HPLC Results Research Articles

Rapid Detection of 6-Benzylaminopurine Residuals Using Surface-Enhanced Raman Scattering

The rapid analysis of 6-benzylaminopurine (6-BAP) residue by surface-enhanced Raman spectroscopy with a portable Raman spectrometer is reported. The results showed that the Raman signals of 6-BAP can be significantly enhanced when mixed with a gold nanoparticle colloid substrate. The typical Raman shifts of 6-BAP extraction were at 1002, 1318, and 1336 cm–1 and the intensities of normalized characteristic band at 1002 cm–1 (I1002 to I738) showed high correlation with 6-BAP concentrations. The concentration linear range was 0.1–5.0 μg/mL. The coefficient of determination (R2) was about 0.99, and calculated RSDs below 10%. The SERS data matched well with the HPLC results.

Graphene oxide encapsulated 3D porous chalcopyrite (CuFeS2) nanocomposite as an emerging electrocatalyst for agro-hazardous (methyl paraoxon) detection in vegetables

Methyl paraoxon (MOX) is a highly toxic organophosphate pesticide. It is recently reported that, MOX can enter the human body through ingestion, inhalation, or by dermal penetration. Due to its high non-degradability, it can bind to the tissues of fruits and vegetables. When it is consumed, it can imposes sub-chronic and chronic diseases, by the inhibition of acetylcholinesterase in human metabolism. Therefore, for the first time, we reported a detection of non-enzymatic electrochemical sensor based on 3D porous phase graphene oxide sheets encapsulated chalcopyrite (GOS@CuFeS2) nanocomposite. Hence, the development of reliable sensors for the real-time detection of pesticides is imperative to overcome practical limitations encountered in conventional methodologies. As synthesized GOS@CuFeS2 nanocomposite film screen-printed carbon modified electrode (SPCE) displays excellent electrocatalytic ability towards MOX. Under optimized working conditions, the modified electrode provides linear response range from 0.073 to 801.5 μM. The detection limit was obtained as 4.5 nM. The sensor displayed outstanding sensitivity as 17.97 μA μM−1 cm−2. This composite could be a promising electrode modifier for electrocatalysis. Finally, the GOS@CuFeS2 nanocomposite modified electrode shows greater real-time practicality in vegetable real samples. The obtained moral parameters from the developed method were compared with the authenticated HPLC results.

Photolysis of Indigo Carmine solution by planar vacuum-ultraviolet (147 nm) light source

Advanced oxidation processes (AOPs) are an attractive method to decompose dye-containing wastewaters, because they avoid issues of secondary pollution. In particular, a vacuum-ultraviolet (VUV) process is the simplest method, because an oxidation accelerator or a catalyst is not required. Conventional VUV sources with wavelengths of 185 nm or 172 nm have been used. We predicted that a shorter wavelength VUV process would have a higher ability to decompose dyes in wastewater. We developed a new planar light source that could emit a resonance line at 147 nm and a broad molecular line at 172 nm. The irradiance was 8.7 mW/cm2 at a distance of 20 mm from the emission surface to the UV power meter. We then conducted photocatalytic experiments of an Indigo Carmine solution at 3.33 × 10−4 mol/L to confirm the decomposition abilities of the developed light source, an excimer lamp of 172 nm and a Hg lamp of 254 nm. From the HPLC results, changes in Indigo Carmine concentration with the developed light source were equivalent to those with the excimer lamp. However, the residual ratio of total organic carbon (TOC) with the developed light source was lower than those with the other lamps. Therefore, a wavelength of 147 nm is superior to the conventional wavelength of 172 nm for Indigo Carmine decomposition. In addition, the developed light source emits VUV from only one side, which is a flat emitting surface. Consequently, the developed light source would have reduced manufacturing and maintenance costs compared to current VUV processing equipment.

