Bioactivity responses to changes in mucus-associated bacterial composition between healthy and bleached Porites lobata corals.

This study aims to investigate how bioactivities of the coral surface mucus layer (SML) respond to changes in mucus-associated bacterial communities between bleached and healthy Porites lobata corals in Nha Trang Bay, Vietnam. The findings suggested that significant shifts in the mucus-associated bacterial communities were related to changes in coral health states from bleached to healthy P. lobata colonies (p < 0.05), while bacterial compositions were not significantly different across seasons and locations (p > 0.05). Of which 8 genera, Shewanella, Fusibacter, Halodesulfovibrio, Marinifilum, Endozoicomonas, Litoribacillus, Algicola, and Vibrio were present only in the SML of bleached coral while absent in the SML of the healthy one. As compared with the bleached SML, the healthy SML demonstrated stronger antibacterial activity against a coral bleaching pathogen, V. coralliilyticus, higher antitumor activity against HCT116 cell accompanied with increased induction of cleaved PARP and accelerated cell nucleic apoptosis and cycle arrest at S and G2/M phases exhibiting several typical characteristics, cell shrinkage, lost cell contact, and apoptotic body formation. Moreover, putative compounds detected at 280 nm in the healthy SML were obviously higher than those in the bleached one, probably they could be bioactive molecules responsible for competitively exclusion of pathogens, Algicola and Vibrio, from the healthy SML.

Ionic conduction in ammonia functionalised closo-dodecaborates MB12H11NH3 (M = Li and Na).

Metal hydroborates and their derivatives have been receiving attention as potential solid-state ion conductors for battery applications owing to their impressive electrochemical and mechanical characteristics. However, to date only a fraction of these compounds has been investigated as solid-state electrolytes. Here, MB12H11NH3 (M = Li and Na) hydroborates are synthesized and investigated as electrolyte materials for all-solid-state batteries. The room temperature α-NaB12H11NH3 was structurally solved in P212121 (a = 7.1972(3) Å, b = 9.9225(4) Å, and c = 14.5556(5) Å). It shows a polymorphic structural transition near 140 °C to cubic Fm3̄m. LiB12H11NH3 and NaB12H11NH3 exhibit cationic conductivities of σ(Li+) = 3.0 × 10-4 S cm-1 and σ(Na+) = 1.2 × 10-4 S cm-1 at 200 °C. Hydration is found to improve ionic conductivity of the hydroborates. It is presumed that modest ionic conductivities could be due to a lack of significant re-orientational dynamics in the crystal structure resulting from the presence of the bulky -NH3 group in the anion.

Enhancement of anti-TNFα monoclonal antibody production in CHO cells through the use of UCOE and DHFR elements in vector construction and the optimization of cell culture media.

Recently, there has been a high demand for anti-tumor necrosis factor-α monoclonal antibodies (mAbTNFα) in the treatment of rheumatoid arthritis and other autoimmune diseases. Thus, efficient strategies and stable high-producing cell lines need to be established to increase antibody production. In this study, we describe an efficient approach to establish a mAbTNFα high-producing clone through the optimization of expression vectors and cell culture media. The ubiquitous chromatin opening element (UCOE) and dihydrofolate reductase (DHFR)-based vectors encoding mAbTNFα were introduced into the CHO-DG44 cells using lipofection. Clones were obtained by selecting transfected cells with G418, amplifying them by treatment with methotrexate, and isolating them by limiting dilution. Different media formulated with commercial feeds and media were also screened to develop an improved medium. The antibody produced by the selected clone was purified, characterized, and compared to standard adalimumab. Using our established protocol, a cell clone obtained from stable mAbTNFα-expressing cell pools showed a 3.8-fold higher antibody titer compared to stable cell pools. Furthermore, the highest antibody yield of selected clones cultured in fed-batch mode using improved medium was 2450 ± 30 µg/mL, which was 13.2-fold higher than that of stable cell pool cultivated in batch mode using a basal medium. The purified antibody had primary chemical and biological characteristics similar to those of adalimumab. Therefore, the use of UCOE and DHFR vectors in combination with the optimization of cell culture media may help in establishing stable and high-producing CHO cell lines for therapeutic antibody production.

