Development of microemulsions to topically deliver 5-aminolevulinic acid in photodynamic therapy

The aim of this study was to obtain and to characterize microemulsions containing 5-aminolevulinic acid (5-ALA) and to investigate the influence of these systems in drug skin permeation for further topical photodynamic therapy (PDT). 5-ALA was incorporated in water-in-oil (W/O), bicontinuous (Bc), and oil-in-water (O/W) microemulsions obtained by the titration of ethyl oleate and PEG-8 caprylic/capric glycerides:polyglyceryl-6 dioleate (3:1) mixtures with water. Selected systems were characterized by conductivity, viscosity, size of the droplets, and drug release. The stability of the drug in the microemulsions was also assessed. Moreover, the in vitro and in vivo skin permeation of 5-ALA was investigated using diffusion cells and confocal scanning laser microscopy (CSLM), respectively. Despite the fact that the O/W microemulsion decreased the 5-ALA diffusion coefficient and retarded the drug release, it also significantly increased the in vitro drug skin permeation when compared to other 5-ALA carriers. It was observed by CSLM that the red fluorescence of the skin increased homogeneously in the deeper skin layers when the 5-ALA microemulsion was applied in vivo, probably due to the formation of the photoactive protoporphyrin IX. The microemulsion developed carried 5-ALA to the deeper skin layers, increasing the red fluorescence of the skin and indicating the potentiality of the system for topical 5-ALA-PDT. (C) 2010 Elsevier B.V. All rights reserved.

Positive impact of omega-3 fatty acid supplementation in a dog with drug-resistant epilepsy: A case study

Epilepsy is the most common neurological disorder in both dogs and humans. Although the pharmacological options for treatment of epilepsies have increased. it has been reported that two-thirds of dogs with epilepsy are refractory to antiepileptic drug therapy. To our knowledge, there are no experimental Studies in the literature that show an effect of omega-3 supplementation oil epilepsy in dogs. Our case study describes the effectiveness of daily intake of a moderate amount of fish oil in a case of canine epilepsy. (C) 2009 Elsevier Inc. All rights reserved.

Postpartum ovarian activity of Santa Ines lactating ewes fed diets containing soybean hulls as a replacement for coastcross (Cynodon sp.) hay

The Santa Ines, a Brazilian hair sheep, has a non-seasonal breeding activity. Data regarding the duration of the postpartum anestrous period in Santa Ines lactating ewes is lacking and the objective of this trial was to determine the effects of replacing neutral detergent fibre (NDF) provided by coastcross (Cynodon sp.) hay with NDF contained in soybean hulls (SH) on the postpartum ovarian activity-as measured by the serum progesterone (P(4)) concentration. Fifty-six lactating ewes (body weight 56.1 +/- 6.8 kg) were individually penned and used in a randomized complete block design with 14 blocks and four treatments. The SH NDF replaced 33 (SH33), 67 (SH67), or 100% (SH100) of the NDF contributed by coastcross hay in the control diet (SHO). This resulted in a SH inclusion at rates of 0, 25, 54, and 85% of the dietary dry matter (DM). Blood samples were collected twice weekly from the 14th to 84th day postpartum and the serum P(4) concentrations determined by radioimmunoassay (RIA). It was estimated that the 1st postpartum ovulation occurred 6 days before the date that a serum P(4) >= 1 ng/ml concentration was first recorded. The mean body condition score (BCS; 0-5 scale) was 3.0 +/- 0.19 on day 14 postpartum and the mean BCS at day 56 postpartum increased linearly (P<0.01) with the inclusion levels of SH (3.09, 3.24, 3.34, and 3.36, respectively). Treatments did not differ significantly in the induction of postpartum days to the resumption of ovarian luteal activity (34.1 +/- 15.3 days postpartum). On days 25, 50, and 75 postpartum 36.80, and 100% of the ewes had resumed ovarian activity, respectively. Nonesterified fatty acid concentration decreased quadratically (P<0.05) with the SH inclusion, with values of 0.323, 0.244, 0.204, and 0.216 mequiv./l for the SHO, SH33, SH67, and SH100 treatments being recorded, respectively. Replacement of the NDF provided by coastcross hay with the NDF from the SH did not influence the duration of the postpartum anestrous period in Santa Ines lactating ewes. Considering a 150-day gestation period and the 34 days postpartum anestrous demonstrated in the present study, the current production system of a lambing interval of 8 months (3 lambing events in 2 years) may not be optimizing the production potential and a system in which the lambing interval is shortened by at least 1 month may be feasible. (C) 2008 Elsevier B.V. All rights reserved.