Antibacterial and anticancer potential of marine endophytic actinomycetes Streptomyces coeruleorubidus GRG 4 (KY457708) compound against colistin resistant uropathogens and A549 lung cancer cells

The aim of the current study is to identify bioactive compound from marine endophytic actinomycetes (MEA) isolated from Gulf of Mannar region, Southeast coast of India. Among the isolated actinomycetes, strain GRG 4 exhibited excellent ability to inhibit isolated colistin resistant (CR) Pseudomonas aeruginosa (P. aeruginosa) and Klebsiella pneumoniae (K. pneumoniae), which is a emerging threat to the world. The strain was identified as Streptomyces coeruleorubidus GRG 4 (KY457708), based on morphological, biochemical, phenotypic and genotypic characters. The bioactive metabolites present in the methanolic extract were partially purified by TLC and preparative HPLC. The active HPLC fraction 2 showed 15, 20 mm zone of inhibition against both CR P. aeruginosa and K. pneumoniae respectively. Analytical HPLC and FT-IR results of fraction 2 showed with carbonyl group. Both GC-MS and LC-MS results confirmed that the fraction 2 contained chemical constituents of Bis (2-Ethylhexyl) Phthalate (BEP). The compromised structure with loosely integrated and ruptured cell wall of BEP treated CR bacteria were observed by confocal laser scanning microscope (CLSM) and scanning electron microscope (SEM) at 75 μg/mL of minimum inhibitory concentration (MIC) dose. Further, cytotoxic effect of BEP against A549 human lung cancer cells revealed complete inhibition by cell proliferation and apoptosis was observed at 100 μg/mL in 24 h treatment. In addition, irreversible ROS dependent oxidative damage was clearly observed at the IC50 concentration of BEP. The toxicity of BEP was also studied against Vibrio fischeri (V. fischeri) and found to be highly toxic after 15 and 30 min of treatment. Based on the results it could be concluded that the identified compound BEP is a potent inhibitor for CR bacteria and A549 lung cancer cells.

Identification of dyes in Egyptian textiles of the first millennium ad from the collection Fill-Trevisiol

High-performance liquid chromatography coupled to a diode-array detector (HPLC-DAD) is used to investigate 30 samples which were removed from 27 ancient Egyptian fabrics of the Fill-Trevisiol collection. Attention is focused in this paper on fabrics of the Roman and Byzantine periods, with red and deep violet–blue wool weft threads which are Z-spun. The following dyes are identified in fabrics which date to the Roman period (first–fifth c. ad): mollusc purple, madder, and indigo/woad (Indigofera species and other, Isatis tinctoria L.). The results for the Byzantine (fifth–seventh c. ad) fabrics are richer in terms of identified dyes: apart from the three aforementioned dyes, the use of kermes (Kermes vermilio Planchon) and cochineal is revealed as the two coccid dyes have been detected in six Egyptian–Byzantine fabrics. Moreover, a yellow dye (probably Reseda luteola L.) is identified in one sample. Finally, samples taken from two fabrics, which date to the Islamic period, were dyed using indigo/woad and lac (Kerria lacca, Kerr). Mixing madder and indigo/woad to imitate true purple was a common practice in ancient Egypt and this is confirmed by the HPLC results in several samples. Semi-quantitative results are obtained from the HPLC peak areas and lead to the following conclusions. Madder dyes which were rich or poor in alizarin, compared to purpurin, and could have been therefore obtained from Rubia tinctorum L. and Rubia peregrina L., respectively are detected in Byzantine fabrics. Only alizarin-rich madder dyes are identified in some samples from Roman fabrics, but in some other samples the relative alizarin-to-purpurin ratio cannot be measured with confidence. Cochineal is found in two samples. The biological source of cochineal (Porphyrophora hamelii Brandt) is possible to be chemically identified only in one sample. The HPLC results of the molluscan dye detected in a Roman fabric are compared with the relative compositions of extracts of the three Mediterranean molluscs leading to the speculation that the Roman purple sample was probably dyed using Hexaplex trunculus L. The molluscan dye was furthermore detected in a Byzantine sample where it was combined with madder. Finally, it is reported that the standard (hydrochloric) acid hydrolysis method which is commonly applied to extract dyes from archaeological samples does not have any noticeable effect on the relative composition of the molluscan dye.

Electrocatalytic Oxidation of 5-(Hydroxymethyl)furfural Using High-Surface-Area Nickel Boride.

