In-vitro release profile of microencapsulated α-tocopherol under simulated gastrointestinal conditions

Alpha-tocopherol, the most common form of Vitamin E in nature, is a well-known antioxidant compound for its effective inhibition of lipid oxidation both in food and biological systems. Additionally, due to its preventive action against reactive oxygen species (ROS), α-tocopherol has been associated with risk decreasing of diseases associated with oxidative stress, such as cardiovascular disease and cancer [1]. The recommended ingestion of Vitamin E varies among the countries and according to criteria such as sex and age. In the USA, the recommended daily allowance (RDA) for an adult is 15 mg/day, whereas in Europe it is 4-15 and 3-12 mg α-tocopherol/day for man and women, respectively. Although α-tocopherol is naturally present in several foods, such as vegetable oils and tree nuts, owing to its antioxidant capacity it is frequently included in food supplements and used in the food industry to extend the shelf-life of several products. Nevertheless, due to α-tocopherol instability and sensitivity towards oxygen and light and its poor aqueous solubility, it is generally administered in the acetate or succinate form. However, these forms are considered to have a lower intestinal absorption compared to α-tocopherol [2]. To overcome these problems, the encapsulation of α-tocopherol in protective matrixes to avoid its oxidation and increase shelf life has been suggested. In fact, during the last years, encapsulation technology has been increasingly important in the food industry as it permits the formation of a physical barrier between the external medium and sensitive core materials, being also used for controlled release of active molecules, formulation stability enhancement, and flavor and taste masking. In this context, it is important to assess α-tocopherol release pattern from microparticles as it can restrain its different applicability. In this work, α-tocopherol microspheres were produced using alginate as a polymeric matrix. This polymer, a linear polysaccharide obtained from brown algae consisting of β-mannuronic acid and α-guluronic acid units, was chosen due to its biocompatibility, biodegradability and non-toxicity. Moreover, it presents a high stability at acidic pH, being easily swollen under mild alkali conditions. Alginate microspheres (ME) loaded with α-tocopherol were produced using a NISCO Var J30 atomization unit, following a previously optimized methodology. The produced microspheres were evaluated for encapsulation efficiency and α-tocopherol release profile by measuring the absorbance at 297 nm using a spectrophotometer. The encapsulation efficiency was calculated both by directly measuring the maximum content released after ME disruption and by quantifying the nonencapsulated α-tocopherol present in the CaCl2 coagulation solution and in the wash solution. α-Tocopherol in-vitro release profiles were determined under simulated gastric (pH 1.2) and intestinal (pH 7.4) media during a period of 24 hours. Additionally, a sample of ME were mixed in gastric media during 1h and then transferred to intestinal media until a total of 24h to simulate gastrointestinal conditions. During the testing period samples of the supernatant were periodically taken to determine the amount of released α-tocopherol. Results evidence a very low % of α-tocopherol release under acidic conditions while an almost complete release is achieved when ME were submitted to simulated intestinal conditions suggesting that the proposed approach can constitute an interesting solution to protect α- tocopherol, allowing for its release in the intestine after ingestion. The next steps in this work in progress will include the evaluation of release profiles of ME added to different food matrices.The authors thank FCT for the financial support LSRE (PEst-C/EQB/LA0020/2011 strategic project).info:eu-repo/semantics/publishedVersio

Study of the encapsulation efficiency and release profiles of alginate microspheres containing α-tocopherol

Vitamin E, a lipophilic natural antioxidant comprising different vitamers (α, β, γ and δ-tocopherols and tocotrianols), is frequently used in food supplements and added in different products, such as foodstuffs and cosmetics, to prevent lipid oxidation processes. Due to solubility and stability issues, α-tocopherol is generally administrated as succinate or acetate derivatives, which have lower bioavailability. In this work, alginate microspheres containing α-tocopherol were produced and evaluated for encapsulation efficiency and release profiles at different pH conditions during 24h. In vitro release tests showed that alginate microspheres maintain its integrity under simulated gastric conditions. By the contrary, under simulated intestinal conditions, an almost complete release is achieved during 24h, with a major portion (70%) being released after 2h.Financial support was provided by FCT and FEDER under Programme COMPETE (Project PEst-C/EQB/LA0020/2013).info:eu-repo/semantics/publishedVersio

Desenvolvimento de um processo de microencapsulação baseado em quitosano para proteção do α-tocoferol

