Comparison of metabolite levels in
\r\n\tThe WHO classification in 2007; was based on the histogenesis and cell origin of the tumor. In the latest classification made in 2016; to better characterize the tumor and obtain better data on its prognosis; The combination of molecular and genetic biomarkers and histopathological features of the tumor was used. Despite all current treatment approaches, the median survival time is around 12 months in most GBM patients. Compared with the situation of some types of successfully treated cancers; the survival time of GBM patients is not at an acceptable level today. In the treatment of CNS tumors; surgery, chemotherapy, and radiation treatments (x-rays, gamma rays, electron and proton beams) are used. The therapeutic potential of chemotherapy; New strategies are needed to increase drug concentration at the diseased site, as this largely depends on the ability of the chemotherapeutic agent to achieve effective concentrations at tumor localization. Based on our better understanding of the genetic and molecular characteristics of CNS tumors; Targeted therapies, including vaccines, and treatment protocols such as immunotherapy are promising developments.
\r\n\r\n\tThis book supposes to be written by many authors who have an internationally honored place in their field to share their ideas about the treatment of CNS tumors. Surgery, Radiotherapy, Chemotherapy and Antiangiogenic Therapy Protocols, Immunotherapy, Molecular Therapy, Specific target-agents therapy with Nanoparticles and Gene Therapy for CNS tumors among the book chapters.
\r\n\tIn these sections; there are many practical pieces of information that can help the students who graduated from the Medicine Faculty and specialist doctors who are interested in Neurosurgery.
Metabolic flux analysis (MFA) aims at the quantitation of the carbon flow through a metabolic network by measuring the enrichment and position of labels in the various measurable metabolites after feeding a labeled precursor
There are two strategies for 13C MFA: one is dynamic-labeling strategy (time course experiments), and the other is steady-state-labeling strategy. The dynamic-labeling strategy has an advantage in studying small partial networks and it is particularly effective for the analysis of secondary metabolism [1]. In this approach, a specific labeled advanced precursor of a pathway is pulse fed, and after a given time the incorporation is measured in the products of the pathway involved. In a steady-state-labeling strategy, the organisms are permanently growing in a medium containing a very early substrate for primary metabolism (e.g., a labeled sugar of pyruvate) and the diffusion of the label through all pathways is monitored by measuring the incorporation and position of the label in all measurable metabolites. This approach is usually utilized in studies of central carbon metabolism. In fact, the limiting factor in flux analyses in plants is the detection limits for the various metabolites, as the level of primary metabolites in plants is many folds higher than of secondary metabolites, the dynamic range analytical tools hamper the analysis of minor compounds. Therefore, often specific and selective extraction methods are used for the dynamic approach, whereas for the steady-state approach one uses the more general metabolomics analytical protocols.
\nIn
In this study, the fate of [1-13C] glucose fed to the intact
Label pattern of TCA cycle, amino acids, loganic acids, chlorogenic acid, and 4-
Murashige & Skoog medium (including vitamins) and gelrite (strength: 550–850 g/cm2) were purchased from Duchefa Biochemie. D (+)-Glucose (>99.0%) was obtained from Fluka Chemie (Buchs, Germany), whereas [1-13C]-D-glucose (>99.0%, with > 99% atom 1-13C) was from Campro Scientific (Veenendaal, The Netherlands). Jasmonic acid (JA) was from Sigma-Aldrich Chemie (Steinheim, Germany).
A stock solution of 10 mg/ml of JA 40% EtOH was prepared, filter sterilized, and used for elicitation. After 5 days of submerging the plant roots with 5 ml of [1-13C] glucose solution (1%, w/v), 11 μl of the JA stock solution was aseptically spiked into each tube. The control samples received only the same volume of 40% EtOH. The plants were harvested at 0, 6, 24, and 72 h after treatment; young leaves, old leaves, stems, and roots of
1H NMR spectra were recorded in CH3OH-
The signal intensity of carbons at certain positions of a given metabolite was obtained from peak height in the 13C-dimension spectra abstracted from the 2D HSQC spectra. The signal height of CH3OH-
Two batches of
After 10–12 days, seeds in both batches germinated and produced their first pair of leaves. In the first 3 weeks after germination, there were no significant differences of height, leaf pairs, and leaf size between plantlets grown in MS medium and in the soil (Figure 2). However, in the following days, the plantlets in MS medium provided one more pair of leaves than those in soil did, but the leaf size was much smaller than that of plantlets grown in the soil (Figure 2A and B). Moreover, the soil plantlets grew higher than those grown in MS medium (Figure 2C). Plantlets in MS medium entered flowering time around 100 days after sowing, whereas those in soil flowered at 75 days. The plantlets grown in soil had a higher growth rate and a larger biomass than those grown in MS medium.
Comparison of the number of leaf pairs (A), leaf size (B), and height (C) of
Metabolic differences between the plants grown in soil and MS medium were observed by 1H NMR (Figure 3). The 1H NMR spectra showed that qualitatively metabolites of plants grown in soil or MS medium were similar, but the levels varied (Table 1). Plants grown in soil produced higher levels of organic acids and sugars (malate, fumaric acid, glucose, and sucrose) than those grown in MS medium, indicating a low function/reduced level of carbon fixation in the leaves of the MS-grown plants. Also, secondary metabolites (such as secologanin, vindoline, quercetin, and kaempherol) were found in higher levels in soil-grown plants than the plants grown in MS medium. On the other hand, plants cultured in MS medium displayed significantly higher levels of arginine, glutamine, and asparagine but relatively low level of glucose and sucrose. The levels of threonine, glutamate, quinic acid, and lactic acid were also higher in plants grown in MS medium than those in soil.
Compounds | \nSignal intensity | \nSignal intensity ratio (S/M) | \n|
---|---|---|---|
\n | Soil (S) | \nMedium (M) | \n\n |
Vindoline | \n0.42 | \n0.22 | \n1.91 | \n
Threonine | \n0.27 | \n0.34 | \n0.79 | \n
Lactic acid | \n0.14 | \n0.22 | \n0.64 | \n
Alanine | \n0.54 | \n0.51 | \n1.06 | \n
Arginine | \n0.82 | \n4.27 | \n0.19 | \n
Quinic acid | \n0.21 | \n0.30 | \n0.70 | \n
Glutamate | \n1.25 | \n2.14 | \n0.58 | \n
Glutamine | \n2.94 | \n7.08 | \n0.42 | \n
Malate | \n5.96 | \n1.82 | \n3.27 | \n
Asparagine | \n0.03 | \n0.45 | \n0.07 | \n
β-glucose | \n0.67 | \n0.13 | \n5.15 | \n
α-glucose | \n0.42 | \n0.08 | \n5.25 | \n
Sucrose | \n1.34 | \n0.39 | \n3.44 | \n
Chlorogenic acid | \n0.12 | \n0.11 | \n1.09 | \n
Fumaric acid | \n0.10 | \n0.07 | \n1.43 | \n
Catharanthine | \n0.20 | \n0.17 | \n1.18 | \n
4- | \n0.15 | \n0.12 | \n1.25 | \n
Quercetin-3- | \n0.10 | \n0.04 | \n2.50 | \n
Kaempherol | \n0.13 | \n0.08 | \n1.63 | \n
Secologanin | \n0.13 | \n0.03 | \n4.33 | \n
Comparison of metabolite levels in
1H NMR spectrum of the crude extracts of
Some groups of metabolites have a close correlation with plant growth and biomass, such as the tricarboxylic acid cycle (TCA) intermediates succinate, citrate, or malate, as well as amino acids [11]. Both glutamine and asparagine are the major compounds for nitrogen fixing, transport, and storage in plants [12]. With the much more abundant nitrogen source in the medium than in the soil, the high levels of the amino acids in the medium grown plants could be explained. Meyer et al. [11] reported a negative correlation to the plant biomass with glutamine, which is in line with our findings. Sucrose starvation may lead to the presence of a large excess of asparagine in plant cells [13]. In the present study, the plants cultured on solid MS medium require an aseptic jar with a cap, which limits the space to grow, and also affects air exchange, CO2 availability, and accumulation of volatiles in the head space if compared with plants grown in soil. Despite the uptake of carbohydrates from the medium through the roots, the growth was less than the plants grown in soil which are dependent of carbon fixation by leaves. The limited availability of CO2 in the sterile closed containers may thus be a reason for lower biomass production.
