as pyruvate and glycerol, and thus [1-13C]-pyruvate will similarly introduce coupling between C1\u2032-C2\u2032-C3\u2032 and C2\u2032-C3\u2032. We, therefore, reasoned that the asymmetry of 13C -labeled pyruvate would provide selective labeling in both the ribose and base moieties of nucleotides that would be impossible with symmetric carbon sources.

", + "

Unlike the previous result with the use of [1, 3-13C]-glycerol (Johnson et al. ##REF## ; Thakur et al. ##REF## , ##REF## ), the use of [3-13C]-pyruvate affords selective 13C enrichment of the ribose C1\u2032 and C5\u2032, of the purine C2 and C8, and of the pyrimidine C5 without any of the residual couplings described above. NMR T1 relaxation experiments and 13C-methylene TROSY or HSQC on a site selectively 13C -labeled 27-nt A-site RNA demonstrate that accurate T1 relaxation parameters and higher resolution spectra can be obtained. Thus, the technology to incorporate site specific labels into both the base and ribose moieties of purine and pyrimidine nucleotides promises to expand the scope of NMR studies to regions hitherto inaccessible.

" ], "methods": [ - "DPPH Method", - "

The most effective and popular method of\nantioxidant evaluation is the DPPH radical scavenging method. Better\nactivity is represented by the discoloration of DPPH solution. From\nstock solution of extracts, different concentrations ranging from\n12.5 to 200 \u03bcg/mL were prepared. To 100 \u03bcL of the extract,\n3.0 mL of DPPH solution was added and incubated at room temperature.\nAfter 20 min of incubation, absorbance was measured at 515 nm against\nmethanol as blank. Absorbance of standard (ascorbic acid) was also\nmeasured. Percentage inhibition was measured by the following equation. ##REF## ##FORMU##

" + "Materials and methods", + "Bacterial strains", + "

The mutant strains DL323 (CGSC # 7538, F-, sdh-1, &lambda-, mdh-2, rph-1) (Hansen and Juni ##REF## ; LeMaster and Kushlan ##UREF## ) and K10-15-16 (CGSC # 4858 Hfr fhuA22, zwf-2, relA1, T2R, pfk-10) (Fraenkel ##REF## ) (referred to here as K10zwf) and the wild-type K12 strain (Clowes and Hayes ##UREF## ; Soupene et al. ##REF## ) (CGSC # 4401:F+) used in this work were obtained from the Yale Coli Genetic Stock Center (CGSC). Dr. Paliy kindly provided the wild-type K12 NCM3722 (Soupene et al. ##REF## ).

", + "Isotopes", + "

Isotopic labeled compounds were purchased from Cambridge Isotope Laboratory (Andover, MA) and Isotec-Sigma-Aldrich (Miamisburg, OH): [3-13C]-pyruvate (99%), 15N-(NH4)2SO4 (99%).

", + "Media for <italic>E</italic>. <italic>coli</italic> growth", + "

Luria\u2013Bertani (LB) media was prepared as described (Sambrook and Russell ##UREF## ) and LeMaster-Richards (LMR) minimal media was prepared as described (LeMaster and Richards ##REF## ; Dayie and Thakur ##REF## ; Thakur et al. ##REF## , ##REF## ). The LMR media contains 176\u00a0mM KH2PO4, 25\u00a0mM NaOH, 10\u00a0\u03bcl H2SO4, 12.6\u00a0mM (NH4)2SO4, 2\u00a0mM MgSO4, 10\u00a0\u03bcM FeSO4 and 0.2% trace metals, supplemented with the [3-13C]-pyruvate and 15N-(NH4)2SO4 enriched nitrogen source.

", + "Method for <italic>E</italic>. <italic>coli</italic> growth optimization", + "

The growth evaluation of the E. coli mutant (DL323) was performed as described previously (Thakur et al. ##REF## , ##REF## ). Briefly, a 5\u00a0ml starter culture in LeMaster media supplemented with unlabeled carbon sources was inoculated from a single colony of DL323 E. coli grown on LB agar plates at 37\u00b0C overnight (~16\u00a0h). The overnight culture was pelleted, cells were washed twice in 1\u00d7 phosphate-buffered saline (PBS), and re-suspended in fresh 5\u00a0ml of LMR medium; 1\u00a0ml of this re-suspension was added to a 50\u00a0ml culture in LMR medium for another overnight growth at 37\u00b0C. The overnight culture was pelleted prior to complete saturation of these cells, the cells were washed twice in 1\u00d7PBS, resuspended in a 50\u00a0ml of LMR medium with no carbon supplements, and then 5\u00a0ml from this resuspension was added to a 500\u00a0ml LMR medium supplemented with [3-13C]-pyruvate and 15N-(NH4)2SO4, and the solution incubated at 37\u00b0C.

", + "P1 nuclease digestion, nucleotide separation and purification", + "

The ribonucleoside monophosphates (rNMPs) were isolated from E. coli (DL323) mutant culture as described earlier (Batey et al. ##REF## ; Thakur et al. ##REF## , ##REF## ). The nucleic acid mixture was digested with P1 nuclease and separated into individual ribo- or deoxy-monophosphates using a cis-diol boronate affinity column chromatography as described (Batey et al. ##REF## ; Thakur et al. ##REF## , ##REF## ). The site specific labeling pattern of rNMPs was verified by NMR prior to re-phosphorylation to rNTPs.

", + "Enzymatic phosphorylation of rNMPs", + "

The detailed procedure for enzymatic phosphorylation of rNMP was described previously (Nyholm et al. ##REF## ; Thakur et al. ##REF## , ##REF## ). Briefly, the individual rAMP, rCMP, rGMP or rUMP and kinases were freshly prepared and pre-incubated at 37\u00b0C for 10\u00a0min prior to mixing. The enzymatic conversion of the labeled rNMPs to rNTP was complete in 5\u00a0h. The progression of the phosphorylation reaction was monitored on a TARGA C18 column using conditions similar to those in the purification step (Supplementary Figure S1). Collected rNTP fractions were pooled and dialyzed overnight and lyophilized.

", + "Site specific enrichment of A-Site RNA and in vitro transcription", + "

A-Site RNA (5\u2032-GGC GUC ACA CCU UCG GGU GAA GUC GCC-3\u2032) was synthesized using in vitro transcription (Milligan et al. ##REF## ; Milligan and Uhlenbeck ##REF## ), with a His-tagged mutant (P266L) T7 RNA polymerase (Guillerez et al. ##REF## ) using a mixture of uniformly labeled guanine and cytosine triphosphates (rGTP and rCTP) (Sigma Aldrich) or site specific labeled rGTP and rCTP. Another set of samples were made using a mixture of uniformly labeled adenosine triphosphates (rATP) (Sigma Aldrich) or site specific labeled rATP. Two terminal 2\u2032-O-methyl modifications in the template strand were introduced to substantially reduce the amount of transcripts with extra nucleotides at the 3\u2032-end (Kao et al. ##REF## ). The optimal transcription conditions were as follows: 10\u00a0mM total NTP and 15\u00a0mM\u00a0Mg2+. The reactions were carried out in a transcription buffer A (40\u00a0mM Tris\u2013HCl, pH 8.1, 1\u00a0mM spermidine, 5\u00a0mM dithiothreitol (DTT), 0.01% Triton X-100, 80\u00a0mg/ml PEG 8000), 300 nM each DNA strand, and 1.5\u00a0\u03bcl of 2.4\u00a0mg/ml T7 RNAP (optimized amount) per 40\u00a0\u03bcl of transcription volume. After three hours of incubation at 310\u00a0K, the RNA from the transcription reactions were purified and dialyzed as described (Dayie ##REF## ). After dialysis, the RNA was lyophilized, and resuspended into NMR buffer (100\u00a0mM KCl, 10\u00a0mM potassium phosphate pH 6.2, 8% or 100% D2O), with or without 3\u00a0mM MgCl2, and a trace of sodium azide). The concentration of each NMR sample in Shigemi tubes in 250\u00a0\u03bcl of buffer was ~0.1\u00a0mM.

", + "NMR experiments", + "

All NMR experiments were run on a four channel Bruker Avance III 600\u00a0MHz spectrometer equipped with actively shielded z-axis gradient triple resonance probe. The experiments were conducted at temperatures ranging from 15\u00b0 to 45\u00b0C. The NMR data sets were processed and the peak positions and intensities were analyzed with Bruker\u2019s TOPSPIN 2.1 as described previously (Dayie and Thakur ##REF## ; Thakur et al. ##REF## , ##REF## ). One dimensional (1D) 13C spectra and two-dimensional non-constant-time (1H, 13C) heteronuclear single quantum correlation (HSQC) spectra (Bodenhausen and Ruben ##UREF## ; Bax et al. ##UREF## ) were acquired to analyze the rNMP fractions extracted from each bacterial strain. The fractional 13C enrichment at each carbon site was quantified directly by 1D proton methods or indirectly using 2-bond (2JHN) HSQC as described previously (Dayie and Thakur ##REF## ; Thakur et al. ##REF## , ##REF## ).

", + "

To demonstrate the usefulness of site selective labeling 2D 1H-13CH2 TROSY (Meissner et al. ##UREF## ; Miclet et al. ##REF## ) spectra were measured using non-constant time evolution in the carbon dimension. Methylene CH2-TROSY experiments (Miclet et al. ##REF## ) were run with the following modifications that dispense with selective decoupling: the WURST-4 decoupling waveform was omitted during the carbon t1 evolution period, and a non-selective 180\u00b0 pulse replaced the selective pulse on the C5\u2032 carbon. To compare the usefulness of the modified CH2-TROSY experiments, the normal CH2-methylene optimized HSQC (Schleucher et al. ##REF## ; Sattler et al. ##UREF## ) were also run under identical conditions: for each data set, 16 scans and 1024 complex points were collected in t2 and 512 complex points were collected in t1 using the Echo-Anti echo method (Palmer et al. ##UREF## ; Kay et al. ##UREF## ) for quadrature detection; proton and carbon carrier was placed at 4.7 and 64\u00a0ppm, respectively, as described (Thakur et al. ##REF## , ##REF## ). The time domain data sets were zero filled in t1 and t2 before Fourier transformation to give a final real matrix size of 2,048\u00a0\u00d7\u00a01,024 points.

", + "

Longitudinal (R1) relaxation rates were measured for ribose C1\u2032 and cytosine nucleobase C5 carbons using TROSY detected experiments (Hansen and Al-Hashimi ##REF## ). To compare the usefulness of the selective labels relative to uniform labels, the R1 relaxation experiments were run under identical conditions: for each data set, 64 scans and 1024 complex points were collected in t2, and 128 complex points were collected in t1 using the Echo-Anti echo method (Palmer et al. ##UREF## ; Kay et al. ##UREF## ) for quadrature detection. Proton and carbon carrier was placed at 4.7 and 93\u00a0ppm respectively. The time domain data sets were zero filled in t1 and t2 before Fourier transformation to give a final real matrix size of 2,048\u00a0\u00d7\u00a01,024 points. For the 2D longitudinal (R1) experiments, relaxation delays of 21.1 (2x), 61.4, 141.1, 301.2, 381.3, 461.3 (2x), 781.6 ms were used. The longitudinal relaxation decay curves were fitted using the Levenberg\u2013Marquardt algorithm by assuming monoexponential decay.

" ], "results": [ "Results", - "

The phytochemical screening of the flower extract indicated the\npresence of carbohydrates, tannins, phenolics, flavonoids, proteins,\nand steroids, and the results are depicted in ##TAB## . The presence of phenolics and flavonoids\ncontents is considered as the main indicators of antioxidant activity\nin herbals.

", - "

The total phenolic content, ##REF## as quantified\nby Folin-Ciocalteu\u2019s method, indicates the highest activity\n(378.5 \u00b1 0.883 mg QE/g) from the alcoholic extract, as compared\nto other extracts. The alcoholic extract of C. paniculatum flower showed the highest flavonoid content and hence high free-radical\nscavenging property (393.23 \u00b1 1.23 mg GAW/g) ( ##TAB## ) compared to other extracts,\nas evaluated by the AlCl3 colorimetric method.

", - "

Antioxidant activity of the extracts was evaluated by four different\nmethods. ##REF## Antioxidants have a high impact\non diseases as they can scavenge the free radicals and thereby modify\nor prevent the diseases. ##REF## DPPH assay carried\nout with the alcoholic extract shows better activity with IC50 of 62.28 \u00b1 0.51 \u03bcg/mL, and the values are presented in ##TAB## and Figure S1 (Supporting Information).

", - "

DPPH is widely used as an evaluation technique due\nto its ease\nof reaction. In the ABTS assay method, radical scavenging ability\nof the extracts was evaluated using a ABTS++ modified solution by\nthe reaction between ABTS and potassium persulfate solution, ##REF## and the IC50 of the extracts was\ncalculated. The alcoholic extract presented the IC50 value\nas 2362.71 \u00b1 9.39 \u03bcg/mL, while the IC50 value\nof the standard ascorbic acid was evaluated as 941.09 \u00b1 8.312\n\u03bcg/mL [ ##TAB## and Figure S2 (Supporting Information)].

", - "

Nitric oxide radical scavenging activity results of\nthe extract\nwere evaluated by comparing with standard gallic acid. Nitric oxide\nis produced by the reaction between aqueous sodium nitroprusside,\noxygen, and nitrite ions, under physiological pH conditions. ##UREF## Antioxidants help scavenge this free radical,\nand this property was compared with the standard. In this method also,\nthe alcoholic extract showed better activity than all other extracts\nwith an IC50 value of 193.09 \u00b1 5.84 \u03bcg/mL, and\nthe results are comparable to the standard (147.11 \u00b1 10.20 \u03bcg/mL);\nthe results are depicted in ##TAB## and Figure S3 (Supporting\nInformation).

", - "

Reducing power ability of the extracts was determined\nusing potassium\nferricyanide. The ability of the extract to reduce potassium ferricyanide\n(Fe III) to potassium ferrocyanide (Fe II) was measured by the reaction\nwith trichloroacetic acid and FeCl3. ##REF## All the extracts exhibited a dose-dependent response. Quercetin\nwas used as the standard, and its response was observed between 1.05\n\u00b1 0.001 and 2.44 \u00b1 0.0005 \u03bcg/mL. A significant increase\nin absorbance indicates a better reducing ability, and the same was\nobserved as the alcoholic extract of C. paniculatum flower ( ##FIG## ).

", - "

An acute oral toxicity\nstudy of the alcoholic extract at a concentration\nof 2000 mg/kg b.w. revealed that the extract is safe up to the abovementioned\nconcentration range. Hepatoprotective investigation was carried out\nwith the CCl4 intoxicated model. The liver enzymes and\nprotein levels were checked for all groups of animals, and the results\nare depicted in ##TAB## and Figure S4 (Supporting Information).

