Application of high-speed counter current chromatography in extraction and separation of alkaloids in natural products / 中国中药杂志

Alkaloids, widespread in plants, have a series of pharmacological activities and have been widely used to treat various diseases. Because alkaloids are usually presented in multicomponent mixtures and are deeply low in content, they are very difficult to extract and separate by traditional methods. High-speed counter current chromatography(HSCCC) is a kind of liquid-liquid chromatography without solid support phase, which has the advantages of large injection volume, low cost, and no irreversible adsorption. Compared with the traditional methods of extraction and separation of alkaloids, HSCCC can ensure the separation of many different alkaloids at one time, with a high recovery and large amount. In this paper, the advantages and disadvantages of HSCCC compared with traditional separation methods were discussed and the solvent system and elution mode of HSCCC used to separate alkaloids in recent years were summarized by referring to the relevant literature to provide some references for the separation of alkaloids by HSCCC.

Improving the thermal stability of Proteus mirabilis lipase based on multiple computational design strategies / 生物工程学报

Proteus mirabilis lipase (PML) features tolerance to organic solvents and great potential for biodiesel synthesis. However, the thermal stability of the enzyme needs to be improved before it can be used industrially. Various computational design strategies are emerging methods for the modification of enzyme thermal stability. In this paper, the complementary algorithm-based ABACUS, PROSS, and FoldX were employed for positive selection of PML mutations, and their pairwise intersections were further subjected to negative selection by PSSM and GREMLIN to narrow the mutation library. Thereby, 18 potential single-point mutants were screened out. According to experimental verification, 7 mutants had melting temperature (Tm) improved, and the ΔTm of K208G and G206D was the highest, which was 3.75 ℃ and 3.21 ℃, respectively. Five mutants with activity higher than the wild type (WT) were selected for combination by greedy accumulation. Finally, the Tm of the five-point combination mutant M10 increased by 10.63 ℃, and the relative activity was 140% that of the WT. K208G and G206D exhibited certain epistasis during the combination, which made a major contribution to the improvement of the thermal stability of M10. Molecular dynamics simulation indicated that new forces were generated at and around the mutation sites, and the rearrangement of forces near G206D/K208G might stabilize the Ca2+ binding site which played a key role in the stabilization of PML. This study provides an efficient and user-friendly computational design scheme for the thermal stability modification of natural enzymes and lays a foundation for the modification of PML and the expansion of its industrial applications.

Difference of Chemical Bonds Between UDMA Bonding Agents with Ethanol Solvent and Acetone Solvent on Dentin Collagen

ABSTRACT Objective: To investigate the difference of chemical bonds between urethane dimethacrylate (UDMA) bonding agents with ethanol solvent and acetone solvent on dentin collagen. Material and Methods: This experimental comparison study used three groups: G1 (Control): UDMA and collagen; G2: UDMA, collagen and ethanol; and G3: UDMA, collagen and acetone. The groups were then pelleted and analysed with FTIR, then the peak value of carbonyl absorption band from each study group was calculated. The result of FTIR analysis and the peak of carbonyl absorption band (P) was calculated using the formula: P = (BC / AB) X 100; AB. BC is measured in centimeters. The study of chemical bond differences between ethanol-solvent UDMA agents compared with acetone-solvent on dentin collagen resulted in a graph of peak of carbonyl absorption bands of UDMA and dentin collagen groups. To determine the chemical bonds of UDMA from the top of the carbonyl ester absorption bands with wavenumber absorption in range 1700-1750 cm-1, the decreasing peak of the carbonyl absorption bands is assumed as more chemical bonds that formed. Data were analysed using Anova one way and Tukey HSD test. Results: There were significant differences between the three study groups (p<0.05). Conclusion: UDMA bonding agents' chemical bonds with acetone solvent are much higher than the chemical bonds between UDMA bonding agents with ethanol solvent on dentin collagen.

Biosynthesis of (R)-2-hydroxy-3-phenylpropionic acid using whole recombinant Escherichia coli cells in an aqueous/n-octane biphasic system / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

(R)-2-hydroxy-3-phenylpropionic acid (PLA) is an ideal antimicrobial compound with broad-spectrum activity against a wide range of Gram-positive bacteria, some Gram-negative bacteria, and fungi. We studied the bioconversion of phenylpyruvate (PPA) to PLA using whole recombinant Escherichia coli cells in a series of buffer/organic solvent systems. Octane was found to be the best organic solvent. The optimum volume ratio of the water phase to the n-octane phase, conversion temperature, substrate concentration, and cell concentration were 6:4, 40 °C, 12.5 g/L, and 30 g/L wet cells, respectively. Under the optimized conditions, the average PLA productivity in the aqueous/ n-octane system was 30.69% higher than that in the aqueous system, and 32.31 g/L PLA was obtained with the use of a stirred reactor (2-L scale). Taken together, our findings indicated that PLA biosynthesis was more efficient in an aqueous/n-octane biphasic system than in a monophasic aqueous system. The proposed biphasic system is an effective strategy for enhancing PLA yield and the biosynthesis of its analogues.

