ALA Concentration Research Articles

Lymphatic Transport of α‐Linolenic Acid and Its Conversion to Long Chain n‐3 Fatty Acids in Rats Fed Microemulsions of Linseed Oil

In the present study we evaluated the uptake of ALA and its conversion to EPA+DHA in rats given linseed oil (LSO) in native form or as a microemulsion in whey protein or in lipoid. In a single oral dose study in which rats maintained on rodent pellets deficient in ω-3 fatty acids were intubated with 0.35g LSO in lipoid, the amount of ALA present in lymph was increased reaching a maximum concentration of 16.23mg/ml at 2.5h. The amount of ALA present in lymph was increased to a maximum level of 10.95mg/ml at 4h in rats given LSO as a microemulsion in whey protein. When LSO was given without emulsification, the amount of ALA present in lymph was found to reach a maximum level of 7.08mg/ml at 6h. A similar result was observed when weaning rats were intubated with 0.15g of LSO per day for a period of 60days. Higher levels of ALA by 41 and 103% were observed in lymph lipids of rats given microemulsions of LSO in whey protein and in lipoid respectively as compared to rats given LSO without pre-emulsification. Very little conversion of ALA to EPA and DHA was observed in lymph lipids but higher amounts of EPA+DHA was observed in liver and serum of rats given LSO in microemulsion form. This study indicated that ALA concentration in lymph lipids was increased when LSO was given in microemulsion form in lipoid and further conversion to EPA and DHA was facilitated in liver and serum.

Exposure of fatty acids after a single oral administration of sacha inchi (Plukenetia volubilis L.) and sunflower oil in human adult subjects

Context: Sacha inchi (Plukenetia volubilis L.) is a potential oilseed crop because it is rich in α-linolenic acid (ALA) (omega-3 fatty acid).Objective: To evaluate the exposure of fatty acids after a single oral administration of sacha inchi or sunflower oil in healthy volunteers.Material and methods: Plasma fatty acids concentrations were assayed by Gas Chromatography with Flame Ionization Detector in 18 adult subjects. After fasting, blood samples were obtained at 0, 0.5, 1, 2, 4, 8 and 24 h after ingestion of 10 or 15 ml of sacha inchi oil or sunflower oil.Results: The proportion ALA/linoleic acid was 1.37 in sacha inchi oil and 0.01 in sunflower oil. ALA, lauric acid, palmitic acid, linolelaidic acid, cis-8,11,14-eicosatrienoic acid, cis-13,16-docosadienoic acid and cis-4,7,10,13,16,19-docosahexaenoic acid (DHA) levels changed over time after sacha inchi oil ingestion but not with sunflower oil. The time at maximal concentration (tmax) for ALA was 2 h after sacha inchi oil ingestion. No ALA in plasma was observed after sunflower oil consumption. The maximal concentration of ALA was 2.84 ± 0.36 mg/ml in women and 0.94 ± 0.57 mg/ml in men, p < 0.05, whereas maximal concentration of DHA was 2.60 ± 0.84 mg/ml in women and 1.00 ± 0.38 mg/ml in men (p > 0.05). There is a trend for higher plasma ALA levels with 15 ml sacha inchi oil. After 2 h of consumption, plasma delta triacylglycerol were reduced with sunflower oil but slightly increased with sacha inchi oil. A reduction in plasma delta triacylglycerol-rich lipoprotein cholesterol was observed with both oils.Conclusion: Consumption of sacha inchi oil increased ALA and DHA in plasma.

Investigation of 5-aminolevolinic acid (ALA) effects on seed germination and seedling growth of Silybum marianum under salinity stress