The electrochemical oxidation of the biorefinery product 5-(hydroxymethyl)furfural (HMF) to 2,5-furandicarboxylic acid (FDCA), an important platform chemical for the polymer industry, is receiving increasing interest. FDCA-based polymers such as polyethylene 2,5-furandicarboxylate (PEF) are sustainable candidates for replacing polyethylene terephthalate (PET). Herein, we report the highly efficient electrocatalytic oxidation of HMF to FDCA, using Ni foam modified with high-surface-area nickel boride (Nix B) as the electrode. Constant potential electrolysis in combination with HPLC revealed a high faradaic efficiency of close to 100 % towards the production of FDCA with a yield of 98.5 %. Operando electrochemistry coupled to ATR-IR spectroscopy indicated that HMF is oxidized preferentially via 5-hydroxymethyl-2-furancarboxylic acid rather than via 2,5-diformylfuran, which is in agreement with HPLC results. This study not only reports a low-cost active electrocatalyst material for the electrochemical oxidation of HMF to FDCA, but additionally provides insight into the reaction pathway.

In vivo synthesis of csypyrone derivatives by exploring the substrate diversity of start units of type Ⅲ polyketide synthase CsyB

As a novel fungal type Ⅲ polyketide synthase, CsyB from Aspergillus oryzae can sequentially accept one molecular short chain fatty acyl CoA as start unit, one molecular malonyl-CoA and one molecular acetoacetyl-CoA as extend unit to produce the short chain csypyrone B1-3. On the basis of crystal structure of CsyB, a fatty acyl CoA binding tunnel of a length of about 16 Å is located in its active center that is proposed to accept diversified start units. In order to examine the substrate diversity of CsyB, CsyB gene was introduced and expressed in Escherichia coli that contained a number of precursors of long chain fatty acyl CoA in vivo. The results of HPLC revealed that a series of long chain csypyrone derivatives were detected in the recombinant strain in comparison with the control strain. These new csypyrone compounds were preliminarily analyzed by UV-visible spectroscopy and LC-HRMS. Three hydroxylated csypyrones were intensively determined by 1D and 2D NMR experiments, especially the position of the hydroxyl group in these compounds. These results demonstrate that CsyB exhibits a broad substrate specificity, which not only can accept the long chain saturated or unsaturated fatty acyl CoA as substrate, but also accept hydroxylated long chain fatty acyl CoA.

Biogenic manganese oxide: An efficient peroxymonosulfate activation catalyst for tetracycline and phenol degradation in water

From the view of environment and easy availability, to develop the most efficient manganese oxide for peroxymonosulfate (PMS) activation is of great importance for the degradation of recalcitrant organic pollutants. In this study, biogenic manganese oxide (BioMnOx) exhibited an unprecedented efficiency than the most efficient 3D α-Mn2O3 prepared by chemical method. 100% of phenol degradation at 40 min and 99.4% of tetracycline removal at 60 min were achieved over BioMnOx with PMS, which was 3-fold faster than 3D α-Mn2O3. BioMnOx also has an excellent long-term stability and good performance toward the pollutants degradation at a wide pH range of 3.0–9.0. Most importantly, 1O2 was identified as the primary reactive species in BioMnOx/PMS system based on the trapping experiment and EPR analysis. The PMS activation over BioMnOx should follow a self-decomposition and energy quenching mechanism instead of electron transfer process as confirmed by the XPS analysis. Finally, the degradation pathways of tetracycline and phenol by 1O2 over BioMnOx were proposed according to HPLC and HPLC-MS results, which are greatly different from that by OH oxidation in the literature.

Prodrugs for Nitroreductase Based Cancer Therapy- 1: Metabolite Profile, Cell Cytotoxicity and Molecular Modeling Interactions of Nitro Benzamides with Ssap-NtrB

