Geographical Discrimination in Curcuminoids Content of Turmeric Assessed by Rapid UPLC-DAD Validated Analytical Method.

A fast and reliable ultra-performance liquid chromatography-diode array detection method was developed and validated for the quantitative assessment of turmeric extracts from different geographical locations. Acclaim RSLC PolarAdvantage II column (2.2 µm, 2.1 × 100 mm) was used to analyze individual curcuminoids (curcumin, demethoxycurcumin and bisdemethoxycurcumin) from turmeric samples. The detection was done on ultraviolet absorbance at 425 nm and the column temperature was maintained at 45 °C. A mobile phase consisting of acetonitrile and water was found to be suitable for separation, at a flow rate of 1 mL/min with linear gradient elution. Linearity, specificity, precision, recovery and robustness were measured to validate the method and instrument. Under the described conditions, curcuminoids were collected within one minute. The calibration curve of each curcuminoid showed good linearity (correlation coefficient > 0.999). The relative standard deviations (RSD) of intra-day, inter-day precision and repeatability were less than 0.73%, 2.47% and 2.47%, respectively. In the recovery test, the accuracy ranged from 98.54%-103.91% with RSD values of less than 2.79%. The developed method was used for quantification of individual curcuminoids of turmeric samples. Analysis of turmeric samples from Nepal and South Korea revealed that curcuminoid content was related to geographical location. Turmeric cultivated in warmer climates were found to have higher curcumionoid content than turmeric samples from cooler climates, the southern part of Nepal was found to have two times higher content of curcuminoids than turmeric from the north.

Resonant energy transfer under the influence of the evanescent field from the metal.

We present a quantum framework based on a density matrix of a dimer system to investigate the quantum dynamics of excitation energy transfer (EET) in the presence of the evanescent field from the metal and the phonon bath. Due to the spatial correlation of the electric field in the vicinity of the metal, the spectral density of the evanescent field is similar to that of a shared phonon bath. However, the EET dynamics under the influence of the evanescent field is an open and a new problem. Here we use a thin metallic film to investigate the effect of the evanescent field on the excitation energy transfer in a dimer system based on a density matrix approach. Our results indicate that a thin metallic film enhances the energy transfer rate at the expense of absorbing energy during the process. Since the spectral density of the evanescent field is affected by the geometry of the medium and the distance of a dimer system from the medium, our results demonstrate the possibility to tune EET based on material geometry and distances. Our model also serves as an expansion to quantum heat engine models and provides a framework to investigate the EET in light harvesting molecular networks under the influence of the evanescent field.

Constituents of Cryptotaenia japonica Inhibit Melanogenesis via CREB- and MAPK-Associated Signaling Pathways in Murine B16 Melanoma Cells.

Melanin plays an important role in protecting the skin against ultraviolet light and is responsible for skin color. However, overproduction of melanin is related to several skin disorders, such as age spots, freckles, café au lait spots, Becker's nevus and other hyperpigmentation syndromes. The aim of this study was to identify the effects of kaempferol-7-O-ß-d-glucuronide (K7G) and tilianin, isolated from Cryptotaenia japonica, on melanogenesis and their mechanisms of action in murine B16 melanoma cells. The α-melanocyte-stimulating hormone (α-MSH)-induced melanin production was significantly inhibited by K7G and tilianin in a dose-dependent manner. The effects of these compounds on the signaling pathway of melanogenesis were examined. K7G and tilianin downregulated the expression of microphthalmia-associated transcription factor (MITF) and melanocyte-specific enzymes, i.e., tyrosinase and TRP1. These compounds also inhibited the phosphorylation of cyclic adenosine monophosphate (cAMP)-response element binding protein (CREB) in a dose-dependent manner. In addition, these compounds increased the phosphorylation of extracellular signal-regulated kinase (ERK) but decreased the phosphorylation of c-Jun N-terminal kinase (JNK) in B16 cells. Based on the above results, the anti-melanogenic effects of these compounds are caused by suppression of the MAPK signaling pathway through the down-regulation of α-MSH-induced CREB accumulation. This finding suggests that K7G and tilianin may be good candidates for further research to develop therapeutic agents for hyperpigmentation diseases.

Evaluation of in vitro and in vivo biological activities of Cheilanthes albomarginata Clarke.