Samples from different organs (upper and lower leaves, stems, and roots) were measured by proton and carbon NMR. After feeding the plants with [1-13C] glucose for 5 days, the incorporation of 13C label was found in some primary and secondary metabolites detected in all organs of the
2-D [13C, 1H] HSQC spectrum of CH3OH-
Figure 5 shows the 13C-dimension HSQC spectra and 1H NMR spectra of the non-labeled sample and the 13C-enriched sample determined in CH3OH-
Compounds | \nRatio of metabolite levels in labeled and non-labeled samples, (L0/C0), based on 1H NMR | \n|||
---|---|---|---|---|
\n | Upper leaf | \nLower leaf | \nStem | \nRoot | \n
β-Glucose | \n1.04 | \n0.76 | \n1.01 | \n0.42 | \n
α-Glucose | \n0.97 | \n0.67 | \n1.03 | \n0.39 | \n
Sucrose | \n0.95 | \n0.62 | \n0.82 | \n0.65 | \n
Threonine | \n1.25 | \n0.97 | \n0.88 | \n1.05 | \n
Alanine | \n1.12 | \n0.92 | \n0.96 | \n0.82 | \n
Arginine | \n1.16 | \n0.75 | \n0.82 | \n0.74 | \n
Glutamate | \n1.13 | \n0.95 | \n0.89 | \n0.70 | \n
Glutamine | \n0.78 | \n0.60 | \n0.89 | \n0.81 | \n
Aspartate | \n1.28 | \n0.83 | \n0.87 | \n1.22 | \n
Asparagine | \n0.88 | \n0.85 | \n0.77 | \n0.93 | \n
Malic acid | \n1.63 | \n0.85 | \n0.98 | \n0.95 | \n
Fumaric acid | \n0.90 | \n0.77 | \n0.67 | \nnd | \n
Vindoline | \n1.07 | \n0.89 | \n1.20 | \nnd | \n
Chlorogenic acid | \n1.54 | \n0.85 | \n0.75 | \nnd | \n
4- | \n1.16 | \n0.90 | \n1.00 | \nnd | \n
Quercetin | \n1.50 | \n1.33 | \nnd | \nnd | \n
Kaempferol | \n1.30 | \n1.00 | \nnd | \nnd | \n
Secologanin | \n1.50 | \n1.00 | \nnd | \nnd | \n
Loganic acid | \n1.10 | \n1.00 | \n0.94 | \n0.89 | \n
Comparison of metabolite levels in different organs between labeled and non-labeled
nd, not detected.
Superimposed 1H NMR spectra and 13C-dimension HSQC spectrum of labeled and non-labeled
The signals in the HSQC spectra of the enriched samples were identified (Figure 4). The carbon position of 13C incorporation into a metabolite was investigated by calculating 13C signal intensity ratios between the same carbons of the metabolite in labeled and non-labeled samples (Table 3).
Compound | \nChemical Shift | \nPeak height | \nRelative intensity to CH3OH- | \nRelative enrichment ratio ( | \n|||
---|---|---|---|---|---|---|---|
\n | H | \nC | \nControl | \nLabeled | \nControl ( | \nLabeled ( | \n\n |
Glucose | \n4.58 | \n97.04 | \n3.2E+07 | \n1.3E+08 | \n0.17 | \n2.89 | \n17.31 | \n
Alanine | \n1.49 | \n16.98 | \n8.0E+06 | \n2.1E+07 | \n0.04 | \n0.47 | \n11.26 | \n
Glutamine | \n2.05 | \n27.11 | \n4.6E+07 | \n3.9E+07 | \n0.23 | \n0.88 | \n3.75 | \n
\n | 2.38 | \n31.83 | \n1.6E+08 | \n1.7E+08 | \n0.80 | \n3.72 | \n4.63 | \n
\n | 3.71 | \n55.02 | \n3.0E+08 | \n2.3E+08 | \n1.56 | \n5.12 | \n3.29 | \n
Glutamate | \n2.14 | \n27.74 | \n2.1E+06 | \n7.0E+06 | \n0.01 | \n0.16 | \n14.81 | \n
\n | 2.46 | \n34.44 | \n6.3E+07 | \n1.5E+08 | \n0.32 | \n3.36 | \n10.45 | \n
\n | 3.72 | \n55.67 | \n9.8E+06 | \n3.9E+07 | \n0.05 | \n0.86 | \n17.14 | \n
Arginine | \n1.72 | \n24.9 | \n1.7E+07 | \n6.7E+06 | \n0.09 | \n0.15 | \n1.77 | \n
\n | 1.92 | \n28.53 | \n2.1E+07 | \n8.1E+06 | \n0.11 | \n0.18 | \n1.68 | \n
\n | 3.24 | \n41.38 | \n5.9E+07 | \n2.1E+07 | \n0.30 | \n0.47 | \n1.56 | \n
Aspartate | \n2.64 | \n37.21 | \n1.0E+07 | \n7.0E+06 | \n0.05 | \n0.16 | \n3.05 | \n
Asparagine | \n2.83,2.96 | \n35.23 | \n1.2E+08 | \n4.0E+07 | \n0.62 | \n0.90 | \n1.44 | \n
\n | 3.96 | \n52.21 | \n1.7E+08 | \n4.7E+07 | \n0.89 | \n1.05 | \n1.19 | \n
Threonine | \n1.34 | \n20.47 | \n8.8E+06 | \n6.2E+06 | \n0.05 | \n0.14 | \n3.08 | \n
Malate | \n2.35,2.72 | \n43.4 | \n6.0E+07 | \n8.5E+07 | \n0.31 | \n1.91 | \n6.21 | \n
Chlorogenic acid | \n7.07 | \n123.12 | \n1.2E+07 | \n7.2E+06 | \n0.06 | \n0.16 | \n2.53 | \n
4- | \n7.09 | \n124.16 | \n1.7E+06 | \n2.5E+06 | \n0.01 | \n0.06 | \n6.63 | \n
Loganic acid | \n1.07 | \n12.69 | \n2.5E+06 | \n5.6E+06 | \n0.01 | \n0.13 | \n9.96 | \n
\n | 7.03 | \n146.1 | \n6.5E+05 | \n5.2E+06 | \n0.00 | \n0.12 | \n34.49 | \n
Vindoline | \n0.49 | \n7.43 | \n6.0E+06 | \n2.4E+06 | \n0.03 | \n0.05 | \n1.76 | \n
The chemical shifts, peak height, and relative enrichment ratio of the same carbon signals in metabolites in labeled and non-labeled
Among amino acids, the signals corresponding to C at δ 16.98, C-3 of alanine, exhibited a high 13C relative enrichment ratio. Glycolysis introduces the C-1 or C-6 of glucose into alanine C-3 [8]. Carbon signals at δ 20.47 of threonine and at δ 37.21 of aspartate also showed a relatively high labeling. The carbons of arginine and asparagine were apparently less labeled.
\nGlutamate (C-3 at δ 27.74, C-4 at δ 34.44, and C-5 at δ 55.67) and glutamine (C-3 at δ 27.11, C-4 at δ 31.83, and C-5 at δ 55.02) showed clear high 13C incorporation. The relative enrichment ratios of C-3 and C-2 of glutamine were lower than that of C-4, which indicate the entry of a diluting flux of C4 compounds into the TCA cycle [17]. For glutamate, however, C-4 had a lower relative enrichment ratio than C-3 and C-2. Non-symmetrical enrichment ratios of C-2 and C-3 imply that there might be a form of channeling that converts oxoglutarate C-4 to oxaloacetate C-2 or C-3 [18].
\nIn plant cells, the labeling of amino acids alanine, glutamate, and aspartate is found to reflect that of the corresponding α-oxoacids: pyruvate, α-oxoglutarate, and oxaloacetate, respectively [19]. The organic acid malate showed a sixfold increased intensity for the carbon signal at δ 43.40.
\nBesides primary metabolites, secondary metabolites also exhibited clear 13C incorporation. Two phenylpropanoids, chlorogenic acid and its isomer 4-
Based on 1H NMR spectra, relative levels of primary and secondary metabolites in different organs were calculated by normalizing the integral of signal peaks to the internal standard (TSP). Table 4 showed that leaves, especially upper leaves, contained higher levels of amino acids, phenylpropanoids, iridoids, and vindoline, than stems and roots. In roots, phenylpropanoids and vindoline, which are biosynthesis dependent on chloroplasts, were not detected, whereas iridoids displayed a much lower level in roots while glucose and sucrose had relatively higher levels than in other organs.
