Uptake of antigen encapsulated in polyethylcyanoacrylate nanoparticles by D1-dendritic cells

Polyethylcyanoacrylate (PECA) nanoparticles were prepared by interfacial polymerization of a water-in-oil microemulsion. Nanoparticles were isolated from the polymerization template by sequential ethanol washing and centrifugation. A nanocapsule preparation yielding the original particle size and distribution following redispersion in an aqueous solution was achieved by freeze-drying the isolated nanoparticles in a solution of 5% w/v sugar. The cytotoxicity and uptake of nanocapsules by dendritic cells was investigated using a murine-derived cell line (D1). PECA nanoparticles were found to adversely effect cell viability at concentrations greater than 10 mug/ml of polymer in the culture medium. In comparison to antigen in solution, cell uptake of antigen encapsulated within nanoparticles was significantly higher at both 4 and 37 degreesC. Following a 24 h incubation period, the percentage of cells taking-up antigen was also increased when antigen was encapsulated in nanoparticles as compared to antigen in solution. The uptake of nanoparticles and the effect of antigen formulation on morphological cell changes indicative of cell maturation were also investigated by scanning electron microscopy (SEM). SEM clearly demonstrated the adherence of nanoparticles to the cell surface. Incubation of D1 dendritic cells with nanoparticles containing antigen also resulted in morphological changes indicative of cell maturation similar to that observed when the cells were incubated with lipopolysaccharide. In contrast, cells incubated with antigen solution did not demonstrate such morphological changes and appeared similar to immature cells that had not been exposed to antigen.

Extrusion of mixtures of starch and D-limonene encapsulated with beta-cyclodextrin: Flavour retention and physical properties

d-Limonene was encapsulated with beta-cyclodextrin to improve its retention during pre-added flavour starch extrusion. The objective of this work was to determine the effect of processing condition on the flavour retention and extrudate properties. Corn starch containing five levels of beta-cyclodextrin-d-limonene capsules (0-5%) were extruded at five different maximum barrel temperatures (133-167 degrees C) and screw speeds (158-242 rpm) using a twin screw extruder. The effect of these parameters on the flavour retention, expansion, texture, colour difference (Delta E), Water Absorption Index, Water Solubility Index, and residence time distribution (RTD) were investigated. Barrel temperature and capsule level predominantly influenced flavour retention and extrudate properties, while screw speed primarily affected extruder performances such as torque, die pressure, specific mechanical energy and RTD. (c) 2005 Elsevier Ltd. All rights reserved.

Synthesis and spectroscopic observation of dendrimer-encapsulated gold nanoclusters

We report on the observation of the excitation/ emission spectrum of a dendrimer-encapsulated gold nanocluster; the synthesis of Au-PAMAM was based on reduction of HAuCl4 center dot 3H(2)O co-dissolved in methanol together with fourth-generation OH-terminated PAMAM.

Survival of probiotics encapsulated in chitosan-coated alginate beads in yoghurt from UHT- and conventionally treated milk during storage

Survival of the microencapsulated probiotics, Lactobacillus acidophilus 547, Bifidobacterium bifidum ATCC 1994, and Lactobacillus casei 01, in stirred yoghurt from UHT- and conventionally treated milk during low temperature storage was investigated. The probiotic cells both as free cells and microencapsulated cells (in alginate beads coated with chitosan) were added into 20 g/100 g total solids stirred yoghurt from UHT-treated milk and 16 g/100 g total solids yoghurt from conventionally treated milk after 3.5 h of fermentation. The products were kept at 4 degrees C for 4 weeks. The survival of encapsulated probiotic bacteria was higher than free cells by approximately 1 log cycle. The number of probiotic bacteria was maintained above the recommended therapeutic minimum (10(7) cfu g(-1)) throughout the storage except for R bifidum. The viabilities of probiotic bacteria in yoghurts from both UHT- and conventionally treated milks were not significantly (P > 0.05) different. (c) 2004 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.