BACKGROUND: The Cheilanthes albomarginata Clarke (CA), a fern belonging to Pteridaceae family, is found mainly in India, Nepal, Pakistan and Bhutan at an altitude of 1300-2700 m. It grows mostly in the rock crevices on slopes. Juice from the rhizome of CA has been used to treat peptic ulcer. In this study, the biological activities (antioxidant, anti-inflammatory, anti-adipogenic and anti-obesity) of the extracts of CA were investigated. The total phenolic content of each extract was quantified. This is the first report regarding the study of biological activities on CA. METHODS: In the current study, the crude methanol and fractionated extract of the aerial part of CA were investigated for the antioxidant tests which were namely DPPH assay, hydrogen peroxide scavenging assay and nitrite scavenging assay. Their phenolic contents were measured by the Folin-Ciocalteu's method.In vitro anti-inflammatory and anti-adipogenic assays were evaluated against the RAW 264.7 macrophage cells and 3 T3-L1 cells respectively. The crude methanol extract and phenolic fraction (combination of ethyl acetate and butanol fraction) were studied for the in vivo anti-obesity test using male Sprague Dawley rats. RESULTS: The ethyl acetate fraction showed the strongest DPPH radical scavenging (82.54 ± 0.48%), hydrogen peroxide scavenging (3.41 ± 0.21 mg/ml) and nitrite scavenging activity (61.39%). The highest phenolic content was found in the ethyl acetate fraction followed by the butanol fraction. The ethyl acetate fraction showed the highest in vitro anti-inflammatory and anti-adipogenic activities. From the in vivo study on rats, the crude methanol extract and phenolic fraction showed plasma triglyceride lowering activity as well as reduction of weight of adipose tissue in high fat diet induced obese rats. CONCLUSION: The current study suggests that the ethyl acetate and butanol extracts of CA are potential source for antioxidant, anti-inflammatory and anti-adipogenic remedies. In addition to that the results of in vivo studies evidenced the possibility of CA as a source of anti-obesity drug remedies.

Sources, effects and present perspectives of heavy metals contamination: Soil, plants and human food chain.

Heavy metal (HM) poisoning of agricultural soils poses a serious risk to plant life, human health, and global food supply. When HM levels in agricultural soils get to dangerous levels, it harms crop health and yield. Chromium (Cr), arsenic (As), nickel (Ni), cadmium (Cd), lead (Pb), mercury (Hg), zinc (Zn), and copper (Cu) are the main heavy metals. The environment contains these metals in varying degrees, such as in soil, food, water, and even the air. These substances damage plants and alter soil characteristics, which lowers crop yield. Crop types, growing circumstances, elemental toxicity, developmental stage, soil physical and chemical properties, and the presence and bioavailability of heavy metals (HMs) in the soil solution are some of the factors affecting the amount of HM toxicity in crops. By interfering with the normal structure and function of cellular components, HMs can impede various metabolic and developmental processes. Humans are exposed to numerous serious diseases by consuming these affected plant products. Exposure to certain metals can harm the kidneys, brain, intestines, lungs, liver, and other organs of the human body. This review assesses (1) contamination of heavy metals in soils through different sources, like anthropogenic and natural; (2) the effect on microorganisms and the chemical and physical properties of soil; (3) the effect on plants as well as crop production; and (4) entering the food chain and associated hazards to human health. Lastly, we identified certain research gaps and suggested further study. If people want to feel safe in their surroundings, there needs to be stringent regulation of the release of heavy metals into the environment.

Cinnamomum verum extract inhibits NOX2/ROS and PKCδ/JNK/AP-1/NF-κB pathway-mediated inflammatory response in PMA-stimulated THP-1 monocytes.

BACKGROUND: Cinnamomum verum J. Presl (Cinnamon) is widely used in the food and pharmaceutical industries. C. verum exhibits various biological activities. However, it is unclear whether C. verum can inhibit NOX, a major source of ROS generation, and exert anti-inflammatory and antioxidant effects in PMA-stimulated THP-1 cells. PURPOSE: This study investigates the anti-inflammatory and antioxidant effects of C. verum in PMA-stimulated THP-1 cells. METHODS: The MeOH extract of C. verum was analyzed using UPLC-QTOF/MS. Anti-inflammatory and antioxidant effects of C. verum extract were examined by DCF-DA staining, immunofluorescence staining, RT-PCR, and immunoblotting in PMA-stimulated THP-1 cells. RESULTS: C. verum and its components, cinnamic acid and coumarin, significantly attenuated the expression of IL-1ß, IL-8, CCL5, and COX-2 in PMA-stimulated THP-1. C. verum decreased ROS levels via NOX2 downregulation, as well as ameliorated plasma membrane translocation of PKCδ and decreased JNK phosphorylation. Besides, C. verum suppressed the nuclear translocation of AP-1 and NF-κB, which modulates diverse pro-inflammatory genes. CONCLUSION: C. verum effectively inhibits inflammation and oxidative stress during monocyte-macrophage differentiation and downregulates inflammatory mediators via NOX2/ROS and PKCδ/JNK/AP-1/NF-κB signaling.

