Fermentable Fibers Enhance Aspects of Innate and Adaptive Immunity in Piglets infected with Salmonella Typhimurium.

OBJECTIVE: To test the hypothesis that fermentable fiber prevents Salmonella typhimurium infection-associated symptoms by enhancing innate and adaptive immune system in neonatal pigs. METHODS: Two-d-old piglets (n=120) were randomized to receive either a nutritionally complete sow milk replacer formula (CON), or supplemented with methylcellulose (MCEL-non-fermentable), soy polysaccharides (SPS-moderately fermentable), or fructooligosaccharides (FOS-highly fermentable). On d7, piglets received an oral gavage of S. typhimurium-798, and continued receiving the same diets up to 48h post-infection. Ileal mucosal samples were obtained for further analyses. RESULTS: A reduction in chloride secretion was observed in FOS when compared to other diets (p<0.0003). The number of ileal sulfo-acidomucins was higher (p<0.05) in FOS before infection compared with other diets. NFkB was inhibited in FOS following infection (p<0.05), when compared with CON. IL-1ß expression was increased at 4h post-infection (p<0.05) in CON; however, this response was attenuated in the fiber groups. IL-6 expression was higher (p<0.05) in CON post- infection, higher in SPS at 24h (p<0.05), but unchanged in MCEL and FOS when compared to pre-infection values. FOS had a higher expression of neutrophil-chemoattractant IL-8 before infection (p<0.05) compared to other groups. CONCLUSION: The reduction in chloride secretion, proinflammatory cytokines expression and NFkB activation, and increased number of sulfo-acidomucins, and IL-8 expression in the fiber groups, indicates that the degree of fermentability impacts the innate and adaptive immune system, and could be the mechanisms by which dietary fibers reduce S. typhimurium infection-associated-symptoms in neonatal pigs and apply these results to infants.

Physical properties and biological effects of mineral trioxide aggregate mixed with methylcellulose and calcium chloride.

OBJECTIVES: Methylcellulose (MC) is a chemical compound derived from cellulose. MTA mixed with MC reduces setting time and increases plasticity. This study assessed the influence of MC as an anti-washout ingredient and CaCl2 as a setting time accelerator on the physical and biological properties of MTA. MATERIAL AND METHODS: Test materials were divided into 3 groups; Group 1(control): distilled water; Group 2: 1% MC/CaCl2; Group 3: 2% MC/CaCl2. Compressive strength, pH, flowability and cell viability were tested. The gene expression of bone sialoprotein (BSP) was detected by RT-PCR and real- time PCR. The expression of alkaline phosphatase (ALP) and mineralization behavior were evaluated using an ALP staining and an alizarin red staining. RESULTS: Compressive strength, pH, and cell viability of MTA mixed with MC/CaCl2 were not significantly different compared to the control group. The flowability of MTA with MC/CaCI2 has decreased significantly when compared to the control (p<.05). The mRNA level of BSP has increased significantly in MTA with MC/CaCl2 compared to the control (p<.05). This study revealed higher expression of ALP and mineralization in cells exposed to MTA mixed with water and MTA mixed with MC/CaCl2 compared to the control (p<.05). CONCLUSIONS: MC decreased the flowability of MTA and did not interrupt the physical and biological effect of MTA. It suggests that these cements may be useful as a root-end filling material.