Bioactive chemical constituents, in vitro anti-proliferative activity and in vivo toxicity of the extract of Curcuma singularis Gagnep rhizomes.

ETHNOPHARMACOLOGICAL RELEVANCE: Curcuma singularis Gagnep is a Vietnamese medicinal plant which has been commonly used as a medicinal remedy in traditional and folk medicines for improving health as well as for treating some diseases, like rheumatoid arthritis, kidney failure. However, pharmacological effects, including anti-cancer activity and the safety of this plant has not been fully investigated. AIM OF THE STUDY: This study aimed to investigate the in vitro anti-growth activity of an extract derived from Curcuma singularis rhizome extract (CSE) against cell lines as well as determine its phytochemical composition. The other goal of our study was to assess the safety of CSE in rats. MATERIALS AND METHODS: The main constituents in the extract were identified and quantitatively analyzed. The in vitro cytotoxicity of CSE was evaluated in several cancer and normal cell lines. The apoptotic activity of CSE and the expression of the apoptosis-related genes were investigated in AGS cells to clarify the underlying molecular mechanisms. The in vivo toxicity of CSE was assessed via acute and subacute oral studies on Sprague-Dawley rats, respectively according to the guidelines 425 and 407 of the Organization for Economic Cooperation and Development (OECD). The drug-related toxicity signs, mortality, body and organ weights were recoreded during the experimental period. In addition, the selected hematological and biochemical parameters, and histological alterations were determined at the end of the subacute toxicity test. RESULTS: Germacrone, ar-turmerone, and curcumol were three sesquiterpene components found in the extract. CSE showed cytotoxic effects in different cancer cells, but had minimal effects on normal cells. Apoptosis in AGS cells was caused by CSE in a concentration-dependent pattern through increase of Bax/Bcl-2 ratio, and release of cytochrome c, which leads to activation of caspase-3/-7, caspase-9, as well as cleavage of PARP. In the acute toxicity test, no signs of toxicity and no mortality were recorded in rats at both doses of 1000 and 5000 mg/kg. In the subacute toxicity study, CSE showed no drug-related adverse effects on water and food consumption, body and organ weights. CSE at a dose of 1000 mg/kg slightly increased WBC and platelet values in female rats, while it increased WBC values in male rats in all tested doses. The decrease of total cholesterol and triglyceride levels were found in female rats treated CSE at doses of 250 or 500 mg/kg. In addition, the increase of serum ALT and AST levels in rats treated at the dose of 1000 mg/kg were noted. No significant changes in histopathological structures of kidneys, spleen, heart and lungs, except liver tissue with minor modifications was found. CONCLUSIONS: Our findings indicated that CSE exhibited in vitro anti-proliferative effects on AGS cells by mainly activating the caspase-dependent mitochondrial apoptotic pathway. CSE also showed in vivo toxicity signals at the dose of 1000 mg/kg with proven minor hepatic injuries, which should be avoided the high dose for prolonged use. Curcuma singularis rhizomes may be used as a chemotherapeutic agent for the treatment of gastric cancer with in vitro anti-cancer investigation and in vivo biological safety evaluation.

Investigation of bioactive chemical constituents and anti-cancer activity of ethanol extract of Curcuma singularis Gagnep rhizomes.