Rubbing time and bonding performance of one-step adhesives to primary enamel and dentin

Abstract Objectives: This study investigated whether increasing the concentration of acidic monomers in one-step adhesives would allow reducing their application time without interfering with the bonding ability to primary enamel and dentin. Material and methods: Experimental one-step self-etch adhesives were formulated with 5 wt% (AD5), 20 wt% (AD20), or 35 wt% (AD35) acidic monomer. The adhesives were applied using rubbing motion for 5, 10, or 20 s. Bond strengths to primary enamel and dentin were tested under shear stress. A commercial etch-and-rinse adhesive (Single Bond 2; 3M ESPE) served as reference. Scanning electron microscopy was used to observe the morphology of bonded interfaces. Data were analysed at p<0.05. Results: In enamel, AD35 had higher bond strength when rubbed for at least 10 s, while application for 5 s generated lower bond strength. In dentin, increased acidic monomer improved bonding only for 20 s rubbing time. The etch-and-rinse adhesive yielded higher bond strength to enamel and similar bonding to dentin as compared with the self-etch adhesives. The adhesive layer was thicker and more irregular for the etch-and-rinse material, with no appreciable differences among the self-etch systems. Conclusion: Overall, increasing the acidic monomer concentration only led to an increase in bond strength to enamel when the rubbing time was at least 10 s. In dentin, despite the increase in bond strength with longer rubbing times, the results favoured the experimental adhesives compared to the conventional adhesive. Reduced rubbing time of self-etch adhesives should be avoided in the clinical setup.

Purification of inulinases by changing the ionic strength of the medium and precipitation with alcohols

ABSTRACT The present study evaluated the purification of inulinase by changing the ionic strength of the medium by addition of NaCl and CaCl2 followed by precipitation with n-propyl alcohol or iso-propyl alcohol. The effects of the concentration of alcohols and the rate of addition of alcohols in the crude extract on the purification yield and purification factor were evaluated. Precipitation caused an activation of enzyme and allowed purification factors up to 2.4-fold for both alcohols. The purification factor was affected positively by the modification of the ionic strength of the medium to 0.5 mol.L-1 NaCl before precipitation with the alcohol (n-propyl or iso-propyl). A purification factor of 4.8-fold and an enzyme yield of 78.1 % could be achieved by the addition of 0.5 mol.L-1 of NaCl to the crude extract, followed by the precipitation with 50 % (v/v) of n-propyl alcohol, added at a flow rate of 19.9 mL/min.

Influence of solvents on the bond strength of resin sealer to intraradicular dentin after retreatment

Abstract This study evaluated the removal of filling material with ProTaper Universal Rotary Retreatment system (PTR) combined with solvents and the influence of solvents on the bond strength (PBS) of sealer to intraradicular dentin after canal reobturation. Roots were endodontically treated and distributed to five groups (n = 12). The control group was not retreated. In the four experimental groups, canals were retreated with PTR alone or in combination with xylol, orange oil, and eucalyptol. After filling material removal, two specimens of each group were analysed by SEM and µCT to verify the presence of filling remnants on root canal walls. The other roots were reobturated and sectioned in 1-mm-thick dentin slices that were subjected to the push-out test. Data were analysed by two-way ANOVA and Tukey’s test (α = 0.05). SEM and µCT analysis revealed that all retreatment techniques left filling remnants on canal walls. The control group (3.47 ± 1.21) presented significantly higher (p < 0.05) PBS than the experimental groups. The groups retreated with PTR alone (2.59 ± 0.99) or combined with xylol (2.54 ± 0.77) and orange oil (2.32 ± 0.93) presented similar bond strength (p > 0.05), and differed significantly from the group with eucalyptol (1.89 ± 0.63). The solvents reduced the PBS of the sealer to dentin and no retreatment technique promoted complete removal of filling material.