Physical properties and biological effects of mineral trioxide aggregate mixed with methylcellulose and calcium chloride

Abstract Objectives: Methylcellulose (MC) is a chemical compound derived from cellulose. MTA mixed with MC reduces setting time and increases plasticity. This study assessed the influence of MC as an anti-washout ingredient and CaCl2 as a setting time accelerator on the physical and biological properties of MTA. Material and Methods: Test materials were divided into 3 groups; Group 1(control): distilled water; Group 2: 1% MC/CaCl2; Group 3: 2% MC/CaCl2. Compressive strength, pH, flowability and cell viability were tested. The gene expression of bone sialoprotein (BSP) was detected by RT-PCR and real­ time PCR. The expression of alkaline phosphatase (ALP) and mineralization behavior were evaluated using an ALP staining and an alizarin red staining. Results: Compressive strength, pH, and cell viability of MTA mixed with MC/CaCl2 were not significantly different compared to the control group. The flowability of MTA with MC/CaCI2 has decreased significantly when compared to the control (p<.05). The mRNA level of BSP has increased significantly in MTA with MC/CaCl2 compared to the control (p<.05). This study revealed higher expression of ALP and mineralization in cells exposed to MTA mixed with water and MTA mixed with MC/CaCl2 compared to the control (p<.05). Conclusions: MC decreased the flowability of MTA and did not interrupt the physical and biological effect of MTA. It suggests that these cements may be useful as a root-end filling material.

Avaliação da eficácia de um novo marcador cirúrgico para capsulorrexe anterior / Effectiveness evaluation of a new surgical marker for anterior capsulorhexis

RESUMO Objetivo: Avaliar a eficácia de um novo marcador cirúrgico para ajudar na confecção da capsulorrexe anterior analisando o seu dimensionamento e formato, comparando com a capsulorrexe confeccionada manualmente de maneira livre. Métodos: Como experimento, 3 residentes (R3) de Oftalmologia do Hospital Universitário Onofre Lopes (HUOL) e 1 oftalmologista em treinamento, voluntários, realizaram (cada um) 10 capsulorrexes em olhos de porco enucleados. Em 5 olhos foi utilizado o marcador e em outros 5, não. Todos os olhos foram fotografados tendo ao lado uma régua para orientar e calibrar um aplicativo para a avaliação morfométrica do procedimento. O diâmetro alvo foi de 5 mm, cujo perímetro correspondente é 15,7 mm e a área 19,652 mm2. Foram avaliados em cada procedimento: os diâmetros máximo, mínimo e médio, o perímetro, a área e o desvio em relação ao diâmetro e quanto ao aspecto ideal. Resultados: No grupo utilizando o marcador o diâmetro médio foi 5,44mm (±0,89) contra 6,37mm (±0,67) (p=0,001), no grupo no qual não se utilizou o marcador; quanto ao perímetro, 17,52mm (±1,92) no grupo utilizando o marcador contra 20,14mm (±2,09) (p<0,001) sem o marcador e quanto a área, 24,73mm2 (±1,92) com o marcador, contra 32,62mm2 (±6,32) (p<0,001), sem o marcador. Em relação ao aspecto da capsulorrexe 1,26mm (±0,12), contra 1,21mm (±0,7) (p=0,09) e em relação ao desvio de curvatura: 0,87 (±0,05), contra 0,9 (±0,04), (p=0,06) respectivamente. Conclusão: O trabalho mostrou que o marcador avaliado é eficaz para auxiliar a confecção da capsulorrexe conduzindo a resultados melhores que o método a mão livre.

ABSTRACT Purpose: To evaluate the effectiveness of a surgical device that intented to help in the preparation of the anterior capsulorhexis analyzing the design and shape, comparing with capsulorhexis made by free hand. Methods: Three ophthalmology residents(R3) at the HUOL and one surgeon in training, participate in this research as volunteers. Each surgeon perform 5 capsulorhexis in porcine eyes using the device, and five others by free hand as a control. All capsulorhexis were photographed having a ruler as reference to guide and calibrate a computer application for morphometric evaluation (Cambuí Labs, Natal, Brazil). All surgeons aimed to produce a circular continuous capsulorhexis of 5 mm diameter that represents 15,7mm in perimeter and 19,652mm2 in area. Each wet-lab capsulorhexis was evaluated in regard to these criteria: diameter (mean, maximum and minimum), perimeter, area, deviation from the ideal diameter and ideal shape. Results: Compare to control groups, capsulorhexis with the aid of the surgical device showed: 5,44mm ±0,89 vs 6,37mm ±0,67, for capsulorhexis diameter (p=0,001); 17,52mm ±1,92 vs 20,14mm ±2,09 for capsulorhexis perimeter (<0.001); 24,73mm2 ±1,92 vs 32,62mm2 ±6,36 for capsulorhexis area (p<0,001). A tendency for better result with the aid of the surgical device was observed for deviation of an ideal diameter or ideal aspect were appreciated: 0,87mm ±0,05 vs 0,9 ±0,04 for deviation of a curve (p=0,06); 1,26mm ±0,12 vs 1,21mm ±0,7 for the capsulorhexis aspect (p=0,09). Conclusion: Capsulorhexis produced with the aid of the surgical device, significantly improved wet-lab capsulorhexis performance.