Curcuma singularis Gagnep is a Vietnamese medicinal plant which has been commonly used in traditional and folk medicines for the treatment of different diseases. The goals of the present study are to investigate chemical composition and anti-proliferative activity of Curcuma singularis rhizome extract (CSE). The in vitro cytotoxicity of CSE was evaluated using WST-1 and LDH assays. The apoptosis induction was determined using nuclei DAPI staining and FACS assays. The main compounds of extract were identified and quantitatively analyzed using the validated HPLC method. The extract showed cytotoxic effects in various liver and breast cancer cells but had minimal effects on normal cells. It induced apoptosis on both Hep3B and SKBR3 cells in a dose-dependent manner. In addition, three sesquiterpene compounds, such as germacrone (3.25 ± 0.32 mg/g), ar-turmerone (1.12 ± 0.24 mg/g), and curcumol (0.31 ± 0.12 mg/g) were found as the main components of CSE. This is the first report on the in vitro cytotoxic effect of Curcuma singularis rhizomes against cancer cells.

Evaluation of in vitro cytotoxicity and in vivo potential toxicity of the extract from in vitro cultivated Anoectochilus roxburghii Lindl.

Anoectochilus roxburghii Lind. (A. roxburghii) has promising anti-oxidant, hyperglycemic, hepatoprotective, and immunomodulatory activities as well as anti-tumor effects. However, the pharmacological actions of in vitro cultured plants remain to be determined. Therefore, the objective of the study was to assess in vitro cytotoxicity and in vivo potential toxicity of an extract derived from in vitro cultivated A. roxburghii, termed as iARE. The total flavonoid content and predominant flavonoid compounds of extract were identified and quantitatively analyzed. The in vitro cytotoxicity of iARE was examined using several cancer and normal cell lines. The apoptotic activity and expression of apoptosis-associated genes were also examined in MCF7 cells to determine the underlying mechanisms related to anti-proliferative effects. In vivo potential toxicity of iARE was assessed following acute and subchronic oral administration in Sprague Dawley rats. Quercetin, kaempferol, and isorhamnetin were three flavonoid components identified in iARE. The extract exerted cytotoxic effects on various cancer cells but not normal fibroblasts. Apoptosis in MCF7 cells was induced by iARE in a concentration-dependent manner associated with increased Bax/Bcl-2 ratio and reduced mitochondrial membrane potential ΔΨm, leading to release of cytochrome c, activation of caspase-3/7 and caspase-9, and cleavage of PARP. In the acute oral toxicity study, no mortality or toxicological signs were observed in rats at 1000 or 5000 mg/kg. In a subchronic oral toxicity study, iARE at a dosage of up to 1000 mg/kg produced no mortality or treatment-related adverse effects on general behavior, food intake, body weight, relative organ weights. No apparent marked changes in the histopathology of the liver and kidney were detected. Data demonstrated that iARE induced in vitro cytotoxic effects in cancer cells are associated with lackof invivo toxicity. Thus, iARE was suggested to be considered as apotential therapeutic candidate for cancer treatment.

Phenolic Extraction of Moringa Oleifera Leaves Induces Caspase-Dependent and Caspase-Independent Apoptosis through the Generation of Reactive Oxygen Species and the Activation of Intrinsic Mitochondrial Pathway in Human Melanoma Cells.

Moringa oleifera Lam. has long been used to treat many diseases, including diabetes, aging, inflammatory, and cancer. Many studies have revealed that the crude extract of Moringa oleifera Lam. leaves possesses anticancer property. Therefore, in this study, the extract of Moringa oleifera leaves was fractionated using different solvents to figure out the most effective fraction for anti-proliferative effect on melanoma cells. Methanol extract (MO-ME), hexane fraction (MO-HE), chloroform fraction (MO-CH), ethyl acetate fraction (MO-EA), and water-soluble fraction (MO-WA) of Moringa oleifera leaves were prepared. Total phenolic and flavonoid contents were determined. The anti-proliferative activity on melanoma cells and normal cells was investigated using WST-1 assay. The apoptotic activity was assessed by testing DNA condensation, DNA fragmentation, and phosphatidylserine (PS) externalization. The expression of apoptosis-related genes, the mitochondrial depolarization, and reactive oxygen species (ROS) were then examined to clarify the underlying molecular mechanisms. In this regard, MO-ME, MO-EA, and MO-CH inhibited the proliferation of both A375 human melanoma cells and A2058 human melanoma cells, but had little effect on WS1 normal human skin fibroblasts and primary normal human dermal fibroblasts (NHDF). Among fractions, the phenolic-rich MO-EA markedly inhibited the growth of A375 cells in a dose- and time-dependent manner. The anti-proliferation was supposed to be mediated via apoptosis, which was demonstrated by the significant increase of condensed chromatin, DNA fragmentation, and PS externalization. The apoptosis was stimulated by enhanced ROS production and reduction of mitochondrial membrane potential. MO-EA activated Bax while reducing Bcl-2 expression, leading to an increase in Bax/Bcl-2 ratio. The mechanisms of cell death involved in activation of Caspase-3/7 and Caspase-9 (Caspase-dependent pathway), activation, and translocation of apoptosis-inducing factor (AIF) into the nucleus (Caspase-independent pathway). Our study indicated that the phenolic-rich fraction exerted significant anticancer effects on melanoma cells in vitro which involved in Caspase-dependent and Caspase-independent apoptosis pathways mediated by mitochondrial ROS. These results provided a fundament for the using of phenolic-rich fraction of Moringa oleifera leaves to treat skin cancer effectively.