Pimarane Diterpenoids from Aerial Parts of Lycopus lucidus and Their Antimicrobial Activity.

The ethyl acetate fraction obtained from aerial parts of L. lucidus was subjected for isolation of new bioactive compounds, which enabled isolation of five new pimarane-type diterpenoids, namely, 3ß, 8ß, 12ß, 18-tetrahydroxy pimar-15-ene (10), 7α, 8ß, 12ß, 18-tetrahydroxy pimar-15-ene (11), 3ß, 8ß, 11ß, 12α, 18-pentahydroxy pimar-15-ene (12), 12ß acetoxy, 8ß, 3ß, 18-trihydroxy pimar-15-ene (13), and 3ß acetoxy, 8ß, 12ß, 18-trihydroxy pimar-15-ene (14), along with nine known compounds. The structures were elucidated by spectroscopic analysis and comparison with literature data. The isolated new pimarane diterpenoids were examined for antimicrobial activity against Gram-negative and Gram-positive bacteria strains. Among them, the compound 3ß, 8ß, 12ß, 18-tetrahydroxy pimar-15-ene (10) was most effective, exhibiting minimum inhibitory concentration (MIC) values of 15.62 µg/mL against Staphylococcus epidermidis, 31.25 µg/mL against Staphylococcus aureus, 62.5 µg/mL against Pseudomonas aeruginosa, and 125 µg/mL against Escherichia coli.

Study of Antibacterial Activity of Root Bark, Leaves, and Pericarp Extracts of Diploknema butyracea and Evaluation of Prospective Antioxidant Activity.

This study was aimed to determine the antibacterial activity of root bark, leaves, and pericarp extract of Diploknema butyracea and to evaluate the prospective antioxidant activity, total flavonoid, polyphenol, and carbohydrate content. The plant parts were collected and extracted by cold maceration, using hexane, ethyl acetate, methanol, and distilled water. Phytochemical screening of different samples of D. butyracea in different solvents revealed the presence of varied extent of alkaloid, saponin, terpenoid, anthraquinones, tannin, cardiac glycoside, flavonoid, carbohydrate, polyphenol, protein and amino acid, resin, and phytosterol. Our study showed that methanolic root bark extract exhibited the potent antimicrobial activity against Staphylococcus aureus, Staphylococcus epidermidis, and Klebsiella pneumonia with an average zone of inhibition of 17.33 mm, 14.33 mm, and 13.0 mm, respectively. Surprisingly, all of the extracts were insensitive to Escherichia coli. The lowest minimum bactericidal concentration (MBC), 4.6 mg/ml, was observed with the aqueous pericarp extract against S. epidermidis and the highest was of 50 mg/ml shown by ethyl acetate pericarp against K. pneumonia. Our results showed that both the polar and nonpolar components present in the different parts of D. butyracea exhibit prominent antibacterial activities against different bacterial strains. The in vitro 2,2'-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity showed that the methanol extract of root barks displayed the most potent antioxidant activity (IC50 : 6.1 µg/ml). The total polyphenol content of the plant part extracts was observed between 19.48 ± 0.23 and 123.48 ± 1.84 µg gallic acid equivalent/mg of dry extract weight. Likewise, flavonoid content ranged from 40.63 ± 1.28 µg to 889.72 ± 3.40 µg quercetin equivalent/mg of dry extract weight and total carbohydrate content ranged from 11.92 ± 0.60 µg to 174.72 ± 0.60 µg glucose equivalent per/mg dry extract weight. Overall, our study showed that the root bark, pericarp, and leaves extract of D. butyracea evinced prominent antibacterial properties against various pathogenic bacterial strains.

Simple solution to preserve plant samples for microbiome analyses.

Culture-independent survey techniques are fundamental tools when assessing plant microbiomes. These methods rely on DNA that is carefully preserved after collecting samples to achieve meaningful results. Immediately freezing samples to -80°C after collection is considered one of the most robust methods for preserving samples before DNA extraction but is often impractical. Preservation solutions can solve this problem, but commercially available products are expensive, and there is limited data comparing their efficacy with other preservation methods. In this study, we compared the impact of three methods of sample preservation on plant microbiome surveys: (1) RNAlater, a proprietary preservative, (2) a home-made salt-saturated dimethyl sulphoxide preservation solution (DESS), and (3) freezing at -80°C. DESS-preserved samples, stored at room temperature for up to four weeks, did not show any significant differences to samples frozen at -80°C, while RNAlater inflated bacterial alpha diversity. Preservation treatments did not distinctively influence fungal alpha diversity. Our results demonstrate that DESS is a versatile and inexpensive preservative of DNA in plant material for diversity analyses of fungi and bacteria.