", - "

The enzyme activities associated\nwith liver functions are good\nbiomarkers for the evaluation of hepatoprotective activity of medicinal\nplants. SGOT and SGPT are enzymes present in the hepatocytes, and\ntheir leakage into the blood stream is observed during cell damage.\nThe level of SGOT is known to increase during cardiac or skeletal\nmuscle damage. ALP is an enzyme which is present in the biliary duct\nlining of liver. The estimation of total bilirubin depicts the depth\nof jaundice and also indicates the severity of liver damage. The decrease\nin the total protein level is an indicator of liver damage caused\nby insignificant protein synthesis.

", - "

In the normal control group,\nthe values of SGOT, SGPT, ALP, total\nand direct bilirubin, and protein content were found to be 59.83 \u00b1\n6.25, 52.83 \u00b1 5.71, 101.5 \u00b1 7.92 IU/L, 0.703 \u00b1 0.05,\n0.163 \u00b1 0.02 mg/dL, and 8.47 \u00b1 0.05 g/dL, respectively.\nHowever, the CCl4 intoxicated control group displayed a\nsignificant increase in these values, like SGOT (145.83 \u00b1 10.91\nIU/L), SGPT (112.17 \u00b1 7.47 IU/L), ALP (324.17 \u00b1 8.70 IU/L),\ntotal bilirubin (1.42 \u00b1 0.11 mg/dL), and direct bilirubin (0.34\n\u00b1 0.06 mg/dL), and a significant decrease in the total protein\ncontent (6.28 \u00b1 0.08 g/dL). CPFA at a concentration of 400 mg/kg\nhas shown a significant decrease in the serum enzymes like SGOT 59.83\n\u00b1 1.70 IU/L, SGPT 58.67 \u00b1 2.63 IU/L, ALP 120.33 \u00b1 2.63\nIU/L, total bilirubin 0.638 \u00b1 0.03 mg/dL, and direct bilirubin\n0.14 \u00b1 0.01 mg/dL and a significant increase in the total protein\ncontent 8.19 \u00b1 0.02 g/dL, as compared to the toxic control group.\nA liver histopathological study also supported the protective effect\nof the extracts compared to the toxic control group. The images are\nrepresented in ##FIG## .

", - "

Column chromatographic analysis of the extract was carried out\nwith the universal solvent system that resulted in 13 different fractions.\nPhytochemical screening of these fractions revealed the presence of\nflavonoids, phenolics, and tannins. The results are depicted in ##TAB## .

", - "

Total phenolic and flavonoid contents\nwere estimated for all the\nabove fractions. Out of 13 fractions, fractions 5 and 9 showed the\nhighest amount of flavonoids and phenolics. The total phenolic content\nof fraction 5 and 9 was 284.91 \u00b1 6.03 and 378.31 \u00b1 3.15\nmg GAE/g, respectively, and their flavonoid content was evaluated\nas 333.82 \u00b1 1.39 and 380.33 \u00b1 1.55 mg QE/g, respectively\n( ##TAB## ).

", - "

Their hepatoprotective\nevaluation was carried out by an in vitro model using\ngoat liver slice culture for fractions\n5 and 9 (highlighted in ##TAB## ) that was estimated to have higher contents of phenolics\nand flavonoids. It was revealed that fraction 9 has much better activity,\nas compared to the toxic control group. [ ##TAB## , Figure S5 (Supporting\nInformation)].

", - "

GC\u2013MS analyses of the fraction 5 and 9 were\ncarried out\nto identify the compounds that are responsible for the exhibited activity.\nThe spectrum of fraction 9 is shown in ##FIG## and fraction 5 in Figure S6 (Supporting Information). From the library search, it was\nrevealed that the peak corresponding to 105.6 could be due to glyceric\nacid, peak of 169.2 could be due to gallic acid, peak of 207.35 could\nbe due to pilocarpine, and peak of 281.39 could be due to pangamic\nacid.

", - "

Further HPTLC analysis was carried out for the alcoholic extract\nof C. paniculatum flower. The active\nfractions 5 and 9, as separated by column chromatography along with\nstandard quercetin were considered for evaluation. The solvent mixture\nchloroform/ethyl acetate/formic acid in the ratio 6:4:1 was used as\nthe mobile phase. Silica gel 60 F 254 HPTLC plates were selected for\nanalysis, and the plates were observed under white light and 254 and\n366 nm. The presence of quercetin was confirmed and quantified. The\nimages are depicted in ##FIG## , and the respective chromatograms of the crude extract, the\nfractions under investigation along with the standard quercetin, are\nshown in Figure S7 (Supporting Information).

", - "

As quantified from HPLC chromatogram\nunder 254 nm, the amount of\nquercetin present in the alcoholic extract, fraction 5, and fraction\n9 was 38.0, 34.6, and 49.6%, respectively.

" + "Incorporation of <sup>13</sup>C into ribose and base of nucleotides", + "

To test the hypothesis that an asymmetric carbon source such as [3-13C]-pyruvate will provide superior site-selective labeling, we compared the growth of wild type and two mutant E. coli strains on [3-13C]-pyruvate (Fig.\u00a0 ##FIG## , Table\u00a0 ##TAB## ). To place our results in context, we outline nucleotide metabolism in E. coli via glycolysis, gluconeogenesis, and the Kreb cycle using pyruvate as the sole carbon source. If glucose is used as the sole carbon source, the conversion of PEP (phosphoenolpyruvate) to pyruvate by pyruvate kinase is an irreversible reaction under physiological conditions (Fig.\u00a0 ##FIG## ). However, under minimal growth conditions in E. coli using pyruvate as the sole carbon source, PEP synthetase (PPS) can convert pyruvate directly to PEP (Fig.\u00a0 ##FIG## ), and PEP carboxylase (PPC) can convert PEP to an oxaloacetate intermediate (Chao et al. ##REF## ; Schilling et al. ##UREF## ). In eukaryotes, pyruvate is converted to PEP via a two-step process with an oxaloacetate intermediate catalyzed by pyruvate carboxylase and PEP carboxykinase. However, pyruvate carboxylase is a mitochondrial protein not operative in E. coli, and so we expect all PEP will be derived from the externally supplied pyruvate without dilution by the TCA cycle for the DL323 E. coli strain. Consequently purine and pyrimidine moieties derived from oxaloacetate and serine should have enrichment levels quite near that of the initial pyruvate. Similarly the labeling pattern in the ribose ring derived from PEP, which is not diluted by the TCA cycle, should label only C1\u2032 and C5\u2032 in 1:2 ratio and all other sites should have minimal label. ##FIG## \n ##TAB## \n

", + "

In agreement with this metabolic analysis (Fig.\u00a0 ##FIG## ) and for DL323 E. coli grown on [3-13C]-pyruvate, the ribose ring is labeled only in the C1\u2032 and C5\u2032 carbon positions at ~42% and 98% respectively (Fig.\u00a0 ##FIG## , Table\u00a0 ##TAB## ). Thus these two positions remain singlet. This labeling pattern is very different from what was observed for DL323 E. coli grown on [1, 3-13C]-glycerol (Johnson and Hoogstraten ##REF## ; Thakur et al. ##REF## , ##REF## ). In this case of growth on glycerol, the ribose ring is labeled in all but the C4\u2032 carbon position, so that the C2\u2032 and C3\u2032 positions suffer from multiplet splitting by carbon\u2013carbon coupling and the C1\u2032 position has some admixture of residual C1\u2032-C2\u2032 coupling (Thakur et al. ##REF## , ##REF## ). ##FIG## \n

", + "

Similar to DL323, K10zwf E. coli grown on [3-13C]-pyruvate leads to exclusive labeling of the C1\u2032 and C5\u2032 carbon position without 13C-13C coupling evident (Fig.\u00a0 ##FIG## ). Importantly, in this case the abundance of 13C label at C1\u2032 is higher than that obtained with DL323 (Table\u00a0 ##TAB## ). Again similar to DL323 and K10zwf E. coli, wildtype K12 grown on [3-13C]-pyruvate also leads to labeling of the C1\u2032 and C5\u2032 carbon positions, but unlike the other two strains, sufficient residual labeling of the C2\u2032 and C3\u2032 carbon atoms leads to 13C-13C coupling (Fig.\u00a0 ##FIG## , ##FIG## ). This residual labeling presumably arises from PEP derived from the mixture of [1, 2, 3-13C]-oxaloacetate and [2, 3, 4-13C]-oxaloacetate intermediates (Fig.\u00a0 ##FIG## ).

", + "

The dramatic improvement of the labeling afforded by using [3-13C]-pyruvate is most visible in the base region. For nucleotides derived from K10zwf and K12 cultures, both the protonated C5 and C6 carbon positions of pyrimidines are substantially labeled (\u226560%) because the flux through the TCA cycle is efficient in these two E. coli strains. This efficient flux also ensures that the purine C4 and C5 positions are labeled. A single pass through the TCA cycle leads to an equal probability of carrying the 13C label into either the pyrimidine C5 or C6 position because the oxaloacetate is generated by [1, 3-13C]-malate or [2, 4-13C]-malate (Fig.\u00a0 ##FIG## ). If the precursor is [1, 3-13C]-oxaloacetate, then the desired label at pyrimidine C5 or C6 is diluted after the first and subsequent rounds of the TCA cycle (Fig.\u00a0 ##FIG## ). This dilution of labeling is observed for the nucleotides derived from the cultures of E. coli K10zwf and K12 strains (Table\u00a0 ##TAB## ). Again, if the precursor is [2, 4-13C]-oxaloacetate, then the second pass through the TCA cycle leads to a [2, 3-13C]-oxaloacetate and subsequent cycles leads to a mixture of [1, 2, 3-13C]-oxaloacetate and [2, 3, 4-13C]-oxaloacetate. The net effect of the TCA cycle is to thoroughly scramble the labels arising from oxaloacetate such that, in addition to the desired selective label at either the pyrimidine C5 or C6, undesired pyrimidine C5-C6 or C4-C5-C6 labeled spin pairs would be obtained. Thus the significant coupling observed between pyrimidine C4 and C5 or C5 and C6 (Fig.\u00a0 ##FIG## ; Supplementary Figure S2) is a consequence of this scrambling by the TCA cycle. These labeling patterns, similar to those observed with [1, 3-13C]-glycerol grown on DL323, have the undesirable consequence of substantive 13C-13C couplings that degrade the resolution and the accuracy of measured relaxation parameters (as shown below). In contrast to K10zwf and K12, E. coli strain DL323 provides a much cleaner labeling pattern that is devoid of unwanted labeling at the pyrimidine C6 and purine C4 and C5 positions (Fig.\u00a0 ##FIG## ; Supplementary Figure S2). For the DL323 strain, the flux through the TCA cycle is reduced essentially to zero, thereby preventing the dilution of the labels arising from oxaloacetate described above. ##FIG## \n

", + "

As expected, all of the C2 and C8 atoms of purine are substantially labeled to >90% for nucleotides derived from DL323 and K12 cultures, but those from K10zwf have these labels reduced to 85 and 57% respectively partly because the scrambling from the TCA cycle apparently feeds more efficiently into the purine biosynthetic pathway for K10zwf than K12 (Figs.\u00a0 ##FIG## , ##FIG## ; Supplementary Figure S2). The origin of these differences is not clear. Nonetheless, some of the scrambled malate or oxaloacetate from the TCA cycle is likely fed back into PEP such that purine C4, purine C5, and pyrimidine C6 positions are labeled in K10zwf and K12 but not DL323 (Fig.\u00a0 ##FIG## ). As a result, K10zwf and K12 provide 13C -13C spin pairs in the purine and pyrimidine base moieties that can potentially hinder accurate extraction of relaxation parameters.

", + "Applications of site selective labels for <sup>13</sup>C NMR study of nucleic acids", + "

Spectra obtained using our selective labeling methodology is of high quality (Figs.\u00a0 ##FIG## , ##FIG## ). A 27\u2013nt A-site ribosomal RNA fragment derived from the 30 S ribosomal subunit (Purohit and Stern ##REF## ; Fourmy et al. ##REF## ; Kaul et al. ##REF## ; Wirmer and Westhof ##REF## ; Schmeing and Ramakrishnan ##REF## ) was transcribed using either the uniformly 13C/15N-labeled GTP and CTP or the site selectively-labeled GTP and CTP derived from DL323 E. coli cells grown on [3-13C]-pyruvate as described above. For the C,G- uniformly 13C\u2014labeled sample, the peaks are not only broader, but they also overlap extensively (Fig.\u00a0 ##FIG## ). These unwanted splittings can also be removed using either constant time experiments, adiabatic band selective decoupling schemes, or maximum entropy reconstruction-deconstruction; as discussed previously, these have their own limitations (Dayie and Thakur ##REF## ; Thakur et al. ##REF## , ##REF## ). In particular, the length of the constant time period (T) limits the acquisition times to multiples of the homonuclear coupling constant. To obtain reasonable digital resolution, large values of T are needed and this leads to significant signal attenuation for RNA molecules larger than 30 nucleotides (Dayie ##REF## ). But with the selective labels, one is not forced to compromise resolution for improved sensitivity. ##FIG## \n ##FIG## \n

", + "

Use of the site selectively labeled NTPs removes the crowding due to the splitting of signals from J-coupling and eliminates the C2\u2032, C3\u2032, and C4\u2032 resonances while preserving only the C1\u2032 and C5\u2032 signals (Fig.\u00a0 ##FIG## ). In addition to improving resolution, sensitivity is also greatly enhanced. For instance, cytosine C5 has a large chemical shielding anisotropy (CSA) and so decays rapidly, and as expected most of the C5 resonances are absent in the spectra of the uniformly labeled sample. These signals are preserved in the spectra of the site selectively labeled sample (Fig.\u00a0 ##FIG## ).

", + "

In the C8 purine and C6 pyrimidine regions, the selective labels once again improve the resolution and sensitivity (Fig.\u00a0 ##FIG## ). By eliminating the C6 pyrimidine signals, the guanosine C8 signals can be readily identified without concerns about potential overlap with cytosine C6, especially for a sample that is labeled with all four nucleotides. These observations underscore the difference selective labeling makes on sensitivity and resolution.