Compounds | Relative levels of metabolites | |||
---|---|---|---|---|
Upper leaf | Lower leaf | Stem | Root | |
β-Glucose | 0.46 | 0.89 | 0.95 | 1.36 |
α-Glucose | 0.34 | 0.56 | 0.62 | 0.83 |
Sucrose | 0.20 | 0.33 | 0.53 | 0.81 |
Threonine | 0.45 | 0.44 | 0.45 | 0.17 |
Alanine | 0.45 | 0.46 | 0.29 | 0.17 |
Arginine | 3.19 | 5.93 | 5.09 | 1.07 |
Glutamate | 3.05 | 2.15 | 2.92 | 0.91 |
Glutamine | 3.72 | 1.74 | 7.91 | 1.38 |
Aspartate | 1.07 | 0.93 | 0.41 | 0.22 |
Asparagine | 2.82 | 1.25 | 1.84 | 0.44 |
Malic acid | 0.32 | 0.40 | 0.22 | 0.09 |
Fumaric acid | 0.11 | 0.11 | 0.02 | nd |
Vindoline | 0.28 | 0.09 | 0.05 | nd |
Chlorogenic acid | 0.25 | 0.37 | 0.03 | nd |
4- | 0.17 | 0.15 | 0.04 | nd |
Quercetin | 0.05 | 0.05 | 0.03 | 0.03 |
Kaempferol | 0.09 | 0.10 | 0.02 | nd |
Secologanin | 0.02 | 0.02 | 0.0006 | 0.0002 |
Loganic acid | 0.07 | 0.14 | 0.11 | 0.05 |
Relative level of metabolites in different organs of
The incorporation of 13C in different organs (upper leaf, lower leaf, stem, and root) was also investigated by comparison of relative enrichment ratios in order to have a clue about the accumulation of label in different organs and its connection with transport and compartmentation of the pathways in the plants (Table 5). From the 13C dimension of HSQC spectra of all organs, 13C signals of labeled samples showed an apparently higher intensity in the amino acid and sugar areas than those of non-labeled ones (Figure 6), which indicated that 13C-isotope was efficiently incorporated into the primary metabolism of intact
Compounds | \n13C chemical shift | \nRelative enrichment ratio (labeled:control) | \n|||
---|---|---|---|---|---|
\n | \n | Upper leaf | \nLower leaf | \nstem | \nRoot | \n
Alanine | \n16.98 | \n9.76 | \n32.38 | \n118.89 | \n84.86 | \n
Threonine | \n20.47 | \n3.55 | \n1.06 | \n1.39 | \n18.91 | \n
Arginine | \n24.9 | \n1.89 | \n1.12 | \n1.67 | \n2.99 | \n
\n | 28.53 | \n1.58 | \n1.07 | \n1.17 | \n1.15 | \n
\n | 41.38 | \n1.52 | \n0.91 | \n0.91 | \n1.66 | \n
Glutamine | \n27.11 | \n2.80 | \n2.34 | \n1.89 | \n6.38 | \n
\n | 31.83 | \n3.21 | \n4.03 | \n3.09 | \n6.70 | \n
\n | 55.02 | \n2.34 | \n1.38 | \n2.01 | \n8.43 | \n
Glutamate | \n27.74 | \n11.79 | \n5.39 | \n3.36 | \n20.65 | \n
\n | 34.44 | \n7.21 | \n3.37 | \n4.63 | \n16.75 | \n
\n | 55.67 | \n15.49 | \n2.72 | \n5.11 | \n25.06 | \n
Asparagine | \n35.23 | \n1.21 | \n0.75 | \n1.09 | \n5.67 | \n
\n | 52.21 | \n1.17 | \n0.95 | \n1.23 | \n4.18 | \n
Aspartate | \n37.21 | \n2.25 | \n3.40 | \n4.07 | \n36.29 | \n
Malate | \n43.4 | \n4.67 | \n4.41 | \n7.82 | \n26.51 | \n
β-glc | \n97.04 | \n30.96 | \n55.55 | \n15.01 | \n32.79 | \n
Vindoline | \n7.43 | \n2.96 | \nnd | \nnd | \nnd | \n
Loganic acid | \n12.69 | \n7.66 | \n3.75 | \n4.26 | \n23.62 | \n
\n | 146.1 | \n27.96 | \n13.77 | \n6.59 | \n24.79 | \n
Chlorogenic acid | \n123.12 | \n2.88 | \n1.42 | \nnd | \nnd | \n
\n | 146.8 | \n93.69 | \nnd | \nnd | \nnd | \n
4- | \n124.16 | \n10.35 | \nnd | \nnd | \nnd | \n
Relative enrichment ratios of the carbons of some metabolites in different organs of
nd, not detected.
13C dimension of HSQC spectra of amino acids (δ 10–55 ppm) and secondary metabolites (δ 105–150 ppm) in different organs of
Based on the 13C dimension of HSQC spectra, leaves had more 13C signals in the area of >δ 100 ppm than stems and roots (Figure 6), even after feeding [1-13C] glucose. Upper leaves had relatively high 13C incorporation for vindoline, chlorogenic acid, and 4-
In stems and roots, no 13C signals of vindoline, chlorogenic acid, and 4-
JA was spiked into the labeled glucose solution at the sixth day after submerging the plant roots in the solution. The control plants were also reared in labeled glucose solution but without JA elicitation. Leaves were harvested at 0, 6, 24, and 72 h (6, 7, and 9 d of incubation with the labeled glucose solution) after elicitation and measured by 1H NMR and HSQC.
\nFor control plants, NMR spectra showed that the enrichments of malic acid and of the amino acids alanine, arginine, glutamate, glutamine, aspartate, and asparagine in the leaves were nearly identical at 6 and 9 d of incubation with the labeled glucose solution (Figure 7), suggesting the establishment of steady state at 6 d. However, the incorporation of label in glucose and threonine increased continuously within the measured period of 9 days. Besides, loganic acid and chlorogenic acid kept the same enrichments while vindoline and 4-
Relative enrichment ratio of primary and secondary metabolites during incubation of
JA elicitation had little effect on the level of most metabolites, except glutamate, glutamine, vindoline, and loganic acid. Although JA induced an increase of glutamate and glutamine levels (Figure 8), their relative enrichment ratio remained unchanged compared with the controls. At the same time, the enrichment of alanine at C-3 showed an increase without levels changing compared to the controls. Vindoline levels showed an increase and reached the highest level at 72 h (23% higher than the controls) after JA treatment (Figure 8). However, the relative enrichment ratio of the C-18 signal of vindoline was lower in JA-elicited samples than in the controls, especially at 6 h (Figure 7). The level of loganic acid decreased with time (Figure 8), leading to a dramatical decrease of its enrichment at both C-3 and C-10 from 6 to 72 h. The levels of chlorogenic acid and 4-
Relative levels of metabolites in
Metabolic flux analysis is the quantification of all intracellular fluxes in an organism, which is thus an important cornerstone of metabolic engineering and systems biology. This study reports a comprehensive 13C-labeling-based metabolomics of a plant system. [1-13C] glucose was efficiently absorbed via the root system and recycled in the whole plant of
Skin application of vitamin K shows several beneficial effects, such as suppression of pigmentation and alleviation of bruising [1, 2, 3], prophylactically limiting the occurrence of acneiform side effects in patients receiving the monoclonal antibody cetuximab [4, 5, 6] and promoting wound healing [7].
Despite these potentially beneficial effects, vitamin K is unstable in the presence of light [8, 9]. Indeed, application of vitamin K on the skin can result in photodegradation without appropriate shielding of the application site, e.g., the face and hands, from light. Furthermore, in Europe, warnings have been issued regarding the use of vitamin K in cosmetics. The Scientific Committee on Consumer Safety has also reported phototoxicity of vitamin K in skin cells [10, 11]. As a result, the use of vitamin K as an external preparation for the skin is limited.
The molecules in the vitamin K family contain 2-methyl-1,4-naphthoquinone as the basic skeleton. Vitamin K molecules can be classified as phylloquinone (PK, vitamin K1) with a phytyl side chain at the 3-position, menaquinone (MK-n, vitamin K2) with an isoprenyl side chain consisting of n isoprenyl groups, and menadione (MD, vitamin K3) with no side chain at the 3-position. PK and MK-4 are widely used as pharmaceutical treatments for vitamin K deficiency and osteoporosis.