Characterization of putative haematopoietic cells from bovine yolk sac.

The yolk sac is an extra-embryonic membrane that plays an important role in early embryonic survival. It is the production site for blood cells during embryonic mammalian development and is a likely source of stem cells. The aim of this study was to identify and characterize the putative haematopoietic cells from the yolk sac of bovine embryos at different stages of gestation. The yolk sac regresses according to gestational age and embryos are characterized into groups (I-V) according to the crown-rump measurement. Groups I-III survived in culture longer and exhibited the formation of cell clusters, whereas groups IV and V could not be maintained in culture for an extended period of time. Flow-cytometry analysis revealed that groups I-III had similar characteristics, including high expression levels of the haematopoietic markers CD34, CD90 and CD117. In groups IV and V, decreases were observed in the expression levels of CD117 and CD34. Cells were found to be capable of survival post-cryopreservation and exhibited varying abilities to form colonies in a methylcellulose matrix, depending on gestational age. Cytological analysis revealed the presence of blood cells (lymphocytes and monocytes). Quantitative PCR analysis demonstrated the presence of the haematopoietic progenitor genes GATA3 and LMO2, but not RUNX1. Thus, we have successfully isolated and characterized haematopoietic cells from the bovine embryo yolk sac at varying gestational ages. This study is crucial for the understanding of the development of the haematopoietic system and the embryonic function of this organ. Copyright © 2015 John Wiley & Sons, Ltd.

Spatio-temporal morphology changes in and quenching effects on the 2D spreading dynamics of cell colonies in both plain and methylcellulose-containing culture media.

To deal with complex systems, microscopic and global approaches become of particular interest. Our previous results from the dynamics of large cell colonies indicated that their 2D front roughness dynamics is compatible with the standard Kardar-Parisi-Zhang (KPZ) or the quenched KPZ equations either in plain or methylcellulose (MC)-containing gel culture media, respectively. In both cases, the influence of a non-uniform distribution of the colony constituents was significant. These results encouraged us to investigate the overall dynamics of those systems considering the morphology and size, the duplication rate, and the motility of single cells. For this purpose, colonies with different cell populations (N) exhibiting quasi-circular and quasi-linear growth fronts in plain and MC-containing culture media are investigated. For small N, the average radial front velocity and its change with time depend on MC concentration. MC in the medium interferes with cell mitosis, contributes to the local enlargement of cells, and increases the distribution of spatio-temporal cell density heterogeneities. Colony spreading in MC-containing media proceeds under two main quenching effects, I and II; the former mainly depending on the culture medium composition and structure and the latter caused by the distribution of enlarged local cell domains. For large N, colony spreading occurs at constant velocity. The characteristics of cell motility, assessed by measuring their trajectories and the corresponding velocity field, reflect the effect of enlarged, slow-moving cells and the structure of the medium. Local average cell size distribution and individual cell motility data from plain and MC-containing media are qualitatively consistent with the predictions of both the extended cellular Potts models and the observed transition of the front roughness dynamics from a standard KPZ to a quenched KPZ. In this case, quenching effects I and II cooperate and give rise to the quenched-KPZ equation. Seemingly, these results show a possible way of linking the cellular Potts models and the 2D colony front roughness dynamics.