Salinity is a major environmental stress which adversely affects germination and seedling establishment in a wide variety of crops. However, studying of seeds with different plant growth regulators may helpful to alleviate the deleterious effects of salinity and enhance germination and seedling growth in salt stress environment. In present experiment we investigated the effects of 5-Aminolevolinic Acid (ALA) on germination and seedling growth parameters of milk thistle grown at different salinity levels. The experiment was arranged in factorial experiment in randomized complete block design with three replications. Results revealed that all parameters were significantly affected by salinity conditions and concentrations of ALA. In general, all parameters decreased with increasing NaCl concentrations, however the amount of these parameters increased with increasing ALA concentration. In addition, different between 0.25 and 0.50mM was not significant for germination percentage, shoot length, seedling fresh weight, seedling dry weight and leaf length. Interaction effects showed that highest value of root length and leaf length were produced from the 0.25 and 0.50mM ALA under the control treatment of salinity level. In contrast, the highest amount of seedling fresh weight were obtained from 0.25 and 0.50mM concentrations of ALA under control treatment of salinity. In general, our results suggest that ALA at low concentrations (0.25-1mM) has the potential to improve salt tolerance in milk thistle seedlings through concentrations treatment. We hope ALA may be useful in helping to solve serious problems occurring on a global scale, such as the desertification of green lands and salt damage to farm lands.

Bioconversion of α-Linolenic Acid into n-3 Long-Chain Polyunsaturated Fatty Acid in Hepatocytes and Ad Hoc Cell Culture Optimisation

This study aimed to establish optimal conditions for a cell culture system that would allow the measurement of 18∶3n-3 (ALA) bioconversion into n-3 long-chain polyunsaturated fatty acid (n-3 LC-PUFA), and to determine the overall pathway kinetics. Using rat hepatocytes (FaO) as model cells, it was established that a maximum 20∶5n-3 (EPA) production from 50 µM ALA initial concentration was achieved after 3 days of incubation. Next, it was established that a gradual increase in the ALA concentration from 0 up to 125µM lead to a proportional increase in EPA, without concomitant increase in further elongated or desaturated products, such as 22∶5n-3 (DPA) and 22∶6n-3 (DHA) in 3 day incubations. Of interest, ALA bioconversion products were observed in the culture medium. Therefore, in vitro experiments disregarding the medium fatty acid content are underestimating the metabolism efficiency. The novel application of the fatty acid mass balance (FAMB) method on cell culture system (cells with medium) enabled quantifying the apparent enzymatic activities for the biosynthesis of n-3 LC-PUFA. The activity of the key enzymes was estimated and showed that, under these conditions, 50% (Km) of the theoretical maximal (Vmax = 3654 µmol.g−1 of cell protein.hour−1) Fads2 activity on ALA can be achieved with 81 µM initial ALA. Interestingly, the apparent activity of Elovl2 (20∶5n-3 elongation) was the slowest amongst other biosynthesis steps. Therefore, the possible improvement of Elovl2 activity is suggested toward a more efficient DHA production from ALA. The present study proposed and described an ad hoc optimised cell culture conditions and methodology towards achieving a reliable experimental platform, using FAMB, to assist in studying the efficiency of ALA bioconversion into n-3 LC-PUFA in vitro. The FAMB proved to be a powerful and inexpensive method to generate a detailed description of the kinetics of n-3 LC-PUFA biosynthesis enzymes activities in vitro.

QUANTIFICATION OF 5-AMINOLEVULINIC ACID IN SWINE MANURE EXTRACT BY HPLC-FLUORESCENCE

A modified high performance liquid chromatography (HPLC)-fluorescence method for determination of the 5-aminolevulinic acid (ALA) concentration in swine waste was studied. The method included digestion of the sample by heating in aqueous HCl and derivatization by acetyl acetone/formaldehyde reagent. The derivative was isolated and detected by fluorescence with excitation at 370 nm and emission at 460 nm. Chromatographic separation of ALA was carried out using the optimum mobile phase of a methanol/water/glacial acetic acid (49.5:49.5:1, v/v/v). The method produced a limit of detection of the ALA concentration of 0.10 µg/mL and a limit of quantification of the ALA concentration of 0.31 µg/mL. The method was used for determination of the ALA concentration in concentrated swine manure extract (CSME) fermented for 0, 7, 14, 21, and 28 d. The ALA concentrations in the CSME fermented for 0, 7, and 14 d were low and ranged from 0.216 to 0.399 µg/mL, but increased to a level ranging between 1.240 and 1.295 µg/mL at 21 and 28 d of fermentation. The results of the study confirmed the presence of ALA in the swine waste. The ability to quantify the ALA concentration in swine waste developed by the study was confirmed.