Enhancement of Resonant Energy Transfer Due to an Evanescent Wave from the Metal.

The high density of evanescent modes in the vicinity of a metal leads to enhancement of the near-field Förster resonant energy transfer (FRET) rate. We present a classical approach to calculate the FRET rate based on the dyadic Green's function of an arbitrary dielectric environment and consider the nonlocal limit of material permittivity in the case of the metallic half-space and thin film. In a dimer system, we find that the FRET rate is enhanced due to shared evanescent photon modes bridging a donor and an acceptor. Furthermore, a general expression for the FRET rate for multimer systems is derived. The presence of a dielectric environment and the path interference effect enhance the transfer rate, depending on the combination of distance and geometry.

Psychosocial problems among individuals with substance use disorders in drug rehabilitation centers, Nepal.

BACKGROUND: Substance use is associated with numerous undesirable short and long term consequences. The individual not only suffer from physical and psychological problems but also loses the ability to interact with family, peers and society. The present study aims to identify the psychosocial problems and associated factors among individuals with substance use disorders. METHODS: The study was conducted using descriptive cross sectional research design in different drug rehabilitation centers of Nepal. Probability proportional to size sampling technique was used to select the drug rehabilitation centers and individuals diagnosed with substance use disorder but free from any substance withdrawal features, admitted in rehabilitation centers within 3 months and clients above 15 years of age were included in the study (N = 204). A standard tool Drug Use Screening Inventory-Revised was used to assess the psychosocial problems and the scores ranged from 0 to 100. The data were analyzed using descriptive and inferential statistics. Linear regression analysis was conducted to identify the predictors of psychosocial problems. RESULTS: The mean and standard deviation of overall psychosocial problems scores was 60.42 ± 19.44. Among different domains, more problems were found in substance use domain (75.00 ± 21.43) followed by school performance (64.60 ± 25.53), behavior pattern (64.53 ± 24.44), peer relationship (64.49 ± 24.91), social competence (61.30 ± 22.60), psychiatric disorder (56.83 ± 23.39), family system (48.28 ± 23.72) and work adjustment (45.90 ± 29.88) domains. Types of substance use, mode of substance use and age of initiation of substance use were the significant predictors of overall and most of the domains of psychosocial problems. CONCLUSION: Substance use poses significant impact on individual's psychological and social wellbeing. Individuals using injecting drugs, initiating substances early in life, using substances many times in a day and using both licit and illicit substances had more psychosocial problems. Hence, treatment centers should take this into account.

Development of UPLC Fingerprint with Multi-Component Quantitative Analysis for Quality Consistency Evaluation of Herbal Medicine "Hyangsapyeongwisan".

Hyangsapyeongwisan (HSPWS), known as traditional herbal medicine, has been used in the treatment of gastric disease. Standardization of HSPWS is a necessary step for the establishment of a consistent biological activity for the production and manufacturing of HSPWS herbal preparations. A simple, sensitive and accurate method using ultra-performance liquid chromatography (UPLC) fingerprinting with a diode array detector (DAD) was developed and validated for systematic quality evaluation of HSPWS. Separation conditions were optimized using a Halo C18 2.7 µm, 4.6 × 100 mm column with a mobile phase of 0.1% phosphoric acid and acetonitrile, and detection wavelengths of 215, 250 and 350 nm. Validation of the analytical method was evaluated by tests of linearity, precision, accuracy and robustness. All calibration curves of components showed good linearity (R(2) > 0.9996). The limit of detection (LOD) and limit of quantification (LOQ) were within the ranges of 0.004-0.134 and 0.012-0.406 µg/mL, respectively. The relative standard deviation (RSD) values of intra- and inter-day testing were within the range of 0.01-3.84%. The result of the recovery test was 96.82-104.62% with an RSD value of 0.14-3.84%. Robustness values of all parameters as well as the stability test of analytical solutions were within the standard limit. It showed that the developed method was simple, specific, sensitive, accurate, precise, reproducible and robust for the quantification of active components of HSPWS. Chromatographic fingerprinting with quantitative analysis of marker compounds in HSPWS prepared by different methods and commercial formulation was also evaluated successfully.