Este trabalho foi realizado no âmbito da unidade curricular de Projeto da Licenciatura em Engenharia Biomédica do Instituto Politécnico de Bragança. Teve como principal objetivo desenvolver um processo de microencapsulação para proteção do α-tocoferol, a principal forma da vitamina E. O α-tocoferol é um antioxidante que possui um papel importante na proteção do organismo contra certos tipos de cancro e do envelhecimento da pele. Contudo, apresenta instabilidade à temperatura, oxigénio e luz, sendo importante a sua microencapsulação para garantir a sua proteção. Os estudos preliminares levados a cabo no âmbito da unidade curricular supramencionada consideram a utilização de duas matrizes poliméricas (quitosano e alginato), optando-se neste trabalho por apresentar os resultados obtidos com o quitosano e na perspetiva do desenvolvimento do processo de microencapsulação. Numa última etapa, o comportamento das microesferas produzidas foi testado para diferentes condições de pH

Fluoride content in children's dentifrices marketed in Lima, Peru

The aim of the present study was to determine the concentration of total fluoride (TF) and total soluble fluoride (TSF) in children's dentifrices marketed in the city of Lima, Peru. Three samples of 23 dentifrices (4 without fluoride and 19 with fluoride) were purchased in different pharmacies in Lima, Peru. The TF and TSF concentrations found in the dentifrices were determined by ion-selective electrode, expressed in ppm F (μg F/g of dentifrice). The TF concentration in the majority of the fluoride toothpastes matched that shown on the label, except for one declared as 1450 ppm F by the manufacturer, whereas only 515.1 ppm F was found. The concentration of TSF found in the fluoride toothpastes ranged from 457.5 to 1134.8 ppm F. All the dentifrices were formulated with silica, but one also presented calcium carbonate. In conclusion, 83% of the children's dentifrices marketed in Lima, Peru, were fluoridated, but only 53% contained a TSF concentration greater than 1000 ppm F, the minimum concentration required to provide an anticaries effect33COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESnão te

Colimetry of marine waters off Fortaleza (Ceará State, Brazil) and detection of enteropathogenic Escherichia coli strains

Bacteriological analyses of seawater from three main beaches in Fortaleza, Brazil were performed during 1997. Thirty-six samples per beach were collected for a total of 108 samples. For Meireles Beach, 44% of the samples had MPN total coliforms values of at least 1100 or over 2400/100 ml, followed by Formosa and Diários beaches showing lower counts. For fecal coliforms the highest numbers were demonstrated for Formosa, followed by Meireles and Diários beaches in this descending order: 13.0%, 11.1% and 8.3%, respectively. Escherichia coli strains were identified in 76.8% of the 108 samples. Among 295 strains of E.coli, 21 belonged to serogroups O25, O26, O91, O112, O119, O158 and O164. Strains from serogroup O26 were tested using PCR, ELISA and Vero cells to detect Verotoxins VT1 and VT2 and all strains were negative. No LT and ST, as determined by ELISA and suckling mice assays, were detected among the 295 strains. All strains of E. coli were sensitive to ampicillin, cephalothin, gentamicin, tetracycline, sulfametox-trimethoprim, chloramphenicol and ciprofloxacin. Although the E. coli strains were not toxigenic, their presence in high numbers could be of public health significance

Optical Density Of Bone Repair After Implantation Of Homogenous Demineralized Dentin Matrix In Diabetic Rabbits