5-aminolaevulinic acid/photo-dynamic therapy and gefitinib in non-small cell lung cancer cell lines: a potential strategy to improve gefitinib therapeutic efficacy.

Often, non-small cell lung cancers (NSCLC) respond only poorly to the tyrosine kinase inhibitor (TKI) gefitinib, which targets the epidermal growth factor receptor (EGFR), these poor responders EGFRs lacking activating mutations. In this study, we have attempted to improve TKI response of NSCLC cell lines (A549 and H1299) devoid of EGFR mutations, by combination of gefitinib and 5-ALA/photodynamic therapy (PDT). Cells of the two lines were incubated with gefitinib (from 0.5 to 50mm, for 48h) then irradiated at doses ranging from 4 to 20J/cm(2) ; 5-ALA concentration and incubation time were kept constant (1mm for 3h). We analysed cell viability, colony-forming efficiency, cell cycle parameters, proteasome and NF-κB activity and expression patterns of specific proteins, after individual or combined treatments. Effects (antagonistic, additive or synergistic) of combination treatment were evaluated using a predictive model (combination index) for expected interactive effects and results are consistent with mutual potentiation exceeding simple additivity. Investigation of molecular mechanisms underlying cytotoxic effects indicated that combination treatment impaired proteasome function, inhibited NF-κB transcriptional activity and hampered AKT pro-survival signalling. The results of this study show that poor response of cells devoid of EGFR activating mutations to TKIs, can be overcome by combining gefitinib with 5-ALA/photodynamic therapy (PDT).

Immunocytochemical studies on the nuclear ubiquitous casein and cyclin-dependent kinases substrate following 5-aminolevulinicacid-mediated photodynamic therapy on MCF-7 cells

Recent data indicates that nuclear ubiquitous casein and cyclin-dependent kinases substrate (NUCKS) may play role in tumor growth. In present study authors examined whether photodynamic therapy with 5-aminolevulinic acid (5-ALA) induces NUCKS expression in breast cancer cell line, MCF-7. In the experiment concentration of 5-ALA was 6.5mM. Excitation wavelength was 630 ± 20 nm, total light dose of light 5 or 10 J/cm(2) and irradiance 60 mW/cm(2) was used. Cells were collected at established time points and Western blot and immunocytochemical studies were performed using antibody against NUCKS. Studies proved strong cytotoxic effects in cells following PDT with 6.5mM of precursor and 10 J/cm(2). Western blot analysis revealed the strongest expression of NUCKS at 7h after PDT. At next time points, 18 and 24h, expression of NUCKS decreased and became similar to that of control group. Further immunocytochemical studies showed very strong expression of NUCKS following PDT with 5-ALA and light irradiation of 5 J/cm(2). Early, at 0 h, that expression was predominantly seen in nuclei, while at 7h expression of NUCKS was observed in disseminated manner within entire cells in both nuclei and cytoplasm, with prevalence of cytoplasmic staining. Authors suggest that NUCKS is involved in cellular responses following PDT, and since parallel induction of NUCKS and proapoptotic marker Bax and inhibition of anti-apoptotic Bcl-2 was observed, this protein might also be involved in induction of apoptosis following PDT.

Abstract P406: N-3 Fatty Acids Measured in Saliva do not Reflect Short-term Changes in Intake but may be Associated with Medium or Long-term Intake