Process optimization for production and purification of a thermostable, organic solvent tolerant lipase from Acinetobacter sp. AU07

ABSTRACT The purpose of this study was to isolate, purify and optimize the production conditions of an organic solvent tolerant and thermostable lipase from Acinetobacter sp. AU07 isolated from distillery waste. The lipase production was optimized by response surface methodology, and a maximum production of 14.5 U/mL was observed at 30 ºC and pH 7, using a 0.5% (v/v) inoculum, 2% (v/v) castor oil (inducer), and agitation 150 rpm. The optimized conditions from the shake flask experiments were validated in a 3 L lab scale bioreactor, and the lipase production increased to 48 U/mL. The enzyme was purified by ammonium sulfate precipitation and ion exchange chromatography and the overall yield was 36%. SDS-PAGE indicated a molecular weight of 45 kDa for the purified protein, and Matrix assisted laser desorption/ionization time of flight analysis of the purified lipase showed sequence similarity with GDSL family of lipases. The optimum temperature and pH for activity of the enzyme was found to be 50 ºC and 8.0, respectively. The lipase was completely inhibited by phenylmethylsulfonyl fluoride but minimal inhibition was observed when incubated with ethylenediaminetetraacetic acid and dithiothreitol. The enzyme was stable in the presence of non-polar hydrophobic solvents. Detergents like SDS inhibited enzyme activity; however, there was minimal loss of enzyme activity when incubated with hydrogen peroxide, Tween 80 and Triton X-100. The kinetic constants (Km and Vmax) revealed that the hydrolytic activity of the lipase was specific to moderate chain fatty acid esters. The Vmax, Km and Vmax/Km ratio of the enzyme were 16.98 U/mg, 0.51 mM, and 33.29, respectively when 4-nitrophenyl palmitate was used as a substrate.

Effectiveness of the waveone and protaper D systems for removing gutta-percha with or without a solvent / Eficiência dos sistemas waveone e protaper D na remoção de guta-percha com ou sem uso solvente

Endodontic retreatment requires complete removal of the filling material and access to the apical foramen. The purpose of this study was to evaluate the effectiveness of the WaveOne reciprocating system and compare it to the ProTaper D rotary system, with or without the use of a solvent, in removing filling material from root canals. The time required for each filling removal technique employed was also determined and compared. Forty extracted human mandibular premolars with a single, straight, flattened canal were prepared and filled. They were divided into four groups (n = 10): Group 1: ProTaper D NiTi rotary instruments; Group 2: ProTaper D NiTi rotary instruments, with a solvent; Group 3: WaveOne primary instrument; and Group 4: WaveOne primary instrument, with a solvent. The teeth were then split along their long axis and photographed using an operating microscope with 5X magnification. The amount of remaining filling material was assessed with Image Tool software. The results were compared using the KruskalWallis test (p <0.05). There was no significant difference between groups regarding the amount of residual filling material (p > 0.05). Operative time was significantly longer in Group 3 than in groups 1, 2 and 4 (p < 0.05). The WaveOne system and the ProTaper D system were equally effective, with or without a solvent. The time required to remove the filling material from the canals was significantly longer in Group 3 than in the other groups.

No retratamento endodôntico, a completa remoção do material obturador e o acesso ao forame apical são necessários para permitir a limpeza do sistema de canais. O propósito deste estudo foi avaliar a eficácia do sistema reciprocante WaveOne e comparála ao sistema rotatório ProTaper Universal, com ou sem o uso de solvente, na remoção do material obturador.O tempo necessário a cada técnica empregada foi determinado e comparado. Quarenta prémolares inferiores humanos extraídos com canal único, reto e achatado foram preparados e obturados. Foram então divididos em quatro grupos (n = 10) de acordo com o sistema utilizado, como segue. Grupo 1: ProTaper D Niti; Grupo 2: Sistema ProTaper D com solvente; Grupo 3: Sistema WaveOne instrumento Primary; e Grupo 4: Sistema WaveOne instrumento Primary com solvente, sendo o tempo registrado. Os dentes foram clivados longitudinalmente e fotografados utilizando microscópio operatório com aumento de 5 vezes.A quantidade de material remanescente foi avaliada com o uso do software Image Tool 3.0. Os resultados foram comparados utilizando o teste de KruskalWallis( p < 0.05). Em relação aos resultados, não houve diferença significativa entre os grupos quanto à quantidade de material obturador residual (p > 0.05). O tempo operatório no Grupo 3 foi significativa mente maior do que nos grupos 1, 2 e 4 (p < 0.05).O sistema WaveOne foi tão efetivo quanto o ProTaper D, com ou sem solvente.tempo necessário à desobturação dos canais no Grupo 3 (WaveOne sem solvente) foi significativamente maior do que nos demais grupos.

