A comparison between the enzymatic oxidation method and headspace gas chromatography with a flame ionization detector in the determination of postmortem blood ethanol.

Ethanol is the most commonly encountered substance in forensic toxicology. Determining blood alcohol concentration (BAC) in autopsies accounts for the majority of work in forensic diagnosis. The most common method to assess BAC is the enzymatic oxidation method because of its low cost, easy operation, and high throughput. Still, the elevated lactate and lactate dehydrogenase (LDH) levels in postmortem blood may affect accuracy. This study uses headspace gas chromatography with a flame ionization detector (HS-GC/FID) to assess the interference of lactate and LDH levels on BAC in 110 autopsied blood samples determined by the enzymatic oxidation method. The results showed that lactate and LDH levels in postmortem blood were higher than in normal blood. There was a weak correlation between the lactate levels and BAC difference (r = 0.23, p < 0.05) and a strong correlation between LDH levels and BAC difference (r = 0.67, p < 0.001). The differentiation of BAC between the enzymatic oxidation method and HS-GC/FID was significant (p < 0.001), confirming the interference significantly. All postmortem blood samples with lactate and LDH levels higher than regular lead to a positive error in determining BAC by enzymatic oxidation method. The study results suggest that the HS-GC/FID method should be used to determine BAC in postmortem blood samples instead of the enzymatic oxidation method to avoid mistakes in forensic diagnosis.

Anti-inflammatory and Antiphytopathogenic Fungal Activity of 2,3-seco-Tirucallane Triterpenoids Meliadubins A and B from Melia dubia Cav. Barks with ChemGPS-NP and In Silico Prediction.

Two new rearranged 2,3-seco-tirucallane triterpenoids, meliadubins A (1) and B (2), along with four known compounds, 3-6, were isolated from the barks of Melia dubia Cav. Compound 2 exhibited a significant inflammatory inhibition effect toward superoxide anion generation in human neutrophils (EC50 at 5.54 ± 0.36 µM). It bound to active sites of a human inducible nitric oxide synthase (3E7G) through interactions with the residues of GLU377 and PRO350, which may benefit in reducing the neutrophilic inflammation effect. The ChemGPS-NP interpretation combined with bioactivity assay and in silico prediction results suggested 2 to be an agent for targeting iNOS with different mechanisms as compared to a selected set of current approved drugs. Moreover, compounds 1 and 2 showed remarkable inhibition against the rice pathogenic fungus Magnaporthe oryzae in a dose-dependent manner with IC50 values of 137.20 ± 9.55 and 182.50 ± 18.27 µM, respectively. Both 1 and 2 displayed interactions with the residue of TYR223, a key active site of trihydroxynaphthalene reductase (1YBV). The interpretation of 1 and 2 in the ChemGPS-NP physical-chemical property space indicated that both compounds are quite different compared to all members of a selected set of reference compounds. In light of demonstrated biological activity and in silico prediction experiments, both compounds possibly exhibited activity against phytopathogenic fungi via a novel mode of action.

Study on the effect of processing methods on the total polyphenol, 2,3,5,4'-tetrahydroxystilben-2-O-ß-D-glucoside, and physcion contents in Fallopia multiflora Thunb. Haraldson root

Abstract This study investigated the changes in the ingredients in Fallopia multiflora Thunb. Haraldson (FMT) root after processing it with different methods such as soaking, stewing, and steaming or combined methods. The total polyphenol, 2,3,5,4'-tetrahydroxystilben-2-O-ß-D-glucoside (THSG), and physcion contents in FMT products after processing were determined using high-performance liquid chromatography (HPLC) and ultraviolet-visible (UV-VIS) methods. The results demonstrated that the processing method and time significantly affected the contents of polyphenol, THSG, and physcion. The physcion and total polyphenol content increased or decreased during processing depending upon the processing time, while the THSG content gradually decreased with an increase in the processing time. The content of physcion (a substance that can cause liver toxicity) was analysed, and the suitable conditions for processing of the FMT products were determined as initial soaking in rice swill for 24 h and subsequent stewing with black beans and water for 12 h

Chemical Composition Analysis and Antioxidant Activity of Coffea robusta Monofloral Honeys from Vietnam.