Background: N-3 fatty acids in blood are adequate biomarkers of intake compared to dietary questionnaires because they do not rely on memory or self-reported information, and are not subject to interviewer bias. However, obtaining blood samples is an invasive approach, particularly among children. Measuring fatty acids in saliva is a non-invasive alternative that has not been explored yet in detail in population studies. Objective: To evaluate if n-3 fatty acids measured in saliva would be adequate biomarkers of intake. Methods: We selected 60 families for a 4-week intervention trial among participants in the Bogotá School Children Cohort. They were randomly assigned to receive a month’s supply of either soybean oil (high in α-linolenic acid [ALA], n=30) or sunflower oil (low in ALA, n=30). We obtained saliva and finger-prick blood samples in the children at baseline and follow-up. Fatty acids were analyzed by gas liquid chromatography in saliva and blood. To measure short-term responsiveness, we compared means of ALA in saliva by intervention arm at follow-up. To assess validity, we compared n-3 fatty acids concentrations in saliva and blood at baseline with the use of Spearman correlation coefficients. Results: Mean (SD) age of study participants was 14.5 (1.5) years; 37% (22 of 60) were male. Treatment groups did not differ at baseline with regard to socio-demographic and anthropometric characteristics, or fatty acids levels. Children assigned to soybean oil had a non-statistically significantly higher mean (SD) ALA concentration in saliva at follow-up (0.62 [0.66]), compared to children assigned to sunflower oil (0.47 [0.44]; p=0.45). Follow-up concentrations of long-chain n-3 fatty acids (EPA and DHA) in saliva did not differ significantly between intervention arms. To assess validity, we compared saliva and blood fatty acids concentrations at baseline (n=60) and found significant correlations for ALA (r=0.44, p=0.0005), EPA (r=0.27, p=0.04), and DHA (r=0.34, p=0.008). Conclusions: N-3 fatty acids measured in saliva do not reflect accurately short-term changes in ALA intake. However, moderate correlations between n-3 fatty acids in saliva and blood suggest that measuring fatty acids in saliva may be a valid proxy for intake. It is possible that fatty acids in saliva may have a slower turnover than those in blood and could be better biomarkers for medium or long-term intake of n-3 fatty acids.

Quantification of 5-aminolevulinic acid by CE using dynamic pH junction technique

An online dynamic pH junction preconcentration method was developed for quantification of 5-aminolevulinic acid (ALA) by CE with the separation time less than 6 min. The optimal dynamic pH junction of ALA was carried out between pH 9.3 borate buffer (BGE, 40 mM) and pH 2.5 phosphate buffer (sample matrix, 40 mM) when 4.1 cm of sample plug was hydrodynamically injected into an uncoated fused-silica capillary (48.5 cm in length, id of 50 μm). If a 24 kV separation voltage was applied, the calibration curve of ALA peak area (200 nm) showed good linearity (R(2) = 0.9991) ranging from 0.01 to 6.5 mg/mL. The reproducibility of this system was excellent with RSDs (n = 10) of 2.5% for peak area response and 0.6% for migration time at ALA concentration of 0.5 mg/mL. The LOD was evaluated as 1.0 μg/mL (S/N > 3). Compared to conventional CE procedure, the sensitivity was successfully improved over 50-fold. The analytical results of pharmaceutical formulations show a good agreement with those by HPLC (r = 0.94).

Effect of α-Lipoic Acid and α-Tocopherol Acetate Enriched Broiler Diet on Oxidative Stability and Quality of Broiler Leg Meat and Meat Products

The present study was conducted to evaluate the effect of inclusion of two dietary antioxidants alpha lipoic acid (ALA 25, 75 and 150 mg/kg diet) and alpha-tocopheryl acetate (ATA 200 mg/kg diet) on growth performance, susceptibility to oxidation and quality of leg chicken meat and meat products during storage. A total of 180 broiler birds were randomly divided into 6 groups with 3 replicates of 10 hens each and fed on different diet plan. Body weight gain of broiler birds was decreased with increasing alpha lipoic acid supplementation in feed and Birds in (T2) containing minimum dose of alphalipoic acid (25 mg/kg feed gained more weight 1948.25g whereas T4 containing maximum dose of alpha lipoic acid (150 mg/kg of feed) gained minimum weight i.e. 1691.25 g. The inclusion of alpha-tocopheryl acetate improved body weight gain. The maximum deposition of ALA took place in T4 which contain maximum dose of ALA. The thiobarbituric acid reactive substances (TBARS) assay and DPPH assay of the broiler leg meat were as in T4 (0.15 ± 0.02 MDA/kg of meat, 76.69 ± 0.14%) and in T5 were (0.25 ± 0.08 MDA/kg of meat, 44.77 ± 0.09%) accordingly. The (HPLC) data revealed that ALA and ATA concentration were highest in T4 (96.54 ± 0.28mg/g of meat, 159.40 ± 0.20 mg/g of meat) and lowest in T5 (17.19 ± 0.12mg/g, 35.86 ± 0.08 mg/g of meat). Sensory results showed that nuggets and patties made of T5 containing oxidize oil were least preferred and T4 got highest score. In a nutshell, 150 mg/kg level feed dietary supplementation of ALA with constant level of ATA can enhanced the antioxidant potential, lipid stability and nutritional qualities of broiler leg meat and meat products.