", + "

A non-constant time version of the 13CH2 TROSY experiment (Miclet et al. ##REF## ) that enables the rescue of the slowest relaxing multiplet component was next performed to demonstrate the usefulness of these site selective labels. Compared to uniform labeling, the site-selective labels shows two clear advantages of improved resolution and sensitivity (Fig.\u00a0 ##FIG## A, B) of a spectrum of a 27-nt A-Site RNA fragment labeled only in G and C. Compared to the normal CH2-methylene optimized experiment (Schleucher et al. ##REF## ; Sattler et al. ##UREF## ), 14 out of the expected 18 GC C5\u2032 resonances are visible in the TROSY experiment but most of these are either overlapped, or weak, or absent in the uniformly labeled sample. Thus, it is anticipated that these and other new experiments that incorporate the 13CH2 TROSY module can be designed to probe RNA-ligand interactions at very high resolution using the site specific labels described here. ##FIG## \n

", + "Site selective labeling affords more accurate relaxation rate measurements using non-constant time non-selective pulse experiments", + "

To place the results of the relaxation in context, it is instructive to examine how spins are dipolar coupled to each magnetization of interest in a relaxation measurement. For instance, the longitudinal relaxation of the 13C5 spin during a relaxation period T for a uniformly labeled RNA sample has the following time dependency (Yamazaki et al. ##UREF## ; Dayie and Wagner ##UREF## ; Boisbouvier et al. ##REF## ; Hansen and Al-Hashimi ##REF## ): ##FORMU## \u0394Ci are the deviations of each carbon\u2019s longitudinal magnetization from its equilibrium magnetization, \u0394H5 is the deviation of the H5 proton magnetization from its equilibrium magnetization, and the autorelaxation and cross relaxation rates are given by RC5-X(z) and RX(x\u00a0\u2192\u00a0C5) respectively. Similar equations can be written for the ribose C1\u2032 spin and all its dipolar coupled neighbors. These rates are given as follows (Eldho and Dayie ##REF## ): ##FORMU## \n ##FORMU## \n ##FORMU## \n ##FORMU## \n ##FORMU## and \u03941\u00a0=\u00a0\u03c3xx\u00a0\u2212\u00a0\u03c3zz and \u03942\u00a0=\u00a0\u03c3yy\u00a0\u2212\u00a0\u03c3zz.

", + "

Here X\u00a0=\u00a0C4 or C6. The physical constants \u03b3H and \u03b3C are the gyromagnetic ratios of 1H and 13C respectively (\u03b3C\u00a0=\u00a06.7283\u00a0\u00d7\u00a0107 rad (Ts)\u22121, \u03b3H\u00a0=\u00a026.75\u00a0\u00d7\u00a0107 rad (Ts)\u22121), rHC is the length of the internuclear 1H-13C bond vector, rCC is the length of the internuclear 13C-13C bond vector, and ##FORMU## is Planck\u2019s constant divided by 2\u03c0 (1.054592\u00a0\u00d7\u00a010\u221227). An asymmetric chemical shielding tensor can be decomposed into an isotropic and two axially symmetric anisotropic terms with the symmetry axes arbitrarily chosen as the two principal axes along X and Y, as discussed previously for carbonyl backbone motional analysis (Dayie and Wagner ##UREF## ). More general expressions for anisotropic motion can be found in the literature (Werbelow ##UREF## ). Because each term J(\u03c9X\u00a0\u2212\u00a0\u03c9C) in ( ##FORMU## ) and ( ##FORMU## ) is proportional to J(0) which scales linearly with molecular weight, it is clear that the dipole\u2013dipole contribution cannot be neglected for uniformly labeled RNA samples.

", + "

We expected therefore for the case of a uniformly labeled cytosine or uridine, the H5, C4, C6 spins will contribute to the longitudinal relaxation of the 13C5 nucleus, and H1\u2032 and C2\u2032 spins will contribute to the relaxation of the 13C1\u2032 nucleus. For the site selectively labeled nucleotides, only H5 will contribute to the longitudinal relaxation of the 13C5 nucleus, and only H1\u2032 will contribute to the relaxation of the 13C1\u2032 nucleus. Thus, the labeling pattern of rNMPs derived from DL323 grown on [3-13C]-pyruvate is particularly attractive for relaxation studies of the ribose C1\u2032 and the base C5 (as well as C2 and C8 positions that are not discussed further here). Each of these positions is essentially singlet. The isolation of the C1\u2032 ribose and the C5 base positions from labeled adjacent neighbors means that the interference described above that arises from strong 13C-13C magnetic interactions in the base and ribose rings is no longer a hindrance for extracting accurate relaxation parameters. Therefore as a third demonstration of the usefulness of the site selectively labeled versus the uniformly labeled nucleotides for quantifying dynamics in RNA, R1 experiments were carried on the 27\u2013nt A-site ribosomal RNA fragment transcribed using uniformly 13C/15N-labeled GTP and CTP and site selectively-labeled GTP and CTP derived from DL323 E. coli cells grown on [3-13C]-pyruvate. Examples of 13C R1 decay curves for both ribose C1\u2032 and base carbon C5 sites are depicted in Fig.\u00a0 ##FIG## and Supplementary Figure S4. The decay curves fit well to a mono-exponential function for the C1\u2032 position of a site selective ATP- labeled sample (Fig.\u00a0 ##FIG## A), but not as well for a uniform ATP-labeled sample (Fig.\u00a0 ##FIG## B). The discrepancy between R1 measured for uniform (2.0\u00a0\u00b1\u00a00.2\u00a0s\u22121) and site-selective (1.8\u00a0\u00b1\u00a00.08\u00a0s\u22121) labeled sample is 0.2\u00a0s\u22121 (Fig.\u00a0 ##FIG## A). Similarly, the fit to a monoexponential decay function is only applicable to the site selectively CTP-labeled RNA sample (Fig.\u00a0 ##FIG## C). The fit to the uniformly CTP-labeled sample is poor. Again fit to a monoexponential decay function is only applicable to the C5 position of the site selectively CTP-labeled RNA sample. This is discrepancy, also worse for the C5 carbons with relaxation rates of 3.4 \u00b1\u00a01.0\u00a0s\u22121 for the uniformly labeled sample and 2.9\u00a0\u00b1\u00a00.4\u00a0s\u22121 for the selectively labeled sample, is partly because of the expected dipolar coupled interactions, but also because of substantial decrease in signal-to noise for the uniformly labeled sample (Supplementary Figure S4). These observations suggest that the contributions from the cross-relaxation terms in ( ##FORMU## ), ( ##FORMU## ), and ( ##FORMU## ) are not negligible for the uniformly labeled sample. ##FIG## \n

" ], "discussion": [ "Discussion", - "

The\nliver, the major metabolic organ of our body, may have toxicity\ndue to various drugs like alcohol, anabolic steroids, nonsteroidal\nanti-inflammatory drug, and so on. ##REF## Oxidative stress can be attributed to the major reason\nbehind this toxicity. Oxidative stress occurs due to the imbalance\nbetween free radicals generated and antioxidants present in the body. ##REF## Many medicinal plants display significant implications\ntoward traditional medicine for the treatment of hepatic diseases\nincluding hepatic disorders. Further these plants are being evaluated\nfor in vivo pharmacological activities to identify\npotent candidates. The generation of free radicals as products of\nmany biomolecular reactions plays major roles in the emergence of\ncancer and other health disturbances. It is also known that CCl4 can induce the levels of various enzymes like ALT, AST, ALP,\nand \u03b3-GT in animal models. CCl4 is also known to\ncause acute hepatocyte injuries that further result in the leakage\nof various hepatocyte enzymes. Many hepatoprotective agents from different\nnatural sources are reported to possess the ability to protect against\nsuch injuries by restoring the levels of the above enzymes along with\nretaining the levels of triglycerides, cholesterol, low-density lipoprotein, and high-density lipoprotein in\nthe serum to normalcy. Earlier reports have also shown the potential\nof polyphenolic compounds to play key roles in establishing such protective\nactivity. ##REF## Several mechanisms were reported\nwith animal models for the above observed activity with hepatoprotective\nagents that have strong ability to trap various metal ions like zinc,\ncalcium, and iron. ##REF## Herein, we have investigated\nthe hepatoprotective effects of C. paniculatum plant bioactive compounds against CCl4-induced hepatotoxicity\nin rats.

", - "

Internal antioxidant deficiency can be overcome by\nsupplementing\nwith external sources. Since polyphenolics are known to have high\nantioxidant potentials, their natural sources from plants have gained\nimmense significance for them to be explored for extracting and isolating\nthese polyphenolic antioxidant constituents. ##UREF## Among these antioxidants, flavonoids have a high health promoting\nrole through their antioxidant mechanism. Due to their high free-radical\nscavenging property, the flavonoids display significant roles toward\nmanaging various diseases and disorders. Among the flavonoids, quercetin,\nrutin, apigenin, catechin, and so on are also reported for their anti-hepatotoxic\nproperties. ##UREF## Enzymatic and non-enzymatic\ncomponents of the oxidative stress can be resolved through the plant\ncell defense system. A non-enzymatic system controls the cellular\nresponses against the free radicals, whereas enzymatic responses directly\nscavenge the free radicals and hence control the antioxidant defense\nsystem. ##REF## The qualitative chemical evaluation\nresults obtained from our studies support the presence of various\nclasses of chemical compounds like carbohydrates, proteins, flavonoids,\ntannins, phenolics, and steroids. The alcoholic extract of CPF exhibited\nhigh phenolics and flavonoids that influence the apparent antioxidant\nactivity compared to other extracts. Herein, the antioxidant activity\nis evaluated by various methods including DPPH, nitric oxide radical\nscavenging, ABTS, and reducing power assay. The IC50 value\nfor the alcoholic extract was represented as 62.28 \u00b1 0.51, 2362.71\n\u00b1 9.39 51, and 193.09 \u00b1 5.84 \u03bcg/mL, respectively,\nfor DPPH, ABTS, and nitric oxide radical scavenging methods. The alcoholic\nextracts were further subjected to hepatoprotective activity evaluation\nusing CCl4-induced hepatotoxic models. The evaluation was\ncarried out by determining SGOT, SGPT, ALP, direct bilirubin, total\nbilirubin, and total protein content and also through histopathology\nof liver. The results obtained revealed a marked decrease in liver\nenzyme levels along with an increase in the total protein content\nwhen compared to toxic control groups that were similar to standard\ngroups administered with silymarin. The total bilirubin in the serum\nserves as a hepatic functional marker which is associated with hepatic\ndisorder along with acute disruption of hepatocellular architecture\nand function. Moreover, the toxicant-induced liver injuries also result\nin increased levels of bilirubin. The above findings are further supported\nby the results of histopathology analysis. In the toxic control group,\nthe liver histopathological characters were showing disarranged hepatic\ncellular architecture with cell necrosis, fatty degeneration, and\ncentral vein crowding. This was almost reframed to the normal liver\narchitecture by 200 mg/kg and 400 mg/kg body weight alcoholic extract\ntreatment. In order to identify the unique constituents of the extract,\nthe fractions displaying significant activities were separated by\ncolumn chromatography using a universal solvent system (hexane, ethyl\nacetate, and methanol), and the fractions were tested for the presence\nof phenolics and flavonoid contents by both qualitative and quantitative\ntechniques. Fractions 5 and 9 which showed better phenolic and flavonoid\ncontents were then considered for in vitro hepatoprotective\nanalyses using goat liver slice culture, and the activity was compared\nwith the alcoholic extract of the plant. Of the two fractions, fraction\n9 showed better activity. GC\u2013MS analyses followed by NIST library\nsearch revealed that the constituents responsible for the above activity\nmay be glyceric acid, gallic acid, pilocarpine, or pangamic acid,\nas shown in ##FIG## . However, fraction 5 revealed the presence of quinovic acid as the\nactive ingredient, and the GC\u2013MS spectra of the same is given\nin Figure S6 (Supporting Information).\nFurther HPTLC analysis revealed the presence of 34.6 and 49.6% quercetin\nin the active fractions 5 and 9, respectively, which was not revealed\nthrough GC\u2013MS.

" + "

Previous labeling technologies using wild type E. coli strains and symmetric carbon sources spurred the development of new solution NMR methods to characterize the structure and dynamics of small to medium sized uniformly labeled RNA molecules (Batey et al. ##REF## ; Hall ##REF## ; F\u00fcrtig et al. ##REF## ; Latham et al. ##REF## ; Dayie ##REF## ; Lu et al. ##REF## ). A number of alternative labeling approaches have been proposed since then, and it is important therefore to discuss the advantages and limitations of these different methods. First, uniform labeling introduces direct one-bond and residual dipolar couplings that prevents accurate measurement of 13C relaxation rates (Johnson et al. ##REF## ; Dayie and Thakur ##REF## ; Thakur et al. ##REF## , ##REF## ) and decreases the resolution and sensitivity of NMR experiments. For instance, growth of wildtype E. coli strain K12 on [1-13C]-acetate yields 13C label at the ribose C1\u2032, C2\u2032, and C3\u2032 atomic sites but very little label at C4\u2032 and C5\u2032 (Hoffman and Holland ##REF## ). As a result, C2\u2032 and C3\u2032 have substantial multiplet structure and even C1\u2032 has residual multiplet structure (Supplementary Figure S3). In addition, the protonated aromatic carbons (purine C2 and C8 and pyrimidine C5 and C6) are not labeled, making this particular labeling pattern less useful for routine NMR applications. In contrast, growth of wildtype E. coli strain K12 on [2-13C]-acetate yields 13C label at all five ribose carbon positions of C1\u2032, C2\u2032, C3\u2032, C4\u2032 and C5\u2032 (Hoffman and Holland ##REF## ). Since each site is highly enriched, all five positions have substantial multiplet structure (Supplementary Figure S3). Also for the aromatic carbons, the protonated carbon C5 and C6 of pyrimidines and C2 and C8 of purines are labeled at very high level (Hoffman and Holland ##REF## ; Supplementary Figure S3). Similarly, growth of wildtype E. coli strain K12 on [2-13C]-glucose yields 13C label only at C1\u2032, C2\u2032, and C4\u2032. No label is observed at C3\u2032 and C5\u2032. For the aromatic carbons, the protonated carbon C5 and C6 of pyrimidine and C2 and C8 of purine are also labeled at very high level (Hoffman and Holland ##REF## ). These second set of labeling patterns are therefore very useful for NMR applications that require substantial coupling for transfer of coherences, but problematic for applications that require suppression of these coupling networks (Supplementary Figure S3 E).

", + "

To remove these coupling networks, alternate 13C-12C labeling schemes have been shown to be important for tackling these difficulties in RNA molecules (Johnson et al. ##REF## ; Johnson and Hoogstraten ##REF## ; Dayie and Thakur ##REF## ; Thakur et al. ##REF## , ##REF## ). However, some of the earlier schemes had their unique problems. For instance, the labeling pattern of rNTPs derived from DL323 grown on [1,3-13C]-glycerol is not attractive for relaxation studies because the ribose C2\u2032 position is doublet, the C1\u2032 retains some residual doublet arising from 13C2\u2032-13C1\u2032 isotopomers, and the C4\u2032 has no label (Thakur et al. ##REF## , ##REF## ). In the base region the pyrimidine C5 site has multiplet structure, again precluding its use for accurate relaxation measurements (Thakur et al. ##REF## , ##REF## ). The use of G6PDH mutant strains rather than DL323 using 2-13C-glycerol was also recommended in previous work (Johnson et al. ##REF## ).