Integrated farming system producing zero emissions and sustainable livelihood for small-scale cattle farms: Case study in the Mekong Delta, Vietnam.

This study proposes an integrated cattle breeding and cultivation system that provides zero emission and sustainable livelihood for the community in rural areas. The proposed integrated farming system improves agricultural productivity and environmental and sanitation conditions, minimizes the amount of waste, and increases the family income up to 41.55%. Several waste types can be recycled and transformed into valuable products, such as energy for cooking, organic fertilizer for crops, and cattle feed for breeding. Wastewater effluent from the biogas tank can be treated by biochar and results show that it then meets the standards for irrigation purposes. Also, the waste flow from cattle breeding supplies enough nutrients to cultivate plants, and the plants grown supply are adequate food for the 30 cows living on the farm. This research shows that the use of an integrated farming system could achieve zero-emission goal. Thereby, it provides a sustainable livelihood for cattle breeding family farms. The proposed integrated cattle breeding and cultivation system improves agricultural productivity, environmental and increases the farmer income up to 41.55%.

Mitochondria-mediated Caspase-dependent and Caspase-independent apoptosis induced by aqueous extract from Moringa oleifera leaves in human melanoma cells.

Malignant melanoma is a very aggressive and serious type of cutaneous cancer. Previous studies indicated the anti-cancer activity of aqueous extract of Moringa oleifera Lam. leaves (MOE) against a variety of cell lines. However, there has not been much research about the effect of MOE on melanoma. Therefore, this study was about to investigate the anti-proliferation mediated by apoptosis of MOE on human melanoma cell lines. Furthermore, the related molecular mechanisms of the apoptosis were also examined. An aqueous extract of Moringa oleifera leaves was prepared and the anti-proliferative activity on melanoma cells and normal cells was tested using WST-1 assay. The apoptotic hallmarks including DNA condensation and phosphatidylserine (PS) externalization were assessed. The expression of apoptosis-related genes and the depolarization of mitochondrial membrane potential were then examined to clarify the underlying molecular mechanisms. MOE inhibited cell growth of A375 cells and A2058 cells in a dose-dependent manner but had little effect on human normal fibroblasts. The cell growth inhibition was induced by apoptosis which was expressed via chromatin condensation and PS externalization. MOE decreased mitochondrial membrane potential. Additionally, MOE increased Bax/Bcl-2 ratio, activated Caspase-3/7, Caspase-9, PARP and AIF translocation, leading to apoptotic cell death. Our study indicated that MOE exerted significant anti-cancer effects on melanoma cells in vitro which involved mitochondria-mediated Caspase-dependent and Caspase-independent apoptosis pathways. These results provided a scientific approach for using Moringa oleifera leaves as an alternative therapy to treat skin cancer.

Carbapenemase Genes and Multidrug Resistance of Acinetobacter Baumannii: A Cross Sectional Study of Patients with Pneumonia in Southern Vietnam.