Directed Enzyme Prodrugs Therapy (DEPT) as an alternative method against conventional cancer treatments, in which the non-toxic prodrugs is converted to highly cytotoxic derivative, has attracted an ample attention in recent years for cancer therapy studies. The metabolite profile, cell cytotoxicity and molecular modeling interactions of a series of nitro benzamides with Ssap-NtrB were investigated in this study. A series of nitro-substituted benzamide prodrugs (1-4) were synthesized and firstly investigated their enzymatic reduction by Ssap-NtrB (S. saprophyticus Nitroreductase B) using HPLC analysis. Resulting metabolites were analyzed by LC-MS/MS. Molecular docking studies were performed with the aim of investigating the relationship between nitro benzamide structures (prodrugs 1-4) and Ssap-NtrB at the molecular level. Cell viability assay was conducted on two cancer cell lines, hepatoma (Hep3B) and colon (HT-29) cancer models and healthy cell model HUVEC. Upon reduction of benzamide prodrugs by Ssap-NtrB, the corresponding amine effectors were tested in a cell line panel comprising PC-3, Hep3B and HUVEC cells and were compared with the established NTR substrates, CB1954 (an aziridinyl dinitrobenzamide). Cell viability assay resulted in while prodrugs 1, 2 and 3 had no remarkable cytotoxic effects, prodrug 4 showed the differential effect, showing moderate cytotoxicity with Hep3B and HUVEC. The metabolites that obtained from the reduction of nitro benzamide prodrugs (1-4) by Ssap-NtrB, showed differential cytotoxic effects, with none toxic for HUVEC cells, moderate toxic for Hep3B cells, but highly toxic for PC3 cells. Amongst all metabolites of prodrugs after Ssap-NtrB reduction, N-(2,4- dinitrophenyl)-4-nitrobenzamide (3) was efficient and toxic in PC3 cells as comparable as CB1954. Kinetic parameters, molecular docking and HPLC results also confirm that prodrug 3 is better for Ssap-NtrB than 1, 2 and 4 or known cancer prodrugs of CB1954 and SN23862, demonstrating that prodrug 3 is an efficient candidate for NTR based cancer therapy.

In situ reduced silver nanoparticles embedded molecularly imprinted reusable sensor for selective and sensitive SERS detection of Bisphenol A

Sensitive and selective detection of target analyte is very important in many fields such as commodity inspection and quality monitoring. In this work, we constructed a MIPs@AgNPs surface-enhanced Raman scattering (SERS) sensor by in-situ preparation of silver nanoparticles (AgNPs) inside molecularly imprinted polymers (MIPs) matrix for bisphenol A (BPA) detection. In-situ formed AgNPs are evenly distributed and closely packed in the matrix, which is beneficial for hot spot formation and analyte-AgNPs affinity. Our MIPs@AgNPs sensor shows good BPA selectivity against structurally related molecules such as bisphenol AF (BPAF) and diethylstilbestrol (DES). Excellent sensitivity of this MIPs@AgNPs sensor is demonstrated with the detection limit of at least 5 × 10−8 mol L−1 for BPA. This sensor is reliable for the quantitative detection of BPA in real samples, validated with results of HPLC. Regeneration of this kind of SERS sensor can be easily realized by solvent rinsing and NaBH4 reduction. Together with the advantages of ease-of-fabrication, selective recognition, sensitivity and reusability, this kind of MIPs@AgNPs SERS sensor will have a promising practical applications.

MPEG-PLA and PLA-PEG-PLA nanoparticles as new carriers for delivery of recombinant human Growth Hormone (rhGH)

mPEG-PLA and PLA-PEG-PLA copolymeric nanoparticles with three different PLA to PEG ratios are synthesized and used for encapsulation of recombinant human Growth hormone (rhGH). The structure and composition of the synthesized copolymers were analyzed by 1H NMR and GPC techniques. Moreover, morphology, encapsulation efficiency (EE), cytotoxicity, release profile and stability of the encapsulated rhGH were measured. Structural analysis of the prepared copolymers showed that they were successfully synthesized with approximately expected molecular weight and relatively low size distribution. It was also revealed that by increasing amounts of PLA/PEG ratio, EE content and size of nanoparticles were increased. Release profile evaluation of rhGH from both formulations indicated that copolymeric nanoparticles of Di-B2 and Tri-B2 exhibited the best results among the synthesized nanospheres, by having initial burst release of 17.5% and 28% and then slow and constant release of rhGH up to 65% and 77% of the encapsulated drug, respectively. Furthermore, results of HPLC, SDS-PAGE and CD analyses showed stability of rhGH during encapsulation and release from nanoparticles. Finally, the results showed that these two formulations provided safe and efficient sustained release of rhGH for more than a month and they have the potential to do further studies under in vivo conditions.