Effect of the time-point of acid etching on the persistence of sealer residues after using different dental cleaning protocols

Abstract: Endodontic sealer residues remaining within the pulp chamber dentin after root canal obturation and cleaning with various solvents may compromise the appearance and the durability of dental restorations. Acid etching is routinely performed prior to application of dentine adhesive systems, but is effect on residual sealer material and the optimal time-point for performing etching, are unknown. Here, we evaluated the effect of acid etching on the dentin surface when performed either immediately or 7 days after removal of the endodontic sealer with two solvents, i.e., 95% ethanol or xylol. Forty crowns fragments from bovine incisors were impregnated with sealer and divided into 4 groups (n = 10 each), according to the dentin cleaning protocol and to the acid etching time-point: G1, 95.0% ethanol and immediate acid etching; G2, xylol and immediate acid etching; G3, 95.0% ethanol and acid etching after 7 days; and G4, xylol and acid etching after 7 days. Scanning electron microscopy (SEM) images (2000 ×) were obtained from each specimen and the number of open dentinal tubules counted and compared. Another 40 fragments were similarly prepared, and SEM images were obtained (500 ×) to score and compare the persistence of sealer residues on the dentin. G4 showed the most open dentinal tubules and the least epoxy resin-based sealer residues on the dentin surface (p < 0.05). The least epoxy resin-based sealer residues was obtained when acid etching, using 37% phosphoric acid, was performed after 7 days after cleaning the dentin with xylol.

Dissolution of a mineral trioxide aggregate sealer in endodontic solvents compared to conventional sealers

Abstract The aim of this study is to evaluate the solubility of a Mineral Trioxide Aggregate sealer (MTA-Fillapex) compared with five other sealers, calcium hydroxide (Sealapex), resin (Realseal), zinc oxide-eugenol (Tubli-Seal), and two epoxy resins (AH-26 and AH-Plus), in chloroform and eucalyptoil in static and ultrasonic environments. Samples of each sealer were prepared (n = 180) and then divided into 12 groups that were immersed in solvents for 5 and 10 min in static and ultrasonic environments. The mean weight loss was determined, and the values were compared using Student's t-test, One-way ANOVA, and Tukey's HSD post-hoc test (p < 0.05). In chloroform, MTA-Fillapex, AH-26, and Sealapex displayed moderate solubility with no significant difference in dissolution (p = 0.125); however, their dissolution was significantly lower than that of AH-Plus (p < 0.001), which was almost fully dissolved after 10 minutes. Realseal was significantly less soluble than all sealers (p < 0.001). In eucalyptoil, MTA-Fillapex showed low solubility, as all of the sealers did, but Tubli-Seal was significantly more soluble than other sealers (p < 0.001). Using ultrasonic activation resulted in a significantly higher dissolution rate in chloroform for all sealers except MTA-Fillapex after 10 min (p = 0.226). In eucalyptoil, ultrasonic activation significantly increased the dissolution rate of all sealers except MTA-Fillapex after 5 and 10 min, Sealapex at 10 min, and AH-Plus at 5 min (p > 0.05). In conclusion, MTA-Fillapex was not sufficiently dissolved in either solvent. Ultrasonic activation had limited effectiveness on MTA-Fillapex dissolution, whereas it significantly increased the efficiency of solvents in dissolving a number of endodontic sealers.

Antitumor and anti-angiogenic potentials of isolated crude saponins and various fractions of Rumexhastatus D. Don