BACKGROUND: Acinetobacter baumannii (Ab) is an opportunistic bacterial pathogen found in hospital-acquired infections including nosocomial pneumonia, especially multidrug-resistant Ab. This study aims to survey the drug resistance profiles of Ab isolated from patients in Thong Nhat Dong Nai General Hospital and assess the relationship between genotypes and antibiotic resistance; Methods: Ninety-seven Ab strains isolated from 340 lower respiratory tract specimens among pneumonia patients were used to screen the most common local carbapenemase genes. Antimicrobial susceptibility testing results and demographic data were collected and minimum inhibitory concentrations (MIC) of colistin were also determined; Results: Over 80% and 90% of Ab strains were determined as carbapenem-resistant and multidrug-resistant (MDR), respectively. Most of the strains carried carbapenemase genes, including blaOXA-51, blaOXA-23-like, blaOXA-58-like, and blaNDM-1, with proportions of 97 (100%), 76 (78.4%), 10 (10.3%), 6 (6.2%), respectively. Amongst these genes, blaOXA-23-like was the only gene which significantly influenced the resistance (p < 0.0001); and Conclusions: The severity of Ab antibiotic resistance is urgent and specifically related to carbapenemase encoding genes. Therefore, screening of MDR Ab and carbapenemase for better treatment options is necessary.

Taccakhanhhoaensis V.S. Dang & Vuong (Taccaceae), a new species from southern Vietnam.

Taccakhanhhoaensis V.S. Dang & Vuong (Taccaceae) is described as a new species from Hon Ba Nature Reserve in southern Vietnam. This species is morphologically similar to T.chantrieri and T.ampliplacenta but differs from its allies by several salient characters: size of leaves and petioles, inflorescent much shorter leaves, number of flowers, stigma lobes, buds colour. A description, conservation assessment, together with photographs and a key to the species of Tacca in Vietnam are presented.

Directing the Viedma ripening of ethylenediammonium sulfate using "Tailor-made" chiral additives.

Both enantiomers of trans-cyclohexane-1,2-diammonium sulfate and trans-1,2-diphenylethylenediammonium sulfate were used as "tailor-made" additives to direct the mirror-symmetry breaking in the attrition-enhanced deracemization (i.e. Viedma ripening) of conglomerate crystals of ethylenediammonium sulfate (EDS). Isothermal titration calorimetry (ITC) shows chiral recognition of (1R,2R)- and (1S,2S)-1,2-diphenylethylenediamine to EDS crystals where the enthalpy of adsorption of the (1R,2R)-isomer on l-EDS crystals is higher in comparison to that on d-EDS crystals. These results are consistent with a "rule of reversal" mechanism driving the chiral outcome of the Viedma ripening of EDS.

Evaluation of microscopic observation drug susceptibility assay for diagnosis of multidrug-resistant tuberculosis in Viet Nam.