Vitamin K (quinone form) delivered intracellularly is converted into vitamin K hydroquinone (VKH) by two-electron reduction. VKH functions as a cofactor for γ-glutamyl carboxylase (GGCX), which converts the glutamic acid (Glu) residue of vitamin K-dependent protein into the γ-carboxyglutamic acid (Gla) residue as a post-translational modification. Subsequently, VKH is oxidized to vitamin K epoxide (VKO). In addition, VKO is reduced to vitamin K (quinone form) to form the vitamin K cycle. Therefore, it is necessary to deliver sufficient VKH to the target site to achieve efficacy.
The active forms of PK and MK-4 are phyllohydroquinone (PKH) and menahydroquinone-4 (MKH), respectively. However, these compounds cannot be used as preparations because they show extreme instability via oxidation. Therefore, PK and MK-4 that are stable against oxidation are used clinically. However, as described above, vitamin K (quinone form) is extremely unstable upon exposure to light. Thus, strict control of lighting is required during formulation, distribution, storage at medical institutions, and administration to patients. To achieve full efficacy of vitamin K, formulations with low photodegradation and phototoxicity are needed for effective topical delivery of VKH. The concepts underlying the delivery system of VKH using Vitamin K are shown in Figure 1.
Schematic illustration of the vitamin K cycle.
Analysis of the PK photolysis reaction by Hangarter et al. showed that charge transfer from the β,γ-double bond of the isoprenyl side chain to the quinone moiety initiates intramolecular proton transfer from the side chain and yields a 1,3-quinone methide (meta-quinone methide) as a mixture of singlet and triplet species diradical in polar solvents, subsequently forming 1,2-quinone methide (ortho-quinone methide), which can be used to generate PK chromenol [9]. Chromenol levels tend to increase with irradiation time, and this compound is expected to be the final product of photodegradation of vitamin K [9, 12, 13].
Vitamin K is also expected to cause two types of phototoxicity during the abovementioned series of photodegradation processes. After acquiring the excited state via light absorption, some chemicals cause oxidative damage to biological components, such DNA and proteins, through the generation of free radicals (type I reaction) by the electron rearrangement reaction and the generation of singlet oxygen from triplet oxygen (ground state) by the energy rearrangement reaction (type II reaction) [14, 15].
We have previously confirmed that irradiation of PK and MK-4 with UVA increases singlet oxygen generation, intracellular reactive oxygen species (ROS) generation, and cytotoxicity in HaCaT human keratinocytes. Thus, vitamin K has phototoxic properties [12, 13]. Moreover, 1,3-quinone methide diradical and 1,2-quinone methide, which are produced during the photodegradation of vitamin K, are highly reactive and show phototoxicity via type I reactions. Additionally, singlet oxygen generation through a type II reaction is an early-stage phototoxic reaction that generates additional secondary ROS [16]. Singlet oxygen is also known to promote the peroxidation of skin surface lipids, resulting in the induction of skin inflammation [17]. Because no enzymes are known to scavenge singlet oxygen in the body, singlet oxygen is thought to exhibit extremely strong cytotoxicity. Figure 2 shows a schematic diagram of the processes of vitamin K photodegradation and phototoxicity.
Schematic diagram of vitamin K photodegradation and phototoxicity.
Vitamin K photodegradation and phototoxicity are derived from its quinone structure [9]. Therefore, as far as it has a quinone structure, the photoreaction is unavoidable. We have previously synthesized VKH derivatives without a quinone structure in which the two hydroxyl groups of the VKH are protected by ester bonds. In addition, we have previously reported cationic VKH derivatives which have
Structure of the VKH derivatives and hydrolytic of VKH derivatives to VKH.
Here, we assessed their photostability and phototoxicity in order to further development of a vitamin K skin application.
The ethanol solutions of PK, MK-4, phyllohydroquinone derivatives (PKH-DMG and PKH-SUC), and menahydroquinone-4 derivatives (MKH-DMG and MKH-SUC) in quartz cells were exposed to artificial sunlight (12000 lx) from the vertical direction at 25°C with and without shading. The residual concentrations were determined by liquid chromatography tandem mass spectrometry [12, 13].
All samples were photodegraded according to the apparent first-order rate equation by artificial sunlight irradiation, and their apparent first order rate constants (
Compounda | Irradiation conditions | ||
---|---|---|---|
PK | Sunlight | 5.532 | 0.125 |
Shadingb | -c | -c | |
PKH-DMG | Sunlight | 0.140 | 4.950 |
Shadingb | -c | -c | |
PKH-SUC | Sunlight | 1.219 | 0.569 |
Shadingb | 0.577 | 1.201 | |
MK-4 | Sunlight | 8.239 | 0.084 |
Shadingb | -c | -c | |
MKH-DMG | Sunlight | 0.167 | 4.150 |
Shadingb | -c | -c | |
MKH-SUC | Sunlight | 2.796 | 0.248 |
Shadingb | 0.883 | 0.785 |
Apparent first order rate constants (
The initial concentration was 1 μM in ethanol.
During irradiation, the compound was covered with aluminum foil to provide shade.
No decomposition.
The half-lives of PK and MK-4 irradiated with artificial sunlight were 0.125 and 0.08 h, respectively. Moreover, the half-lives of PKH-SUC and MKH-SUC were approximately 5- and 3-fold more stable than those of PK and MK-4, respectively, although the stability was not greatly improved. In contrast, the half-life of PKH-DMG was approximately 40-fold greater than that of PK, and the half-life of MKH-DMG was approximately 50-fold greater than that of MK-4, supporting that high light stability could be ensured against artificial sunlight. Note that no formation of chromenol was observed from irradiating the VKH derivatives.
The wavelength distribution of sunlight is wide from ultraviolet to infrared. To examine the wavelength characteristics of photodegradation, the photostability of quinone-type vitamin K and VKH derivatives was evaluated after irradiation with monochromatic light at 279, 341, 373, 404, or 435 nm. Table 2 shows the photodegradation rate (
Compounda | Wavelength (nm) | ||
---|---|---|---|
PK | 279 | 0.549 | 1.262 |
341 | 0.359 | 1.933 | |
373 | 0.094 | 7.390 | |
404 | 0.026 | 26.260 | |
435 | 0.021 | 32.459 | |
PKH-DMG | 279 | 0.146 | 4.750 |
341 | -b | -b | |
373 | -b | -b | |
404 | -b | -b | |
435 | -b | -b | |
PKH-SUC | 279 | 0.137 | 5.047 |
341 | 0.070 | 9.889 | |
373 | 0.076 | 9.169 | |
404 | 0.078 | 8.860 | |
435 | 0.079 | 8.828 | |
MK-4 | 279 | 0.533 | 1.301 |
341 | 0.422 | 1.643 | |
373 | 0.151 | 4.583 | |
404 | 0.049 | 15.800 | |
435 | 0.035 | 19.738 | |
MKH-DMG | 279 | 0.146 | 4.750 |
341 | -b | -b | |
373 | -b | -b | |
404 | -b | -b | |
435 | -b | -b | |
MKH-SUC | 279 | 0.110 | 6.323 |
341 | 0.069 | 10.036 | |
373 | 0.059 | 11.792 | |
404 | 0.061 | 11.296 | |
435 | 0.068 | 10.253 |
The rate constants (
The initial concentration was 1 μM in ethanol.
No decomposition.
The above results clearly confirmed that the VKH derivative is more stable to sunlight than vitamin K (quinone form) and has a narrow wavelength range for photodegradation.
To confirm whether the VKH derivatives had phototoxic properties, singlet oxygen generation, intracellular ROS generation, and cytotoxicity after irradiation with UVA were evaluated in HaCaT cells [12, 13].
Figure 4 shows the amounts of singlet oxygen produced by each compound (200 μM) irradiated with UVA (15 J/cm2) in phosphate-buffered saline (PBS). Ketoprofen was used as a positive control, and sulisobenzone was used as a negative control. Vitamin K (quinone form) showed singlet oxygen generation depending on UVA irradiation energy, whereas VKH derivatives showed almost no singlet oxygen generation.
Singlet oxygen generation in aqueous solutions of various compounds (200 μM) exposed to UVA (15 J/cm2). Data represent means ± standard deviations (n = 3).
Analysis of intracellular ROS generation and cell viability following UVA irradiation at the time of MK-4, MKH-DMG, or MKH-SUC addition (50 μM) in HaCaT cells is shown in Table 3. MK-4 irradiated with UVA (5 J/cm2) increased intracellular ROS generation and decreased cell viability, whereas the MKH derivative did not. Similar trends were observed with PK and PKH derivatives [12].