Effect of extrusion parameters on flavour retention, functional and physical properties of mixtures of starch and D-limonene encapsulated in milk protein

The purpose of this research was to investigate the retention of flavour volatiles encapsulated in water-insoluble systems during high temperature–short time extrusion process. A protein precipitation method was used to produce water-insoluble capsules encapsulating limonene, and the capsules were added to the extruder feed material (cornstarch). A twin-screw extruder was used to evaluate the effect of capsule level of addition (0–5%), barrel temperature (125–145 °C) and screw speed (145–175 r.p.m.) on extruder parameters (torque, die pressure, specific mechanical energy, residence time distribution) and extrudate properties [flavour retention, texture, colour, density, expansion, water absorption index, water solubility index (WSI)]. Capsule level had a significant effect on extrusion conditions, flavour retention and extrudate physical properties. Flavour retention increased with the increase in capsule level from 0% to 2.5%, reached a maximum value at capsule level of 2.5% and decreased when the capsule level increased from 2.5% to 5%. The die pressure, torque, expansion ratio, hardness and WSI exhibited the opposite effect with the presence of capsules.

Retinal and lenticular ultrastructure in the aestivating salamanderfish, Lepidogalaxias salamandroides (Galaxiidae, Teleostei) with special reference to a new type of photoreceptor mosaic

The salamanderfish, Lepidogalaxias salamandroides (Galaxiidae, Teleostei) is endemic to southwestern Australia and inhabits shallow, freshwater pools which evaporate during the hot summer months. Burrowing into the substrate in response to falling water levels allows these fish to aestivate for extended periods of time while encapsulated in a mucous cocoon even when the pools contain no water. Only a few minutes after a major rainfall, these fish emerge into relatively clear water which subsequently becomes laden with tannin, turning the water black and reducing the pH to approximately 4.3. As part of a large study of the visual adaptations of this unique species, the retinal and lenticular morphology of the aestivating salamanderfish is examined at the level of the light and electron microscopes. The inner retina is highly vascularised by a complex system of vitreal blood vessels, while the outer retina receives a blood supply by diffusion from a choriocapillaris. This increased retinal blood supply may be an adaptation for reducing the oxygen tension during critical periods of aestivation. Large numbers of Muller cells traverse the thickness of the retina from the inner to the outer limiting membranes. The ganglion cells are arranged in two ill-defined layers, separated from a thick inner nuclear layer containing two layers of horizontal cells by a soma-free inner plexiform layer. The photoreceptors can be divided into three types typical of many early actinopterygian representatives; equal double cones, small single cones and large rods (2:1:1). These photoreceptors are arranged into a unique regular square mosaic comprising a large rod bordered by four equal double cones with a small single cone located at the corner of each repeating unit. The double cones may optimise perception of mobile prey which it tracks by flexion of its head and neck and the large rods may increase sensitivity in the dark tannin-rich waters in which it lives. Each single cone also possesses a dense collection of polysomes and glycogen (a paraboloid) beneath its ellipsoid, the first such finding in teleosts. The retinal pigment epithelium possesses melanosomes, pha,oocytes and a large number of mitochondria. The anatomy of the retina and the photoreceptor mosaic is discussed in relation to the primitive phylogeny of this species and its unique life history.

Embracing ligands. A synthetic strategy towards new nitrogen-thioether multidentate ligands and characterization of the cobalt(III) complexes

The synthesis of the hexadentate ligand 2,2,9,9-tetra(methyleneamine)-4,7-dithiadecane (EtN(4)S(2)amp) is reported. The ligand is of a type in which bifurcations of the chain occur at atoms other than donor atoms. The cobalt(III) complex [Co(EtN(4)S(2)amp)](3+) (1) was isolated and characterized. The synthetic methodology also results in a number of by-products, notably 2,9,9-tris(methyleneamine)-9-methylenehydroxy-4,7-dithiadecane (Et(HO)N(3)S(2)amp) and an eleven-membered pendant arm macrocyclic ligand 6,10-dimethyl-6,10-bis(methyleneamine)-1,4-dithia-8-azaacycloundec-7- ene (dmatue). The complexes [Co(Et(HO)N(3)S(2)amp)](3+) (2), in which the alcohol is coordinated to the metal ion, and [Co(dmatue)Cl](2+) (4) were isolated and characterized. Et(HO)N(3)S(2)amp also undergoes complexation with cobalt(III) to produce two isomers endo-[Co(Et(HO) N(3)S(2)amp)Cl](2+) (endo-3) and exo-[Co(Et(HO) N(3)S(2)amp)Cl](2+) (exo-3), both with an uncoordinated alcohol group. endo- 3 has the alcohol positioned cis, and exo-3 trans, to the sixth metal coordination site. Reaction of 1 with isobutyraldehyde, paraformaldehyde and base in dimethylformamide results in the encapsulated complex [Co(1,5,5,9,13,13-hexamethyl-18,21-dithia-3,7,11,15-tetraazabicyclo[7.7.6]docosa- 3,14-diene)](ClO4)(3) . 2H(2)O ([Co(Me(6)docosadieneN(4)S(2))](3+) ( 5). All complexes have been characterized by single crystal X-ray study. The low-temperature (11 K) absorption spectrum of 1 has been measured in Nafion films with spin-allowed (1)A(1g) --> T-1(1g) and (1)A(1g) --> T-1(2g) and spin forbidden (1)A(1g) --> T-3(1g) and (1)A(1g) --> T-3(2g) bands observed. The octahedral ligand-field parameters were determined (10Dq = 22570 cm(-1), B = 551 cm(-1); C = 3500 cm(-1)). For 5 10Dq and B were determined (20580 cm(-1); 516 cm(-1), respectively) and compared with those for similar expanded cavity complexes [Co(Me(8)tricosatrieneN(6))](3+) and [Co(Me(5)tricosatrieneN(6))](3+).