BACKGROUND: Cancer, being the foremost challenge of the modern era and the focus of world-class investigators, gargantuan research is in progress worldwide to explore novel therapeutic for its management. The exploitation of natural sources has been proven to be an excellent approach to treat or minify the excessive angiogenesis and proliferation of cells. Similarly, based the ethnomedicinal uses and literature survey, the current study is designed to explore the anti-tumor and anti-angiogenic potentials of Rumex hastatus. Anti-tumor and anti-angiogenic activities were carried out using potato-disc model and chorioallantoic membrane (CAM) assay respectively. Moreover, R. hastatus was also assessed for antibacterial activity against Agrobacterium tumefaciens (tumor causing bacterial strain). The positive controls used in anti-tumor, anti-angiogenic and antibacterial activities were vincristine sulphate, dexamethasone and cefotaxime respectively. RESULTS: The crude saponins (Rh.Sp), methanolic extract (Rh.Cr) and other solvent extracts like n-hexane (Rh.Hex), chloroform (Rh.Chf), ethylacetate (Rh.EtAc) and aqueous fraction (Rh.Aq) exhibited notable anti-tumor and anti-angiogenic activities. In potato tumor assay, the chloroform and saponin fractions were observed to be the most effective showing 86.7 and 93.3 % tumor inhibition at 1000 µg/ml with IC50 values 31.6 and 18.1 µg/ml respectively. Similarly, these two samples i.e., chloroform and saponins also excelled among the entire test samples in anti-angiogenic evaluation exhibiting 81.6 % (IC50 = 17.9 µg/ml) and 78.9 % (IC50 = 64.9 µg/ml) at 1000 µg/ml respectively. In contrast, the antibacterial investigations revealed a negligible potential against A. tumefaciens. CONCLUSION: Based on our results we can claim that R. hastatus possesses both anti-tumor and anti-angiogenic potentials. In all of the solvent fractions, Rh.Chf and Rh.Sp were most effective against tumor and angiogenesis while having negligible activity against A. tumefaciens. It can be concluded that Rh.Chf and Rh.Sp might be potential targets in the isolation of natural product having anti-neoplastic action.

Thermostable, alkaline and detergent-tolerant lipase from a newly isolated thermophilic Bacillus stearothermophilus.

Lipases are the enzymes of choice for laundry detergent industries, owing to their triglyceride removing ability from the soiled fabric, which eventually reduces the usage of phosphate-based chemical cleansers in the detergent formulation. In this study, a novel thermo-alkaline lipase-producing strain identified as Bacillus stearothermophilus was isolated from the soil samples of olive oil mill. Enhanced lipase production was observed at 55°C, pH 11 and after 48 h of incubation. Among the substrates tested, xylose (a carbon source), peptone (a nitrogen source) and olive oil at a concentration of 1% were suitable substrates for enhancing lipase production. MgSO4 and Tween-80 were suitable substrates for maximizing lipase production. The enzyme was purified to homogeneity by a single CM-Sephadex column chromatography and revealed molecular mass of 67 kDa. The enzyme (BL1) was active over a wide range of pH from 9.0 to 13.0, with an optimum at pH 11.0, exhibited maximal activity at 55°C and retained more than 70% of its activity after incubation at 70°C or pH 13 for 0.5 h or 24 h, respectively. The enzyme hydrolyzed both short and long-chain triacylglycerols at comparable rates. BL1 was studied in a preliminary evaluation for use in detergent formulation solutions. This novel lipase showed extreme stability towards non-ionic and anionic surfactants after pre-incubation for 1 h at 40°C, and good stability towards oxidizing agents. Additionally, the enzyme showed excellent stability and compatibility with various commercial detergents, suggesting its potential as an additive in detergent formulations.

Sandfly (Diptera: Psychodidae: Phlebotominae) species diversity in an urban area of the municipality of Tapachula, Chiapas, Mexico

Monitoring phlebotomine sandflies in urban areas is key for epidemiological studies in susceptible populations. This paper describes sandfly fauna that were present in an urban area of the municipality of Tapachula, Chiapas, Mexico, and were captured with Shannon and CDC light traps. During February and March of 2014, 1,442 sandflies were captured, specifically Lutzomyia cruciata (Coquillet) (98.8%), Lutzomyia cayennensis cayennensis (Floch and Abonnenc) (0.8%), Lutzomyia chiapanensis (Dampf) (0.3%) and Lutzomyia atulapai (De León) (0.1%). Lu. cruciata was the most abundant and the most frequently trapped species. This is the first record of its remarkable ability to adapt to urban green areas. The three other species trapped represent new records of geographic distribution for the study region. These results indicate the need to establish measures for reducing both human contact with this vector and the risk of possible sites of infection.

BAY 41-2272 activates host defence against local and disseminated Candida albicans infections

In our previous study, we have found that 5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridine-3-yl]-pyrimidin-4-ylamine (BAY 41-2272), a guanylate cyclase agonist, activates human monocytes and the THP-1 cell line to produce the superoxide anion, increasing in vitro microbicidal activity, suggesting that this drug can be used to modulate immune functioning in primary immunodeficiency patients. In the present work, we investigated the potential of the in vivo administration of BAY 41-2272 for the treatment of Candida albicans and Staphylococcus aureus infections introduced via intraperitoneal and subcutaneous inoculation. We found that intraperitoneal treatment with BAY 41-2272 markedly increased macrophage-dependent cell influx to the peritoneum in addition to macrophage functions, such as spreading, zymosan particle phagocytosis and nitric oxide and phorbol myristate acetate-stimulated hydrogen peroxide production. Treatment with BAY 41-2272 was highly effective in reducing the death rate due to intraperitoneal inoculation of C. albicans, but not S. aureus. However, we found that in vitro stimulation of peritoneal macrophages with BAY 41-2272 markedly increased microbicidal activities against both pathogens. Our results show that the prevention of death by the treatment of C. albicans-infected mice with BAY 41-2272 might occur primarily by the modulation of the host immune response through macrophage activation. .