Can excreted thiocyanate be used to detect cyanide exposure in live reef fish?

Cyanide fishing, where a solution of sodium or potassium cyanide is used to stun reef fish for easy capture for the marine aquarium and live fish food trades, continues to be pervasive despite being illegal in many countries and destructive to coral reef ecosystems. Currently, there is no easy, reliable and universally accepted method to detect if a fish has been exposed to cyanide during the capture process. A promising non-invasive technique for detecting thiocyanate ions, the metabolic byproduct excreted by exposed fish, has been reported in the literature. In an effort to validate this method, four cyanide exposure studies on Amphiprion ocellaris (common clownfish) were carried out over three years. Fish were either exposed to the same (25 ppm) or twice the concentration (50 ppm) as the previsouly published method. Over 100 water samples of fish exposed to cyanide were analyzed by reverse phase HPLC with a C30 column treated with polyethylene glycol and UV detector operating at 220 nm. No thiocyanate was detected beyond the analytical standards and positive controls prepared in seawater. As an alternate means of detecting thiocyanate, water samples and thiocyanate standards from these exposures were derivatized with monobromobimane (MBB) for LC-MS/MS analysis. Thiocyanate was detected in standards with concentrations as low as 0.6 μg/L and quantified to 1 μg/L, but thiocyanate could not be detected in any of the water samples from fish exposed to cyanide with this method either, confirming the HPLC results. Further, we calculated both the mass balance of thiocyanate and the resultant plausible dosage of cyanide from the data reported in the previously published method. These calculations, along with the known lethal dosage of cyanide, further suggests that the detection of thiocyanate in aquarium water is not a viable method for assessing fish exposure to cyanide.

Biodegradation of Lignin Monomers Vanillic, p-Coumaric, and Syringic Acid by the Bacterial Strain, Sphingobacterium sp. HY-H.

Many bacterial strains have been demonstrated to biodegrade lignin for contaminant removal or resource regeneration. The goal of this study was to investigate the biodegradation amount and associated pathways of three lignin monomers, vanillic, p-coumaric, and syringic acid by strain Sphingobacterium sp. HY-H. Vanillic, p-coumaric, and syringic acid degradation with strain HY-H was estimated as 88.71, 76.67, and 72.78%, respectively, after 96h. Correspondingly, the same three monomers were associated with a COD removal efficiency of 87.30, 55.17, and 67.23%, and a TOC removal efficiency of 82.14, 61.03, and 43.86%. The results of GC-MS, HPLC, FTIR, and enzyme activities show that guaiacol and o-dihydroxybenzene are key intermediate metabolites of the vanillic acid and syringic acid degradation. p-Hydroxybenzoic acid is an important intermediate metabolite for p-coumaric and syringic acid degradation. LiP and MnP play an important role in the degradation of lignin monomers and their intermediate metabolites. One possible pathway is that strain HY-H degrades lignin monomers into guaiacol (through decarboxylic and demethoxy reaction) or p-hydroxybenzoic acid (through side-chain oxidation); then guaiacol demethylates to o-dihydroxybenzene. The p-hydroxybenzoic acid and o-dihydroxybenzene are futher through ring cleavage reaction to form small molecule acids (butyric, valproic, oxalic acid, and propionic acid) and alcohols (ethanol and ethanediol), then these acids and alcohols are finally decomposed into CO2 and H2O through the tricarboxylic acid cycle. If properly optimized and controlled, the strain HY-H may play a role in breaking down lignin-related compounds for biofuel and chemical production.