Photodynamic treatment induces cell death by apoptosis or autophagy depending on the melanin content in two B16 melanoma cell lines

Photodynamic therapy (PDT) is now a well-established treatment modality for cutaneous carcinomas and is based on the administration of a light-activated drug followed by illumination of the pathological area. The treatment of metastatic melanoma remains a therapeutic challenge. To define the possible role of melanin in relative phototoxicity of 5-aminolevulinic acid (5-ALA), a photosensitizer used in PDT invivo, we studied cell death in two variants (with or without melanin, B16F10 and B16G4F cells, respectively) of a melanoma cell line. Concentrations of 5-Ala up to 10mM induced similar cytostatic effects in the B16G4F and B16F10 cells. PDT and high 5-ALA concentrations induced photocytotoxicity in both melanoma cell lines (at 10mM for B16F10 cells and at 5mM for B16G4F cells). Cell death corresponded to p53-dependent apoptotic signaling in pigmented B16F10 cells, whereas an autophagic response leading to a caspase-independent death was detected in non-pigmented B16G4F cells. Therefore, the PDT-induced cell death pathway appeared to correlate with melanin synthesis capacity in melanoma cells. To reduce the cytotoxicity of 5-ALA without irradiation, a low drug concentration could be used. Consequently, in combination with current therapeutics, a moderate concentration of 5-ALA and PDT may constitute a supplementary promising approach to eliminate metastatic melanoma.

Effect of 5-aminolevulinic acid on yield and quality of lettuce in sunlit greenhouse

The role of 5-aminolevulinic acid (ALA) as a precursor of chlorophyll and heme is well documented. Low concentration of exogenous ALA has been found to regulate plant growth and increase crop yield, but there is little information on how ALA influences the yield and quality of lettuce in sunlit greenhouse. Here, we report the effects of ALA on photosynthetic rate, yield and quality of lettuce in sunlit greenhouse.5-aminolevulinic acid and 5-aminolevulinic acid with nitrogen fertilizer (ALA+N) were applied by foliage and soil. The results showed that application of ALA improved the photosynthetic rate of lettuce leaves by 23.9 to 34.7% and by 35.3 to 41.6%. Moreover, exogenous ALA increased vitamin C and soluble sugar content, reduced nitrate and crude fiber content and lead to better quality and taste of lettuce. Keywords : 5-aminolevulinic acid, lettuce, plant growth, promotive effects, yield, vegetable quality

Dietary supplementation with 5-aminolevulinic acid modulates growth performance and inflammatory responses in broiler chickens

The objective of this study was to investigate the effect of dietary supplementation with 5-aminolevulinic acid (5-ALA) on the immune system, inflammatory response, and growth performance of broiler chickens. The levels of cluster of differentiation 3 (CD3) mRNA in the spleens of chickens gradually increased with dietary 5-ALA concentration, while the expression levels of interleukin (IL)-2 decreased. Mitogen-induced proliferation of splenic mononuclear cells and blood mononuclear cell phagocytosis in chickens fed 0.001 and 0.01% 5-ALA-supplemented diets were significantly greater than in chickens fed a basal diet (control). Plasma thiobarbituric acid reactive substance (TBARS) concentration gradually increased along with 5-ALA supplement concentration. These results provide the first evidence that the use of dietary 0.001 and 0.01% 5-ALA supplementation induces the T-cell immune system via mild oxidative stress in chickens. Three hours after Escherichia coli lipopolysaccharide-induced immune stimulation, the levels of mRNA encoding pro-inflammatory cytokines, such as IL-6 and tumor necrosis factor-like ligand 1A (TL1A), in chickens fed a 0.001% 5-ALA-supplemented diet were significantly lower than those in chickens exposed to other treatments. The plasma caeruloplasmin concentration in chickens fed a 0.001% 5-ALA-supplemented diet was significantly lower than in controls or in chickens fed diets supplemented with other concentrations of 5-ALA 24 h after injection of LPS. In addition, BW at 21 and 50 d of age was significantly higher in chickens fed a 0.001% 5-ALA-supplemented diet than in control chickens. The findings suggest that supplementation of diets with 0.001% 5-ALA could prevent the catabolic changes induced by immunological stimulation. These results show that 5-ALA might be useful as an immunomodulator to stimulate T-cells via mild oxidative stress in growing broiler chickens, thereby improving the growth performance.