Monofloral honey samples (Coffea robusta) from Vietnam were determined for their chemical compositions. This is the first report on the chemical composition and antioxidant activity of coffee honey from Vietnam. These samples were characterized by their high contents of total and reducing sugars, total phenolic contents, and total flavonoid contents. The contents of seven phenolic acids (PAs) were quantified by high performance liquid chromatography (HPLC) and analyzed with the assistance of principle component analysis (PCA) to differentiate the honey samples into groups. The hydroxymethylfurfural (HMF) (0.048-2.933 mg/kg) and free acid contents (20.326-31.163 meq/kg) of coffee honey were lower in Nepal, which reflected the freshness of the honey when conducting this survey. The coffee honey had total sugar and reducing sugar contents 831.711 g/kg and 697.903 g/kg, respectively. The high level of total phenolic (0.642 mg GAE/g) and flavonoid (0.0341 mg GE/g) contents of coffee honey contributed to their antioxidant activity of this honey sample. Among the coffee honey tested, the IC50 of DPPH radical-scavenging activities value was 1.134-17.031 mg/mL, while the IC50 of ABTS radical-scavenging activities value was 115.381-213.769 mg/mL. The phenolic acids composition analysis displayed that gallic acid appeared in high concentrations in all studied honey samples, ranging from 0.037-1.015 mg/kg, and ferulic acid content ranged from 0.193 to 0.276 mg/kg. The content of trigonelline and caffeine in coffee honey samples ranged from 0.314-2.399 mg/kg and 8.946-37.977 mg/kg. The data in this article highlight the relevance of coffee honey as a healthy substance.

Inhibition of α-Glucosidase, Acetylcholinesterase, and Nitric Oxide Production by Phytochemicals Isolated from Millettia speciosa-In Vitro and Molecular Docking Studies.

The phytochemical constituents from the roots of Millettia speciosa were investigated by chromatographic isolation, and their chemical structures were characterized using the MS and NMR spectroscopic methods. A total of 10 compounds, including six triterpenoids, two flavonoids, and two phenolic compounds, were identified from the roots of M. speciosa. Out of the isolated compounds, eight showed inhibitory effects on NO production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, with IC50 values ranging from 43.9 to 449.5 µg/mL. Ursane-type triterpenes significantly suppressed NO production compared to the remaining compounds. In addition, these compounds also exhibited remarkable inhibitory effects on α-glucosidase. Among the tested compounds, 4, 5, and 10 exhibited excellent α-glucosidase inhibition, with IC50 values ranging from 1.1 to 2.2 µg/mL. Almost all of the test compounds showed little or no acetylcholinesterase inhibition, except for 5, which showed moderate anti-acetylcholinesterase activity in vitro. The molecular docking study of α-glucosidase inhibition by 3-5 and 10 was conducted to observe the interactions of these molecules with the enzyme. Compounds 4, 5, and 10 exhibited a better binding affinity toward the targeted receptor and the H-bond interactions located at the entrance of the enzyme active site pocket in comparison to those of 3 and the positive control acarbose. Our findings evidence the pharmacological potential of this species and suggest that the phytochemicals derived from the roots of M. speciosa may be promising lead molecules for further studies on the development of anti-inflammatory and anti-diabetes drugs.

Constituents from the Fruiting Bodies of Trametes cubensis and Trametes suaveolens in Vietnam and Their Anti-Inflammatory Bioactivity.

It is reported that various fungi have been used for medicine and edible foods. The tropical Trametes genus is popular and well-known in Vietnam for its health effects and bioactivities. In this study, the fruiting bodies of the edible fungi T. cubensis and T. suaveolens were collected in Vietnam. The preliminary bioactivity screening data indicated that the methanol extracts of the fruiting bodies of T. cubensis and T. suaveolens displayed significant inhibition of superoxide anion generation and elastase release in human neutrophils. Therefore, the isolation and characterization were performed on these two species by a combination of chromatographic methods and spectrometric analysis. In total, twenty-four compounds were identified, and among these (1-3) were characterized by 1D-, 2D-NMR, and HRMS analytical data. In addition, the anti-inflammatory potentials of some purified compounds were examined by the cellular model for the inhibition of superoxide anion generation and elastase release in human neutrophils. Among the isolated compounds, (5,14), and (19) displayed significant anti-inflammatory potential. As the results suggest, the extracts and isolated compounds from T. cubensis and T. suaveolens are potential candidates for the further development of new anti-inflammatory lead drugs or natural healthy foods.

A novel flavonoid from Fissistigma cupreonitens, 5­hydroxy­7,8­dimethoxyflavanone, competitively inhibited the efflux function of human P-glycoprotein and reversed cancer multi-drug resistance.