Safety and feasibility of the use of a bevacizumab-methylcellulose mixture as an adjunct to glaucoma surgery: a pilot study.

Bevacizumab, a monoclonal anti-vascular endothelial growth factor antibody, has been suggested as a potential healing therapeutic following glaucoma surgery. Here, we aimed to improve the bioavailability of bevacizumab when used as an adjunct therapy to non-penetrating deep sclerectomy (DS) by using a bevacizumab-methylcellulose mixture (BMM). Ten previously non-operated eyes in ten patients diagnosed with primary open angle glaucoma underwent DS with a subconjunctival injection of 0.3 ml of BMM (bevacizumab 3.75 mg incorporated into 4% methylcellulose) at the surgical site. Bevacizumab release was evaluated in vitro using size-exclusion high performance liquid chromatography (HPLC). Intraocular pressure (IOP), bleb morphology, corneal endothelial cell count (CECC), and complications were evaluated at 6 months after surgery. Using HPLC, bevacizumab was detected in BMM for up to 72 h. Moreover, all surgical blebs remained expanded with hyaline material during the first week. A significant IOP reduction (mean ± SD= -10.3 ± 5.4 mmHg, P<0.001) and diffuse blebs were observed at the final follow-up period. Although CECC was slightly reduced (-7.4%), no complications were observed. In conclusion, bevacizumab was released from BMM, and the use of this innovative mixture yielded good results following DS with no complications. Further studies are required to determine its efficacy prior to establishing BMM as an adjunct treatment for penetrating and non-penetrating glaucoma surgeries.

Safety and feasibility of the use of a bevacizumab-methylcellulose mixture as an adjunct to glaucoma surgery: a pilot study / Segurança e viabilidade do uso de uma mistura de bevacizumabe-metilcelulose como terapia adjuvante à cirurgia anti-glaucomatosa: um estudo piloto

ABSTRACT Bevacizumab, a monoclonal anti-vascular endothelial growth factor antibody, has been suggested as a potential healing therapeutic following glaucoma surgery. Here, we aimed to improve the bioavailability of bevacizumab when used as an adjunct therapy to non-penetrating deep sclerectomy (DS) by using a bevacizumab-methylcellulose mixture (BMM). Ten previously non-operated eyes in ten patients diagnosed with primary open angle glaucoma underwent DS with a subconjunctival injection of 0.3 ml of BMM (bevacizumab 3.75 mg incorporated into 4% methylcellulose) at the surgical site. Bevacizumab release was evaluated in vitro using size-exclusion high performance liquid chromatography (HPLC). Intraocular pressure (IOP), bleb morphology, corneal endothelial cell count (CECC), and complications were evaluated at 6 months after surgery. Using HPLC, bevacizumab was detected in BMM for up to 72 h. Moreover, all surgical blebs remained expanded with hyaline material during the first week. A significant IOP reduction (mean ± SD= -10.3 ± 5.4 mmHg, P<0.001) and diffuse blebs were observed at the final follow-up period. Although CECC was slightly reduced (-7.4%), no complications were observed. In conclusion, bevacizumab was released from BMM, and the use of this innovative mixture yielded good results following DS with no complications. Further studies are required to determine its efficacy prior to establishing BMM as an adjunct treatment for penetrating and non-penetrating glaucoma surgeries.