Effect of intake of linoleic acid and α‐linolenic acid levels on conversion into long‐chain polyunsaturated fatty acids in backfat and in intramuscular fat of growing pigs

A study was conducted to determine the effect of two levels of linoleic acid (LA) intake at either high or low α-linolenic acid (ALA) intake on their conversion and subsequent deposition into long-chain (20-22 C-atoms) polyunsaturated fatty acids (LC PUFA) in muscle and backfat in growing pigs. In a 2 × 2 factorial arrangement, 32 gilts from 8 litters were assigned to one of four dietary treatments, varying in LA and ALA intakes. Low ALA and LA intakes were 0.15 and 1.31 g/(kg BW(0.75) /day), respectively, and high ALA and LA intakes were 1.48 and 2.65 g/(kg BW(0.75) /day) respectively. There was a close positive relation between intake of ALA and the concentration of ALA in backfat and in intramuscular fat. Dietary ALA did not affect the concentration of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), but increased docosapentaenoic acid (DPA) in backfat. High ALA intake did not significantly affect DHA but significantly increased EPA, 20:3 n-3 and DPA concentrations in intramuscular fat. The n-3 LC PUFA proportion in backfat was increased from approximately 1-3%, which may be useful to enrich meat with these fatty acids. The effect of ALA intake on n-3 LC PUFA was suppressed by LA intake. Dietary ALA suppressed the concentration of n-6 LC PUFA in blood plasma by more than 50%. When compared at equal incremental dose, the inhibiting effect of ALA on blood arachidonic acid was stronger than the stimulating effect of LA as precursor.

基于ALA-PDT中PpIX亚细胞分布位点光漂白实验研究

随着光动力学疗法(photodynamic therapy，PDT)基础研究的不断深入和临床应用的广泛开展，光敏剂的光漂白现象日渐成为PDT研究的热点问题。实验在ALA的终浓度为10 mM/ml，辐射光波长为410 nm，辐照光功率为5 mW/cm2等条件下采用荧光探针标记技术在自行设计的光动力疗法反应室进行了PDT实验研究。Cell Counting Kit-8 (CCK-8)法用来检测细胞的活性，通过获得的荧光光谱研究了亚细胞的光漂白情况。结果表明：以每种细胞器光照前的平均荧光强度为基准，则光照60 min后线粒体、溶酶体、内质网的荧光强度下分别降低了95%、91.5%和51.2%。线粒体区域光漂白速率较快，其荧光强度的变化基本表征了HL60细胞活性的变化，推断线粒体是光动力效应的主要作用靶点之一。 With the development of the fundamental research of photodynamic therapy (PDT) and its clinical applica-tion, the photobleaching properties of photosensitizers have become a hotspot of research. The experimental research of ALA mediated PDT was carried out in the own designed reaction chamber when the final concentration of ALA is 10 mM/ml, the wavelength and the output power of the light source are 410 nm and 5 mW/cm2 separately. Cell Counting Kit-8 (CCK-8) was used to evaluate the cell viability while the photobleaching of PpIX in the subcellular structure of HL60 was detected by fluorescence spectroscopy. The results showed that after 60 min irradiation the fluorescence in-tensity of mitochondria, lysosomes and endoplasmic reticulum had decreased by 95%, 91.5% and 51.2% respectively compared to that before irradiation. The rate of photobleaching in mitochondria which can characterize the activity of HL60 was significantly higher than others, this suggest that mitochondria might be one of main therapeutic targets of photodynamic therapy.