Compounda | UVA irradiation intensity (J/cm2) | ROS generation (%control without UVA) | Cell viabilityb (%control without UVA) |
---|---|---|---|
Control | 0 | 100 ± 6.19 | 100 ± 6.04 |
5 | 275 ± 22.0 | 86.6 ± 5.44 | |
MK-4 | 0 | 99.2 ± 6.87 | 104 ± 2.40 |
5 | 1150 ± 131 | 11.3 ± 2.17 | |
MKH-DMG | 0 | 126 ± 6.63 | 104 ± 1.93 |
5 | 282 ± 4.40 | 101 ± 2.00 | |
MKH-SUC | 0 | 125 ± 4.54 | 93.9 ± 4.79 |
5 | 271 ± 15.5 | 88.8 ± 6.07 |
Percent of intracellular ROS generation and cell viability in HaCaT cells in aqueous solutions of vitamin K and VKH derivatives with or without UVA irradiation (5 J/cm2).
Cells were treated with PBS containing 50 μM MK-4, MKH-DMG, or MKH-SUC. Data represent means ± standard deviations (n = 3).
Cell viability was measured at 24 h after UVA irradiation.
As mentioned above, the photodegradation and phototoxicity of vitamin K (quinone form) is charge transfer from the β, γ-double bond of the isoprenyl side chain to the quinone moiety initiates intramolecular proton transfer from the side chain. Since VKH derivatives in which the two hydroxyl groups were protected by ester bond, it is considered that the charge transfer from the isoprenyl side chain that triggers a photochemical reaction was suppressed. These results strongly supported that VKH derivatives without a quinone structure did not show the same photodegradation and phototoxicity as vitamin K (quinone form) and may therefore be applied topically to the skin.
To confirm whether VKH derivatives function as VKH prodrugs in skin-derived cells, the delivery properties of VKH to HaCaT cells were evaluated [12, 13]. Table 4 shows the area under the curve (AUC) of intracellular VKO up to 72 h after the administration of MK-4 and VKH derivatives to HaCaT cells. VKO was used as an index of VKH because it is stoichiometrically produced from VKH after functioning as a cofactor for GGCX.
Compounda | AUCVKO(0–72h) (nmol × h/mg of protein) |
---|---|
PK | 1.176 ± 0.056 |
PKH-DMG | 0.872 ± 0.138 |
PKH-SUC | 26.967 ± 2.030 |
MK-4 | 10.543 ± 0.795 |
MKH-DMG | 10.786 ± 1.696 |
MKH-SUC | 17.304 ± 1.068 |
Area under the curve over 72 h of VKO treated with PK, PKH derivatives, MK-4 or MKH derivatives in HaCaT cells.
Cells were treated with medium containing 5 μM compounds. Data represent means ± standard deviations (n = 3).
The AUCVKO (0-72h) values of PKH-DMG and PKH-SUC were 0.741- and 22.9-fold higher than that of PK, respectively. Additionally, the AUCVKO (0-72h) values of MKH-DMG and MKH-SUC were 1.02- and 1.64-fold higher than that of MK-4, respectively.
Based on these findings, vitamin K (quinone form) and VKH derivatives are converted to VKH in HaCaT cells and function as cofactors for GGCX. Thus, VKH derivatives can function as prodrugs of VKH. Furthermore, VKH derivatives could deliver VKH at concentrations equal to or higher than vitamin K (quinone form).
Although many studies have supported the application of vitamin K for the treatment of skin pathologies, this compound is difficult to use as an external preparation to the skin owing to its photo-instability and phototoxic properties. The photodegradation and phototoxicity of vitamin K are derived from its quinone structure. Avoiding chromenol formation may suppress photodegradation via singlet oxygen and radical formation. Moreover, VKH derivatives in which the quinone structure is protected by ester bonds do not show chromenol formation and can be used to overcome the photo-instability and phototoxicity associated with vitamin K while promoting VKH delivery to skin cells. Thus, VKH derivatives may be used for application to sites where shading may be difficult, as alternatives to vitamin K (quinone form).
This work was funded by JSPS KAKENHI (grant number: 20 K16066).
The authors declare no conflict of interest.
IntechOpen implements a robust policy to minimize and deal with instances of fraud or misconduct. As part of our general commitment to transparency and openness, and in order to maintain high scientific standards, we have a well-defined editorial policy regarding Retractions and Corrections.
",metaTitle:"Retraction and Correction Policy",metaDescription:"Retraction and Correction Policy",metaKeywords:null,canonicalURL:"/page/retraction-and-correction-policy",contentRaw:'[{"type":"htmlEditorComponent","content":"IntechOpen’s Retraction and Correction Policy has been developed in accordance with the Committee on Publication Ethics (COPE) publication guidelines relating to scientific misconduct and research ethics:
\\n\\n1. RETRACTIONS
\\n\\nA Retraction of a Chapter will be issued by the Academic Editor, either following an Author’s request to do so or when there is a 3rd party report of scientific misconduct. Upon receipt of a report by a 3rd party, the Academic Editor will investigate any allegations of scientific misconduct, working in cooperation with the Author(s) and their institution(s).
\\n\\nA formal Retraction will be issued when there is clear and conclusive evidence of any of the following:
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\\n\\n1.2. REMOVALS AND CANCELLATIONS
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\\n\\nA Statement of Concern detailing alleged misconduct will be issued by the Academic Editor or publisher following a 3rd party report of scientific misconduct when:
\\n\\nIntechOpen believes that the number of occasions on which a Statement of Concern is issued will be very few in number. In all cases when such a decision has been taken by the Academic Editor the decision will be reviewed by another editor to whom the author can make representations.
\\n\\n3. CORRECTIONS
\\n\\nA Correction will be issued by the Academic Editor when:
\\n\\n3.1. ERRATUM
\\n\\nAn Erratum will be issued by the Academic Editor when it is determined that a mistake in a Chapter originates from the production process handled by the publisher.
\\n\\nA published Erratum will adhere to the Retraction Notice publishing guidelines outlined above.
\\n\\n3.2. CORRIGENDUM
\\n\\nA Corrigendum will be issued by the Academic Editor when it is determined that a mistake in a Chapter is a result of an Author’s miscalculation or oversight. A published Corrigendum will adhere to the Retraction Notice publishing guidelines outlined above.
\\n\\n4. FINAL REMARKS
\\n\\nIntechOpen wishes to emphasize that the final decision on whether a Retraction, Statement of Concern, or a Correction will be issued rests with the Academic Editor. The publisher is obliged to act upon any reports of scientific misconduct in its publications and to make a reasonable effort to facilitate any subsequent investigation of such claims.
\\n\\nIn the case of Retraction or removal of the Work, the publisher will be under no obligation to refund the APC.
\\n\\nThe general principles set out above apply to Retractions and Corrections issued in all IntechOpen publications.
\\n\\nAny suggestions or comments on this Policy are welcome and may be sent to permissions@intechopen.com.
\\n\\nPolicy last updated: 2017-09-11
\\n"}]'},components:[{type:"htmlEditorComponent",content:'IntechOpen’s Retraction and Correction Policy has been developed in accordance with the Committee on Publication Ethics (COPE) publication guidelines relating to scientific misconduct and research ethics:
\n\n1. RETRACTIONS
\n\nA Retraction of a Chapter will be issued by the Academic Editor, either following an Author’s request to do so or when there is a 3rd party report of scientific misconduct. Upon receipt of a report by a 3rd party, the Academic Editor will investigate any allegations of scientific misconduct, working in cooperation with the Author(s) and their institution(s).
\n\nA formal Retraction will be issued when there is clear and conclusive evidence of any of the following:
\n\nPublishing of a Retraction Notice will adhere to the following guidelines:
\n\n1.2. REMOVALS AND CANCELLATIONS
\n\n2. STATEMENTS OF CONCERN
\n\nA Statement of Concern detailing alleged misconduct will be issued by the Academic Editor or publisher following a 3rd party report of scientific misconduct when:
\n\nIntechOpen believes that the number of occasions on which a Statement of Concern is issued will be very few in number. In all cases when such a decision has been taken by the Academic Editor the decision will be reviewed by another editor to whom the author can make representations.
\n\n3. CORRECTIONS
\n\nA Correction will be issued by the Academic Editor when:
\n\n3.1. ERRATUM
\n\nAn Erratum will be issued by the Academic Editor when it is determined that a mistake in a Chapter originates from the production process handled by the publisher.
\n\nA published Erratum will adhere to the Retraction Notice publishing guidelines outlined above.