BACKGROUND: Early diagnosis of tuberculosis (TB) and multidrug resistant tuberculosis (MDR TB) is important for the elimination of TB. We evaluated the microscopic observation drug susceptibility (MODS) assay as a direct rapid drug susceptibility testing (DST) method for MDR-TB screening in sputum samples METHODS: All adult TB suspects, who were newly presenting to Pham Ngoc Thach Hospital from August to November 2008 were enrolled into the study. Processed sputum samples were used for DST by MODS (DST-MODS) (Rifampicin (RIF) 1 µg/ml and Isoniazid (INH) 0.4 µg/ml), MGIT culture (Mycobacterial Growth Indicator Tube) and Lowenstein Jensen (LJ) culture. Cultures positive by either MGIT or LJ were used for proportional DST (DST-LJ) (RIF 40 µg/ml and INH 0.2 µg/ml). DST profiles on MODS and LJ were compared. Discrepant results were resolved by multiplex allele specific PCR (MAS-PCR). RESULTS: Seven hundred and nine TB suspects/samples were enrolled into the study, of which 300 samples with DST profiles available from both MODS and DST-LJ were analyzed. Cording in MODS was unable to correctly identify 3 Mycobacteria Other Than Tuberculosis (MOTT) isolates, resulting in 3 false positive TB diagnoses. None of these isolates were identified as MDR-TB by MODS. The sensitivity and specificity of MODS were 72.6% (95%CI: 59.8, 83.1) and 97.9% (95%CI: 95.2, 99.3), respectively for detection of INH resistant isolates, 72.7% (95%CI: 30.9, 93.7) and 99.7% (95%CI: 98.1, 99.9), respectively for detecting RIF resistant isolates and 77.8% (95%CI: 39.9, 97.1) and 99.7% (95%CI: 98.1, 99.9), respectively for detecting MDR isolates. The positive and negative predictive values (PPV and NPV) of DST-MODS were 87.5% (95%CI: 47.3, 99.6) and 99.3% (95%CI: 97.5, 99.9) for detection of MDR isolates; and the agreement between MODS and DST-LJ was 99.0% (kappa: 0.8, P < 0.001) for MDR diagnosis. The low sensitivity of MODS for drug resistance detection was probably due to low bacterial load samples and the high INH concentration (0.4 µg/ml). The low PPV of DST-MODS may be due to the low MDR-TB rate in the study population (3.8%). The turnaround time of DST-MODS was 9 days and 53 days for DST-LJ. CONCLUSION: The DST-MODS technique is rapid with low contamination rates. However, the sensitivity of DST-MODS for detection of INH and RIF resistance in this study was lower than reported from other settings.

Simultaneous Determination of Berberine and Palmatine in “Huong Lien Hoan” by HPLC

Berberine and Palmatine were extracted from the powdered Huong Lien pills with a mixture of methanol and hydrochloric acid (100:1) and subjected to HPLC to simultaneously determine the alkaloids. The HPLC technique was carried out on a Lichrosorb RP-8 (10 m, 250x4 mm) column using a UV detector at =345 nm and a mixture of 3,4g potassium dihydrogen phosphate and 1,7g sodium lauryl sulfate dissolved in 1000 ml of water-acetonitrile (1:1) as a mobile phase. The experimental results proved that the proposed HPLC method was rapid, specific, accurate and precise.

Variation of essential oil content in Japanese Perpermin SK – 33 according to period of development and term of conservation

This paper presented a study on the dynamic of essential oil accumulation in Japanese mint SK33 (M. arrvensis L.) and the influence of storage time on its quantity and quality. This mint was cultivated at Co Nhue experimental station, Tu Liem district, Ha Noi City between 2000 and 2002. The results showed that during the vegetation and development process, the essential oil content in the plant gradually increased and achieved maximum at first branch flowering stage and reduced to minimum at the end of flowering stage. The menthol content in essential oil of Japanese mint SK33 reached the highest value at full flowering stage (81.79%). The best harvest time for this mint is from the beginning flowering stage to full flowering period. After harvesting the whole plants can be stored during 20 days without the change in content and quality of essential oil

Chemical components of essential oil of Mentha piperita L. collected from Sapa – Lao Cai and now cultivated in Co Nhue, Tu Liem district – Ha Noi

Mentha piperita L.Mentha piperita L. was collected from Sapa, Lao Cai province and cultivated at Co Nhue – Tu Liem – Ha Noi. The content of the essential oil in fresh branches of this peppermint was about 0.16%. Its green yield is 1.85-2.10 Kg/m2. First time in Vietnam, the chemical composition of peppermint oil was analyzed by GC/MS and 43 constituents were identified. The main constituents were L-menthol (47.60%), menthol (24.10%), menthofuran (6.07%) and 1.8 cineol (5.55%), pulegone (4.22%). Base on these data. The quality of Sa Pa peppermint oil is good as one from native origin. Peppermint collected from Sa Pa enrich the gene source of medicinal plant in general and particularly the gene source of essential oil plant group of Vietnam