RESUMO O bevacizumabe (um agente anti-fator de crescimento endotelial vascular) tem sido sugerido como potencial modulador cicatricial na cirurgia do glaucoma. Este estudo objetivou melhorar a biodisponibilidade do bevacizumabe, investigando a viabilidade de uma nova mistura de bevacizumabe-metilcelulose (BMM) como terapia adjuvante para a esclerectomia profunda não-penetrante (DS). Dez olhos sem cirurgias prévias de 10 pacientes com glaucoma primário de ângulo aberto foram submetidos à DS associada à uma injeção subconjuntival de 0,3 ml da mistura de bevacizumabe-metilcelulose (bevacizumabe 3,75 mg incorporado em metilcelulose 4%) no sítio cirúrgico. A liberação de bevacizumabe foi avaliada in vitro através de cromatografia líquida de alta performance por exclusão de tamanho (HPLC). A pressão intraocular (PIO), a morfologia da ampola de filtração, a contagem de células endoteliais da córnea (CECC) e as complicações foram estudadas aos seis meses de seguimento. O bevacizumabe foi detectado a partir da mistura de bevacizumabe-metilcelulose por meio do HPLC até 72 horas. Além disso, todas as ampolas cirúrgicas permaneceram expandidas com material hialino durante a primeira semana. Uma redução significativa da pressão intraocular (média ± DP= -10,3 ± 5,4 mmHg, P<0,001) e ampolas difusas foram observadas ao final do período de seguimento. Embora a contagem de células endoteliais da córnea se mostrou discretamente diminuída (-7,4%), nenhuma complicação foi observada. Neste estudo, o bevacizumabe foi liberado da mistura de bevacizumabe-metilcelulose e o uso desta nova mistura se associou com bons resultados cirúrgicos e nenhuma complicação. Estudos futuros serão necessários para determinar sua eficácia, antes de se estabelecer a mistura de bevacizumabe-metilcelulose como um tratamento adjuvante às cirurgias penetrantes e não-penetrantes para o glaucoma.

Biopolymeric antimicrobial films: study of the influence of hydroxypropyl methylcellulose, tapioca starch and glycerol contents on physical properties.

Mixture design methodology was applied to study the effect of different levels of tapioca starch (TS), hydroxypropyl methylcelullose (HPMC), and glycerol (Gly) on the physical properties of biopolymeric films supporting potassium sorbate (KS; 0.3% w/w) with the goal of contributing to the development of materials for preventing food surface contamination. Mechanical properties, water vapour permeability (WVP), solubility in water (S) and colour attributes were evaluated on the films. HPMC addition produced an increase of elastic modulus (Ec), stress at break (σb) and S. It also decreased the yellow index (YI) values and the strain at break (εb). The study was deepened using the formulation containing 2.67 g/100g of TS, 0.67 g/100g of HPMC, 1.67 g/100g Gly and 0.3g/100g KS, observing that it behaved as an effective antimicrobial barrier against Zygosaccharomyces bailii external contamination. Microstructural analysis allowed us to conclude that HPMC incorporation to a TS network decreased roughness of the films and it also increased permeability to oxygen (PO2).

Physical properties of emulsion-based hydroxypropyl methylcellulose films: effect of their microstructure.

The initial characteristics of emulsions and the rearrangement of the oil droplets in the film matrix during film drying, which defines its microstructure, has an important role in the physical properties of the emulsion-based films. The objective of this work was to study the effect of the microstructure (two droplet size distributions) and stability (with or without surfactant) of HPMC oil-in-water emulsions over physical properties of HPMC emulsion-based edible films. HPMC was used to prepare sunflower oil-in-water emulsions containing 0.3 or 1.0% (w/w) of oil with or without SDS, as surfactant, using an ultrasonic homogenizer. Microstructure, rheological properties and stability of emulsions (creaming) were measured. In addition, microstructure, coalescence of oil droplets, surface free energy, optical and mechanical properties and water vapor transfer of HPMC films were evaluated. Image analysis did not show differences among droplet size distributions of emulsions prepared at different oil contents; however, by using SDS the droplet size distributions were shifted to lower values. Volume mean diameters were 3.79 and 3.77 µm for emulsions containing 0.3 and 1.0% without surfactant, respectively, and 2.72 and 2.71 µm for emulsions with SDS. Emulsions formulated with 1.0% of oil presented higher stability, with almost no change during 5 and 3 days of storage, for emulsions with and without SDS, respectively. Internal and surface microstructure of emulsion-based films was influenced by the degree of coalescence and creaming of the oil droplets. No effect of microstructure over the surface free energy of films was found. The incorporation of oil impaired the optical properties of films due to light scattering of light. Addition of oil and SDS decreased the stress at break of the emulsion-based films. The replace of HPMC by oil and SDS produce a lower "amount" of network structure in the films, leading to a weakening of their structure. The oil content and SDS addition had an effect over the microstructure and physical properties of HPMC-based emulsions which lead to different microstructures during film formation. The way that oil droplets were structured into the film had an enormous influence over the physical properties of HPMC films.