This research evaluated the bone repair process after implantation of homogenous demineralized dentin matrix (HDDM) in surgical defects in the parietal bone of rabbits with alloxan-induced diabetes, using a polytetrafluorethylene (PTFe) barrier for guided bone regeneration. Thirty-six rabbits were used and divided into four groups: control (C, n = 12), diabetic (D, n = 12, left parietal bone), diabetic with PTFe (DPTFe, same 12 rabbits, right parietal bone), and diabetic with PTFe associated to HDDM (D-PTFe+HDDM, n = 12). Bone defects were created in the parietal bone of the rabbits and the experimental treatments were performed, where applicable. The rabbits were sacrificed after 15, 30, 60 and 90 days. The bone defects were examined radiographically and by optical density (ANOVA and Tukey test, p < .05). The radiographic findings showed that the D-PTFe+HDDM group presented greater radiopacity and better trabecular bone arrangement when compared to that of the C, D and D-PTFe groups. The statistical analysis showed significant differences in the optical density of the newly formed bone among the studied groups. It was possible to conclude that HDDM was biocompatible in diabetic rabbits.223275280Abreu, P.P., Morosolli, A., Araújo, M.M., Carvalho, V.A.P., Gomes, M.F., Effects of autogenous demineralized dentin matrix on dental socket wound healing process in human (2004) Braz Oral Res, 18, p. 52. , SupplCarvalho, V.A.P., Tosello, D.O., Salgado, M.A.C., Gomes, M.F., Histomorphometric analysis of homogenous demineralized dentin matrix as osteopromotive material in rabbit mandibules (2004) Int J Oral Maxillofac Implants, 19 (5), pp. 679-686Gomes, M.F., Anjos, M.J.S., Nogueira, T.O., Catanzaro-Guimarães, S.A., Autogenous demineralized dentin matrix for tissue engineering applications: Radiographic and histomorphometric studies (2002) Int J Oral Maxillofac Implants, 17 (4), pp. 488-497Gomes, M.F., Silva, M.J.S., Nogueira, T.O., Catanzaro-Guimarães, S.A., Histologic evaluation of the osteoinductive property of autogenous demineralized dentin matrix on surgical bone defects in rabbit skulls using human amniotic membrane for guided bone regeneration (2001) Int J Oral Maxillofac Implants, 16 (4), pp. 563-571Guyton, A.C., Hall, J.E., (2006) Insulin, Glucagon, and Diabetes mellitus, pp. 827-840. , Guyton AC, Hall JE. Textbook of Medical Physiology. 11th ed, Elsevier;Fiorellini, J.P., Nevins, M.L., Norkin, A., Weber, H.P., Karimbux, N.Y., The effect of insulin therapy on osseointegration in a diabetic rat model (1999) Clin Oral Implants Res, 10 (5), pp. 362-368Lu, H., Kraut, D., Gerstenfeld, L.C., Graves, D.T., Diabetes interferes with the bone formation by affecting the expression of transcription factors that regulate osteoblast differentiation (2003) Endocrinology, 144 (1), pp. 346-352Nevins, M.L., Karimbux, N.Y., Weber, H.P., Giannobile, W.V., Fiorellini, J.P., Wound healing around endosseous implants in experimental diabetes (1998) Int J Oral Maxillofac Implants, 13 (5), pp. 620-629Claro, F.A., Lima, J.R., Salgado, M.A.C., Gomes, M.F., Porous Polyethylene for tissue engineering applications in diabetic rats treated with calcitonin: Histomorphometric analysis (2005) Int J Oral Maxillofac Implants, 20 (2), pp. 211-219Catanzaro-Guimarães, S.A., Catanzaro Guimarães, B.P., Garcia, R.B., Alle, N., Osteogenic potential of autogenic demineralized dentin implanted in bony defects in dogs (1986) Int J Oral Maxillofac Surg, 15 (2), pp. 160-169Bessho, K., Tagawa, T., Murata, M., Purification of rabbit bone morphogenetic protein derived from bone, dentin, and wound tissue after tooth extraction (1990) J Oral Maxillofac Surg, 48 (2), pp. 162-169Nakashima, M., Induction of dentine in amputated pulp of dogs by recombinant human bone morphogenetic proteins-2 and -4 with collagen matrix (1994) Arch Oral Biol, 39 (12), pp. 1085-1089Catanzaro-Guimarães SA. Possibility to reinforce bone repair with decalcified dentin matrix. In: Gesellschaft für orale Implantologie (ed.). Jahrbuch für Orale Implatologie. Berlin: Quintessenz1993. p. 33-

Retention and loss of RNA interference pathways in trypanosomatid protozoans

RNA interference (RNAi) pathways are widespread in metaozoans but the genes required show variable occurrence or activity in eukaryotic microbes, including many pathogens. While some Leishmania lack RNAi activity and Argonaute or Dicer genes, we show that Leishmania braziliensis and other species within the Leishmania subgenus Viannia elaborate active RNAi machinery. Strong attenuation of expression from a variety of reporter and endogenous genes was seen. As expected, RNAi knockdowns of the sole Argonaute gene implicated this protein in RNAi. The potential for functional genetics was established by testing RNAi knockdown lines lacking the paraflagellar rod, a key component of the parasite flagellum. This sets the stage for the systematic manipulation of gene expression through RNAi in these predominantly diploid asexual organisms, and may also allow selective RNAi-based chemotherapy. Functional evolutionary surveys of RNAi genes established that RNAi activity was lost after the separation of the Leishmania subgenus Viannia from the remaining Leishmania species, a divergence associated with profound changes in the parasite infectious cycle and virulence. The genus Leishmania therefore offers an accessible system for testing hypothesis about forces that may select for the loss of RNAi during evolution, such as invasion by viruses, changes in genome plasticity mediated by transposable elements and gene amplification (including those mediating drug resistance), and/or alterations in parasite virulence