Análise das propriedades químicas, biológicas e antimicrobianas de solventes endodônticos / Analysis of chemical, biological and antimicrobial properties of endodontic solvents

O objetivo deste estudo foi avaliar as propriedades químicas, biológicas e antimicrobianas dos solventes endodônticos, Citrol, Eucaliptol, d-Limoneno, Xilol e Endosolv E. Dentre os testes químicos, foram avaliados a microdureza dentinária, onde utilizamos blocos de dentina bovina que foram expostos aos solventes por 2 períodos, 5 e 15 min e submetidos ao teste de microdureza. Outro teste químico foi a capacidade de dissolução dos materiais obturadores pelo teste de imersão nos solventes em 3 períodos, 2, 5 e 15 min. Para essa avaliação foram, confeccionados corpos de prova dos cimentos, AH Plus, Acroseal, Sealer 26, Endofill, MTA Fillapex e RealSeal SE e dos cones de guta-percha estandardizado Dentsply (DP), ProTaper e Resilon. Um terceiro teste, foi realizado para avaliar a capacidade de desobturação dos canais radiculares e o efeito da agitação ultrassônica dos solventes endodônticos após a desobturação. Sessenta incisivos centrais superiores foram divididos em 6 grupos sendo um para cada solvente (n=10) mais um grupo controle (solução fisiológica). Todos dentes foram instrumentados e obturados pela mesma técnica e escaneados no Micro-CT. Os dentes foram então desobturados utilizando ProTaper Retratamento/ProTaper Universal como técnica para todos grupos, associada a um dos solventes e foram escaneados novamente para a avaliação do volume de material remanescente após a desobturação. Cada um dos dentes foi desobturado e preenchido com o mesmo solvente utilizado na desobturação. Cada solvente foi agitado pelo ultrassom por 1 min e novamente foram escaneados e avaliados os volumes restantes dos materiais nos canais radiculares. Na avaliação biológica, utilizamos o teste de citotoxicidade com células de camundongo NIH-3T3 pelo ensaio MTT. As culturas celulares foram plaqueadas e submetidas aos solventes diluídos nas concentrações de 0.5 a 2.5%, e avaliados quanto à viabilidade celular. Por fim o último teste foi o antimicrobiano, avaliado pelo teste de...

The aim of this study was to evaluate the chemical, biological and antimicrobial properties of the Endodontic solvents, Citrol, Eucalyptol, d-Limonene, Xylene and Endosolv E. First chemical properties analysis were evaluated by dentin microhardness, where we used bovine dentin blocks that were exposed to the solvents in 2 periods, 5 and 15 min and subjected to the microhardness test. Second test was to evaluate the ability of endodontic solvents to dissolve root-filling materials by immersion test in, 2, 5 and 15 minutes. Specimens of sealers, AH Plus, Acroseal, Sealer 26, Endofill, MTA Fillapex an RealSeal SE and guttapercha Dentsply (DP), ProTaper and Resilon points were prepared and evaluated in the three periods. Next test was conducted to evaluate the ability of solvents to desobturate root canals filled and analyse the effect of the ultrasonic passive agitation of solvents after canal unfill procedure. Sixty maxillary central incisors were prepared and obturated by same technique and randomly divided into 6 groups, one for each solvent (n=10) and a control group (saline solution). Teeth were scanned in Microcomputed Tomography (Micro-CT), unfilled with ProTaper Retratament/Protaper Universal with one of the solvents and scanned again. The residual root-fillings volumes were analysed and all teeth root canal were filled with the same solvent used in unfilling process and were passive-ultrasonically agitated (PUl) for 1 min. All teeth were scanned again in Micro-CT and data were analysed. Biological properties assessments were evaluated by cytotoxicity test with NIH-3T3 mouse cells by MTT assay. Cell cultures were subjected to diluted solvents at concentrations of 0.5 to 2.5%, and cell viability were analysed. Last evaluation in our study was the antimicrobial test. A total of 60 bovine dentin specimens infected intraorally were exposed for 5 min in direct contact to one of the solvents evaluated. After that, the dentin blocks with biofilms were...