Quantitative assessment of traditional Oriental herbal formulation Samhwangsasim-tang using UPLC technique.

A specific and reliable ultra-performance liquid chromatography-diode array detection method has been developed and validated for the quantitative assessment of a traditional Oriental herbal formulation, Samhwangsasim-tang (SST). A Halo reversed-phase amide column (2.7 µm, 4.6 × 150 mm) was used to separate marker compounds; detection was conducted by ultraviolet absorbance at 250 nm. The column temperature was maintained at 45°C. A mobile phase consisting of acetonitrile (A) and 0.1% trifluoroacetic acid in water (B) was found to be suitable for the separation, at a flow rate of 1.8 mL/min with gradient elution. Linearity, specificity, precision and recovery were calculated to validate the method and instrumentation. Under the described conditions, all marker compounds (rhaponticin, berberine, palmatine, baicalin, baicalein and wogonin) were collected within 25 min. All calibration curves of components showed good linearity (correlation coefficient > 0.9996). The limit of detection and limit of quantification ranged from 0.08-3.05 and 0.23-8.12 µg/mL, respectively. The relative standard deviation (RSD) and repeatability values of intra-day and inter-day precision were less than 2.30, 2.99 and 1.82%, respectively. In the recovery test, the accuracy ranged from 97.56-103.30% with RSD values less than 2.63%. The developed method was simple, specific, sensitive, accurate, precise and reproducible for the quantification of the active chemical constituents of SST. The simultaneous analysis of the contents of marker compounds in different SST samples prepared by different extraction procedures and different commercial products was successfully evaluated.

Development of simultaneous analysis for marker constituents in Hwangryunhaedok-tang () and its application in commercial herbal formulas.

This study was performed to develop methods for the chromatographic determination of biomarkers in Hwangryunhaedok-tang (HHT) and the quantitative evaluation of commercial HHT. To develop an analytical method, an RP-amide column (2.7 µm, 4.6 × 100 mm, Halo: Supelco, Bellefonte, PA) was used with a gradient solvent system of mixed acetonitrile and 0.1 % phosphoric acid/water and an ultra performance liquid chromatography-diode array detector. The method was validated by specificity, linearity, accuracy (recovery) and precision tests (repeatability, intra and inter-day). The correlation coefficients (R (2)) of biomarkers were calculated as 0.9998-1.000 and their ranges were as follows: geniposide (62.5-1,000.0 µg/ml), berberine (31.3-500.0 mg/ml), palmatine (31.3-500.0 µg/ml), baicalin (125.0-1,500.0 µg/ml), baicalein (15.6-250.0 µg/ml) and wogonin (5.2-125.0 µg/ml), respectively. The limit of detection was 0.34-4.01 µg/ml, and the limit of quantification was 1.02-12.16 µg/ml. The intra-day and inter-day precision of six components were revealed as 0.02-2.48 % as a relative standard deviation (RSD). The repeatability value of biomarkers in three different concentrations of HHT was 0.29-2.98 % (RSD value) and recovery was 95.72-104.90 %. Among several extraction methods tested, biomarker content was higher with the 20 times extraction (20TE) and mixture of extract powder (MEP) methods than with any other method, and some differences among diverse pharmaceutical medicines were revealed. The validation data indicated that the method developed is suited to the determination of six marker compounds in HHT. The content of biomarkers by simultaneous analysis was evaluated in 20TE, MEP, USA formula and Taiwan formula.

Ixeris dentata NAKAI Reduces Clinical Score and HIF-1 Expression in Experimental Colitis in Mice.

Ixeris dentata (ID) is an herbal medicine used in Asian countries to treat indigestion, pneumonia, hepatitis, contusions, and tumors; however, its effect on intestinal inflammation is unknown. Thus, we investigated the effect of ID in the dextran sulfate sodium (DSS) model of colitis in female BALB/c mice; animals were evaluated after seven days of DSS treatment. DSS-treated mice showed considerable clinical signs, including weight loss, reduced colon length, colonic epithelial injury, infiltration of inflammatory cells in the colon tissue, and upregulation of inflammatory mediators. However, administration of ID attenuated body weight loss, colon shortening, and the increase in disease activity index score. ID also significantly decreased the colonic mucosal injury and the number of infiltrating mast cells. Moreover, ID inhibited the expressions of cyclooxygenase-2 and hypoxia-inducible factor-1 α in colon tissue. Taken together, the results provide experimental evidence that ID might be a useful therapy for patients with ulcerative colitis.