\n\n3.2. CORRIGENDUM
\n\nA Corrigendum will be issued by the Academic Editor when it is determined that a mistake in a Chapter is a result of an Author’s miscalculation or oversight. A published Corrigendum will adhere to the Retraction Notice publishing guidelines outlined above.
\n\n4. FINAL REMARKS
\n\nIntechOpen wishes to emphasize that the final decision on whether a Retraction, Statement of Concern, or a Correction will be issued rests with the Academic Editor. The publisher is obliged to act upon any reports of scientific misconduct in its publications and to make a reasonable effort to facilitate any subsequent investigation of such claims.
\n\nIn the case of Retraction or removal of the Work, the publisher will be under no obligation to refund the APC.
\n\nThe general principles set out above apply to Retractions and Corrections issued in all IntechOpen publications.
\n\nAny suggestions or comments on this Policy are welcome and may be sent to permissions@intechopen.com.
\n\nPolicy last updated: 2017-09-11
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He previously worked as a post-doctoral fellow at the Ben-Gurion University of Negev, Israel; University of the Free State, South Africa; and Central University of Technology Bloemfontein, South Africa. He obtained his Ph.D. in Organic Chemistry from Nagaoka University of Technology, Japan. He has published more than seventy-four journal articles and attended several national and international conferences as speaker and chair. Dr. Kendrekar has received many international awards. He has several funded projects, namely, anti-malaria drug development, MRSA, and SARS-CoV-2 activity of curcumin and its formulations. He has filed four patents in collaboration with the University of Central Lancashire and Mayo Clinic Infectious Diseases. His present research includes organic synthesis, drug discovery and development, biochemistry, nanoscience, and nanotechnology.",institutionString:"Visiting Scientist at Lipid Nanostructures Laboratory, Centre for Smart Materials, School of Natural Sciences, University of Central Lancashire",institution:null},{id:"428125",title:"Dr.",name:"Vinayak",middleName:null,surname:"Adimule",slug:"vinayak-adimule",fullName:"Vinayak Adimule",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/428125/images/system/428125.jpg",biography:"Dr. Vinayak Adimule, MSc, Ph.D., is a professor and dean of R&D, Angadi Institute of Technology and Management, India. He has 15 years of research experience as a senior research scientist and associate research scientist in R&D organizations. He has published more than fifty research articles as well as several book chapters. He has two Indian patents and two international patents to his credit. Dr. Adimule has attended, chaired, and presented papers at national and international conferences. He is a guest editor for Topics in Catalysis and other journals. He is also an editorial board member, life member, and associate member for many international societies and research institutions. His research interests include nanoelectronics, material chemistry, artificial intelligence, sensors and actuators, bio-nanomaterials, and medicinal chemistry.",institutionString:"Angadi Institute of Technology and Management",institution:null},{id:"284317",title:"Prof.",name:"Kantharaju",middleName:null,surname:"Kamanna",slug:"kantharaju-kamanna",fullName:"Kantharaju Kamanna",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/284317/images/21050_n.jpg",biography:"Prof. K. Kantharaju has received Bachelor of science (PCM), master of science (Organic Chemistry) and Doctor of Philosophy in Chemistry from Bangalore University. He worked as a Executive Research & Development @ Cadila Pharmaceuticals Ltd, Ahmedabad. He received DBT-postdoc fellow @ Molecular Biophysics Unit, Indian Institute of Science, Bangalore under the supervision of Prof. P. Balaram, later he moved to NIH-postdoc researcher at Drexel University College of Medicine, Philadelphia, USA, after his return from postdoc joined NITK-Surthakal as a Adhoc faculty at department of chemistry. Since from August 2013 working as a Associate Professor, and in 2016 promoted to Profeesor in the School of Basic Sciences: Department of Chemistry and having 20 years of teaching and research experiences.",institutionString:null,institution:{name:"Rani Channamma University, Belagavi",country:{name:"India"}}},{id:"158492",title:"Prof.",name:"Yusuf",middleName:null,surname:"Tutar",slug:"yusuf-tutar",fullName:"Yusuf Tutar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/158492/images/system/158492.jpeg",biography:"Prof. Dr. Yusuf Tutar conducts his research at the Hamidiye Faculty of Pharmacy, Department of Basic Pharmaceutical Sciences, Division of Biochemistry, University of Health Sciences, Turkey. He is also a faculty member in the Molecular Oncology Program. He obtained his MSc and Ph.D. at Oregon State University and Texas Tech University, respectively. He pursued his postdoctoral studies at Rutgers University Medical School and the National Institutes of Health (NIH/NIDDK), USA. His research focuses on biochemistry, biophysics, genetics, molecular biology, and molecular medicine with specialization in the fields of drug design, protein structure-function, protein folding, prions, microRNA, pseudogenes, molecular cancer, epigenetics, metabolites, proteomics, genomics, protein expression, and characterization by spectroscopic and calorimetric methods.",institutionString:"University of Health Sciences",institution:null},{id:"180528",title:"Dr.",name:"Hiroyuki",middleName:null,surname:"Kagechika",slug:"hiroyuki-kagechika",fullName:"Hiroyuki Kagechika",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/180528/images/system/180528.jpg",biography:"Hiroyuki Kagechika received his bachelor’s degree and Ph.D. in Pharmaceutical Sciences from the University of Tokyo, Japan, where he served as an associate professor until 2004. He is currently a professor at the Institute of Biomaterials and Bioengineering (IBB), Tokyo Medical and Dental University (TMDU). From 2010 to 2012, he was the dean of the Graduate School of Biomedical Science. Since 2012, he has served as the vice dean of the Graduate School of Medical and Dental Sciences. He has been the director of the IBB since 2020. Dr. Kagechika’s major research interests are the medicinal chemistry of retinoids, vitamins D/K, and nuclear receptors. He has developed various compounds including a drug for acute promyelocytic leukemia.",institutionString:"Tokyo Medical and Dental University",institution:{name:"Tokyo Medical and Dental University",country:{name:"Japan"}}},{id:"94311",title:"Prof.",name:"Martins",middleName:"Ochubiojo",surname:"Ochubiojo Emeje",slug:"martins-ochubiojo-emeje",fullName:"Martins Ochubiojo Emeje",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/94311/images/system/94311.jpeg",biography:"Martins Emeje obtained a BPharm with distinction from Ahmadu Bello University, Nigeria, and an MPharm and Ph.D. from the University of Nigeria (UNN), where he received the best Ph.D. award and was enlisted as UNN’s “Face of Research.” He established the first nanomedicine center in Nigeria and was the pioneer head of the intellectual property and technology transfer as well as the technology innovation and support center. Prof. Emeje’s several international fellowships include the prestigious Raman fellowship. He has published more than 150 articles and patents. He is also the head of R&D at NIPRD and holds a visiting professor position at Nnamdi Azikiwe University, Nigeria. He has a postgraduate certificate in Project Management from Walden University, Minnesota, as well as a professional teaching certificate and a World Bank certification in Public Procurement. Prof. Emeje was a national chairman of academic pharmacists in Nigeria and the 2021 winner of the May & Baker Nigeria Plc–sponsored prize for professional service in research and innovation.",institutionString:"National Institute for Pharmaceutical Research and Development",institution:{name:"National Institute for Pharmaceutical Research and Development",country:{name:"Nigeria"}}},{id:"436430",title:"Associate Prof.",name:"Mesut",middleName:null,surname:"Işık",slug:"mesut-isik",fullName:"Mesut Işık",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/436430/images/19686_n.jpg",biography:null,institutionString:null,institution:{name:"Bilecik University",country:{name:"Turkey"}}},{id:"268659",title:"Ms.",name:"Xianquan",middleName:null,surname:"Zhan",slug:"xianquan-zhan",fullName:"Xianquan Zhan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/268659/images/8143_n.jpg",biography:"Dr. Zhan received his undergraduate and graduate training in the fields of preventive medicine and epidemiology and statistics at the West China University of Medical Sciences in China during 1989 to 1999. He received his post-doctoral training in oncology and cancer proteomics for two years at the Cancer Research Institute of Human Medical University in China. In 2001, he went to the University of Tennessee Health Science Center (UTHSC) in USA, where he was a post-doctoral researcher and focused on mass spectrometry and cancer proteomics. Then, he was appointed as an Assistant Professor of Neurology, UTHSC in 2005. He moved to the Cleveland Clinic in USA as a Project Scientist/Staff in 2006 where he focused on the studies of eye disease proteomics and biomarkers. He returned to UTHSC as an Assistant Professor of Neurology in the end of 2007, engaging in proteomics and biomarker studies of lung diseases and brain tumors, and