Why fear snakes and spiders? Evidence for a learning-based account of fear-relevant stimulus-processing

Fear-relevant stimuli, such as snakes, spiders and heights, preferentially capture attention as compared to nonfear-relevant stimuli. This is said to reflect an encapsulated mechanism whereby attention is captured by the simple perceptual features of stimuli that have evolutionary significance. Research, using pictures of snakes and spiders, has found some support for this account; however, participants may have had prior fear of snakes and spiders that influenced results. The current research compared responses of snake and spider experts who had little fear of snakes and spiders, and control participants across a series of affective priming and visual search tasks. Experts discriminated between dangerous and nondangerous snakes and spiders, and expert responses to pictures of nondangerous snakes and spiders differed from those of control participants. The current results dispute that stimulus fear relevance is based purely on perceptual features, and provides support for the role of learning and experience.

Occurrence and histopathogenesis of a didymozoid trematode (Gonapodasmius epinepheli) in pond-reared orange-spotted grouper, Epinephelus coioides

A didymozoid trematode encapsulated in the gills of orange-spotted grouper, Epinephelus coioides Hamilton, was observed in October 1997 and September 1999 among pond-reared fish in the Philippines. Capsule prevalence was 33% and 18% and mean intensity 2 and 1, respectively. The opaque-white and yellowish capsules were found only on the first gill arch and were attached lengthwise along the posterior surface of the primary gill filaments. When the capsules were opened, long thread-like worms were revealed, which were identified as Gonapodasmius epinepheli Abdul-Salam, Sreelatha and Farah. The parasites were encapsulated between the basement membrane of the epithelium and the efferent artery of the gill filament. The response of the host included mild hyperplasia of the interlamellar epithelium and an increase in the number of mucous cells. (C) 2001 Elsevier Science B.V. All rights reserved.

Role of myofibroblasts in tumour encapsulation of hepatocellular carcinoma in haemochromatosis

Background/Aims: Hepatocellular carcinoma is a carcinoma malignancy and a major complication of untreated haemochromatosis. Encapsulation of liver tumours has been associated with a better prognosis and longer disease-free periods following resection, This study investigated the source of the tumour capsule in patients with haemochromatosis and coexisting hepatocellular carcinoma and examined potential factors influencing development. Methods: Five haemochromatosis patients with encapsulated hepatocellular carcinoma were studied. Myofibroblasts were identified using combined immunohistochemistry and in situ hybridisation for a-smooth muscle actin and procollagen alpha (1)(I) mRNA, respectively. Immunohistochemistry was also performed for transforming growth factor (TGF)-beta (1), platelet-derived growth factor (PDGF)-beta receptor and malondialdehyde. Results. Procollagen alpha (1)(I) mRNA co-localised to alpha -smooth muscle actin positive myofibroblasts. The number of myofibroblasts was maximal within the capsule and decreased away from the tumour. TGF-beta (1) protein was expressed in iron-loaded cells in non-tumour liver at the interface of tumour capsule. PDGF-beta receptor expression was observed in mesenchymal cells in the tumour capsule and in portal tracts. Malondialdehyde adducts were observed in the tumour, non-tumour tissue and in the capsule. Conclusions: This study provides evidence that myofibroblasts are the cell type responsible for collagen production within the tumour capsule surrounding hepatocellular carcinoma in haemochromatosis, The production of TGF-beta (1) by iron-loaded hepatic cells at the tumour capsule interface may perpetuate the myofibroblastic phenotype, resulting in, the formation of the tumour capsule.