Responses of sugarcane, maize, and soybean to phosphorus and vesicular-arbuscular mycorrhizal fungi

The presence of vesicular-arbuscular mycorrhizal (VAM) fungi in long-term cane-growing fields associated with yield decline led to the supposition that VAM fungi may be responsible for the poor yields. A glasshouse trial was established to test the effectiveness of a species of VAM fungi, Glomus clarum, extracted from one of these North Queensland fields on the growth of sugarcane (Saccharum interspecific hybrid), maize (Zea mays), and soybean (Glycine max) for 6 phosphorus (P) rates (0, 2.7, 8.2, 25, 74, 222 mg/kg). For maize and soybean plants that received VAM (+ VAM), root colonisation was associated with enhanced P uptake, improved dry weight (DW) production, and higher index tissue-P concentrations than those without VAM (-VAM). By comparing DW responses of maize and soybean for different P rates, savings in fertiliser P of up to 160 and 213 kg/ha, respectively, were realised. Sugarcane plants were generally less responsive. Apart from a 30% DW increase with VAM when 2.7 mg P/kg was added, DW of +VAM plants was equivalent to, or worse than in the case of 222 mg P/kg, DW of -VAM plants. For all 3 host species, colonisation was least at the highest P application, presumably from excessive P within the plant tissue. Critical P concentrations for the 3 host species were below those reported elsewhere, and for soybean and sugarcane, the critical concentration for +VAM plants was lower than that of -VAM plants. There are 3 implications that arise from this study. First, VAM fungi present in cane-growing soils can promote the growth of maize and soybean, which are potential rotation crops, over a range of P levels. Second, the mycorrhizal strain taken from this site did not generally contribute to a yield decline in sugarcane plants. Third, application of P fertiliser is not necessary for sugarcane when acid-extractable P is

Quantitative trait loci for agronomic and seed quality traits in an F-2 and F-4 : 6 soybean population

Molecular breeding is becoming more practical as better technology emerges. The use of molecular markers in plant breeding for indirect selection of important traits can favorably impact breeding efficiency. The purpose of this research is to identify quantitative trait loci (QTL) on molecular linkage groups (MLG) which are associated with seed protein concentration, seed oil concentration, seed size, plant height, lodging, and maturity, in a population from a cross between the soybean cultivars 'Essex' and 'Williams.' DNA was extracted from F-2 generation soybean leaves and amplified via polymerase chain reaction (PCR) using simple sequence repeat (SSR) markers. Markers that were polymorphic between the parents were analyzed against phenotypic trait data from the F-2 and F-4:6 generation. For the F-2 population, significant additive QTL were Satt540 (MLG M, maturity, r(2)=0.11; height, r(2)=0.04, seed size, r(2)=0.061, Satt373 (MLG L, seed size, r(2)=0.04; height, r(2)=0.14), Satt50 (MLG A1, maturity r(2)=0.07), Satt14 (MLG D2, oil, r(2)=0.05), and Satt251 (protein r(2)=0.03, oil, r(2)=0.04). Significant dominant QTL for the F-2 population were Satt540 (MLG M, height, r(2)=0.04; seed size, r(2)=0.06) and Satt14 (MLG D2, oil, r(2)=0.05). In the F-4:6 generation significant additive QTL were Satt239 (MLG I, height, r(2)=0.02 at Knoxville, TN and r(2)=0.03 at Springfield, TN), Satt14 (MLG D2, seed size, r(2)=0.14 at Knoxville, TN), Satt373 (MLG L, protein, r(2)=0.04 at Knoxville, TN) and Satt251 (MLG B I, lodging r(2)=0.04 at Springfield, TN). Averaged over both environments in the F-4:6 generation, significant additive QTL were identified as Satt251 (MLG B 1, protein, r(2)=0.03), and Satt239 (MLG 1, height, r(2)=0.03). The results found in this study indicate that selections based solely on these QTL would produce limited gains (based on low r(2) values). Few QTL were detected to be stable across environments. Further research to identify stable QTL over environments is needed to make marker-assisted approaches more widely adopted by soybean breeders.

