[Enhancing effects of chemical permeation enhancer propylene glycol and ultrasonic technology on transdermal permeation of Baimai Ointment].

This study aimed to improve the transdermal permeation quantity of Baimai Ointment by investigating the enhancing effects of physical and chemical permeation promoting methods on transdermal permeation of Baimai Ointment. The improved Franz diffusion cell method was used for in vitro transdermal experiment. The abdominal skin of mice was used, and the skin was treated with 3% propylene glycol in the chemical enhancement group. Ultrasonic technology was introduced in the physical enhancement group. The conditions of ultrasonic technology were optimized by single factor trial. Taking Q_(EF) and ER as the indexes of penetration promotion performance, the enhancing effects of the two methods were compared. The results showed that the promotion performance of 3% propylene glycol for ammonium glycyrrhizinate, nardosinone and curcumin of the chemical enhancement group were 1.74, 1.60, and 3.73 times higher than those of the blank group, respectively. The overall permeation efficiency of the Baimai Ointment was significantly improved. The comprehensive promoting effect on each component was curcumin>ammonium glycyrrhizinate>nardosinone. In the physical enhancement group, the penetration promoting effect of ultrasonic power 1.0 W was better than that of 2.0 W and 0.5 W, ultrasonic time 5 min was better than 3 min and 8 min, and the ultrasonic frequency 1 MHz was better than 3 MHz. Therefore, the optimal ultrasonic condition was 1.0 W-5 min-1 MHz. Under this condition, in terms of the transdermal permeation for ammonium glycyrrhizinate, the Q_(EF) and ER of the ultrasonic technology were better than those of 3% propylene glycol. In terms of the transdermal permeation for nardosinone and curcumin, the QEF and ER of 3% propylene glycol were better than those of the ultrasonic technology. Therefore, 3% propylene glycol combined with ultrasonic technology can be used to promote permeation of Baimai Ointment that contains both water-soluble and fat-soluble components in the clinical application. This study provides a theoretical basis for the clinical application of Baimai Ointment and other transdermal preparations.

In situ microemulsion-gel obtained from bioadhesive hydroxypropyl methylcellulose films for transdermal administration of zidovudine.

This study aims to develop in situ microemulsion-gel (ME-Gel) obtained from hydroxypropyl methylcellulose (HPMC) films for transdermal administration of Zidovudine (AZT). Firstly, HPMC films containing propylene glycol (PG) and eucalyptus oil (EO) were obtained and characterized. Later, a pseudo-ternary phase diagram composed of water, EO, tween 80 and PG was obtained and one microemulsion (ME) with a similar proportion of the film components was obtained. ME was transformed in ME-Gel by the incorporation of HPMC. Finally, HPMC films were hydrated with Tween 80 solution to yield in situ ME-Gel and its effect on AZT skin permeation was compared with HPMC film hydrated with water (F5hyd). The results showed that the ME and ME-Gel presented a droplet size of 16.79 and 122.13 µm, respectively, polydispersity index (PDI) < 0.39 and pH between 5.10 and 5.40. The incorporation of HPMC resulted in viscosity about 2 times higher than the use of ME. The presence of AZT did not alter the formulation properties. The in situ ME-Gel promoted a two-fold increase in the permeated amount of AZT compared to F5hyd. The results suggest that it was possible to obtain an ME-Gel in situ from HPMC films and that its effect on transdermal permeation of AZT was significant.

Formulation Development and Evaluation of Diphenhydramine Nasal Nano-Emulgel.

The aim of present study is to formulate diphenhydramine nasal nano-emulgels, having lipophilic nano-sized interior droplets, with better penetration for targeted controlled delivery to mucous membrane. Different diphenhydramine (DPH) nasal nano-emulgels were developed having propylene glycol and olive oil (as permeation enhancers) by using RSM for optimization and then evaluated for physico-chemical characteristics and thermal stability. In-vitro drug release through cellophane membrane was conducted and results were analyzed statistically. Further, gelation, mucoadhesive stress, and ex-vivo and histopathological studies were performed on optimized formulation by using goat nasal membrane. Among all formulations, E2 showed maximum DPH release at higher concentration olive oil (4%) and lower concentration propylene glycol (PG) (25%) within 4 h. All formulations have followed first-order kinetics and drug release mechanism was Fickian diffusion. Analysis of variance (ANOVA) and multiple linear regression analysis (MLRA) were used to compare results among formulations and 3D surface plots were constructed also. Optimized formulation showed immediate prolong gelation in artificial nasal mucosa and excellent mucoadhesive property (72.5 ± 1.5 dynes/cm2). Approximately 97.1% optimized formulation was permeated through membrane within 4 h, having a high flux rate (33.19 ± 0.897 µg/cm2/min) with diffusion coefficient (0.000786 ± 4.56 × 10-5 cm2/min) while drug contents remained on mucosal membrane for 24 h. Histopathologically, change on intra-mucosal surface of excised membrane was observed due to passage of drug through it. In summary, combination of PG and olive oil in nasal DPH nano-emulgel can be utilized successfully for targeted controlled delivery. The optimized formulation has excellent permeability and prolonged residence time on mucosal surface, which prove its good anti-histaminic activity in case of allergic rhinitis.