initiating the studies of predictive, preventive, and personalized medicine (PPPM) in cancer. In 2010, he was promoted to Associate Professor of Neurology, UTHSC. Currently, he is a Professor at Xiangya Hospital of Central South University in China, Fellow of Royal Society of Medicine (FRSM), the European EPMA National Representative in China, Regular Member of American Association for the Advancement of Science (AAAS), European Cooperation of Science and Technology (e-COST) grant evaluator, Associate Editors of BMC Genomics, BMC Medical Genomics, EPMA Journal, and Frontiers in Endocrinology, Executive Editor-in-Chief of Med One. He has\npublished 116 peer-reviewed research articles, 16 book chapters, 2 books, and 2 US patents. His current main research interest focuses on the studies of cancer proteomics and biomarkers, and the use of modern omics techniques and systems biology for PPPM in cancer, and on the development and use of 2DE-LC/MS for the large-scale study of human proteoforms.",institutionString:null,institution:{name:"Xiangya Hospital Central South University",country:{name:"China"}}},{id:"40482",title:null,name:"Rizwan",middleName:null,surname:"Ahmad",slug:"rizwan-ahmad",fullName:"Rizwan Ahmad",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/40482/images/system/40482.jpeg",biography:"Dr. Rizwan Ahmad is a University Professor and Coordinator, Quality and Development, College of Medicine, Imam Abdulrahman bin Faisal University, Saudi Arabia. Previously, he was Associate Professor of Human Function, Oman Medical College, Oman, and SBS University, Dehradun. Dr. Ahmad completed his education at Aligarh Muslim University, Aligarh. He has published several articles in peer-reviewed journals, chapters, and edited books. His area of specialization is free radical biochemistry and autoimmune diseases.",institutionString:"Imam Abdulrahman Bin Faisal University",institution:{name:"Imam Abdulrahman Bin Faisal University",country:{name:"Saudi Arabia"}}},{id:"41865",title:"Prof.",name:"Farid A.",middleName:null,surname:"Badria",slug:"farid-a.-badria",fullName:"Farid A. Badria",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/41865/images/system/41865.jpg",biography:"Farid A. Badria, Ph.D., is the recipient of several awards, including The World Academy of Sciences (TWAS) Prize for Public Understanding of Science; the World Intellectual Property Organization (WIPO) Gold Medal for best invention; Outstanding Arab Scholar, Kuwait; and the Khwarizmi International Award, Iran. He has 250 publications, 12 books, 20 patents, and several marketed pharmaceutical products to his credit. He continues to lead research projects on developing new therapies for liver, skin disorders, and cancer. Dr. Badria was listed among the world’s top 2% of scientists in medicinal and biomolecular chemistry in 2019 and 2020. He is a member of the Arab Development Fund, Kuwait; International Cell Research Organization–United Nations Educational, Scientific and Cultural Organization (ICRO–UNESCO), Chile; and UNESCO Biotechnology France",institutionString:"Mansoura University",institution:{name:"Mansoura University",country:{name:"Egypt"}}},{id:"329385",title:"Dr.",name:"Rajesh K.",middleName:"Kumar",surname:"Singh",slug:"rajesh-k.-singh",fullName:"Rajesh K. Singh",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/329385/images/system/329385.png",biography:"Dr. Singh received a BPharm (2003) and MPharm (2005) from Panjab University, Chandigarh, India, and a Ph.D. (2013) from Punjab Technical University (PTU), Jalandhar, India. He has more than sixteen years of teaching experience and has supervised numerous postgraduate and Ph.D. students. He has to his credit more than seventy papers in SCI- and SCOPUS-indexed journals, fifty-five conference proceedings, four books, six Best Paper Awards, and five projects from different government agencies. He is currently an editorial board member of eight international journals and a reviewer for more than fifty scientific journals. He received Top Reviewer and Excellent Peer Reviewer Awards from Publons in 2016 and 2017, respectively. He is also on the panel of The International Reviewer for reviewing research proposals for grants from the Royal Society. He also serves as a Publons Academy mentor and Bentham brand ambassador.",institutionString:"Punjab Technical University",institution:{name:"Punjab Technical University",country:{name:"India"}}},{id:"142388",title:"Dr.",name:"Thiago",middleName:"Gomes",surname:"Gomes Heck",slug:"thiago-gomes-heck",fullName:"Thiago Gomes Heck",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/142388/images/7259_n.jpg",biography:null,institutionString:null,institution:{name:"Universidade Regional do Noroeste do Estado do Rio Grande do Sul",country:{name:"Brazil"}}},{id:"336273",title:"Assistant Prof.",name:"Janja",middleName:null,surname:"Zupan",slug:"janja-zupan",fullName:"Janja Zupan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/336273/images/14853_n.jpeg",biography:"Janja Zupan graduated in 2005 at the Department of Clinical Biochemistry (superviser prof. dr. Janja Marc) in the field of genetics of osteoporosis. Since November 2009 she is working as a Teaching Assistant at the Faculty of Pharmacy, Department of Clinical Biochemistry. In 2011 she completed part of her research and PhD work at Institute of Genetics and Molecular Medicine, University of Edinburgh. She finished her PhD entitled The influence of the proinflammatory cytokines on the RANK/RANKL/OPG in bone tissue of osteoporotic and osteoarthritic patients in 2012. From 2014-2016 she worked at the Institute of Biomedical Sciences, University of Aberdeen as a postdoctoral research fellow on UK Arthritis research project where she gained knowledge in mesenchymal stem cells and regenerative medicine. She returned back to University of Ljubljana, Faculty of Pharmacy in 2016. She is currently leading project entitled Mesenchymal stem cells-the keepers of tissue endogenous regenerative capacity facing up to aging of the musculoskeletal system funded by Slovenian Research Agency.",institutionString:null,institution:{name:"University of Ljubljana",country:{name:"Slovenia"}}},{id:"357453",title:"Dr.",name:"Radheshyam",middleName:null,surname:"Maurya",slug:"radheshyam-maurya",fullName:"Radheshyam Maurya",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/357453/images/16535_n.jpg",biography:null,institutionString:null,institution:{name:"University of Hyderabad",country:{name:"India"}}},{id:"418340",title:"Dr.",name:"Jyotirmoi",middleName:null,surname:"Aich",slug:"jyotirmoi-aich",fullName:"Jyotirmoi Aich",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000038Ugi5QAC/Profile_Picture_2022-04-15T07:48:28.png",biography:"Biotechnologist with 15 years of research including 6 years of teaching experience. Demonstrated record of scientific achievements through consistent publication record (H index = 13, with 874 citations) in high impact journals such as Nature Communications, Oncotarget, Annals of Oncology, PNAS, and AJRCCM, etc. Strong research professional with a post-doctorate from ACTREC where I gained experimental oncology experience in clinical settings and a doctorate from IGIB where I gained expertise in asthma pathophysiology. A well-trained biotechnologist with diverse experience on the bench across different research themes ranging from asthma to cancer and other infectious diseases. An individual with a strong commitment and innovative mindset. Have the ability to work on diverse projects such as regenerative and molecular medicine with an overall mindset of improving healthcare.",institutionString:"DY Patil Deemed to Be University",institution:null},{id:"349288",title:"Prof.",name:"Soumya",middleName:null,surname:"Basu",slug:"soumya-basu",fullName:"Soumya Basu",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000035QxIDQA0/Profile_Picture_2022-04-15T07:47:01.jpg",biography:"Soumya Basu, Ph.D., is currently working as an Associate Professor at Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Pune, Maharashtra, India. With 16+ years of trans-disciplinary research experience in Drug Design, development, and pre-clinical validation; 20+ research article publications in journals of repute, 9+ years of teaching experience, trained with cross-disciplinary education, Dr. Basu is a life-long learner and always thrives for new challenges.