Cultivo in vitro de Crambe abyssinica Hochst. : germinação, micropropagação, estabilidade genética e anatomia foliar

Crambe (Crambe abyssinica) pertence à família Brassicaceae, originário da Etiópia e principalmente destinado à produção de forragem (30 a 32% de proteína bruta). Atualmente, tem sido bastante cultivado visando à extração de óleo vegetal. Com os atuais incentivos à busca de fontes de energias renováveis, o cultivo de crambe vem ganhando papel de destaque na produção de biodiesel por suas diversas vantagens, como: (a) rápido ciclo de vida (colhida em torno de 90 dias); (b) alta produção de biomassa; (c) alta produtividade de sementes (1000 e 1500 kg ha-1); (d) menor custo de produção em relação a outras fontes oleaginosas, como, canola, girassol e soja; (e) um percentual de óleo total na semente entre 32 e 38%, superando, por exemplo, a soja; (f) potencial de fitorremediação, eficiente na descontaminação de arsênio, cromo e outros metais pesados; e (g) elevado percentual de ácido erúcico (50 a 60%) sendo útil na indústria de plástico e lubrificante. Devido aos poucos trabalhos realizados com crambe, abre-se um vasto campo de investigações científicas que tenham como objetivo desenvolver as potencialidades dessa cultura e, consequentemente, melhorar os aspectos agronômicos e tecnológicos para seu emprego na indústria de biodiesel. Nesse contexto, as técnicas de cultivo in vitro foram importantes tanto para a propagação massal, quanto como ferramenta para uma possível aplicação de outras técnicas biotecnológicas, contribuindo para uma produção homogênea, fiel e em larga escala. Portanto, este trabalho teve como objetivo geral avaliar as condições mais favoráveis à germinação, estabelecimento in vitro e micropropagação de Crambe abyssinica Hochst., além de verificar possíveis alterações genéticas e anatômicas, possibilitando a regeneração e produção de plântulas viáveis. Para a germinação e estabecimento in vitro de crambe, as condições mais favoráveis foram em meio B5 ou WPM, na presença ou ausência de pericarpo e na presença de luz. Na micropropagação dessa espécie, uma frequência satisfatória de regeneração de brotos foi obtida a partir de segmentos apicais utilizados como explante em meio contendo 5 μM de BAP (6- benzilaminopurina), e o alongamento foi satisfatório com 1 μM de GA3 (ácido giberélico). Os marcadores moleculares ISSR (Inter-Simples Sequence Repeats) utilizados para a análise da estabilidade genética indicaram que o segmento apical de crambe é um explante confiável para a micropropagação de plantas geneticamente verdadeiras (true-to-tipe), ou seja, mantém a estabilidade genética. As diversas fontes de citocininas e concentrações utilizadas neste trabalho não promoveram mudanças, no sentido de alterar a organização e/ou a espessura em relação ao controle, e as alterações observadas na estrutura e espessura das folhas dos tratamentos de aclimatização prejudicaram o processo de estabelecimento da plântula ex vitro. Contudo, existe a necessidade de um enraizamento e aclimatização eficiente para completa propagação in vitro de crambe. Portanto, este protocolo de regeneração de plantas in vitro de crambe pode ser útil no processo de criação e desenvolvimento de novas cultivares em um tempo mais curto e no melhoramento genético usando explantes apicais.

Chemical composition and antioxidante activity of Tunisian Origanum glandulosum Desf. essential oil and volatile oil obtain by supercritical CO2 extraction

Dried flowers and leaves of Origanum glandulosum Desf. were submitted to hydrodistillation (HD) and supercritical fluid extraction with CO2 (SFE). The essential oils isolated by HD and volatile oils obtained by SFE were analysed by GC and GC/MS. Total phenolics content and antioxidant effectiveness were performed. The main components of the essential oils from Bargou and Nefza were: p-cymene (40.4% and 39%), thymol (38.7% and 34.4%) and γ- terpinene (12.3% and 19.2%), respectively. The major components obtain by SFE in the volatile oil, from Bargou and Nefza, were: p-cymene (32.3% and 36.2%), thymol (41% and 40%) and γ-terpinene (20.3% and 13.3%). Total phenolic content, expressed in gallic acid equivalent (GAE) g kg-1 dry weight, varied from 12 to 27 g kg-1 dw, and the ability to scavenge the DPPH radicals, expressed by IC50 ranged from 44 to143 mg L-1.