Flavour retention during high temperature short time extrusion cooking process: a review

Research on the stability of flavours during high temperature extrusion cooking is reviewed. The important factors that affect flavour and aroma retention during the process of extrusion are illustrated. A substantial number of flavour volatiles which are incorporated prior to extrusion are normally lost during expansion, this is because of steam distillation. Therefore, a general practice has been to introduce a flavour mix after the extrusion process. This extra operation requires a binding agent (normally oil), and may also result in a non-uniform distribution of the flavour and low oxidative stability of the flavours exposed on the surface. Therefore, the importance of encapsulated flavours, particularly the beta -cyclodextrin-flavour complex, is highlighted in this paper.

Ultrastructure of Hepatozoon boigae (Mackerras, 1961) nov comb. from brown tree snakes, Boiga irregularis, from northern Australia

Intraerythrocytic bodies identified as haemogregarine gamonts were found in 29% of 97 brown tree snakes (Boiga irregularis) examined during a haematological survey of reptiles in Australasia during 1994-1998. The morphological characteristics of the parasites were consistent with those of Haemogregarina boigae Mackerras, 1961, although the gamonts were slightly larger and lacked red caps but contained distinctive polar grey capsules. Gamonts did not distend host cells but laterally displaced their nuclei. They were contained within parasitophorous vacuoles and possessed typical apicomplexan organelles, including a conoid, polar rings, rhoptries and micronemes. Schizonts producing up to 30 merozoites were detected in endothelial cells of the lungs of 11 snakes. The absence of erythrocytic schizogony suggests the parasites belong to the genus Hepatozoon. Electron microscopy also revealed the presence of curious encapsulated organisms in degenerating erythrocytes. These stages did not possess apical complex organelles and were surrounded by thick walls containing circumferential junctions and interposed strips reminiscent of oocyst sutures.

Fusion of interleukin-2 to subunit antigens increase their antigenicity in vitro due to an interleukin-2 receptor beta-mediated antigen uptake mechanism

Subunit vaccines, based on one or more epitopes, offer advantages over whole vaccines in terms of safety but are less antigenic. We investigated whether fusion of the cytokine interleukin-2 (IL-2) to influenza-derived subunit antigens could increase their antigenicity. The fusion of IL-2 to the subunit antigens increased their antigenicity in vitro. Encapsulation of the subunit antigen in liposomes also increased its antigenicity in vitro, yet encapsulation of the subunit IL-2 fusion did not. The use of anti-IL-2 receptor beta (IL-2Rbeta) antibody to block the receptor subunit on macrophages suggested that the adjuvancy exerted by IL-2 in our in vitro system is due to, at least in part, a previously unreported IL-2Rbeta-mediated antigen uptake mechanism.

Tumor progression in hepatocellular carcinoma: Relationship with tumor stroma and parenchymal disease

Background: Encapsulation in hepatocellular carcinoma is associated with decreased invasiveness and improved survival in several series. Although active fibrogenesis by myofibroblasts has been demonstrated in the capsule, it is unclear if the capsule results from a general increase in peritumoral fibrosis, or an inherently less invasive tumor phenotype. The relationship between collagen deposition within tumor stroma, presence of cirrhosis and invasiveness also needs clarification. Methods: We performed immunohistochemistry for collagens I, III, IV and VI on sections of encapsulated and non-encapsulated hepatocellular carcinoma, arising in cirrhotic and non-cirrhotic livers. Staining was graded semi-quantitatively in tumor stromal elements and adjacent parenchymal sinusoids. The relationship of this staining with encapsulation, cirrhosis, and vascular invasion was analyzed. Results: Formation of a discrete capsular layer was associated with reduced vascular invasion, but not with a pervasive increase in peritumoral fibrosis. Increased collagen I content of tumor stroma and adjacent parenchymal sinusoids was associated with non-encapsulated tumors and vascular invasion. The presence of cirrhosis had little effect on capsule composition. Conclusions: Encapsulation of hepatocellular carcinoma reflects reduced invasiveness, rather than increased peritumoral collagen synthesis, which may instead enhance invasion. Increased intratumoral collagen I protein is also associated with increased tumor invasiveness. Pre-existing cirrhosis has little effect on tumor progression, possibly because the characteristics of cirrhosis are overwhelmed by tumor-induced changes in the adjacent parenchyma.(C) 2003 Blackwell Publishing Asia Pty Ltd.