", + "

Recently de novo biosynthetic methods have been proposed that potentially resolve these problems (Schultheisz et al. ##REF## , ##REF## ). However, the new method requires 29 enzymes for making ATP and GTP and 18 enzymes for making CTP and UTP. Of these enzymes only a few are commercially available, which makes this method excellent for adaptation by companies but not by individual laboratories. Additionally, the de novo method requires selectively labeled glucose and serine which are not readily available commercially, or prohibitively expensive, or both. For instance the de novo method of labeling ribose C1\u2032 and C5\u2032 in the context of purine C2, C5, and C8, indicated above by our method, requires 13C-2,6-glucose that is not commercially available and 13C-3-serine that costs ~$4000/g; the biomass method is able to make these labels readily.

", + "

Of greater interest the main advantage of using the E. coli strain DL323 grown on a non-symmetric carbon source such as [3-13C]-pyruvate is that it provides isolated two-spin systems that are ideal for relaxation measurements as well as improving resolution and sensitivity. For example in the ribose region, the C1\u2032 and C5\u2032 labels are completely isolated from C2\u2032 and C4\u2032 spins respectively. Similarly in the base region, the purine C2 and C8 and pyrimidine C5 spins are isolated from the network of carbon spins within the aromatic ring. For instance in a uniformly labeled sample, adenine C2 has substantial 2JC2C5 coupling of 11\u00a0Hz to the C5 carbon, and the purine C8 carbons have 2JC4C8 coupling of\u00a0~9\u00a0Hz to the C5 carbon (Wijmenga and van Buuren ##UREF## ). In addition, the carbon atoms C4 (149\u2013154\u00a0ppm)/C5 (116\u2013120\u00a0ppm)/C6 (156\u2013161\u00a0ppm)/C8 (131\u2013142\u00a0ppm) resonate within a chemical shift range that precludes effectively decoupling C8 from the C4, C5, or C6 carbon atoms. These complications again prevent accurate relaxation measurements using CPMG experiments. Formate supplementation has been proposed as a potential cost-effective solution (Latham et al. ##REF## ; Dayie and Thakur ##REF## ; Thakur et al. ##REF## , ##REF## ). However, the C8 is labeled to ~90% and C2 to only ~25%, limiting the general usefulness of formate labeling. Thus the confounding effects of scalar coupling from adjacent labeled sites are eliminated using the current method proposed in this work, unlike the case for previous labels.

" ], "conclusion": [ "Conclusions", - "

The implication on the\npharmacological properties from the flower\nextract of C. paniculatum, family Verbenaceae,\nis reported for the first time in the literature. Various antioxidant\nactivities carried out following various assays including DPPH, NO\nradical scavenging, ABTS activity, and reducing ability with the alcoholic\nextract showing significant activity compared to other extracts. The\ninvestigations by various enzymatic studies and histopathological\nsections with the alcoholic extract revealed excellent hepatoprotective\nactivity. The best two fractions of the extract, as determined by\ncolumn chromatographic analysis when subjected to in vitro hepatoprotective investigations, revealed fraction 9 to possess\nefficient hepatoprotective activity against carbon tetrachloride-induced\nliver toxicity in goat liver slice culture. Further analyses revealed\nthat the hepatoprotective activities of C. paniculatum flower may be attributed to the presence of the following compounds\nthat were isolated from fraction 9\u2014(i) glyceric acid, (ii)\ngallic acid, (iii) pilocarpine, (iv) pangamic acid, and (v) quercetin\neither individually or in combination of the above.

" + "

Any RNA sequence can be site specifically labeled with site alternate 13C-enriched isotopes efficiently prepared from site-specifically labeled rNTPs using in vitro transcription by T7 RNA polymerase. We have presented an efficient method for preparing these ribonucleotides containing site specific distributions of 13C enrichment that has significant advantages in terms of cost, flexibility, resolution and sensitivity over previously reported procedures. These labels will be particularly beneficial to investigators using heteronuclear NMR spectroscopy to study the structure and dynamics of RNA and RNA complexes of increasingly large sizes, currently an important and active area of research.

" ], "front": [ - "

##GRAPH##

", - "

The aim of the presented\nwork involves the isolation, characterization,\nand evaluation of hepatoprotective potential of Clerodendrum\npaniculatum flower extracts. For this purpose, petroleum\nether, chloroform, ethyl acetate, alcohol, and water extracts of C. paniculatum flower were screened for the flavonoid\nand phenolic content and quantified. Various antioxidant activity\nassays including 2,2\u2032-diphenyl-1-picrylhydrazyl (DPPH), nitric\noxide (NO) radical scavenging, 2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic\nacid) (ABTS), and reducing ability were carried out. Of the above\nmethods, the alcoholic extract exhibited high antioxidant potential\nand was selected further for the hepatoprotective evaluations. Hepatoprotective\nevaluation of the alcoholic extract was carried out for carbon tetrachloride\n(CCl4)-intoxicated model systems. Enzymes associated with\nliver functions were estimated, and histopathological evaluations\nwere carried out to monitor the liver architecture. Prominently, reduced\nlevels of various associated enzymes along with increased protein\ncontent were observed when the liver specimen was pretreated with\nthe extract. Moreover, the liver architecture was almost comparable\nto that of the normal control group. The column chromatographic analysis\nof the extract revealed 13 fractions to possess high phenolics and\nflavonoid contents. The best two fractions were identified for in vitro hepatoprotective evaluation in the goat liver model.\nFurthermore, the GC\u2013MS analyses of the fractions were carried\nout followed by a library search, to identify the constituents responsible\nfor the hepatoprotective activity which revealed the presence of four\nmajor constituents\u2014pilocarpine, glyceric acid, pangamic acid,\nand gallic acid. An in vitro hepatoprotective study\nof the isolated fractions showed better activity compared to the whole\nalcoholic extract, and the results were comparable to the normal group\ntaken as a control. The investigations with an in vitro model suggest that the isolated fraction with rich flavonoid content\nshowed better hepatoprotective activity. GC\u2013MS analysis of\nthe fractions that displayed good hepatoprotective activity suggested\nthe presence of pilocarpine, glyceric acid, pangamic acid, and gallic\nacid, while HPTLC analysis revealed the presence of quercetin.

" + "

Selective isotopic labeling provides an unparalleled window within which to study the structure and dynamics of RNAs by high resolution NMR spectroscopy. Unlike commonly used carbon sources, the asymmetry of 13C-labeled pyruvate provides selective labeling in both the ribose and base moieties of nucleotides using E. coli variants, that until now were not feasible. Here we show that an E. coli mutant strain that lacks succinate and malate dehydrogenases (DL323) and grown on [3-13C]-pyruvate affords ribonucleotides with site specific labeling at C5\u2032 (~95%) and C1\u2032 (~42%) and minimal enrichment elsewhere in the ribose ring. Enrichment is also achieved at purine C2 and C8 (~95%) and pyrimidine C5 (~100%) positions with minimal labeling at pyrimidine C6 and purine C5 positions. These labeling patterns contrast with those obtained with DL323 E. coli grown on [1, 3-13C]-glycerol for which the ribose ring is labeled in all but the C4\u2032 carbon position, leading to multiplet splitting of the C1\u2032, C2\u2032 and C3\u2032 carbon atoms. The usefulness of these labeling patterns is demonstrated with a 27-nt RNA fragment derived from the 30S ribosomal subunit. Removal of the strong magnetic coupling within the ribose and base leads to increased sensitivity, substantial simplification of NMR spectra, and more precise and accurate dynamic parameters derived from NMR relaxation measurements. Thus these new labels offer valuable probes for characterizing the structure and dynamics of RNA that were previously limited by the constraint of uniformly labeled nucleotides.

", + "Electronic supplementary material", + "

The online version of this article (doi:10.1007/s10858-011-9581-6) contains supplementary material, which is available to authorized users.

", + "Keywords" ], "body": [ - "Experimental\nSection", - "Chemicals and Reagents", - "

The chemicals and reagents used\nfor the experiments carried out for the present investigation were\npurchased from Spectrochem, SRL, Acros-Organics, RANKEM, and Fisher\nScientific and used as received without further purification. ABTS,\nDPPH, and Folin-Ciocalteu\u2019s reagents were obtained from Sigma-Aldrich.

", - "Plant Collection", - "

C. paniculatum flowers were collected from Malappuram, Kerala during August to\nOctober. The plant was identified and authenticated by Kottakkal Ayurveda\nresearch center, Kerala. The voucher specimen was deposited in the\ndepartment for further reference.

", - "Preparation of Plant Extracts", - "

The flowers weighing\n\u223c300 g were collected and dried under shade. Successively,\nsolvent extraction was carried out using petroleum ether, chloroform,\nethyl acetate, alcohol, and water as solvent medium following the\nhot continuous percolation method using a conventional Soxhlet apparatus.\nThe extracts were concentrated, and the percentage yield was calculated.

", - "Preliminary Phytochemical Screening", - "

Preliminary qualitative\nanalysis was carried out for detecting the presence of various classes\nof compounds like alkaloids, phenolics, flavonoids, saponins, glycosides,\nand steroids. Specific color change or any precipitate formation was\nconsidered as a positive response. ##UREF## , ##UREF##

", - "Determination\nof Total Phenolic and Flavonoid Contents", - "

The Folin\u2013Ciocalteau\nmethod was used for the quantitative\ndetermination of phenolics in each extract. 1 mL of the extract was\nmixed with 1 mL of Folin-Ciocalteu\u2019s reagent. Then, 3 mL of\nsodium carbonate solution was added. The whole mixture was incubated\nfor 2 h, and absorbance was measured at 725 nm. ##UREF##

", - "

The total flavonoid content was estimated using the\naluminium chloride colorimetric method. 0.5 mL of the extract was\nmixed with 2 mL of water and 0.15 mL of NaNO2 followed\nby the addition of 0.15 mL of 6% AlCl3 solution. After\n6 min, 2 mL of 4% sodium hydroxide solution was added, and volume\nwas maintained to 5 mL using distilled water. Absorbance was measured\nat 510 nm. ##REF##

", - "<italic>In Vitro</italic> Assays", - "ABTS Method", - "

In this method, the ABTS radical was generated\nby the oxidation of ABTS with potassium persulfate. For preparing\nABTS solution, 7 mM aqueous ABTS solution and 2.45 mM potassium per\nsulfate were mixed together. The solution was incubated in the dark\nat 29 \u00b0C for 14 h. The working solution was prepared by diluting\nthe previously prepared solution with phosphate buffer (pH 7.4). 50\n\u03bcL of the extract was mixed with 3 mL of the freshly prepared\nsolution and was allowed to stand for 20 min. After the incubation\ntime, absorbance was measured at 734 nm. The percentage inhibition\nwas measured using the above equation. ##UREF##

", - "Nitric Oxide Radical Inhibitory Assay", - "

Different concentrations\nof plant extracts were mixed with 1.5 mL of 5 mM sodium nitroprusside\nin phosphate buffered saline (pH 7.4). The solution was mixed together\nand incubated at 25 \u00b0C for 3 h. After the incubation time, 1.5\nmL of the Griess reagent (2% phosphoric acid, 1% sulfanilamide, and\n0.1% N-1-naphthyl ethylene diamine dihydrochloride)\nwas added to the reaction mixture. Absorbance of the reaction mixture\nwas measured at 546 nm with reference to standard gallic acid. The\npercentage inhibition was also measured. ##UREF## , ##REF##

", - "Reducing Power Assay", - "

In this method, better antioxidant\nactivity was represented by an increase in the absorbance of the reaction\nmixture. Potassium ferricyanide, ferric chloride, and trichloroacetic\nacid produce a colored complex with the antioxidant compounds, and\nits absorbance can be measured at 700 nm. 2.5 mL of phosphate buffer\n(0.2 M, pH 6.6) and 2.5 mL of potassium ferricyanide were added to\n10 mg/mL samples. The resulting mixture was incubated at 50 \u00b0C\nfor 20 min. After the incubation period, 2.5 mL of trichloroacetic\nacid was added to the mixture, and the upper layer of the resulting\nmixture was collected by the 10 min centrifugation process. To 5 mL\nof above liquid, 5 mL of distilled water and 1 mL of ferric chloride\n(0.1% w/v) were added, and absorbance was measured at 700 nm. ##REF## , ##REF##

", - "Statistical Analysis", - "

All the procedures were carried\nout in triplicates, and the results were expressed as the mean \u00b1\nstandard deviation. The statistical analyses were carried out using\nSPSS software version 20.

", - "<italic>In Vivo</italic> Animal Experiments", - "Selection\nof Animals", - "

Female Wistar Albino rats weighing\n100\u2013130 g were used for the hepatoprotective analysis of the\nalcoholic extract. Standard laboratory conditions were followed for\nanimal maintenance, and they were fed with standard food and water\nad libitum. The experimental protocol was approved by the Al Shifa\nCollege of Pharmacy Institutional Animal Ethical Committee (IAEC)\n(regd. no. 1195/Re/S/08/CPCSEA).

", - "Acute Toxicity Study", - "

C. paniculatum flower alcoholic\nextract\u2019s acute toxicity was conducted on\nalbino rats, according to the OECD guidelines no. 425. After 12 h\nof fasting, the extract (2000 mg/kg) was administered orally. The\nanimals were observed for 14 days to check mortality, any behavioral\nchanges, any discomforts, and so on. ##REF##

", - "CCl<sub>4</sub>-Induced Hepatoprotective\nStudy", - "

Five\ngroups of six animals were selected, and the experiment was conducted\nfor 14 days.

", - "

Doses: the animals of the vehicle group (group\nI) received CMC (5 ml/kg b.w.) orally, and CCl4 in liquid\nparaffin was administered to animals of group II\u2013V via subcutaneous route for 14 days. Standard dose (silymarin,\n50 mg/kg b.w.) was administered to group III, and group IV animals\nreceived CPFA1 (200 mg/kg b.w.) by oral route. Group V animals received\nthe CPFA 2 (400 mg/kg b.w.) extract. After 14 days of treatment, all\nthe animals were sacrificed by cervical decapitation, and blood samples\nwere collected by cardiac puncture. Serum was collected by centrifugation\nat 3000 rpm and examined for enzyme analysis such as SGOT, SGPT, ALP,\ntotal and direct bilirubin, and total protein content. Data were analyzed\nby one-way ANOVA followed by Dunnett\u2019s test. ##REF## \u2212 ##UREF##

", - "Statistical\nAnalysis", - "

The results of the in\nvivo experiments conducted were expressed as mean \u00b1\nS.E.M. SPSS software version 20 was used for statistical analysis.

", - "Histopathology", - "

The livers were excised and washed with\nnormal saline. Liver fragments were fixed in 10% buffered formalin\nfollowed by paraffin embedding. Liver sections of 0.5 \u03bcm thickness\nwere taken and stained with the hematoxylin\u2013eosin dye. The\nsections were mounted and microscopically observed for studying histological\nchanges if any. ##UREF##

", - "Isolation of Compound Using\nColumn Chromatography", - "

Around\n1 g of the alcoholic extract of the plant was loaded into the column,\nand gradient elution was carried out using hexane, ethyl acetate,\nand methanol as the mobile phase. ##REF## , ##REF## The solvent\nwas passed through the column at 1 mL per minute under gravity to\nfractionate the sample extract. Each fraction was collected in a test\ntube and was numbered subsequently. The fractions obtained were subjected\nto the qualitative chemical test for identification of tannins, phenolics,\nand flavonoids. ##UREF## , ##UREF## , ##UREF## The quantitative estimation for phenolics and flavonoid was carried\nout, ##UREF## , ##REF## and the best two fractions were selected\nfor an in vitro hepatoprotective activity study using\ngoat liver slice culture.