Utilization of photocatalytic hydrogen evolved (Zn,Sn)(O,S) nanoparticles to reduce 4-nitrophenol to 4-aminophenol

Abstract To reduce 4-nitrophenol (4-NP) to 4-aminophenol (4-AP) using sodium borohydrate (NaBH4) in the presence of catalyst has been widely accepted in catalysis field. Many works have claimed that the reduction reaction cannot occur without using NaBH4 as a reducing agent. Herein, we reported the hydrogen evolved Sn(II)-doped Zn(O,S) photocatalyst was synthesized by using precipitation method at 90 °C and then utilized to completely reduce 30 ppm 4-NP to 4-AP under 4 × 6 W UV lamp illumination (0.088 mW/cm2) in 3 h without using NaBH4 as a hydrogen source. It was observed that pure Zn(O,S) nanoparticles did not exhibit a 4-NP reduction, however Zn(O,S) with Sn(II) cation doping showed a 4-NP reduction behavior, as indicated by UV–vis absorbance and HPLC results. The role of Sn(II) doped into Zn(O,S) is considerably important to enhance the amount of photo-induced hydrogen ions for reducing 4-NP to 4-AP on catalyst surfaces.

Antiproliferative and Antioxidant Activities of Two Extracts of the Plant Species Euphorbia dendroides L.

Background: These days, the desire for naturally occurring antioxidants has significantly increased, especially for use in foodstuffs, cosmetics, and pharmaceutical products, to replace synthetic antioxidants that are regularly constrained due to their carcinogenicity. Methods: The study in hand aimed to appraise the antioxidant effect of two Euphorbia dendroides extracts using reducing power, anti-peroxidation, and DPPH (1,1 Diphenyl 2 Pycril Hydrazil) scavenging essays, in addition to the anticancer activity against two tumor cell lines, namely C6 (rat brain tumor)cells, and Hela (human uterus carcinoma)cell lines. Results: The results indicated that the ethyl acetate extract exhibited antiradical activity of 29.49%, higher than that of n-butanol extract (18.06%) at 100 µg/mL but much lower than that of gallic acid (78.21%).The ethyl acetate extract exhibits better reducing capacity and lipid peroxidation inhibitory activity compared to n-butanol extract but less than all tested standards. Moreover, the ethyl acetate extract was found to have an antiproliferative activity of more than 5-FU (5-fluoro-Uracil) against C6 cells at 250 µg/mL with IC50 and IC75 of 113.97, 119.49 µg/mL, respectively, and good cytotoxic activity against the Hela cell lines at the same concentration. The HPLC-TOF-MS (high performance liquid chromatography-Time-of-flight-Mass Spectrometry) analyses exposed the presence of various compounds, among which Gallic and Chlorogenic acids functioned as major compounds. Conclusions: The two extracts exhibited moderate anticancer abilities and behaved somewhat as average antioxidant agents. Based on the total phenolics and flavonoids contents, as well as HPLC results, it could be concluded that antiproliferative and antioxidant activities depend upon the content of different phenolics and flavonoids.

Electricity generation from banana peels in an alkaline fuel cell with a Cu2O-Cu modified activated carbon cathode

Low-cost and highly active catalyst for oxygen reduction reaction is of great importance in the design of alkaline fuel cells. In this work, Cu2O-Cu composite catalyst has been fabricated by a facile laser-irradiation method. The addition of Cu2O-Cu composite in activated carbon air-cathode greatly improves the performance of the cathode. Our results indicate the enhanced performance is likely attributed to the synergistic effect of high conductivity of Cu and the catalytic activity of Cu2O towards the oxygen reduction reaction. Furthermore, an alkaline fuel cell equipped with the composite air-cathode has been built to turn banana peels into electricity. Peak power density of 16.12Wm−2 is obtained under the condition of 3M KOH and 22.04gL−1 reducing sugar, which is higher than other reported low-temperature direct biomass alkaline fuel cells. HPLC results indicate the main oxidation products in the alkaline fuel cell were small organic acids.

Interference Effect of Alcohol on Nessler’s Reagent in Photocatalytic Nitrogen Fixation

Photocatalytic ammonia synthesis is another important reaction to mimic natural nitrogen fixation, which has attracted more and more attention. In recent reports, sacrificial agents are often used to promote charge separation, and high-activity photocatalysts are discovered by using Nessler’s reagent method as a detection technique of ammonia production. However, there is an open question on the rationality and accuracy of the ammonia production amount in the presence of the sacrificial agent and Nessler’s reagent detection method. In this report, P25 TiO2 is employed as a model photocatalyst and alcohol as sacrificial agent, and both Nessler’s reagent and cation exchange chromatography are employed as ammonia detection methods. The different ammonia production amount was found by the different detection method. HPLC and 1H NMR results indicate that carbonyl compounds (formaldehyde, acetaldehyde, and acetone) are produced in the reaction. When the carbonyl compound was added to the ammonia standard soluti...