Evaluation of the ability to metabolize 1, 2-propanediol by heterofermentative bacteria of the genus Lactobacillus

Background: New directions of research on lactic acid bacteria include investigation of metabolic pathways for the synthesis and/or metabolism of 1,2-propanediol, commonly used in the food and chemical industry, medicine, pharmacy and cosmetology as well as agriculture. The objective of this study was to compare the capacity of strains representing three diverse heterofermentative species belonging to the genus Lactobacillus to synthesize and/or transform 1,2-PD as well as to suggest new directions of research aimed at commercial use of this metabolite. Results: The novel strain of Lactobacillus buchneri A KKP 2047p, characterized as exhibiting an unusual trait for that species in the form of capacity to metabolize 1,2-PD, grew poorly in a medium containing 1,2-PD as a sole carbon source. The supplementation with glucose facilitated rapid growth of bacteria and use of 1,2-PD for the synthesis of propionic acid. A similar observation was noted for Lactobacillus reuteri. On the other hand, Lactobacillus diolivorans effectively metabolized 1,2-PD which was the sole carbon source in the medium, and the addition of glucose inhibited the synthesis of propionic acid. The experiments also investigated the effect of cobalamin as a diol dehydratase coenzyme involved in the propionic acid synthesis from 1,2-PD whose addition promoted the yield of the reaction in the case of all tested strains. Conclusions: All tested isolates showed the ability to effectively metabolize 1,2-PD (in the presence of cobalamin) and its conversion to propionic acid, which reveals that investigated bacteria meet the essential requirements of microorganisms with a potential application.

Simultaneous Saccharification and Fermentation of Sugar Beet Pulp with Mixed Bacterial Cultures for Lactic Acid and Propylene Glycol Production.

Research into fermentative production of lactic acid from agricultural by-products has recently concentrated on the direct conversion of biomass, whereby pure sugars are replaced with inexpensive feedstock in the process of lactic acid production. In our studies, for the first time, the source of carbon used is sugar beet pulp, generated as a by-product of industrial sugar production. In this paper, we focus on the simultaneous saccharification of lignocellulosic biomass and fermentation of lactic acid, using mixed cultures with complementary assimilation profiles. Lactic acid is one of the primary platform chemicals, and can be used to synthesize a wide variety of useful products, including green propylene glycol. A series of controlled batch fermentations was conducted under various conditions, including pretreatment with enzymatic hydrolysis. Inoculation was performed in two sequential stages, to avoid carbon catabolite repression. Biologically-synthesized lactic acid was catalytically reduced to propylene glycol over 5% Ru/C. The highest lactic acid yield was obtained with mixed cultures. The yield of propylene glycol from the biological lactic acid was similar to that obtained with a water solution of pure lactic acid. Our results show that simultaneous saccharification and fermentation enables generation of lactic acid, suitable for further chemical transformations, from agricultural residues.

Perfil eletroforético do colostro de ovelhas suplementadas com propileno glicol e cobalto associado à vitamina B12 no final da gestação / Electrophoretic profile of the colostrum of ewes supplemented with propylene glycol and cobalt associated with vitamin B12 in late pregnancy

Este trabalho teve por objetivo avaliar o proteinograma do colostro de ovelhas submetidas a administração de propileno glicol e de cobalto associado à vitamina B12 no final da gestação. Dezoito ovelhas da raça Santa Inês, prenhas e com idade variando entre 18 meses a cinco anos foram distribuídas, por amostragem probabilística em três grupos experimentais, aproximadamente 30 dias antes da data prevista para o parto. No Grupo 1 (G1/n=6) foram administrados 30mL de propileno glicol P.A. via oral diariamente; no Grupo 2 (G2/n=6) foi administrado 1mg de cloreto de cobalto em solução a 1% via oral diariamente e 2mg de vitamina B12, via intramuscular semanalmente e no Grupo 3 (G3/n=6): grupo controle. Logo após o parto procedeu-se a colheita de 30mL de colostro, que foram acondicionados em recipientes apropriados e encaminhados ao laboratório. Após homogeneização, adicionou-se a cada 1.000µL de colostro, 75µL de solução de renina, que foi mantido em banho-maria a 37ºC por aproximadamente 20 minutos e centrifugado a 21.000G durante dez minutos em centrífuga refrigerada. Posteriormente, a fração intermediária, correspondente ao soro do colostro, foi aliquotada e mantida em ultrafreezer a -80oC para posterior determinação das proteínas. A determinação da proteína total do soro colostral foi realizada empregando-se reagente comercial. A separação das proteínas foi realizada utilizando-se a técnica de eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE). Foram identificadas as proteínas IgA, lactoferrina, albumina, IgG de cadeia pesada (IgGCP), ß-caseína, IgG de cadeia leve (IgGCL), ß-lactoglobulina and α-lactoalbumina, não havendo influência da administração dos suplementos na fase final da gestação sobre as concentrações protéicas do colostro.(AU)