", - "<italic>In Vitro</italic> Hepatoprotective\nStudy", - "

The\nfresh goat livers were obtained from the local market. The liver was\nremoved and transferred to presterilized Krebs Ringer Herpes (KRH)\nmedium. The liver was cut into thin slices ranging from 4 to 6 mg\nusing a sharp blade and was used for the study. Each set weighing\n100 mg contains 20\u201325 slices. Tissues were washed with 10 mL\nof KRH medium in every 10 min over a period of 1 h. The slices were\npreincubated at 37 \u00b0C for 60 min in cotton-plugged beakers containing\n10 mL of KOH. The liver slices were further divided into individual\nculture for respective treatment. All the cultures were incubated\nat constant temperature in a water bath at 37 \u00b0C for 2 h. The\ncells were isolated from the culture medium of each set by centrifuging\nat 3000 rpm for 10 min at 4 \u00b0C, and the corresponding supernatants\nwere assayed for the presence of leaked biochemical markers such as\nalanine transaminase (ALT), aspartate transaminase (AST), alkaline\nphosphatase (ALP), and acid phosphatase (ACP). ##UREF## \u2212 ##UREF##

", - "Experimental Setup for\nEvaluation of Hepatoprotective Activity", - "

Group 1: normal control, Group 2: toxic\ncontrol\u2014CCl4 (15.5 mM), Group\n3: standard (silymarin-50 mM), Group 4\u20136:C. paniculatum alcoholic extract\nat the concentration range of 25\u2013100 \u03bcg/mL, Group\n7\u20139: column fraction 5 at the concentration range of\n2.5\u201310 \u03bcg/mL, and Group 10\u201312: column\nfraction 9 at the concentration range of 2.5\u201310 \u03bcg/mL.

", - "GC\u2013MS Analyses", - "

Mass spectra were recorded on\na Schimadzu GCMS-QP2020 Gas chromatograph mass spectrometer. The constituents\npresent in various fractions of the extracts were predicted using\nNIST/EPA/NIH mass spectral library\u20142017. The equipment has\na DB 35-MS capillary standard non-polar poly (dimethylsiloxane) column\nwith dimensions of 30 mm \u00d7 0.25 mm ID \u00d7 0.25 \u03bcm film.\nHelium was used as the carrier gas with a flow rate of 1.0 mL/min.\nThe injector was operated at 280 \u00b0C, and the oven temperature\nwas programmed as follows: 70\u2013280 \u00b0C, gradually increased\nby 10 \u00b0C per/min.

" + "Electronic supplementary material", + "

Below is the link to the electronic supplementary material.\n ##SUPPL## \n

" ], "back": [ - "Supporting Information Available", - "

The Supporting Information is\navailable free of charge at https://pubs.acs.org/doi/10.1021/acsomega.1c03722.

Antioxidant activity and\nGC\u2013MS spectrum of fraction\n5 (PDF)

", - "Supplementary Material", - "

This research\ndid not receive any specific grant from funding agencies in the public,\ncommercial, or not-for-profit sectors.

", - "

The authors declare no\ncompeting financial interest.

", - "Acknowledgments", - "

The authors would like to express their sincere\nthanks to the Management of Vellore Institute of Technology, Vellore\nand Chennai campus for providing necessary support.

" + "

This work was supported in part by the University of Maryland Nano-Biotechnology Award and the National Institutes of Health grant GM077326 to T.K.D. We are grateful to Professors Jon Dinman and Steve Rokita for helpful discussions.

", + "Open Access", + "

This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.

" ], "figure": [ - "

Determination\nof reducing power for various solvent extracts from\nthe flowers of C. paniculatum, where\nCPFP denotes the C. paniculatum flower\npetroleum ether extract, CPFC indicates the C. paniculatum flower chloroform extract, CPFE indicates the C.\npaniculatum flower ethyl acetate extract, CPFA indicates\nthe C. paniculatum flower alcohol extract,\nand CPFW indicates the C. paniculatum flower water extract.

##GRAPH## ", - "

Photographs of hematoxylin/eosin-stained liver sections. (A) Normal\ncontrol group rat representing normal liver architecture, (B) CCl4 intoxicated rat liver showing cell necrosis around central\nvein, loss of cell boundaries, and ballooning degeneration, (C) liver\nsection of the standard group showing less cell necrosis and less\ncentral vein crowding, (D) CPFA (200 mg/kg), and (E) CPFA (400 mg/kg)\nreceived group showed a moderate degree of liver damage and cell inflammation\nand reduced cell crowding.

##GRAPH## ", - "

GC\u2013MS chromatogram of fraction 9.

##GRAPH## ", - "

HPTLC\nplates observed under (a) normal light and (b) 254 and (c)\n366 nm.

##GRAPH## " + "

Major metabolic pathways involved in the production of nucleic acid nucleotides from pyruvate, including key steps in glycolysis, gluconeogenesis and several passes through the tricarboxylic (TCA) cycle as derived from Covert and Palsson ( ##REF## ). With the E. coli strain lacking succinate and malate dehydrogenases (DL323), the oxidative branch of the pentose phosphate pathway remains intact but the TCA cycle is severed in two places such that the oxaloacetate is derived exclusively from carboxylation of phosphoenolpyruvate (PEP) and the resulting label is not diluted by the TCA cycle. Atom labels for the terminal (3) carbon (magenta and thin\ncircle), the central (2) carbon (square), and the terminal (1) carbon (triangle) of pyruvate are highlighted. Positions that are enriched due to the presence of 13CO2 in the growth medium are shown with a circled x. Pyrimidine base derived from the oxaloacetate (OAA) produced by carboxylation of PEP is shown via the aspartate intermediate. This OAA cannot be used as a substrate in the first and subsequent rounds of the TCA cycle because of the two mutations. Consequently OAA derived aspartate amino acid can be produced with 13C labeling at only the C\u03b1 (C6) position if [2-13C]-pyruvate is used. If [3-13C]-pyruvate is used only C\u03b2 (C5) position is labeled. In either case carboxylation of PEP leads to labeling of the C\u03b3 (C4) position. Similarly the labeling pattern of purines from glycine derived from 3PG are shown such that if [2-13C]-pyruvate is used only the C\u03b1 position of Gly and therefore C5 position of the purine ring is labeled. Otherwise if [3-13C]-pyruvate is used the CO of Gly and therefore C4 of purine ring is labeled, and the labeling of the C\u03b2 position of Ser also leads to labeling of the purine C2 and C8 positions

##GRAPH## ", + "

2D non-constant time HSQC spectra of all four labeled nucleotides extracted from K10zwf (blue contours, left shifted) or K12 (red contours) or DL323 (purple contours, right shifted) E. coli strains grown on [3-13C]-pyruvate. Growth on [3-13C]-pyruvate results in label at only C1\u2032 and C5\u2032 for DL323 and K10zwf strains, whereas growth of K12 leads to some residual label at C2\u2032, C3\u2032, and C4\u2032 in addition to the major labels at the C1\u2032 and C5\u2032 carbons. The cytosine (Cyt) and Uracil (Ura) C5 resonances at 96.67 and 102.69\u00a0ppm respectively are folded into the spectrum

##GRAPH## ", + "

Labeling pattern of a mixture of four rNMPs isolated from K10zwf, K12, and DL323 E. coli grown on a [3-13C]-Pyruvate background, and DL323 E. coli grown on a [1, 3-13C]-glycerol background. The direct carbon detection 1D spectrum shows all the labeled carbon positions. A long recycle delay of 5\u00a0s was used to allow for sufficient magnetization recovery and proton decoupling was limited to the acquisition period only. The residual 13C-13C coupling observed in a [1, 3-13C]-glycerol is absent with the [3-13C]-Pyruvate only in DL323; K10zwf and K12 grown on [3-13C]-Pyruvate still retain the residual 13C-13C coupling

##GRAPH## ", + "

NMR spectra showing enhanced resolution afforded by site selective labeling of A-Site RNA bound to paromomycin. The experiments were performed on the A-Site RNA A site-selectively 13C-GTP and CTP labeled and B uniformly 13C-GTP and CTP labeled. 2D non-constant time HSQC spectra of the ribose region. The cytosine (Cyt) and Uracil (Ura) C5 resonances at 96.67 and 102.69\u00a0ppm respectively are folded into the spectrum and are boxed to highlight the reduced signal from the uniformly labeled sample. The C2\u2032, C3\u2032, and C4\u2032 regions are boxed to highlight the absence of labeling in selectively labeled sample. Blown-up views of the C2\u2032, C3\u2032, and C4\u2032 regions boxed in (A) and (B) are depicted in (C) and (D) respectively

##GRAPH## ", + "

NMR spectra showing enhanced resolution afforded by site selective labeling of A-Site RNA bound to paromomycin. The experiments were performed on the A-Site RNA A site-selectively 13C-GTP and CTP labeled and B uniformly 13C-GTP and CTP labeled. 2D non-constant time HSQC spectra of the base region. The cytosine (Cyt) and Uracil (Ura) C6 resonances are boxed to highlight the absence of labeling in selectively labeled sample

##GRAPH## ", + "

CH2-TROSY spectra depicting the C5\u2032 region of A-Site RNA bound to paromomycin. Experiments were performed on the A-Site RNA A site-selectively 13C-GTP and CTP labeled and B uniformly 13C-GTP and CTP labeled. Spectra are plotted at identical contour levels. Notice the dramatic gain in sensitivity and resolution with the site selectively labeled sample

##GRAPH## ", + "

Representative longitudinal R1 relaxation decay curves showing marked deviation from monoexponential decay for uniformly labeled samples. Ribose C1\u2032 R1 relaxation measurements at 25\u00b0C for the A-site RNA labeled with A site selectively-labeled ATP and B uniformly 13C/15N-labeled ATP. Ribose C1\u2032 R1 relaxation measurements at 25\u00b0C for A-site RNA labeled with C site selectively-labeled GTP and CTP and D uniformly 13C/15N-labeled GTP and CTP