5-aminolevulinic acid-incorporated poly(vinyl alcohol) nanofiber-coated metal stent for application in photodynamic therapy

BackgroundThe study investigated the use of combined photodynamic therapy (PDT) and stent placement for the treatment of cholangiocarcinoma (CC). For this purpose, 5-aminolevulinic acid (ALA) was incorporated into poly(vinyl alcohol) (PVA) nanofiber, and coated onto metal stents. Their efficacy was assessed in PDT towards HuCC-T1 CC cells.MethodsFabrication of ALA-PVA nanofiber, and simultaneous coating onto metal stents, was performed through electrospinning. The dark-toxicity, generation of protoporphyrin IX (PpIX), and PDT effect of ALA and ALA-PVA nanofiber were studied in vitro, using HuCC-T1 CC cells.ResultsThe ALA-PVA nanofibers were coated onto metal stents less than 1000 nm in diameter. ALA-only displayed marginal cytotoxicity; ALA-PVA nanofiber showed less cytotoxicity. PpIX generation was not sigficantly different between ALA and ALA-PVA nanofiber treatments. PVA itself did not generate PpIX in tumor cells. ALA and ALA-PVA nanofiber displayed a similar PDT effect on tumor cells. Cell viability was decreased, dose-dependently, until ALA concentration reached 100 μg/mL. Necrosis and apoptosis of tumor cells occurred similarly for ALA and ALA- PVA nanofiber treatments.ConclusionThe ALA-PVA nanofiber-coated stent is a promising candidate for therapeutic use with cholangiocarcinoma.

A temporal survey on protoporphyrin IX fluorescence signal after administration of 5ALA conjugated gold nanoparticles: An in vivo study on CT26 tumors

Introduction: Photodynamic therapy (PDT) is a therapeutic method using a photosensitizer that is activated by proper wavelength of light and molecular oxygen. Effective delivery of photosensitizer to target tumor cells that causes minimal damage to healthy cells is an important requirement in photodynamic therapy of cancer. Protoporphyrin IX (PpIX) induced endogenously by 5-aminolevulinic acid can be used as a photosensitizer to destroy photodynamically tumor cells. In this study biocompatible gold nanoparticles were used as a vehicle to deliver 5-aminolevulinic acid (5ALA) in intratumoral injection for selective and efficient in vivo photodynamic therapy of CT26 tumor model. Accumulation time in equivalent concentration of 5ALA and 5ALA-AU was compared with each other. Results: PpIX accumulation achieved the peak level at 3 and 5 h after intratumoral injection of 5ALA-AU and 5ALA, respectively and was completely eliminated within 24 h. Conclusions: It seems that 5ALA conjugated gold nanoparticles are not efficient on decrease of incubation time of 5ALA.

Modulation of Plasma Levels and Percentages of Incorporation of ω-3 PUFAs in Egg Yolk under the Influence of Supplementation Sources Rich in Omega 3 to Diet of Laying Hens

o Abstract: Hundred forty-four Shaver White laying hens were used over a 4 week experimental period to investigate the effect of 3% of soybean oil, corn oil (MIL), canola oil, flaxseed oil (LIN), salmon oil (SAL) or tuna and sardine oil (SR/AT) added to the diets, upon the fatty acid egg yolk composition, blood plasma levels and incorporation time of each fatty acid into the egg yolk. Hens were allocated into 72 cages and the experimental design was a 6 x 6 randomized factorial model. Hens fed 3% of different oils, responded with increased polyunsaturated fatty acids omega 3 (ω-3 PUFAs), except for corn oil. The addition of flaxseed, soybean or corn oil into the diet increased the PUFAs levels into the egg yolk and in the blood plasma. Adding tuna and sardine oil into the diet increased the concentration of yolk saturated fatty acids. The levels of ω-3 PUFAs were increased in the tuna and sardine oil treatment, while the flaxseed oil increased the plasma fatty acids. The deposition of 349.28 mg/yolk of α-linolenic fatty acids (ALA) was higher in the group fed LIN, while the higher equal to 157.13 mg DHA/yolk was observed in group SR/AT. In the plasma, deposition increased from 0.33% (MIL) for 6.29% ALA (LIN), while that of DHA increase of 0.47% (MIL) for 4.24% (SAL) and 4.48% (SR/AT) and of 0.98% (MIL) for 6.14% (SR/AT) and 8.44% (LIN) of ω-3 PUFAs. The percentage of EPA into the yolk and plasma was higher for the hens fed 3% tuna and sardine oil diet, as well as the levels of yolk DHA. The concentration of DHA into the plasma was higher for the salmon and tuna/sardine oil treatments. The PUFAs yolk decreased during the first eight days of experiment, while the ω-3 PUFAs increased during the same period. The concentration of ALA increased until ten days of experiment, while the percentage of EPA and DHA increased up to the eighth experimental day.