This study aimed to evaluate the proteinogram of the colostrum of ewes submitted to administration of propylene glycol and cobalt associated with vitamin B12 in late pregnancy. Eighteen pregnant Santa Inês ewes 18 months to 5 years old were distributed by probabilistic sampling into three experimental groups, about 30 days before the expected delivery date. In group 1 (G1/n=6), daily oral doses of 30ml propylene glycol PA were administered; Group 2 (G2/n=2) received a daily oral dosage of 1mg cobalt chloride in 1% solution and 2mg of vitamin B12 intramuscularly once a week, and Group 3 (G3/n=6) was the control group. Soon after delivery 30mL of colostrum was harvested from each ewe, which were stored in appropriate containers and sent to the laboratory. After homogenization, we added to each 1000µL of colostrum 75µL solution of rennin, which was kept in a water bath at 37°C for about 20 minutes and centrifuged at 21.000G for 10 minutes in a refrigerated centrifuge. Later, the intermediate fraction, corresponding to colostrum whey, was aliquoted and kept in a -80oC ultrafreezer for subsequent determination of proteins. The determination of the total colostral protein whey was performed using a commercial reagent, observing the linearity test for colostrum. The separation of proteins was performed using the technique of electrophoresis on polyacrylamide gel containing sodium dodecyl sulfate (SDS-PAGE). Lactoferrin, IgA, albumin, IgG heavy chain (IgGCP), ß-casein, IgG light chain (IgGCL), ß-lactoglobulin and α-lactalbumin proteins were identified. There was no influence of the administration of supplements during late pregnancy on the concentration of proteins identified in the colostrum of the ewes.(AU)

Ruminal and blood responses to propylene glycol during frequent feeding.

The objective of the current experiment was to study the responses of ruminal and blood metabolites of Holstein dairy cows to propylene glycol (PG) under different methods of delivery during frequent feeding. By providing the same amount (200 mL or 200 g) of PG, delivery methods for PG were assessed: 1) control treatment: no PG; 2) dietary treatment: 200 g of PG as a dry product (65% purity; corresponded to 308 g of the dry product) mixed into the TMR; 3) oral-drench treatment: 200 mL of liquid PG (100% purity) orally drenched; and 4) rumen-drench treatment: 200 g of PG as a dry product drenched via the rumen cannula to mimic top dressing. Eight multiparous (lactation = 3 +/- 1.1 SD) ruminally cannulated Holstein dairy cows (DIM = 204 +/- 104.5 SD) were fed PG for 4 d (d 11 to 14) in a replicated 4 x 4 Latin square design with an experimental length of 14 d for each period. On the last day of each period, serial blood samples were removed from an indwelling catheter placed in the right jugular vein immediately before and for 4 h after PG administration. Cows were fed at 12x feeding/d for 2 d before entering the serial sampling period to minimize postprandial influences on blood metabolites. Ruminal content was also sampled hourly for 4 h on d 14. Milk was sampled from 2 consecutive milkings on d 13 during each period. Dry matter intake and milk yield were not affected by PG. Percentages of milk lactose were increased by PG delivered by all methods tested in the current experiment. Ruminal concentrations (as percentages of total volatile fatty acids) of acetate were decreased and concentrations of propionate and isovalerate were increased by PG, regardless of the delivery method; however, total volatile fatty acid concentration was not affected by PG. Ruminal concentrations of butyrate were decreased and concentrations of valerate were increased by PG drench, via either an oral or ruminal drench. The degree of reduction in butyrate concentration or increase in valerate concentration was affected by PG dose. Serum insulin peaked more rapidly and at a greater concentration for cows receiving PG via drenching, but not when PG was provided as a part of the TMR. Plasma glucose, however, tended to peak more rapidly at a greater concentration for cows receiving PG, regardless of the delivery method. Propylene glycol for the amount drenched (orally or ruminally) or fed (incorporated into the ration) shifted ruminal fermentation toward a more glucogenic environment. Drenching demonstrated a better efficacy than feeding PG because of the amount of PG that was available to the animal at the time of sampling. Effects of drenching dry PG into the rumen were comparable with orally drenching liquid PG.