##GRAPH## " ], "table": [ - "Phytochemical Screening for the Flower\nExtract of <italic>C. paniculatum</italic>
chemical classpetroleum etherchloroformethyl acetatealcoholawatera
proteins----++
carbohydrates-+++++++
flavonoids---+++++
tannins and phenolics---+++++
steroids+++--
glycosides-----
alkaloids-----
volatile oils-----
##FOOTN## ", - "Quantification of Total Phenolics\nand Flavonoids in Extracts of <italic>C. paniculatum</italic> Flower
sampletotal phenolic\ncontent (mg\u00a0GAE/g)flavonoid content (mg\u00a0QE/g)
petroleum ether81.5\u00a0\u00b1\u00a00.9164.03\u00a0\u00b1\u00a01.21
chloroform114.6\u00a0\u00b1\u00a00.46137.10\u00a0\u00b1\u00a01.85
ethyl acetate148.6\u00a0\u00b1\u00a00.81121.27\u00a0\u00b1\u00a01.20
alcohol378.5\u00a0\u00b1\u00a00.88393.23\u00a0\u00b1\u00a01.33
water108.9\u00a0\u00b1\u00a01.15131.07\u00a0\u00b1\u00a00.96
", - "Antioxidant\nPotential, as Determined\nby DPPH Assay with the Alcoholic Extract of <italic>C. paniculatum</italic> Flower
Sl. no.sampleIC50 (\u03bcg/mL) (mean\u00a0\u00b1\u00a0S.D)
1standard (ascorbic acid)48.74\u00a0\u00b1\u00a00.21
2petroleum ether693.59\u00a0\u00b1\u00a013.51a
3chloroform409.17\u00a0\u00b1\u00a02.16a
4ethyl acetate568.54\u00a0\u00b1\u00a05.27a
5alcohol62.28\u00a0\u00b1\u00a00.51a
6water342.07\u00a0\u00b1\u00a00.95a
##FOOTN## ", - "Antioxidant Activity, as Evaluated\nby ABTS Assay with the Alcoholic Extract of <italic>C. paniculatum</italic> Flower
Sl. no.sampleIC50 (\u03bcg/mL) (mean\u00a0\u00b1\u00a0S.D)
1standard (ascorbic acid)941.09\u00a0\u00b1\u00a08.31
2petroleum ether7986\u00a0\u00b1\u00a02.00a
3chloroform3643.36\u00a0\u00b1\u00a01.37a
4ethyl acetate8628.2\u00a0\u00b1\u00a06.21a
5alcohol2362.71\u00a0\u00b1\u00a09.39a
6water8781.6\u00a0\u00b1\u00a016.31a
##FOOTN## ", - "Antioxidant Activity, as Determined\nby Nitric Oxide Radical Scavenging Assay with the Alcoholic Extract\nof <italic>C. paniculatum</italic> Flower
Sl. no.sampleIC50 (\u03bcg/mL) (mean\u00a0\u00b1\u00a0S.D)
1standard (gallic acid)147.11\u00a0\u00b1\u00a010.20
2petroleum ether4765.42\u00a0\u00b1\u00a06.88a
3chloroform1129.17\u00a0\u00b1\u00a017.74a
4ethyl acetate3970.74\u00a0\u00b1\u00a03.57a
5alcohol193.09\u00a0\u00b1\u00a05.84a
6water1384.17\u00a0\u00b1\u00a013.77a
##FOOTN## ", - "SGOT, SGPT, ALP, Total and Direct\nBilirubin, and Total Protein Levels of All Groups of Animals<xref rid=\"t6fn1\" ref-type=\"table-fn\">a</xref><sup>,</sup><xref rid=\"t6fn2\" ref-type=\"table-fn\">b</xref>
\u00a0\u00a0\u00a0\u00a0Bilirubin (mg/dL)
\u00a0
groupsSGOT (IU/L)SGPT (IU/L)ALP (IU/L)totaldirecttotal protein (g/dL)
vehicle control59.33\u00a0\u00b1\u00a02.55**52.83\u00a0\u00b1\u00a02.33**101.5\u00a0\u00b1\u00a07.92**0.703\u00a0\u00b1\u00a00.02**0.163\u00a0\u00b1\u00a00.01**8.47\u00a0\u00b1\u00a00.02**
toxic control145.83\u00a0\u00b1\u00a04.45112.17\u00a0\u00b1\u00a03.05324.17\u00a0\u00b1\u00a03.551.42\u00a0\u00b1\u00a00.050.34\u00a0\u00b1\u00a00.026.28\u00a0\u00b1\u00a00.03
standard (silymarin)66.33\u00a0\u00b1\u00a02.19**56.67\u00a0\u00b1\u00a02.20**109.83\u00a0\u00b1\u00a03.21**0.752\u00a0\u00b1\u00a00.01**0.17\u00a0\u00b1\u00a00.01**8.36\u00a0\u00b1\u00a00.02**
CPFA (200\u00a0mg/kg)64.67\u00a0\u00b1\u00a02.08**62.83\u00a0\u00b1\u00a00.79**128.50\u00a0\u00b1\u00a02.88**0.787\u00a0\u00b1\u00a00.03**0.18\u00a0\u00b1\u00a00.01**8.12\u00a0\u00b1\u00a00.01**
CPFA (400\u00a0mg/kg)59.83\u00a0\u00b1\u00a01.70**58.67\u00a0\u00b1\u00a02.63**120.33\u00a0\u00b1\u00a02.63**0.638\u00a0\u00b1\u00a00.03**0.14\u00a0\u00b1\u00a00.01**8.19\u00a0\u00b1\u00a00.02**
##FOOTN## ", - "Phytochemical Screening for Phenolics\nand Flavonoids of Isolated Fractions of the Alcoholic Extract of <italic>C. paniculatum</italic><xref rid=\"t7fn1\" ref-type=\"table-fn\">a</xref>
fractionsshinoda testferric\nchloridelead acetate testalkaline reagent test
F1----
F 2----
F 3----
F 4-++-
F 5+++++++++++
F\n6-++++
F 7+++++
F 8++++++
F 9++++++++++++
F 10++++++
F 11++++++++
F 12+++++++++
F 13+++++++
##FOOTN## ", - "Total Phenolic and Flavonoid Contents\nof Fractions Obtained from the Alcoholic Extract of <italic>C. paniculatum</italic>
column fractiontotal phenolic content (mg\u00a0GAE/g)flavonoid content (mg\u00a0QE/g)
F 5284.91\u00a0\u00b1\u00a06.03333.82\u00a0\u00b1\u00a01.39
F 6139.66\u00a0\u00b1\u00a01.93198.39\u00a0\u00b1\u00a02.56
F 7109.93\u00a0\u00b1\u00a03.48115.02\u00a0\u00b1\u00a01.62
F 899.083\u00a0\u00b1\u00a04.4999.74\u00a0\u00b1\u00a01.25
F 9378.31\u00b13.15380.33\u00b11.55
F 1097.38\u00a0\u00b1\u00a02.18116.70\u00a0\u00b1\u00a01.77
F 11138.78\u00a0\u00b1\u00a04.85174.50\u00a0\u00b1\u00a01.604
F 12146.40\u00a0\u00b1\u00a00.50178.56\u00a0\u00b1\u00a01.72
F 13147.99\u00a0\u00b1\u00a01.06192.52\u00a0\u00b1\u00a01.61
", - "<italic>In Vitro</italic> Studies\nof Fractions<xref rid=\"t9fn1\" ref-type=\"table-fn\">a</xref>
groupsSGOT (IU/L)SGPT (IU/L)ALP (IU/L)ACP (IU/L)
vehicle\ncontrol39.33\u00a0\u00b1\u00a03.512**56.00\u00a0\u00b1\u00a02.646**95.67\u00a0\u00b1\u00a02.082**10.52\u00a0\u00b1\u00a01.572**
toxic control191.00\u00a0\u00b1\u00a05.568248.67\u00a0\u00b1\u00a02.082309.00\u00a0\u00b1\u00a06.55745.33\u00a0\u00b1\u00a01.528
standard\n(silymarin 50\u00a0mM)42.67\u00a0\u00b1\u00a04.042**62.00\u00a0\u00b1\u00a01.000**106.67\u00a0\u00b1\u00a02.517**12.33\u00a0\u00b1\u00a02.517**
CPLE\nextract (100\u00a0\u03bcg/mL)150.00\u00a0\u00b1\u00a02.000**210.00\u00a0\u00b1\u00a04.583**266.00\u00a0\u00b1\u00a06.557**41.67\u00a0\u00b1\u00a01.528**
CPLE extract (200\u00a0\u03bcg/mL)128.33\u00a0\u00b1\u00a01.526**180.00\u00a0\u00b1\u00a02.000**221.33\u00a0\u00b1\u00a03.215**36.33\u00a0\u00b1\u00a01.528**
CPLE extract (400\u00a0\u03bcg/mL)101.33\u00a0\u00b1\u00a02.517**157.33\u00a0\u00b1\u00a04.510**181.67\u00a0\u00b1\u00a06.658**33.00\u00a0\u00b1\u00a04.000**
fraction 5 (25\u00a0\u03bcg/mL)132.00\u00a0\u00b1\u00a02.000**156.33\u00a0\u00b1\u00a04.042**172.67\u00a0\u00b1\u00a02.516**40.67\u00a0\u00b1\u00a02.082**
fraction 5 (50\u00a0\u03bcg/mL)124.00\u00a0\u00b1\u00a03.606**142.67\u00a0\u00b1\u00a03.055**161.33\u00a0\u00b1\u00a03.215**31.67\u00a0\u00b1\u00a00.577**
fraction 5 (100\u00a0\u03bcg/mL)96.33\u00a0\u00b1\u00a06.110**132.33\u00a0\u00b1\u00a05.860**145.67\u00a0\u00b1\u00a04.042**30.00\u00a0\u00b1\u00a05.000**
fraction 9 (25\u00a0\u03bcg/mL)95.67\u00a0\u00b1\u00a03.055**124.00\u00a0\u00b1\u00a02.646**147.00\u00a0\u00b1\u00a06.245**27.33\u00a0\u00b1\u00a01.528**
fraction 9 (50\u00a0\u03bcg/mL)70.33\u00a0\u00b1\u00a01.528**110.67\u00a0\u00b1\u00a02.082**132.33\u00a0\u00b1\u00a02.517**26.00\u00a0\u00b1\u00a02.000**
fraction 9 (100\u00a0\u03bcg/mL)56.67\u00a0\u00b1\u00a04.726**83.0\u00a0\u00b1\u00a04.000**118.00\u00a0\u00b1\u00a02.646**20.00\u00a0\u00b1\u00a01.000**
##FOOTN## " + "

\n13C enrichment levels at various carbon positions within ribonucleotides harvested from K12, DL323, and K10zwf E. coli strains grown on [3-13C]-pyruvate

Carbon position labeledDL323 E. coli strainK10zwf E. coli strainK12 E. coli strain
Purinea\n
\u00a0Ade C299\u00a0\u00b1\u00a0185\u00a0\u00b1\u00a0495\u00a0\u00b1\u00a02
\u00a0C893\u00a0\u00b1\u00a0157\u00a0\u00b1\u00a0198\u00a0\u00b1\u00a02
Pyrimidinea\n
\u00a0C598\u00a0\u00b1\u00a0481\u00a0\u00b1\u00a0491\u00a0\u00b1\u00a01
\u00a0C615\u00a0\u00b1\u00a0160\u00a0\u00b1\u00a0464\u00a0\u00b1\u00a01
Ribose
\u00a0C1\u2032a\n42\u00a0\u00b1\u00a0482\u00a0\u00b1\u00a0283\u00a0\u00b1\u00a02
\u00a0C2\u2032b\n49\u00a0\u00b1\u00a0121\u00a0\u00b1\u00a01
\u00a0C3\u2032b\n<2<18
\u00a0C4\u2032b\n39\u00a0\u00b1\u00a0130\u00a0\u00b1\u00a01
\u00a0C5\u2032c\n95\u00a0\u00b1\u00a059595\u00a0\u00b1\u00a01
##FOOTN## " ], "formula": [ - " ##GRAPH## " + "\\documentclass[12pt]{minimal}\n\t\t\t\t\\usepackage{amsmath}\n\t\t\t\t\\usepackage{wasysym} \n\t\t\t\t\\usepackage{amsfonts} \n\t\t\t\t\\usepackage{amssymb} \n\t\t\t\t\\usepackage{amsbsy}\n\t\t\t\t\\usepackage{mathrsfs}\n\t\t\t\t\\usepackage{upgreek}\n\t\t\t\t\\setlength{\\oddsidemargin}{-69pt}\n\t\t\t\t\\begin{document}$$ \\begin{gathered} \\frac{{d(\\Updelta C_{Z}^{5} )}}{dT} = - \\{ R_{C5 - H5} (z) + R_{C5 - C4} (z) + R_{C5 - C6} (z) + \\rho_{others} \\} \\Updelta C_{Z}^{5} - \\hfill \\\\ R_{C6} (C6 \\to C5)\\Updelta C_{Z}^{6} - R_{C4} (C4 \\to C5)\\Updelta C_{Z}^{4} - R_{C5} (H5 \\to C5)\\Updelta H_{Z}^{5} \\hfill \\\\ \\end{gathered} $$\\end{document} ##GRAPH## ", + "\\documentclass[12pt]{minimal}\n\t\t\t\t\\usepackage{amsmath}\n\t\t\t\t\\usepackage{wasysym} \n\t\t\t\t\\usepackage{amsfonts} \n\t\t\t\t\\usepackage{amssymb} \n\t\t\t\t\\usepackage{amsbsy}\n\t\t\t\t\\usepackage{mathrsfs}\n\t\t\t\t\\usepackage{upgreek}\n\t\t\t\t\\setlength{\\oddsidemargin}{-69pt}\n\t\t\t\t\\begin{document}$$ {\\text{R}}_{{{\\text{C5}} - {\\text{H}}}} \\left( {\\text{z}} \\right) = A[{\\text{J}}(\\omega_{\\text{H}} - \\omega_{\\text{C}} ) + 6 {\\text{J}}(\\omega_{\\text{H}} + \\omega_{\\text{C}} )\\left] { + \\left( { 3 {\\text{A}} + B} \\right)} \\right[{\\text{J}}(\\omega_{\\text{C}} )] $$\\end{document} ##GRAPH## ", + "\\documentclass[12pt]{minimal}\n\t\t\t\t\\usepackage{amsmath}\n\t\t\t\t\\usepackage{wasysym} \n\t\t\t\t\\usepackage{amsfonts} \n\t\t\t\t\\usepackage{amssymb} \n\t\t\t\t\\usepackage{amsbsy}\n\t\t\t\t\\usepackage{mathrsfs}\n\t\t\t\t\\usepackage{upgreek}\n\t\t\t\t\\setlength{\\oddsidemargin}{-69pt}\n\t\t\t\t\\begin{document}$$ {\\text{R}}_{{{\\text{C5}} - {\\text{X}}}} ({\\text{z}}) = A^{\\prime } [{\\text{J}}(\\omega_{\\text{X}} - \\omega_{\\text{C}} ) + 6 {\\text{J}}(\\omega_{\\text{X}} + \\omega_{\\text{C}} )] + ( 3 {\\text{A}}^{\\prime } + B)[{\\text{J}}(\\omega_{\\text{C}} )] $$\\end{document} ##GRAPH## ", + "\\documentclass[12pt]{minimal}\n\t\t\t\t\\usepackage{amsmath}\n\t\t\t\t\\usepackage{wasysym} \n\t\t\t\t\\usepackage{amsfonts} \n\t\t\t\t\\usepackage{amssymb} \n\t\t\t\t\\usepackage{amsbsy}\n\t\t\t\t\\usepackage{mathrsfs}\n\t\t\t\t\\usepackage{upgreek}\n\t\t\t\t\\setlength{\\oddsidemargin}{-69pt}\n\t\t\t\t\\begin{document}$$ {\\text{R}}_{\\text{C5}} ({\\text{C5}} \\to {\\text{H5}}) = {\\text{A}}[ 6 {\\text{J}}(\\omega {\\text{H}} + \\omega {\\text{C}}) - {\\text{J}}(\\omega {\\text{H}} - \\omega {\\text{C}})] $$\\end{document} ##GRAPH## ", + "\\documentclass[12pt]{minimal}\n\t\t\t\t\\usepackage{amsmath}\n\t\t\t\t\\usepackage{wasysym} \n\t\t\t\t\\usepackage{amsfonts} \n\t\t\t\t\\usepackage{amssymb} \n\t\t\t\t\\usepackage{amsbsy}\n\t\t\t\t\\usepackage{mathrsfs}\n\t\t\t\t\\usepackage{upgreek}\n\t\t\t\t\\setlength{\\oddsidemargin}{-69pt}\n\t\t\t\t\\begin{document}$$ {\\text{R}}_{\\text{X}} \\left( {{\\text{X}} \\to {\\text{C5}}} \\right) = {\\text{A}}^{\\prime } [ 6 {\\text{J}}(\\omega {\\text{C}} + \\omega {\\text{X}}) - {\\text{J}}(\\omega {\\text{X}} - \\omega {\\text{C}})] $$\\end{document} ##GRAPH## ", + "\\documentclass[12pt]{minimal}\n\t\t\t\t\\usepackage{amsmath}\n\t\t\t\t\\usepackage{wasysym} \n\t\t\t\t\\usepackage{amsfonts} \n\t\t\t\t\\usepackage{amssymb} \n\t\t\t\t\\usepackage{amsbsy}\n\t\t\t\t\\usepackage{mathrsfs}\n\t\t\t\t\\usepackage{upgreek}\n\t\t\t\t\\setlength{\\oddsidemargin}{-69pt}\n\t\t\t\t\\begin{document}$$ {\\text{A}} = \\frac{{\\hbar^{2} \\gamma_{C}^{2} \\gamma_{H}^{2} }}{{4r_{HC}^{6} }},\\;{\\text{A}}^{\\prime } = \\frac{{\\hbar^{2} \\gamma_{C}^{4} }}{{4r_{CC}^{6} }},\\;{\\text{B}} = \\frac{{\\omega_{C}^{2} \\Updelta_{C}^{2} }}{3},\\quad {\\text{and}}\\quad \\Updelta_{C}^{2} = \\Updelta_{1}^{2} + \\Updelta_{2}^{2} - \\Updelta_{1} \\Updelta_{2} $$\\end{document} ##GRAPH## ", + "\\documentclass[12pt]{minimal}\n\t\t\t\t\\usepackage{amsmath}\n\t\t\t\t\\usepackage{wasysym} \n\t\t\t\t\\usepackage{amsfonts} \n\t\t\t\t\\usepackage{amssymb} \n\t\t\t\t\\usepackage{amsbsy}\n\t\t\t\t\\usepackage{mathrsfs}\n\t\t\t\t\\usepackage{upgreek}\n\t\t\t\t\\setlength{\\oddsidemargin}{-69pt}\n\t\t\t\t\\begin{document}$$ \\hbar $$\\end{document} ##GRAPH## " ], - "box": [ - "EXAMPLE FROM ANOTHER ARTICLE", - "Key messages

To understand the disease burden of chronic cough, and the burden among subgroups of the patients with chronic cough in Japan.

There is an unmet need for better interventions and treatments to improve the quality of life, and reduce work productivity and activity impairment, healthcare resource utilisation and experience of anxiety, depression and sleep problems among patients with chronic cough in Japan.