\r\nHer research area is the design and synthesis of small molecule partial agonists of PPAR-γ in lung cancer. She is also using artificial intelligence and deep learning methods to understand the exosomal miRNA’s role in cancer metastasis. Dr. Basu is the recipient of many awards including the Early Career Research Award from the Department of Science and Technology, Govt. of India. She is a reviewer of many journals like Molecular Biology Reports, Frontiers in Oncology, RSC Advances, PLOS ONE, Journal of Biomolecular Structure & Dynamics, Journal of Molecular Graphics and Modelling, etc. She has edited and authored/co-authored 21 journal papers, 3 book chapters, and 15 abstracts. She is a Board of Studies member at her university. She is a life member of 'The Cytometry Society”-in India and 'All India Cell Biology Society”- in India.",institutionString:"Dr. D.Y. Patil Vidyapeeth, Pune",institution:{name:"Dr. D.Y. Patil Vidyapeeth, Pune",country:{name:"India"}}},{id:"354817",title:"Dr.",name:"Anubhab",middleName:null,surname:"Mukherjee",slug:"anubhab-mukherjee",fullName:"Anubhab Mukherjee",position:null,profilePictureURL:"https://intech-files.s3.amazonaws.com/0033Y0000365PbRQAU/ProfilePicture%202022-04-15%2005%3A11%3A18.480",biography:"A former member of Laboratory of Nanomedicine, Brigham and Women’s Hospital, Harvard University, Boston, USA, Dr. Anubhab Mukherjee is an ardent votary of science who strives to make an impact in the lives of those afflicted with cancer and other chronic/acute ailments. He completed his Ph.D. from CSIR-Indian Institute of Chemical Technology, Hyderabad, India, having been skilled with RNAi, liposomal drug delivery, preclinical cell and animal studies. He pursued post-doctoral research at College of Pharmacy, Health Science Center, Texas A & M University and was involved in another postdoctoral research at Department of Translational Neurosciences and Neurotherapeutics, John Wayne Cancer Institute, Santa Monica, California. In 2015, he worked in Harvard-MIT Health Sciences & Technology as a visiting scientist. He has substantial experience in nanotechnology-based formulation development and successfully served various Indian organizations to develop pharmaceuticals and nutraceutical products. He is an inventor in many US patents and an author in many peer-reviewed articles, book chapters and books published in various media of international repute. Dr. Mukherjee is currently serving as Principal Scientist, R&D at Esperer Onco Nutrition (EON) Pvt. Ltd. and heads the Hyderabad R&D center of the organization.",institutionString:"Esperer Onco Nutrition Pvt Ltd.",institution:null},{id:"319365",title:"Assistant Prof.",name:"Manash K.",middleName:null,surname:"Paul",slug:"manash-k.-paul",fullName:"Manash K. Paul",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/319365/images/system/319365.png",biography:"Manash K. Paul is a Principal Investigator and Scientist at the University of California Los Angeles. He has contributed significantly to the fields of stem cell biology, regenerative medicine, and lung cancer. His research focuses on various signaling processes involved in maintaining stem cell homeostasis during the injury-repair process, deciphering lung stem cell niche, pulmonary disease modeling, immuno-oncology, and drug discovery. He is currently investigating the role of extracellular vesicles in premalignant lung cell migration and detecting the metastatic phenotype of lung cancer via machine-learning-based analyses of exosomal signatures. Dr. Paul has published in more than fifty peer-reviewed international journals and is highly cited. He is the recipient of many awards, including the UCLA Vice Chancellor’s award, a senior member of the Institute of Electrical and Electronics Engineers (IEEE), and an editorial board member for several international journals.",institutionString:"University of California Los Angeles",institution:{name:"University of California Los Angeles",country:{name:"United States of America"}}},{id:"311457",title:"Dr.",name:"Júlia",middleName:null,surname:"Scherer Santos",slug:"julia-scherer-santos",fullName:"Júlia Scherer Santos",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/311457/images/system/311457.jpg",biography:"Dr. Júlia Scherer Santos works in the areas of cosmetology, nanotechnology, pharmaceutical technology, beauty, and aesthetics. Dr. Santos also has experience as a professor of graduate courses. Graduated in Pharmacy, specialization in Cosmetology and Cosmeceuticals applied to aesthetics, specialization in Aesthetic and Cosmetic Health, and a doctorate in Pharmaceutical Nanotechnology. Teaching experience in Pharmacy and Aesthetics and Cosmetics courses. She works mainly on the following subjects: nanotechnology, cosmetology, pharmaceutical technology, aesthetics.",institutionString:"Universidade Federal de Juiz de Fora",institution:{name:"Universidade Federal de Juiz de Fora",country:{name:"Brazil"}}},{id:"219081",title:"Dr.",name:"Abdulsamed",middleName:null,surname:"Kükürt",slug:"abdulsamed-kukurt",fullName:"Abdulsamed Kükürt",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/219081/images/system/219081.png",biography:"Dr. Kükürt graduated from Uludağ University in Turkey. He started his academic career as a Research Assistant in the Department of Biochemistry at Kafkas University. In 2019, he completed his Ph.D. program in the Department of Biochemistry at the Institute of Health Sciences. He is currently working at the Department of Biochemistry, Kafkas University. He has 27 published research articles in academic journals, 11 book chapters, and 37 papers. He took part in 10 academic projects. He served as a reviewer for many articles. He still serves as a member of the review board in many academic journals. He is currently working on the protective activity of phenolic compounds in disorders associated with oxidative stress and inflammation.",institutionString:null,institution:{name:"Kafkas University",country:{name:"Turkey"}}},{id:"178366",title:"Dr.",name:"Volkan",middleName:null,surname:"Gelen",slug:"volkan-gelen",fullName:"Volkan Gelen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/178366/images/system/178366.jpg",biography:"Volkan Gelen is a Physiology specialist who received his veterinary degree from Kafkas University in 2011. Between 2011-2015, he worked as an assistant at Atatürk University, Faculty of Veterinary Medicine, Department of Physiology. In 2016, he joined Kafkas University, Faculty of Veterinary Medicine, Department of Physiology as an assistant professor. Dr. Gelen has been engaged in various academic activities at Kafkas University since 2016. There he completed 5 projects and has 3 ongoing projects. He has 60 articles published in scientific journals and 20 poster presentations in scientific congresses. His research interests include physiology, endocrine system, cancer, diabetes, cardiovascular system diseases, and isolated organ bath system studies.",institutionString:"Kafkas University",institution:{name:"Kafkas University",country:{name:"Turkey"}}},{id:"418963",title:"Dr.",name:"Augustine Ododo",middleName:"Augustine",surname:"Osagie",slug:"augustine-ododo-osagie",fullName:"Augustine Ododo Osagie",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/418963/images/16900_n.jpg",biography:"Born into the family of Osagie, a prince of the Benin Kingdom. I am currently an academic in the Department of Medical Biochemistry, University of Benin. Part of the duties are to teach undergraduate students and conduct academic research.",institutionString:null,institution:{name:"University of Benin",country:{name:"Nigeria"}}},{id:"192992",title:"Prof.",name:"Shagufta",middleName:null,surname:"Perveen",slug:"shagufta-perveen",fullName:"Shagufta Perveen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/192992/images/system/192992.png",biography:"Prof. Shagufta Perveen is a Distinguish Professor in the Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia. Dr. Perveen has acted as the principal investigator of major research projects funded by the research unit of King Saud University. She has more than ninety original research papers in peer-reviewed journals of international repute to her credit. 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She has been a Professor since 1996. Currently, she is the Head of the Laboratory of Metabolism, a division of the Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, Moscow, Russian Federation. N.V. Beloborodova has many years of clinical experience in the field of intensive care and surgery. She studies infectious complications and sepsis. She initiated a series of interdisciplinary clinical and experimental studies based on the concept of integrating human metabolism and its microbiota. Her scientific achievements are widely known: she is the recipient of the Marie E. Coates Award \\"Best lecturer-scientist\\" Gustafsson Fund, Karolinska Institutes, Stockholm, Sweden, and the International Sepsis Forum Award, Pasteur Institute, Paris, France (2014), etc. Professor N.V. 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Reasoning in a sustainable way entails, first and foremost, managing the available resources efficiently and strategically, whether they are natural, financial, human or relational. In this way, value is generated by contributing to the growth, improvement and socio-economic development of the communities and of all the players that make up its value chain. In the coming decades, we will need to be able to transition from a society in which economic well-being and health are measured by the growth of production and material consumption, to a society in which we live better while consuming less. In this context, digitization has the potential to disrupt processes, with significant implications for the environment and sustainable development. There are numerous challenges associated with sustainability and digitization, the need to consider new business models capable of extracting value, data ownership and sharing and integration, as well as collaboration across the entire supply chain of a product. In order to generate value, effectively developing a complex system based on sustainability principles is a challenge that requires a deep commitment to both technological factors, such as data and platforms, and human dimensions, such as trust and collaboration. Regular study, research and implementation must be part of the road to sustainable solutions. Consequently, this topic will analyze growth models and techniques aimed at achieving intergenerational equity in terms of economic, social and environmental well-being. It will also cover various subjects, including risk assessment in the context of sustainable economy and a just society.
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