BACKGROUND: P-glycoprotein (P-gp) over-expression plays a vital role in not only systemic drug bioavailability but also cancer multi-drug resistance (MDR). Develop functional inhibitors of P-gp can conquer both problems. PURPOSE AND STUDY DESIGN: The aim of the present study was to research the P-gp modulating effects and MDR reversing ability of a novel flavonoid from Fissistigma cupreonitens, the underlying inhibitory mechanisms were further elucidated as well. METHODS: Calcein-AM, rhodamine 123, and doxorubicin were fluorescent substrates for the evaluation of P-gp inhibitory function and detailed drug binding modes. Docking simulation was performed to reveal the in silico molecular bonding. ATPase assay and MDR1 shift assay were adopted to reveal the ATP consumption and conformational change of P-gp. The MDR reversing effects were demonstrated through cytotoxicity, cell cycle, and apoptosis analyses. RESULTS: 5­hydroxy­7,8­dimethoxyflavanone inhibited the efflux of rhodamine 123 and doxorubicin in a competitive manner, and increased the intracellular fluorescence of calcein at a concentration as low as 2.5 µg/ml. 5­hydroxy­7,8­dimethoxyflavanone slightly changed P-gp's conformation and only stimulated ATPase at very high concentration (100 µg/ml). The docking results showed that 5­hydroxy­7,8­dimethoxyflavanone and verapamil exhibited similar binding affinity to P-gp. The MDR reversing effects were prominent in the vincristine group, the reversal folds were 23.01 and 13.03 when combined with 10 µg/ml 5­hydroxy­7,8­dimethoxyflavanone in the P-gp over-expressing cell line (ABCB1/Flp-In™-293) and MDR cancer cell line (KB/VIN), respectively. CONCLUSION: The present study demonstrated that 5­hydroxy­7,8­dimethoxyflavanone was a novel effective flavonoid in the P-gp efflux inhibition and in vitro cancer MDR reversion.

An overview of the chemical composition and biological activities of essential oils from Alpinia genus (Zingiberaceae).

Alpinia Roxb. is the largest genus of the Zingiberaceae family. A large number of Alpinia species has been used as food and traditional medicines. Alpinia essential oils have been studied for their chemical profiles, in which 1,8-cineole, ß-pinene, α-pinene, ß-myrcene, camphor, γ-terpinene, p-cymene, geraniol, α-fenchyl acetate, ocimene, methyl cinnamate, and ß-caryophyllene have been found to be the major compounds. Essential oils isolated from Alpinia plants have been reported to have antimicrobial, cytotoxic, antioxidant, anti-inflammatory, anti-asthmatic, tyrosinase inhibitory, insecticidal, and larvicidal activities and slimming aromatherapy. In this review, the comprehensive information regarding the volatile components of various Alpinia plants, the bioactivities of Alpinia essential oils and their major compounds are provided.

Triterpenoids and steroids from the fruiting bodies of Hexagonia tenuis and their cytotoxicity.

One new triterpenoid, hexagonin F (1) was characterized from the fruiting bodies of Hexagonia tenuis with the assistance of spectroscopic and spectrometric analytical methods. In addition, two triterpenoids and two steroids were also identified by comparison of their physical and spectroscopic data with those reported. The purified compounds were examined for their cytotoxicity against five tumor cell lines, however, only weak cytotoxicity was demonstrated.

A new triterpenoid and other compounds from lichens Cryptothecia faveomaculata Makhija & Patw.

Ten compounds (1-10) were purified from the extract of lichens C. faveomaculata, and among these one new triterpenoid, cryptothecin A (1) was characterized through mass spectrometric and 2D NMR spectroscopic analyses. Some of the isolated compounds were assessed for their antimicrobial (1, 6, and 8) and cytotoxic activity (1, 4-5, and 7-9), but only weak inhibitory effects were observed.

Chemical Constituents of the Leaves of Peltophorum pterocarpum and Their Bioactivity.

Two new sesquiterpenoids peltopterins A and B (compounds 1 and 2) and fifty-two known compounds were isolated from the methanol extract of P. pterocarpum and their chemical structures were established through spectroscopic and mass spectrometric analyses. The isolates 40, 43, 44, 47, 48, 51 and 52 exhibited potential inhibitory effects of superoxide anion generation or elastase release.

Chemical Constituents from the Stems of Tinospora sinensis and Their Bioactivity.

Fifty-seven compounds were purified from the stems of Tinospora sinensis, including three new compounds characterized as a lignan (1), a pyrrole alkaloid (11), and a benzenoid (17), respectively. Their structures were elucidated and established by various spectroscopic and spectrometric analytical methods. Among the isolates, fifteen compounds were examined for their anti-inflammatory potential in vitro. The results showed that several compounds displayed moderate inhibition of N-formyl-methionyl-leucyl-phenylalanine/cytochalasin B (fMLP/CB)-induced superoxide anion generation and elastase release.