Effects of various glutamine concentrations on gene expression and steviol glycosides accumulation in Stevia rebaudiana Bertoni.

Stevia rebaudiana Bertoni is one of the most important biologically sourced and low-calorie sweeteners that contains a lots of Steviol glycosides. Tissue culture is the best method for propagation of stevia and micro nutrients can affect both morphological traits and steviol glycosides production. In the present study, we investigated the effect of different concentrations of glutamine (10, 20, 30 and 40 g/l) on expression of UGT74G1 and UGT76G1 genes and stevioside and rebaudioside A accumulation in the leaves of stevia under in vitro conditions. The highest level of expression for UGT74G1 (1.000 Total lab unit) was seen at plants grown in MS media without glutamine and the highest gene expression level for UGT76G1 (1.321 Total lab unit) was observed at plants grown in 2% glutamine. Based on HPLC results, the highest amount of stevioside (22.74) was accumulated in plants which were under 3% glutamine treatment and the lowest production level of stevioside (16.19) was resulted under MS (0 glutamine) medium. The highest rebaudioside A (12.19) accumulation was observed under 2% glutamine treatment and the lowest accumulation of rebaudioside A (8.41) was seen at plants grown in MS medium.

Metal oxides as a biostimulator for upregulation of genes involved in the biosynthesis of Rebaudioside- A.

Stevia rebaudiana Bertoni is a kind of perennial medicinal plant with sweetening properties which belongs to Asteraceae family. Its leaves with fundamental glycoside compounds consist of both a sugar part and a non-sugar sector. One of the glycoside compounds is Rebaudioside- A which has a greater importance in business. This experiment was conducted to evaluate the effects of Ag2O, CrO3, PbO, Fe2O3, BaO and TiO2 on the expression pattern of these genes in the Stevia rebaudiana. Rebaudioside- A biosynthesis was repeated 3 times with concentrations of 50, 100 and 200µM. Also, the results of the study pertaining to the expression pattern of these genes showed that metal oxides have led to an increase in the expression of the regulatory genes involved in biosynthesis of Rebaudioside- A. According to the expression profile, it was found that its effect on DXR, HDS, HDR, IDI and CPPS genes is more than other genes. The peak HPLC indicated for stevioside and Rebaudioside- A represents an increase in the production of this active ingredient under the influence of all treatments. In general, the expression profile of these genes and the results of HPLC show that whatever going to the end of the pathway of production of Rebaudioside- A, the activity of the enzymes increases under the influence of these treatments, and eventually a greater amount of Rebaudioside- A will be produced. This process shows that metal oxides will have a significant effect on the biosynthesis of Rebaudioside- A.

Cytotoxicity of propolis nanopreparations in cancer cell monolayers: multimode of action including apoptotsis and nitric oxide production.

Natural products are invaluable resource of anticancer drug discovery. They generally viewed as safe but weak, within the framework of nanotechnology, they can serve as template for potent anticancer drugs. We first evaluated the cytotoxic activity of different propolis extracts (water, 70% ethanol, absolute ethanol and hexane) in many cancer cell lines, then the solid nanoparticles from the organic solvent extracts were prepared and their cytotoxicity was evaluated as well. Finally, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) liposomes were prepared from the most cytotoxic organic solvent extract and their cytotoxicity was also evaluated. All results collectively showed that hexane extract and its solid nanoparticles as well as its liposomal form exhibited high cytotoxic activity. DPPC/DOPC-hexane extract cytotoxicity selectively depends on the cell line and DOPC liposomal form was characterized by reduced IC50 compared with the other preparations/extracts, the average IC50 value is 165.8 ± 3 µg/ml. The antiproliferative activity of propolis was associated to multiple modes of actions including apoptosis and nitric oxide production and as indicated by the HPLC and FTIR results, it is functioning in many propolis ingredients rather than a single component and influenced by the presence of more lipophilic components within the extract and not by the extract mass yield. These results may have an impact on the multidrug resistivity issue.