Aeromonas spp. isoladas de ostras (Crassostrea rhizophorea) coletadas em um criadouro natural, Ceará, Brazil

Between April and October 2002, thirty fortnightly collections of oysters (Crassostrea rhizophorea) from a natural oyster bed at the Cocó River estuary in the Sabiaguaba region (Fortaleza, Ceará, Brazil) were carried out, aiming to isolate Aeromonas spp. strains. Oyster samples were submitted to the direct plating (DP) and the presence/absence (P/A) methods. Aeromonas were identified in 15 (50%) samples analyzed by the DP method and in 13 (43%) analyzed by the P/A method. A. caviae, A. eucrenophila, A. media, A. sobria, A. trota, A. veronii bv. sobria, A. veronii bv. veronii and Aeromonas sp. were isolated. The predominant species was A. veronii (both biovars), which was identified in 13 (43%) samples, followed by A. media in 11 (37%) and A. caviae in seven (23%). From the 59 strains identified, 28 (48%) presented resistance to at least one of the eight antibiotics tested.Foram realizadas 30 coletas quinzenais, entre abril e outubro de 2002, de ostras (Crassostrea rhizophorea) de um criadouro natural, no estuário do rio Cocó (Fortaleza/Ceará/Brasil), objetivando-se isolar cepas de Aeromonas spp. As amostras de ostras foram submetidas aos métodos de plaqueamento direto (PD) e presença/ausência (P/A). Foram identificadas Aeromonas em 15 (50%) amostras analisadas pelo método PD e em 13 (43%) pelo método P/A. Foram isoladas: A. caviae, A. eucrenophila, A. media, A. sobria, A. trota, A. veronii bv. sobria, A. veronii bv. veronii e Aeromonas sp. A espécie predominate foi A. veronii (ambos biovars), identificada em 13 (43%) amostras, seguida de A. media em 11 (37%) e A. caviae em 7 (23%). Das 59 cepas identificadas, 28 (48%) apresentaram resistência a pelo menos um, dos oitos antibióticos testados

An EST-based analysis identifies new genes and reveals distinctive gene expression features of Coffea arabica and Coffea canephora

Background: Coffee is one of the world’s most important crops; it is consumed worldwide and plays a significant role in the economy of producing countries. Coffea arabica and C. canephora are responsible for 70 and 30% of commercial production, respectively. C. arabica is an allotetraploid from a recent hybridization of the diploid species, C. canephora and C. eugenioides. C. arabica has lower genetic diversity and results in a higher quality beverage than C. canephora. Research initiatives have been launched to produce genomic and transcriptomic data about Coffea spp. as a strategy to improve breeding efficiency. Results: Assembling the expressed sequence tags (ESTs) of C. arabica and C. canephora produced by the Brazilian Coffee Genome Project and the Nestlé-Cornell Consortium revealed 32,007 clusters of C. arabica and 16,665 clusters of C. canephora. We detected different GC3 profiles between these species that are related to their genome structure and mating system. BLAST analysis revealed similarities between coffee and grape (Vitis vinifera) genes. Using KA/KS analysis, we identified coffee genes under purifying and positive selection. Protein domain and gene ontology analyses suggested differences between Coffea spp. data, mainly in relation to complex sugar synthases and nucleotide binding proteins. OrthoMCL was used to identify specific and prevalent coffee protein families when compared to five other plant species. Among the interesting families annotated are new cystatins, glycine-rich proteins and RALF-like peptides. Hierarchical clustering was used to independently group C. arabica and C. canephora expression clusters according to expression data extracted from EST libraries, resulting in the identification of differentially expressed genes. Based on these results, we emphasize gene annotation and discuss plant defenses, abiotic stress and cup quality-related functional categories. Conclusion: We present the first comprehensive genome-wide transcript profile study of C. arabica and C. canephora, which can be freely assessed by the scientific community at http://www.lge.ibi.unicamp.br/ coffea. Our data reveal the presence of species-specific/prevalent genes in coffee that may help to explain particular characteristics of these two crops. The identification of differentially expressed transcripts offers a starting point for the correlation between gene expression profiles and Coffea spp. developmental traits, providing valuable insights for coffee breeding and biotechnology, especially concerning sugar metabolism and stress tolerance