Chemical constituents from the fruiting bodies of Phellinus igniarius.

A new tirucallane-type triterpenoid igniarine (1), and four known compounds meshimakobnol A (2), meshimakobnol B (3), ergosterol (4) and ergosterol peroxide (5), were purified from the methanol extracts of the fruiting bodies of Phellinus igniarius (DC. ex Fr.) Quél. The structure of 1 was elucidated using a combination of 1D and 2D NMR techniques and HR-ESI-MS analyses. In addition, the isolated compounds were examined for their cytotoxicity against several tumour cell lines and part of the tested compounds demonstrated weak cytotoxicity.

Characterization of Cyclodepsipeptides from the Mycelium of Isariajaponica from Vietnam.

A new cyclodepsipeptide, isarin (1), was isolated from the insect pathogenic fungus Isaria japonica Yasuda, together with one known compound, beauveriolide 1 (2). The structure of this new compound was characterized using a combination of spectroscopic and spectrometric analyses.

Constituents from the leaves of Clausena lansium and their anti-inflammatory activity.

Five new acyclic amides, clausenalansamides C-G (1-5), clausenaline G (6) and (±)-5-(4-methylphenyl)-γ-valerolactone (7) reported from the natural source for the first time, were characterized from the leaves of Clausena lansium. Their structures were established by spectroscopic and spectrometric methods, and the absolute configurations of new compounds 1-5 were determined by electronic circular dichroism and single-crystal X-ray diffraction analyses. Eighteen compounds were evaluated for their anti-inflammatory activity and only imperatorin (11) and wampetin (12) displayed significant inhibition of fMLP/CB-induced superoxide anion generation with IC50 values of 1.7 ± 0.3 and 6.8 ± 1.1 µM, respectively.

Mechanistic Study of Tetrahydrofuran- acetogenins In Triggering Endoplasmic Reticulum Stress Response-apotoposis in Human Nasopharyngeal Carcinoma.

For past three decades, numerous studies have elucidated the antiproliferative effects of acetogenins in hopes of developing a new class of clinical anticancer agents. However, clear and definitive action mechanisms of acetogenins were less clarified. In the present study, three tetrahydrofuran (THF)-containing acetogenins were found to have potent and selective antiproliferative activity against human nasopharyngeal carcinoma (NPC) cell lines and their methotrexate-resistant counterparts. The THF-containing acetogenins induced G2/M phase arrest, mitochondrial damage and apoptosis, and increased cytosolic and mitochondrial Ca2+ in NPCs. Microarray analysis of NPC-TW01 cells treated with squamostatin A, a non-adjacent bis-THF acetogenin, demonstrated an increased endoplasmic reticulum (ER)-stress response (ESR). Enhanced ESR in squamostatin A-treated cells was confirmed by real-time PCR, Western blot and shRNA gene knockdown experiments. Although our results showed that squamostatin A-induced ESR was independent of extracellular Ca2+, the presence of extracellular Ca2+ enhanced the antiproliferative effect of acetogenins. In vivo analyses demonstrated that squamostatin A showed good pharmacokinetic properties and significantly retarded NPC tumor growth in the xenograft mouse model. Conclusively, our work demonstrates that acetogenins are effective and selective inducers of the ESR that can block NPC proliferation, and illustrate a previously unappreciated antitumor mechanism of acetogenins that is effective against nasopharyngeal malignancies.

3-Methyl-4,5-dihydro-oxepine, polyoxygenated seco-cyclohexenes and cyclohexenes from Uvaria flexuosa and their anti-inflammatory activity.

The phytochemical investigation of the methanolic extract of Uvaria flexuosa (Annonaceae) leaves led to the isolation of seven compounds including, 3-methyl-4,5-dihydro-oxepine (flexuvaroxepine A), four polyoxygenated seco-cyclohexene (flexuvarin A-D) and two polyoxygenated cyclohexene (flexuvarol A-B) derivatives, together with four known flavones. The structures of the isolated compounds were elucidated using different spectroscopic techniques. A plausible biogenetic route of the new compounds was discussed. The anti-inflammatory activity of the isolated compounds was evaluated by superoxide anion generation and elastase release assays. Among the tested compounds, flexuvarol B and chrysin showed the most potent anti-inflammatory effect by inhibiting superoxide anion generation and elastase release from human neutrophils in response to fMLP with IC50 2.25-5.55µM. Moreover, 5-hydroxy-6,7-dimethoxy-flavone showed selective inhibitory activity on superoxide anion generation (IC50=1.19±0.34µM).