Análise das propriedades químicas, biológicas e antimicrobianas de solventes endodônticos / Analysis of chemical, biological and antimicrobial properties of endodontic solvents

O objetivo deste estudo foi avaliar as propriedades químicas, biológicas e antimicrobianas dos solventes endodônticos, Citrol, Eucaliptol, d-Limoneno, Xilol e Endosolv E. Dentre os testes químicos, foram avaliados a microdureza dentinária, onde utilizamos blocos de dentina bovina que foram expostos aos solventes por 2 períodos, 5 e 15 min e submetidos ao teste de microdureza. Outro teste químico foi a capacidade de dissolução dos materiais obturadores pelo teste de imersão nos solventes em 3 períodos, 2, 5 e 15 min. Para essa avaliação foram, confeccionados corpos de prova dos cimentos, AH Plus, Acroseal, Sealer 26, Endofill, MTA Fillapex e RealSeal SE e dos cones de guta-percha estandardizado Dentsply (DP), ProTaper e Resilon. Um terceiro teste, foi realizado para avaliar a capacidade de desobturação dos canais radiculares e o efeito da agitação ultrassônica dos solventes endodônticos após a desobturação. Sessenta incisivos centrais superiores foram divididos em 6 grupos sendo um para cada solvente (n=10) mais um grupo controle (solução fisiológica). Todos dentes foram instrumentados e obturados pela mesma técnica e escaneados no Micro-CT. Os dentes foram então desobturados utilizando ProTaper Retratamento/ProTaper Universal como técnica para todos grupos, associada a um dos solventes e foram escaneados novamente para a avaliação do volume de material remanescente após a desobturação. Cada um dos dentes foi desobturado e preenchido com o mesmo solvente utilizado na desobturação. Cada solvente foi agitado pelo ultrassom por 1 min e novamente foram escaneados e avaliados os volumes restantes dos materiais nos canais radiculares. Na avaliação biológica, utilizamos o teste de citotoxicidade com células de camundongo NIH-3T3 pelo ensaio MTT. As culturas celulares foram plaqueadas e submetidas aos solventes diluídos nas concentrações de 0.5 a 2.5%, e avaliados quanto à viabilidade celular. Por fim o último teste foi o antimicrobiano, avaliado pelo teste de...

The aim of this study was to evaluate the chemical, biological and antimicrobial properties of the Endodontic solvents, Citrol, Eucalyptol, d-Limonene, Xylene and Endosolv E. First chemical properties analysis were evaluated by dentin microhardness, where we used bovine dentin blocks that were exposed to the solvents in 2 periods, 5 and 15 min and subjected to the microhardness test. Second test was to evaluate the ability of endodontic solvents to dissolve root-filling materials by immersion test in, 2, 5 and 15 minutes. Specimens of sealers, AH Plus, Acroseal, Sealer 26, Endofill, MTA Fillapex an RealSeal SE and guttapercha Dentsply (DP), ProTaper and Resilon points were prepared and evaluated in the three periods. Next test was conducted to evaluate the ability of solvents to desobturate root canals filled and analyse the effect of the ultrasonic passive agitation of solvents after canal unfill procedure. Sixty maxillary central incisors were prepared and obturated by same technique and randomly divided into 6 groups, one for each solvent (n=10) and a control group (saline solution). Teeth were scanned in Microcomputed Tomography (Micro-CT), unfilled with ProTaper Retratament/Protaper Universal with one of the solvents and scanned again. The residual root-fillings volumes were analysed and all teeth root canal were filled with the same solvent used in unfilling process and were passive-ultrasonically agitated (PUl) for 1 min. All teeth were scanned again in Micro-CT and data were analysed. Biological properties assessments were evaluated by cytotoxicity test with NIH-3T3 mouse cells by MTT assay. Cell cultures were subjected to diluted solvents at concentrations of 0.5 to 2.5%, and cell viability were analysed. Last evaluation in our study was the antimicrobial test. A total of 60 bovine dentin specimens infected intraorally were exposed for 5 min in direct contact to one of the solvents evaluated. After that, the dentin blocks with biofilms were...