Monoterpene isolated from the essential oil of Trachyspermum ammi is cytotoxic to multidrug-resistant Pseudomonas aeruginosa and Staphylococcus aureus strains

Abstract INTRODUCTION: The aim of this study was to determine whether an herbal extract containing monoterpene exhibited activity against multidrug-resistant Staphylococcus aureus and Pseudomonas aeruginosa isolated from clinical infection samples. METHODS: The essential oil of Trachyspermum ammi (L.) Sprague ex Turrill (Apiaceae) fruit was extracted by hydrodistillation. Fruit residues were treated with hydrochloric acid and re-hydrodistilled to obtain volatile compounds. Compounds in the distilled oil were identified using gas-chromatography (GC) and GC-mass spectrometry (MS). The antibiotic susceptibility of all bacterial isolates was analyzed using both the disc diffusion method and determination of the minimum inhibitory concentration (MIC). The sensitivity of antibiotic-resistant isolates to essential oil was also determined by using the disc diffusion method and MIC determination. RESULTS: Of 26 clinical isolates, 92% were multidrug-resistant (MDR). Aromatic monoterpenes (thymol, paracymene, and gamma-terpinene) were the major (90%) components of the oil. Growth of S. aureus strains was successfully inhibited by the oil, with an inhibitory zone diameter (IZD) between 30-60mm and MIC <0.02μL/mL. The oil had no antimicrobial activity against clinical isolates of P. aeruginosa; rather, it prevented pigment production in these isolates. CONCLUSIONS: This study revealed that the essential oil of Trachyspermum ammi, which contains monoterpene, has good antibacterial potency. Monoterpenes could thus be incorporated into antimicrobial ointment formulas in order to treat highly drug-resistant S. aureus infections. Our findings also underscore the utility of research on natural products in order to combat bacterial multidrug resistance.

Development of a water-in-oil-in-water multiple emulsion system integrating biomimetic aqueous-core lipid nanodroplets for protein entity stabilization. Part I: experimental factorial design

Lipid nanoballoons integrating multiple emulsions of the type water-in-oil-in-water enclose, at least in theory, a biomimetic aqueous-core suitable for housing hydrophilic biomolecules such as proteins, peptides and bacteriophage particles. The research effort entertained in this paper reports a full statistical 23x31 factorial design study (three variables at two levels and one variable at three levels) to optimize biomimetic aqueous-core lipid nanoballoons for housing hydrophilic protein entities. The concentrations of protein, lipophilic and hydrophilic emulsifiers, and homogenization speed were set as the four independent variables, whereas the mean particle hydrodynamic size (HS), zeta potential (ZP) and polydispersity index (PI) were set as the dependent variables. The V23x31 factorial design constructed led to optimization of the higher (+1) and lower (-1) levels, with triplicate testing for the central (0) level, thus producing thirty three experiments and leading to selection of the optimized processing parameters as 0.015% (w/w) protein entity, 0.75% (w/w) lipophilic emulsifier (soybean lecithin) and 0.50% (w/w) hydrophilic emulsifier (poloxamer 188). In the present research effort, statistical optimization and production of protein derivatives encompassing full stabilization of their three-dimensional structure, has been attempted via housing said molecular entities within biomimetic aqueous-core lipid nanoballoons integrating a multiple (W/O/W) emulsion.

Infection by microorganisms on soybean seeds obtained from plants treated with growth regulators

This research, deals with the effects of exogenous growth regulators on infection by microorganisms on soybean (Glycine max cv. Davis) seeds. To study the influence of the chemicals, soybean plants were sprayed with gibberellic acid (GA) 100 ppm, (2-chloroethyl) trimethylammonium chloride (CCC) 2,000 ppm, succinic acid-2,2-dimethy1hydrazide (SADH) 4,000 ppm, indolylacetic acid (IAA) 100 ppm, 2,3,5-triiodobenzoic acid (TIBA) 20 ppm (three applications), and Agrostemin (1g/10 ml/ 3 1). Application of growth regulators did not affect infect ion by microorganisms on soybean seeds. The prominent fungus isolated was Phomopsis sojae. Alternaria and Fusarium spp. were isolated from seeds. The presence of a bacterium on the seeds was observed. The delay in harvest and high humidity increased the number of seeds from which Phomopsis was recovered.