γ- and δ-Lactams from the Leaves of Clausena lansium.

Eight new clausenamides, including three γ-lactams (1-3), four δ-lactams (4-7), and an amide (8), and seven known lactams, including compounds 9-11, which were purified from natural sources for the first time, were characterized from the leaves of Clausena lansium. Their structures were elucidated using spectroscopic methods, and the absolute configurations were determined using electronic circular dichroism and single-crystal X-ray diffraction analyses with Cu Kα radiation. Compound 2 (50 µM) protected 22.24% of cortical neurons against Aß25-35-induced cell death.

Chemical Constituents from the Fruiting Bodies of Hexagonia apiaria and Their Anti-inflammatory Activity.

A chemical investigation of the fruiting bodies of Hexagonia apiaria resulted in the identification of nine compounds including five new triterpenoids, hexagonins A-E (1-5), along with four known compounds. The purified constituents were examined for their anti-inflammatory activity. Among the tested compounds, hexatenuin A displayed the most significant inhibition of superoxide anion generation and elastase release. These triterpenoids may have potentials as anti-inflammatory agents.

Composition of essential oils from Euodia lepta (Spreng.) Merr and Euodia calophylla Guill., grown in Vietnam / Composición de los aceites esenciales de Euodia lepta (Spreng.) Merr y Euodia calophylla Guill., crecidas en Vietnam

The chemical compositions of essential oils obtained by hydrodistillation of the leaves, stems and flowers of Euodia lepta and Euodia callophylla grown in Vietnam and then analysed by gas chromatography-flame ionization detector (GC-FID) and gas chromatography/mass spectrometry (GC-MS) were being reported. The main compounds of the leaves oil of E. lepta were (E)-beta-ocimene (24.4 percent), alpha-pinene (9.8 percent), (Z)-beta-ocimene (6.3 percent) and delta-cadinene (5.2 percent), while the stems oil comprised of spathulenol (26.0 percent), (E)-beta- ocimene (9.9 percent) and (Z)-9-octadecenamide (7.7 percent). However, ciscarane (19.2 percent), alpha-cadinol (10.8 percent), alpha-pinene (10.5 percent) and (E)-beta-ocimene (9.0 percent) were present in the flowers oil of E. lepta. On the other hand, alpha-pinene (8.3 percent), trans-alpha-bergamotene (7.5 percent), (E)-beta-ocimene (7.0 percent) and (E)-nerolidol (6.6 percent) were the major constituents of the leaves oil of E. calophylla. The quantitatively significant compounds of the stems oil were (E,E)-alpha-farnesene (11.9 percent), alpha-terpinolene (11.3 percent) and alpha-pinene (8.2 percent), while alpha-pinene (21.6 percent), limonene (19.0 percent) and sabinene (15.5 percent) were obtained from the flowers oil...

La composición químicas de los aceites esenciales obtenidos por hidrodestilación de las hojas, tallos y flores de Euodia lepta y Euodia callophylla cultivadas en Vietnam, fueron analizados por cromatografía de gases-detector de ionización de llama (GC-FID) y la cromatografía de gases/espectrometría de masas (GC-MS). Los principales compuestos del aceite de hojas de E. lepta fueron (E) -beta-ocimeno (24,4 por ciento), alfa-pineno (9,8 por ciento), (Z)-beta- ocimeno (6,3 por ciento) y delta-cadineno (5.2 por ciento), mientras que los tallos de aceite estaban compuestos de spatulenol (26,0 por ciento), (E) -beta-ocimeno (9,9 por ciento) y (Z) -9- octadecenamida (7,7 por ciento). Sin embargo, cis-carano (19,2 por ciento), alfa-cadinol (10,8 por ciento), alfa-pineno (10,5 por ciento) y (E) -beta-ocimeno (9,0 por ciento) estaban presentes en el aceite de flores de E. lepta. Por otro lado, alfa-pineno (8,3 por ciento), trans-alfa-bergamoteno (7,5 por ciento), (E) - beta-ocimeno (7,0 por ciento) y (E) -nerolidol (6,6 por ciento) fueron los principales constituyentes del aceite de las hojas de E. calophylla. Los compuestos cuantitativamente significativos del aceite de los tallos fueron (E, E)-farneseno -alfa (11,9 por ciento), alfa-terpinoleno (11,3 por ciento) y alfa-pineno (8,2 por ciento), mientras que alfa-pineno (21,6 por ciento), limoneno (19,0 por ciento) y sabineno (15,5 por ciento) se obtuvieron del aceite de las flores...