Comparison of enteric-coated capsules and liquid formulation of Ty21a typhoid vaccine in randomised controlled field trial.

In a randomised, double-blind, controlled field trial in Santiago, Chile, 81,621 schoolchildren aged 5-19 years received three doses, within a week, of attenuated Salmonella typhi oral vaccine Ty21a in enteric-coated capsules or in a new liquid suspension, or placebo. Over 36 months of surveillance, the liquid formulation (76.9% vaccine efficacy) was significantly superior to the enteric-coated capsules (33.2% vaccine efficacy). The liquid formulation protected young children (5-9 years) (efficacy 82.3%) as well as older children (efficacy 69.3%), whereas the capsules significantly protected only older children. The liquid suspension was easy to prepare by mixing lyophilised vaccine with buffer in water and was easily administered, even to the youngest children. Thus, the liquid formulation of Ty21a is preferable to enteric-coated capsules.

Açao da metilcelulose a 2 por cento na câmara anterior do olho do coelho: estudo biomicroscópico, paquimétrico, tonométrico, citológico e histológico / Action of methylcelulose a 2 per cent in the anterior chamber of the eye of rabbit: study biomicroscopic, pachimetric, cytologic and histologic

Utilizaram-se 45 coelhos divididos em dois grupos: o Grupo Po (22 coelhos) e o Grupo Pk (23 coelhos). Todos sofreram substituição parcial do humor aquoso por soluções de Metilcelulose a 2 por cento em solução salina balanceada (SSB) em um olho, e SSB no olho contralateral. No grupo Po foram estudados os comportamentos do diâmetro pupilar, biomicroscópico e tonométrico, antes e depois (2, 4, 8h e 1§, 2§, 3§, 4§, 5§, 10§ e 20§ dia) das injeções das soluções na câmara anterior. No Grupo Pk foi observado o comportamento da espessura corneana antes e depois (4 e 18h) da injeção das soluções na câmara anterior, sendo em seguida realizada a histologia plana do endotélio corneano para avaliação do padrão e densidade celular. Foi colhido humor aquoso de 10 pares de olhos desse grupo para coloração e estudo. Ainda foram selecionados alguns olhos do grupo teste e controle para estudo histológico. Os resultados mostraram que nem a Metilcelulose nem a solução salina balanceada influenciaram no diâmetro pupilar. A pressão ocular dos olhos injetados com Metilcelulose aumentou em média 13,68 mmHg em relação ao olho-controle, porém com grande variabilidade de valores. O pico de pressão ocorreu às 2h, retornando aos níveis basais em 24 h. A biomicroscopia, a citologia e a histologia mostraram que a substância com um maior número de células no humor aquoso, além de uma irite localizada em 36,3 por cento dos casos. A Metilcelulose tende a se condensar e distribuir-se em diferentes locais da câmara anterior antes de desaparecer. Não foi evidenciada nenhuma influência sobre o endotélio corneano. O autor conclui que a Metilcelulose a 2 por cento pode causar alterações tipo inflamatórias de curta duração na câmara anterior do olho de coelho, sem deixar sequela ou dano aparente às estruturas intra-oculares.