Targeting of sebaceous glands by δ‐aminolevulinic acid‐based photodynamic therapy: An in vivo study

Wide application of ALA-PDT for acne is limited due to relative strong side effects, such as pain and erythema. The objective of this study was to establish a protocol for ALA-PDT that would provide specific destruction of sebaceous glands at the lowest concentrations and shortest contact times of ALA. The rhino (hr(rh) hr(rh) ) murine model, an experimental acne model, was used in this study. A freshly prepared hydrophilic ALA hydrochloride ointment (2.5%, 5%, and 20%) was applied to the backs of 16-week-old male rhino mice. The fluorescence intensity (FI) of ALA-induced protoporphyrin IX (ALA-PpIX) on the skin was measured by spectrofluorometry. Skin samples were taken at 1, 2, and 4 hours after ALA application to determine the tissue distribution of ALA-PpIX by fluorescence microscopy. Light irradiation was also performed with a broadband light source (600-1100 nm, 15 J/cm² , 60 mW/cm²) with subsequent histological examination 1 day after treatment. Prominent increases of ALA-PpIX were observed 1 hour after application of 5% and 20% ALA, while no increase was observed with 2.5% ALA until 2 hours. A direct correlation was found between ALA concentration and ALA-PpIX FI. While no fluorescence was detected 1 hour after application of 2.5% or 5% ALA, 20% ALA produced a strong fluorescence in the epidermis, utricle walls, and sebaceous glands. Histological evaluation showed no damage to skin treated with 2.5% ALA-PDT incubated for 1 hour. Damage was still focused within the sebaceous glands with longer incubation times. Increased ALA concentrations resulted in more prominent damage to the epidermis and sebaceous glands, with deeper damage to the dermis when longer incubation times were used. Focused damage of sebaceous glands can be achieved with ALA-PDT when low concentrations of ALA (2.5-5.0%) and short incubation times (to 2 hours) were used.

Effects of 5-Aminolevulinic Acid on Chlorophyll, Photosynthesis, Soluble Sugar and Flavonoids of Ginkgo biloba

The flavonoid content determines the quality of Ginkgo biloba that can be increased by using of plant growth regulators. The objective of study was to observe the effect of 5-aminolevulinic acid (ALA), a key precursor in the biosynthesis of porphyrins and a new plant growth regulator, on photosynthetic rate, chlorophyll and soluble sugar content, flavonoid accumulation, and flavonoid enzyme activity in G. biloba leaves. The ginkgo seedlings were grown in greenhouse conditions under low levels (10 and 100 mg l-1) of foliar application of ALA. Photosynthetic rates of leaves increased significantly at day 4 in response to both ALA concentrations and remained elevated as compared to control for further 12 days. Chlorophyll and soluble sugar contents were significantly increased by day 4 and continued to increase by day 16; however, Chl a/b ratio remained unchanged. Total polyphenols, flavonoids, and anthocyanins, phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS) and chalcone isomerase (CHI) activities were increased from day 4 to 16 after ALA treatment. The increase in chlorophyll and soluble sugar contents, and activities of flavonoid enzymes (PAL, CHS and CHI) were likely to be closely associated with improvement of the accumulation of total polyphenols, flavonoids, anthocyanins and advance of leaf quality by ALA treatment. Foliar treatment with a low concentration of ALA therefore, might provide a useful means of improving pharmacological properties of G. biloba leaves.