Action of growth regulators on flowering and pod production of soybean cultivar Davis

This research deals with the effects of growth regulators on flowering and pod formation in soybean plant (Glycine max cv. Davis). Under greenhouse conditions, soybean plants were sprayed with 2,3,5-triiodobenzoic acid (TIBA) 20 ppm, Agrostemmin (1g/10 ml/3 l) gibberellic acid (GA) 100 ppm, and (2-chloroethyl) trimethylammonium chloride (CCC) 2,000 ppm. Application of TIBA increased number of flowers. 'Davis' soybean treated with CCC and TIBA presented a tendency to produce a lower number of pods.

Action of growth regulators on production of soybean cultivar Davis

This research deals with the effects of exogenous growth regulators on production of soybean plant (Glycine max cv.. Davis) under greenhouse conditions, At the flower anthesis, 2,3,5-triiodobenzoic acid (TIBA) 20 ppm was applied. Other two applications with TiBA, with intervals of four days, were realized. Before flowering, Agrostemin (1 g/10 ml/3 1), gibberellic acid (GA) 100 ppm, and (2-chloroethyl) trimethylammonium chloride (CCC) 2,000 ppm were applied. It was observed that CCC and TIBA reduced stem dry weight. Soybean plants treated with TIBA reduced weight of pods without seeds , seed number and seed weight.

Action of growth regulators on soybean cultivar Davis

The effects of growth regularots on soybean plant (Glycine max) under greenhouse conditions were studied. Before flower ing, Agrostemmin (1 g/10 ml/3 1), gibberellic acid (GA) 100 ppm, and (2-chloroethyl) trimethylammonium chloride (CCC) 2,000 ppm were applied. At the flower anthesis, 2,3,5 - triio dobenzoic acid (TIBA) 20 ppm was applied. Other two applications with TIBA, with intervals of four days, were realized. Treatment with GA increased plant height while CCC presented a tendency to reduce it. Numbers of leaves, internods, and stems were not affected by the growth regulators.

Effects of growth regulators on productivity of soybean (Glycine max cv. Davis) under competition

The effects of growth substances on productivity of 'Davis' soybean maintained under competition was investigated. Before the flowering, Agrostemmin (1 g/10 ml/3 1), gibberellic acid (GA) 100 ppm, and (2-chloroethyl) trimethylammonium chloride (CCC) 2,000 ppm were applied. At the flower anthesis, 2,3,5-triiodobenzoic acid (TIBA) 20 ppm was applied. Other two applications with TIBA, with intervals of four days, were realized. The growth regulators did not affect the productivity of 'Davis' soybean maintened under competition. The competition among plants did not affect the stem dry weight and number of pods, and seeds. The competition reduced weight of pods without seeds, seed weight, and weight of 100 seeds.

Dietary fish oil is antihypertrophic but does not enhance postischemic myocardial function in female mice.

Clinically and experimentally, a case for omega-3 polyunsaturated fatty acid (PUFA) cardioprotection in females has not been clearly established. The goal of this study was to investigate whether dietary omega-3 PUFA supplementation could provide ischemic protection in female mice with an underlying genetic predisposition to cardiac hypertrophy. Mature female transgenic mice (TG) with cardiac-specific overexpression of angiotensinogen that develop normotensive cardiac hypertrophy and littermate wild-type (WT) mice were fed a fish oil-derived diet (FO) or PUFA-matched control diet (CTR) for 4 wk. Myocardial membrane lipids, ex vivo cardiac performance (intraventricular balloon) after global no-flow ischemia and reperfusion (15/30 min), and reperfusion arrhythmia incidence were assessed. FO diet suppressed cardiac growth by 5% and 10% in WT and TG, respectively (P < 0.001). The extent of mechanical recovery [rate-pressure product (RPP) = beats/min x mmHg] of FO-fed WT and TG hearts was similar (50 +/- 7% vs. 45 +/- 12%, 30 min reperfusion), and this was not significantly different from CTR-fed WT or TG. To evaluate whether systemic estrogen was masking a protective effect of the FO diet, the responses of ovariectomized (OVX) WT and TG mice to FO dietary intervention were assessed. The extent of mechanical recovery of FO-fed OVX WT and TG (RPP, 50 +/- 4% vs. 64 +/- 8%) was not enhanced compared with CTR-fed mice (RPP, 60 +/- 11% vs. 80 +/- 8%, P = 0.335). Dietary FO did not suppress the incidence of reperfusion arrhythmias in WT or TG hearts (ovary-intact mice or OVX). Our findings indicate a lack of cardioprotective effect of dietary FO in females, determined by assessment of mechanical and arrhythmic activity postischemia in a murine ex vivo heart model.