Effects of solvent evaporation on water sorption/solubility and nanoleakage of adhesive systems

Objective: To evaluate the influence of solvent evaporation in the kinetics of water diffusion (water sorption-WS, solubility-SL, and net water uptake) and nanoleakage of adhesive systems. Material and Methods: Disk-shaped specimens (5.0 mm in diameter x 0.8 mm in thickness) were produced (N=48) using the adhesives: Clearfil S3 Bond (CS3)/Kuraray, Clearfil SE Bond - control group (CSE)/Kuraray, Optibond Solo Plus (OS)/Kerr and Scotchbond Universal Adhesive (SBU)/3M ESPE. The solvents were either evaporated for 30 s or not evaporated (N=24/per group), and then photoactivated for 80 s (550 mW/cm2). After desiccation, the specimens were weighed and stored in distilled water (N=12) or mineral oil (N=12) to evaluate the water diffusion over a 7-day period. Net water uptake (%) was also calculated as the sum of WS and SL. Data were submitted to 3-way ANOVA/Tukey's test (α=5%). The nanoleakage expression in three additional specimens per group was also evaluated after ammoniacal silver impregnation after 7 days of water storage under SEM. Results: Statistical analysis revealed that only the factor "adhesive" was significant (p<0.05). Solvent evaporation had no influence in the WS and SL of the adhesives. CSE (control) presented significantly lower net uptake (5.4%). The nanoleakage was enhanced by the presence of solvent in the adhesives. Conclusions: Although the evaporation has no effect in the kinetics of water diffusion, the nanoleakage expression of the adhesives tested increases when the solvents are not evaporated. .

Study of reaction parameters and kinetics of esterification of lauric acid with butanol by immobilized Candida antarctica lipase.

Esterification of lauric acid with n-butanol, catalyzed by immobilized Candida antarctica lipase (CAL) in aqueous-organic biphasic solvent system was studied. Effects of various reaction parameters on esterification were investigated, such as type and amount of solvent, amount of buffer, pH, temperature, speed of agitation, amount of enzyme, butanol and lauric acid. The most suitable reaction conditions for esterification were observed at 50ºC and pH 7.0 using 5000 μmoles of lauric acid, 7000 μmoles of butanol, 0.25 ml phosphate buffer, 1 ml of isooctane as the solvent and 50 mg of immobilized enzyme in the reaction medium at agitation speed of 150 rpm. Maximum esterification of 96.36% was acheived in 600 min of reaction time at n-butanol to lauric acid molar ratio of 1: 0.7. Kinetic study for the esterification of lauric acid with n-butanol using immobilized CAL was carried out and the kinetic constants were estimated by using non-linear regression method. The estimated value of Michaelis kinetic constants for butanol (KmBt) and acid (KmAc) were 451.56 (M) and 4.7 × 10-7(M), respectively and the value of dissociation constant (KBt) of the butanol-lipase complex was 9.41 × 107(M). The estimated constants agreed fairly well with literature data.

Airborne agricultural particulate matter induces inflammatory cytokine secretion by respiratory epithelial cells: Mechanisms of regulation by eicosanoid lipid signal mediators.

The purpose of this study was to elucidate the mechanism of the airborne poultry dust (particulate matter, PM)-induced respiratory tract inflammation, a common symptom in agricultural respiratory diseases. The study was based on the hypothesis that poultry PM would induce the release of inflammatory cytokine interleukin-8 (IL-8) by respiratory epithelial cells under the upstream regulation by cytosolic phospholipase A2 (cPLA2) activation and subsequent formation of cyclooxygenase (COX)- and lipoxygenase (LOX)-catalyzed arachidonic acid (AA) metabolites (eicosanoids). Human lung epithelial cells (A549) in culture were treated with the poultry PM (0.1-1.0 mg) for different lengths of time, following which PLA2 activity, release of eicosanoids and secretion of IL-8 in cells were determined. Poultry PM (1.0 mg/ml) caused a significant activation of PLA2 in a time-dependent manner (15-60 min), which was significantly attenuated by the calcium-chelating agents, cPLA2-specific inhibitor (AACOCF3) and antioxidant (vitamin C) in A549 cells. Poultry PM also significantly induced the release of COX- and LOX-catalyzed eicosanoids (prostaglandins, thromboxane A2 and leukotrienes B4 and C4) and upstream activation of AA LOX in the cells. Poultry PM also significantly induced release of IL-8 by the cells in a dose- and time-dependent manner, which was significantly attenuated by the calcium chelating agents, antioxidants and COX- and LOX-specific inhibitors. The current study for the first time revealed that the poultry PM-induced IL-8 release from the respiratory epithelial cells was regulated upstream by reactive oxygen species, cPLA2-, COX- and LOX-derived eicosanoid lipid signal mediators.