Temporal dynamics and degradation activity of an bacterial inoculum for treating waste metal-working fluid.

In order for established bioreactors to be effective for treating chemically mixed wastes such as metal working fluids (MWF) it is essential that they harbour microbial populations that can maintain sufficient active biomass and degrade each of the chemical constituents present. In this study we investigated the effectiveness of a bacterial consortium composed of four species (Clavibacter michiganensis, Methylobacterium mesophilicum, Rhodococcus erythropolis and Pseudomonas putida), assembled on the basis of their apparent ubiquity in waste MWF, degradation ability and tolerance to fluctuating chemistry of the waste. The temporal dynamics of the inoculum and its effects on the fate of individual chemical components of the waste were studied, by regular sampling, over 400 h. Using a complementary approach of culture with chemotaxonomic (FAME) analysis and applying group specific probes (FISH), the inoculum was found to represent a significant component of the community in bioreactors with and without presence of indigenous MWF populations. In addition, the reduction in the COD by the consortium was approximately 85% of the total pollution load, and 30-40% more effectively than any other treatment (indigenous MWF community alone or activated sludge). Furthermore, all the chemical constituents, including the biocide (a formaldehyde release agent) demonstrated > 60% reduction. Many chemical components of the MWF proved to be recalcitrant in the other treatments. The results of this study confirm that assemblage of an inoculum, based on a comprehensive knowledge of the indigenous microbial community, in the target habitat, is a highly effective way of selecting microbial populations for bioaugmentation of bioreactors.

Effect of nutrient limitation on product formation during continuous fermentation of xylose with Thermoanaerobacter ethanolicus JW200 Fe(7).

Thermoanaerobacter ethanolicus JW200 Fe(7) was grown in continuous culture, using xylose as the primary carbon source, with progressively lower concentrations of supplementary yeast extract. This enabled the comparison of metabolic flux to fermentation end-products under carbon-limited and carbon-sufficient (yeast extract-limited) conditions and the determination of process data under fully mass-balanced conditions. Under carbon-limitation, the specific ethanol-formation rate was described by q (p)=40.34 micro +3.74, the specific rate of substrate utilisation for maintenance was 0.31+/-0.02 g x g(-1) x h(-1) and the maximum cell yield on xylose, corrected for maintenance requirements, was 0.15+/-0.04 g x g(-1). Based on the product profiles, these corresponded to a maintenance coefficient of m(ATP)=4.1+/-0.5 mmol x g(-1) x h(-1) and a maximum cell yield of = 14.7+/-0.8 x g x mol(-1). Limitation by a component in yeast extract resulted in incomplete xylose utilisation, increased catabolic flux rates (primarily resulting in increased lactate production, due to limitations in the flux through the phosphoroclastic reaction), a reduction in cell yield = 10.0+/-1.0 g x mol(-1) and an increase in maintenance energy requirements of m(ATP)=7.95+/-0.7 mmol x g(-1). The latter was also reflected in a shift from ethanol to acetate production at lower growth rates. An analysis of ethanol and acetate tolerance indicated that any high-intensity process employing this strain would require a bioreactor design which incorporated continuous ethanol stripping.

Functional genomic, biochemical, and genetic characterization of the Salmonella pduO gene, an ATP:cob(I)alamin adenosyltransferase gene.

Salmonella enterica degrades 1,2-propanediol by a pathway dependent on coenzyme B12 (adenosylcobalamin [AdoCb1]). Previous studies showed that 1,2-propanediol utilization (pdu) genes include those for the conversion of inactive cobalamins, such as vitamin B12, to AdoCbl. However, the specific genes involved were not identified. Here we show that the pduO gene encodes a protein with ATP:cob(I)alamin adenosyltransferase activity. The main role of this protein is apparently the conversion of inactive cobalamins to AdoCbl for 1,2-propanediol degradation. Genetic tests showed that the function of the pduO gene was partially replaced by the cobA gene (a known ATP:corrinoid adenosyltransferase) but that optimal growth of S. enterica on 1,2-propanediol required a functional pduO gene. Growth studies showed that cobA pduO double mutants were unable to grow on 1,2-propanediol minimal medium supplemented with vitamin B(12) but were capable of growth on similar medium supplemented with AdoCbl. The pduO gene was cloned into a T7 expression vector. The PduO protein was overexpressed, partially purified, and, using an improved assay procedure, shown to have cob(I)alamin adenosyltransferase activity. Analysis of the genomic context of genes encoding PduO and related proteins indicated that particular adenosyltransferases tend to be specialized for particular AdoCbl-dependent enzymes or for the de novo synthesis of AdoCbl. Such analyses also indicated that PduO is a bifunctional enzyme. The possibility that genes of unknown function proximal to adenosyltransferase homologues represent previously unidentified AdoCbl-dependent enzymes is discussed.