This study provides novel evidence on the burden of chronic cough in Japan through a population-based survey, using validated instruments.

" - ], - "code": [ - "EXAMPLE FROM ANOTHER ARTICLE", - "\n\n\nLupe.dat$w <\u2013 ifelse(Lupe.dat$case_==1, 1, 5000)\n\n\nHSF.Lupe <\u2013 glm(case_ ~ elevation + popden + landuseC,\n\n\n data = Lupe.dat,\n\n\n weight = w,\n\n\n family = binomial)\n\n\n\n", - "\n\n\nLupe.dat <\u2010 within(Lupe.dat,\n\n\n landuseC1 <\u2010 relevel(landuseC, ref = \"wet\"))\n\n\n HSF.Lupe2 <\u2010 glm(case_ ~ elevation + popden + landuseC1,\n\n\n data = Lupe.dat,\n\n\n weight = w,\n\n\n family = binomial)\n\n\n\n", - "\n\n\nLupe.dat <\u2010 within(Lupe.dat, landuseC <\u2010 relevel(landuseC, ref = \"forest\"))\n\n\nHSF.Lupe3 <\u2010 glm(case_ ~ elevation + popden + landuseC + elevation:landuseC,\n\n\n data = Lupe.dat,\n\n\n weight = w,\n\n\n family = binomial)\n\n\n\n" - ], - "quote": [ - "EXAMPLE FROM ANOTHER ARTICLE", - "

I constantly feel like I am fighting some system or individual to get my medical needs met.

\u2014Keisha Currie

", - "

Generally, there is a neurological exam performed, but it is subjective and by no means inclusive of invisible symptoms.

\u2014Keisha Currie

", - "

Every day you wake up and do an inventory of what body parts are functioning properly, first. Then, you decide if you are going to lie to yourself and others and say, \u201cI\u2019m fine,\u201d when asked how you are doing because you know it is easier.

\u2014Keisha Currie

", - "

As an administrator of a 35,000-member online group of people living with MS, I often hear, \u201cWe are not taken seriously!\u201d, \u201cI am treated as a drug seeker,\u201d and \u201cThe doctor doesn\u2019t believe me because they can\u2019t see how this impacts me because I look good during the visit.\u201d

\u2014Cherie Binns

", - "

Often, African Americans have religious views and will reply, \u201cJust pray about it!\u2019\u201d to imply that God will simply take my MS away. Thankfully, I had developed my own views about this shortly after diagnosis or it would have been more isolating.

\u2014Keisha Currie

", - "

I feel that HCPs can have honest conversations with patients about how their MS is presenting, how management strategies can be beneficial, and long-term planning.

\u2014Keisha Currie

" - ], - "chem": [ - "EXAMPLE FROM ANOTHER ARTICLE", - " ##GRAPH## ", - " ##GRAPH## ", - " ##GRAPH## ", - " ##GRAPH## " - ], + "box": [], + "code": [], + "quote": [], + "chem": [], "supplementary": [ - " ##MEDIA## " + " ##MEDIA## " ], "footnote": [ - "

+++: highly present, ++: moderately\npresent, +: low, and -: absent.

", - "

p < 0.01, as\ncompared with the standard group.

", - "

p < 0.01, as\ncompared with the standard group.

", - "

p < 0.01, as\ncompared with the standard group.

", - "

Values\nare expressed as mean \u00b1\nSEM of six rats in each group. **p < 0.05, as\ncompared with the toxic control group.

CPFA: C. paniculatum flower\nalcoholic extract, SGOT: serum glutamate oxaloacetate transferase,\nSGPT: serum glutamate pyruvate transferase, and ALP: alkaline phosphatase.

", - "

+++: highly present, ++: moderately\npresent, +: low, -: absent, and F represents fractions.

", - "

The values are represented as mean\n\u00b1 S.D (n = 3), **P < 0.05,\nas compared with the toxic control group.

" + "

\naThe percentage label is calculated as an average of two methods: (i) the ratio of the sum of the intensities of satellite peaks to the sum of the intensities of the satellite and center peaks using the 2-bond 15N HSQC without 13C decoupling during acquisition (Dayie and Thakur ##REF## ) and (ii) the ratio of the sum of the intensities of satellite peaks to the sum of the intensities of the satellite and center peaks using the 1D 1H experiment without 13C decoupling during acquisition as described in the text

\nb,cThe percentage label (Plabel) is calculated as in (a) but this time with only method (ii)

" ], "graphic": [ - "", - "", - "", - "", - "", - "" + "", + "", + "", + "", + "", + "", + "", + "", + "", + "", + "", + "", + "", + "" ], "media": [ - "

ao1c03722_si_001.pdf

" + "

Supplementary material 1 (PDF 858 kb)

" ], - "unknown_pub": "[{\"surname\": [\"Anand\", \"Jacobo-Herrera\", \"Altemimi\", \"Lakhssassi\"], \"given-names\": [\"U.\", \"N.\", \"A.\", \"N.\"], \"article-title\": [\"A comprehensive\\nreview on medicinal plants as antimicrobial therapeutics: Potential\\navenues of biocompatible drug discovery\"], \"source\": [\"Metabolites\"], \"year\": [\"2019\"], \"volume\": [\"9\"], \"fpage\": [\"258\"], \"pub-id\": [\"10.3390/metabo9110258\"]}, {\"surname\": [\"Kopilakkal\", \"Rani\", \"Rasheed\", \"Bency\", \"Azeem\"], \"given-names\": [\"R.\", \"S.\", \"S. P.\", \"B. T.\", \"A. K.\"], \"article-title\": [\"Monograph\\nof Clerodendrum paniculatum\"], \"source\": [\"World J. Pharm.\\nRes.\"], \"year\": [\"2019\"], \"volume\": [\"8\"], \"fpage\": [\"448\"], \"pub-id\": [\"10.20959/wjpr20198-15328\"]}, {\"surname\": [\"Ekor\"], \"given-names\": [\"M.\"], \"article-title\": [\"The growing\\nuse of herbal medicines: issues relating to adverse reactions and\\nchallenges in monitoring safety\"], \"source\": [\"Front. Pharmacol.\"], \"year\": [\"2013\"], \"volume\": [\"4\"], \"fpage\": [\"177\"], \"pub-id\": [\"10.3389/fphar.2013.00177\"]}, {\"surname\": [\"Thomford\", \"Senthebane\", \"Rowe\", \"Munro\", \"Seele\", \"Maroyi\", \"Dzobo\"], \"given-names\": [\"N.\", \"D.\", \"A.\", \"D.\", \"P.\", \"A.\", \"K.\"], \"article-title\": [\"Natural Products for\\nDrug Discovery\\nin the 21st Century: Innovations for Novel Drug Discovery\"], \"source\": [\"Int. J. Mol. Sci.\"], \"year\": [\"2018\"], \"volume\": [\"19\"], \"fpage\": [\"1578\"], \"pub-id\": [\"10.3390/ijms19061578\"]}, {\"surname\": [\"Boora\", \"Chirisa\", \"Mukanganyama\"], \"given-names\": [\"F.\", \"E.\", \"S.\"], \"article-title\": [\"Evaluation\\nof nitrite radical scavenging\\nproperties of selected Zimbabwean plant extracts and their phytoconstituents\"], \"source\": [\"J. Food Process\"], \"year\": [\"2014\"], \"volume\": [\"2014\"], \"fpage\": [\"918018\"], \"pub-id\": [\"10.1155/2014/918018\"]}, {\"surname\": [\"Altemimi\", \"Lakhssassi\", \"Baharlouei\", \"Watson\", \"Lightfoot\"], \"given-names\": [\"A.\", \"N.\", \"A.\", \"D.\", \"D.\"], \"article-title\": [\"Phytochemicals:\\nExtraction, isolation, and identification of bioactive compounds from\\nplant extracts\"], \"source\": [\"Plants\"], \"year\": [\"2017\"], \"volume\": [\"6\"], \"fpage\": [\"42\"], \"pub-id\": [\"10.3390/plants6040042\"]}, {\"surname\": [\"Fernando\", \"Soysa\"], \"given-names\": [\"C. D.\", \"P.\"], \"article-title\": [\"Evaluation of Hepatoprotective\\nactivity of Eriocaulon quinquangulare\\nin vitro using porcine liver slices against ethanol induced liver\\ntoxicity and free radical scavenging capacity\"], \"source\": [\"BMC Complementary Altern. Med.\"], \"year\": [\"2016\"], \"volume\": [\"16\"], \"fpage\": [\"74\"], \"pub-id\": [\"10.1186/s12906-016-1044-x\"]}, {\"surname\": [\"Khandelwal\", \"Varunda\"], \"given-names\": [\"K. R.\", \"S.\"], \"source\": [\"Practical\\nPharmacognosy, Techniques and Experiments\"], \"edition\": [\"23\"], \"publisher-name\": [\"Nirali Prakashan publications\"], \"publisher-loc\": [\"New Delhi\"], \"year\": [\"2008\"]}, {\"surname\": [\"Trease\", \"Evans\"], \"given-names\": [\"G. E.\", \"W. C.\"], \"source\": [\"Pharmacognosy\"], \"publisher-name\": [\"ELBS Publication\"], \"publisher-loc\": [\"East Bourne\"], \"year\": [\"2002\"]}, {\"surname\": [\"Maier\", \"Basu\"], \"given-names\": [\"C.\", \"P.\"], \"article-title\": [\"In vitro antioxidant activities and polyphenol contents of seven\\ncommercially available fruits\"], \"source\": [\"Pharmacogn. Res.\"], \"year\": [\"2016\"], \"volume\": [\"8\"], \"fpage\": [\"258\"], \"pub-id\": [\"10.4103/0974-8490.188875\"]}, {\"surname\": [\"Almeida\", \"de Sousa\", \"Arriaga\", \"do Prado\", \"Magalh\\u00e3es\", \"Maia\", \"de Lemos\"], \"given-names\": [\"M. M. B.\", \"P. H. M.\", \"\\u00c2. M. C.\", \"G. M.\", \"C. E.\\nd. C.\", \"G. A.\", \"T. L. G.\"], \"article-title\": [\"Bioactive compounds and antioxidant activity of fresh\\nexotic fruits from northeastern Brazil\"], \"source\": [\"Food\\nRes. Int.\"], \"year\": [\"2011\"], \"volume\": [\"44\"], \"fpage\": [\"2155\"], \"pub-id\": [\"10.1016/j.foodres.2011.03.051\"]}, {\"surname\": [\"Junejo\", \"Gogoi\", \"Islam\", \"Rudrapal\", \"Mondal\", \"Hazarika\", \"Zaman\"], \"given-names\": [\"J. A.\", \"G.\", \"J.\", \"M.\", \"P.\", \"H.\", \"K.\"], \"article-title\": [\"Exploration\\nof antioxidant, antidiabetic\\nand hepatoprotective activity of Diplazium esculentum - A wild edible\\nplant from North Eastern India\"], \"source\": [\"Future J. Pharm.\\nSci.\"], \"year\": [\"2018\"], \"volume\": [\"4\"], \"fpage\": [\"93\"], \"pub-id\": [\"10.1016/j.fjps.2017.10.005\"]}, {\"surname\": [\"Gnanadesigan\", \"Ravikumar\", \"Anand\"], \"given-names\": [\"M.\", \"S.\", \"M.\"], \"article-title\": [\"Hepatoprotective activity\\nofCeriops\\ndecandra(Griff.) Ding Hou mangrove plant against CCl4induced liver\\ndamage\"], \"source\": [\"J. Taibah Univ. Sci.\"], \"year\": [\"2017\"], \"volume\": [\"11\"], \"fpage\": [\"450\"], \"pub-id\": [\"10.1016/j.jtusci.2016.07.004\"]}, {\"surname\": [\"Remya\", \"Balamurali\"], \"given-names\": [\"K.\", \"M. M.\"], \"article-title\": [\"In vivo and in vitro analyses to reveal the potential\\nof Solanum violaceum as efficient hepatoprotective agent\"], \"source\": [\"Eur. J. Mol. Clin. Med.\"], \"year\": [\"2020\"], \"volume\": [\"7\"], \"fpage\": [\"41\"], \"pub-id\": [\"10.1016/S2221-1691(12)60075-6\"]}, {\"surname\": [\"Ecobichon\"], \"given-names\": [\"D. J.\"], \"source\": [\"The Basis of Toxicology\\nTesting\"], \"publisher-name\": [\"CRC Press\"], \"publisher-loc\": [\"New York\"], \"year\": [\"1997\"]}, {\"surname\": [\"Bajpai\", \"Majumder\", \"Park\"], \"given-names\": [\"V. K.\", \"R.\", \"J. G.\"], \"article-title\": [\"Isolation\\nand purification of plant\\nsecondary metabolites using column-chromatographic technique\"], \"source\": [\"Bangladesh J. Pharmacol.\"], \"year\": [\"2016\"], \"volume\": [\"11\"], \"fpage\": [\"844\"], \"pub-id\": [\"10.3329/bjp.v11i4.28185\"]}, {\"surname\": [\"Gangwar\", \"Gautam\", \"Sharma\", \"Tripathi\", \"Goel\", \"Nath\"], \"given-names\": [\"M.\", \"M. K.\", \"A. K.\", \"Y. B.\", \"R. K.\", \"G.\"], \"article-title\": [\"Antioxidant Capacity\\nand Radical Scavenging Effect of Polyphenol\\nRichMallotus philippenensisFruit Extract on Human Erythrocytes: AnIn\\nVitroStudy\"], \"source\": [\"Sci. World J.\"], \"year\": [\"2014\"], \"volume\": [\"2014\"], \"fpage\": [\"279451\"], \"pub-id\": [\"10.1155/2014/279451\"]}, {\"surname\": [\"Chaudhari\", \"Mahajan\"], \"given-names\": [\"G. M.\", \"R. T.\"], \"article-title\": [\"In vitro hepatoprotective activity of Terminalia arjuna\\nstem bark and its flavonoids against CCl\"], \"sub\": [\"4\"], \"source\": [\"Asian J. Plant\\nSci. Res.\"], \"year\": [\"2016\"], \"volume\": [\"6\"], \"fpage\": [\"10\"]}, {\"surname\": [\"Rajopadhye\", \"Upadhye\"], \"given-names\": [\"A.\", \"A. S.\"], \"article-title\": [\"Estimation of Bioactive\\nCompound, Maslinic Acid by\\nHPTLC, and Evaluation of Hepatoprotective Activity on Fruit Pulp ofZiziphus\\njujubaMill. Cultivars in India\"], \"source\": [\"Evidence-Based\\nComplementary Altern.\"], \"year\": [\"2016\"], \"volume\": [\"2016\"], \"fpage\": [\"4758734\"], \"pub-id\": [\"10.1155/2016/4758734\"]}, {\"surname\": [\"Rao\", \"Rao\", \"Nitesh\", \"Raj\", \"Chandrasekhar\", \"Vijayan\", \"Dhanaraj\"], \"given-names\": [\"M.\", \"V.\", \"K.\", \"V.\", \"R.\", \"P.\", \"S.\"], \"article-title\": [\"In vitro and\\nin vivo hepatoprotective\\neffects of the total alkaloid fraction of Hygrophila auriculata leaves\"], \"source\": [\"Indian J. Pharmacol.\"], \"year\": [\"2010\"], \"volume\": [\"42\"], \"fpage\": [\"98\"], \"pub-id\": [\"10.4103/0253-7613.64500\"]}]", + "unknown_pub": "[{\"surname\": [\"Bax\", \"Ikura\", \"Kay\", \"Torchia\", \"Tschudin\"], \"given-names\": [\"A\", \"M\", \"LE\", \"DA\", \"R\"], \"article-title\": [\"Comparison of different modes of two-dimensional reverse correlation NMR for the study of proteins\"], \"source\": [\"J Magn Reson\"], \"year\": [\"1990\"], \"volume\": [\"86\"], \"fpage\": [\"304\"], \"lpage\": [\"318\"], \"pub-id\": [\"10.1016/0022-2364(90)90262-8\"]}, {\"surname\": [\"Bodenhausen\", \"Ruben\"], \"given-names\": [\"G\", \"DJ\"], \"article-title\": [\"Natural abundance nitrogen-\"], \"sup\": [\"15\"], \"source\": [\"Chem Phys Lett\"], \"year\": [\"1980\"], \"volume\": [\"69\"], \"fpage\": [\"185\"], \"lpage\": [\"189\"], \"pub-id\": [\"10.1016/0009-2614(80)80041-8\"]}, {\"mixed-citation\": [\"Clowes R, Hayes W (1968) Experiments in microbial genetics. Wiley, New York\"]}, {\"surname\": [\"Dayie\", \"Wagner\"], \"given-names\": [\"KT\", \"G\"], \"article-title\": [\"Carbonyl carbon probe of local mobility in \"], \"sup\": [\"13\", \"15\"], \"source\": [\"J Am Chem Soc\"], \"year\": [\"1997\"], \"volume\": [\"119\"], \"fpage\": [\"7797\"], \"lpage\": [\"7806\"], \"pub-id\": [\"10.1021/ja9633880\"]}, {\"surname\": [\"Kay\", \"Keifer\", \"Saarinen\"], \"given-names\": [\"LE\", \"P\", \"T\"], \"article-title\": [\"Pure absorption gradient enhanced heteronuclear single quantum correlation spectroscopy with improved sensitivity\"], \"source\": [\"J Am Chem Soc\"], \"year\": [\"1992\"], \"volume\": [\"114\"], \"fpage\": [\"10663\"], \"lpage\": [\"10665\"], \"pub-id\": [\"10.1021/ja00052a088\"]}, {\"surname\": [\"LeMaster\", \"Kushlan\"], \"given-names\": [\"DM\", \"DM\"], \"article-title\": [\"Dynamical mapping of \"], \"italic\": [\"E\", \"coli\"], \"sup\": [\"13\"], \"source\": [\"J Am Chem Soc\"], \"year\": [\"1996\"], \"volume\": [\"118\"], \"fpage\": [\"9255\"], \"lpage\": [\"9264\"], \"pub-id\": [\"10.1021/ja960877r\"]}, {\"surname\": [\"Meissner\", \"Schulte-Herbrueggen\", \"Briand\", \"Sorensen\"], \"given-names\": [\"A\", \"T\", \"J\", \"OW\"], \"article-title\": [\"Double spin-state-selective coherence transfer. Application for two-dimensional selection of multiplet components with long transverse relaxation times\"], \"source\": [\"Mol Phys\"], \"year\": [\"1998\"], \"volume\": [\"96\"], \"fpage\": [\"1137\"], \"lpage\": [\"1142\"]}, {\"surname\": [\"Milecki\"], \"given-names\": [\"J\"], \"article-title\": [\"Specific labelling of nucleosides and nucleotides with \"], \"sup\": [\"13\", \"15\"], \"source\": [\"J Label Compd Radiopharm\"], \"year\": [\"2002\"], \"volume\": [\"45\"], \"fpage\": [\"307\"], \"lpage\": [\"337\"], \"pub-id\": [\"10.1002/jlcr.553\"]}, {\"surname\": [\"Palmer\", \"Cavanagh\", \"Wright\", \"Rance\"], \"given-names\": [\"AG\", \"J\", \"PE\", \"M\"], \"article-title\": [\"Sensitivity improvement in proton -detected two-dimensional heteronuclear correlation NMR spectroscopy\"], \"source\": [\"J Magn Reson\"], \"year\": [\"1991\"], \"volume\": [\"93\"], \"fpage\": [\"151\"], \"lpage\": [\"170\"], \"pub-id\": [\"10.1016/0022-2364(91)90036-S\"]}, {\"surname\": [\"Sambrook\", \"Russell\"], \"given-names\": [\"J\", \"D\"], \"source\": [\"Molecular cloning: a laboratory manual\"], \"year\": [\"2001\"], \"publisher-loc\": [\"Cold Spring Harbor\"], \"publisher-name\": [\"Cold Spring Harbor Laboratory\"]}, {\"surname\": [\"Sattler\", \"Schwendinger\", \"Schleucher\", \"Griesinger\"], \"given-names\": [\"M\", \"MG\", \"J\", \"C\"], \"article-title\": [\"Novel strategies for sensitivity enhancement in heteronuclear multi-dimensional NMR experiments employing pulsed field gradients\"], \"source\": [\"J Biomol NMR\"], \"year\": [\"1995\"], \"volume\": [\"5\"], \"fpage\": [\"11\"], \"lpage\": [\"22\"], \"pub-id\": [\"10.1007/BF00417487\"]}, {\"surname\": [\"Schilling\", \"Edwards\", \"Letscher\", \"Palsson\"], \"given-names\": [\"CH\", \"JS\", \"D\", \"B\\u00d8\"], \"article-title\": [\"Combining pathway analysis with flux balance analysis for the comprehensive study of metabolic systems\"], \"source\": [\"Biotechnol Bioeng\"], \"year\": [\"2001\"], \"volume\": [\"71\"], \"fpage\": [\"286\"], \"lpage\": [\"306\"], \"pub-id\": [\"10.1002/1097-0290(2000)71:4<286::AID-BIT1018>3.0.CO;2-R\"]}, {\"mixed-citation\": [\"Werbelow LG (1994) Relaxation-induced transfer of nuclear spin polarization as a probe of molecular structure and dynamics in mobile phases. In: Tycko R (ed) Understanding chemical reactivity: nuclear magnetic resonance probes of molecular dynamics. Kluwer Academic Publishers, Boston, pp 223\\u2013263\"]}, {\"surname\": [\"Wijmenga\", \"van Buuren\"], \"given-names\": [\"S\", \"B\"], \"article-title\": [\"The use of NMR methods for conformational studies of nucleic acids\"], \"source\": [\"Prog Nucl Magn Reson Spectrosc\"], \"year\": [\"1998\"], \"volume\": [\"32\"], \"fpage\": [\"287\"], \"lpage\": [\"387\"], \"pub-id\": [\"10.1016/S0079-6565(97)00023-X\"]}, {\"surname\": [\"Yamazaki\", \"Muhandiram\", \"Kay\"], \"given-names\": [\"T\", \"R\", \"LE\"], \"article-title\": [\"NMR experiments for the measurement of carbon relaxation properties in highly enriched, uniformly \"], \"sup\": [\"13\", \"15\", \"13\", \"\\u03b1\"], \"source\": [\"J Am Chem Soc\"], \"year\": [\"1994\"], \"volume\": [\"114\"], \"fpage\": [\"8266\"], \"lpage\": [\"8278\"], \"pub-id\": [\"10.1021/ja00097a037\"]}]", "glossary": { "acronym": [ - "CPFP", - "CPFC", - "CPFE", - "CPFA", - "CPFW", - "DPPH", - "ABTS", - "NO", - "GAE", - "QE", - "SGOT", - "SGPT", - "ALP", - "\u03b3-GT", - "CMC", - "b.w.", - "s.c" + "AMP", + "CMP", + "UMP", + "GMP", + "R5P", + "FBP", + "F6P", + "GA3P", + "G6PDH", + "K10zwf", + "Gly", + "Ser", + "noPPP", + "OAA", + "DHAP", + "oPPP", + "rNTPs", + "TIM", + "PEP", + "G6P", + "Ru5P", + "X5P", + "S7P", + "E4P", + "3PG", + "6PG", + "6PGA" ], "definition": [ - "Clerodendrum paniculatum flower petroleum ether extract", - "Clerodendrum paniculatum flower chloroform extract", - "Clerodendrum paniculatum flower\nethyl acetate extract", - "Clerodendrum paniculatum flower\nalcohol extract", - "Clerodendrum paniculatum flower water\nextract", - "2,2-diphenyl-2-picryl\nhydrazyl", - "2,2-azino-bis\n(3-ethyl benzothiazoline-6-sufonic\nacid)", - "nitric\noxide", - "gallic\nacid equivalent", - "quercetin equivalent", - "serum glutamate oxaloacetate transaminase", - "serum glutamate pyruvate transaminase", - "alkaline phospahtase", - "gamma glutamyl\ntransferase", - "carboxy methyl cellulose", - "body weight", - "subcutaneous" + "Adenosine 5\u2032-monophosphate", + "Cytidine 5\u2032-monophosphate", + "Uridine 5\u2032-monophosphate", + "Guanosine 5\u2032-monophosphate", + "Ribose-5-phosphate", + "Fructose-6-bisphosphate", + "Fructose-6-phosphate", + "Glyceraldehyde-3-phosphate", + "Glucose-6-phosphate dehydrogenase", + "Glucose-6-phosphate dehydrogenase mutant", + "Glycine", + "Serine", + "Non-oxidative pentose phosphate pathway", + "Oxaloacetate", + "Dihydroxyacetone phosphate", + "Oxidative pentose phosphate pathway", + "Ribonucleoside triphosphates", + "Triosephosphate isomerase", + "Phosphoenolpyruvate", + "Glucose-6-phosphate", + "Ribulose-5-phosphate", + "Xylulose-5-phosphate", + "Sedoheptulose-7-phosphate", + "Erythrose-4-phosphate", + "3-Phosphoglycerate", + "6-Phosphogluconate", + "6-Phosphogluconate-\u03b4-lactone" ] }, "references": { - "introduction": { - "pmid_ref": [ - "31358053", - "31080832", - "25646037", - "23268465" - ], - "unknown_pub_ref": [ - 0, - 1, - 2, - 3 - ], - "figure_ref": [], - "table_ref": [], - "formula_ref": [], - "box_ref": [], - "code_ref": [], - "quote_ref": [], - "chem_ref": [], - "supplementary_ref": [], - "footnote_ref": [], - "graphic_ref": [], - "media_ref": [] - }, + "introduction": {...}, "methods": {...}, "results": { - "pmid_ref": [ - "34151056", - "34124456", - "28442109", - "29379645", - "28962407" - ], - "unknown_pub_ref": [ - 4 - ], - "figure_ref": [ - 0, - 1, - 2, - 3 - ], - "table_ref": [ - 0, - 1, - 2, - 3, - 4, - 5, - 6, - 7, - 7, - 8 - ], - "formula_ref": [], + "pmid_ref": ["8226637", "19049467", "20730533", "21057854", "20730533", "21057854", "8065453", "8910275", "16433544", "17116475", "19838167", "20309608", "20730533", "21057854", "16034664", "15327312", "8019138", "12913409", "18047338", "17098254"], + "unknown_pub_ref": [11, 10, 14, 3, 3, 12], + "figure_ref": [0, 0, 0, 0, 0, 1, 1, 1, 0, 1, 0, 0, 0, 2, 2, 2, 1, 2, 2, 3, 4, 3, 3, 4, 3, 3, 4, 5, 5, 6, 6, 6, 6, 6, 6], + "table_ref": [0, 0, 0, 0, 0], + "formula_ref": [0, 1, 2, 3, 4, 5, 6, 2, 4, 0, 2, 4], "box_ref": [], "code_ref": [], "quote_ref": [], @@ -375,62 +309,14 @@ a corpus of pre-annotated text for other tasks (e.g. figure caption to graphic, "footnote": {...} }, "references_text": { - "introduction": { - "pmid_ref": [ - "1", - "2", - "4", - "5" - ], - "unknown_pub_ref": [ - "3", - "6", - "7", - "8" - ], - "figure_ref": [], - "table_ref": [], - "formula_ref": [], - "box_ref": [], - "code_ref": [], - "quote_ref": [], - "chem_ref": [], - "supplementary_ref": [], - "footnote_ref": [], - "graphic_ref": [], - "media_ref": [] - }, + "introduction": {...}, "methods": {...}, "results": { - "pmid_ref": [ - "9", - "10", - "11", - "12", - "14" - ], - "unknown_pub_ref": [ - "13" - ], - "figure_ref": [ - "Figure 1", - "Figure 2", - "Figure 3", - "Figure 4" - ], - "table_ref": [ - "Table 1", - "Table 2", - "Table 3", - "Table 4", - "Table 5", - "Table 6", - "Table 7", - "Table 8", - "Table 8", - "Table 9" - ], - "formula_ref": [], + "pmid_ref": ["1993", "2008", "2010a", "b", "2010a", "b", "1994", "1996", "2006", "2006", "2009", "2010", "2010a", "b", "2005", "2004", "1994", "2003", "2007", "2007"], + "unknown_pub_ref": ["2001", "1995", "1994", "1997", "1997", "1994"], + "figure_ref": [ "1", "1", "1", "", "1", "2", "", "2", "1", "2", "1", "1", "1", "3", "3", "", "2", "3", "3", "4", "5", "4", "", "", "4", "4", "5", "6", "", "7", "7", "7", "7", "7", ""], + "table_ref": ["1", "", "1", "1", "1"], + "formula_ref": ["", "", "", "", "", "", "", "3", "5", "1", "3", "5"], "box_ref": [], "code_ref": [], "quote_ref": [], @@ -453,14 +339,12 @@ a corpus of pre-annotated text for other tasks (e.g. figure caption to graphic, "quote": {...}, "chem": {...}, "supplementary": {...}, - "footnote": {...} - }, - "n_references": 43, - "license": "CC BY-NC-ND", + "n_references": 58, + "license": "CC BY-NC", "retracted": "no", - "last_updated": "2022-01-15 12:37:30", - "citation": "ACS Omega. 2021 Sep 29; 6(40):26489-26498", - "package_file": "oa_package/ca/94/PMC8515580.tar.gz" + "last_updated": "2022-01-14 22:48:41", + "citation": "J Biomol NMR. 2011 Oct 30; 51(4):505-517", + "package_file": "oa_package/a8/33/PMC3222825.tar.gz" } ```