Supercritical CO2 interpolymer complex encapsulation improves heat stability of probiotic bifidobacteria

The probiotic industry faces the challenge of retention of probiotic culture viability as numbers of these cells within their products inevitably decrease over time. In order to retain probiotic viability levels above the therapeutic minimum over the duration of the product’s shelf life, various methods have been employed, among which encapsulation has received much interest. In line with exploitation of encapsulation for protection of probiotics against adverse conditions, we have previously encapsulated bifidobacteria in poly-(vinylpyrrolidone)-poly-(vinylacetate-co-crotonic acid) (PVP:PVAc-CA) interpolymer complex microparticles under supercritical conditions. The microparticles produced had suitable characteristics for food applications and also protected the bacteria in simulated gastrointestinal fluids. The current study reports on accelerated shelf life studies of PVP:PVAc-CA encapsulated Bifidobacterium lactis Bb12 and Bifidobacterium longum Bb46. Samples were stored as free powders in glass vials at 30 °C for 12 weeks and then analysed for viable counts and water activity levels weekly or fortnightly. Water activities of the samples were within the range of 0.25–0.43, with an average a _w  = 0.34, throughout the storage period. PVP:PVAc-CA interpolymer complex encapsulation retained viable levels above the recommended minimum for 10 and 12 weeks, for B. longum Bb46 and B. lactis Bb12, respectively, thereby extending their shelf lives under high storage temperature by between 4 and 7 weeks. These results reveal the possibility for manufacture of encapsulated probiotic powders with increased stability at ambient temperatures. This would potentially allow the supply of a stable probiotic formulation to impoverished communities without proper storage facilities recommended for most of the currently available commercial probiotic products.     

Comparison of different methods for release of Bifidobacterium longum Bb46 from the poly(vinylpyrrolidone)-poly(vinylacetate-co-crotonic acid) interpolymer complex matrix,and the effect of grinding on the microparticles

Bifidobacteria have been efficiently encapsulated in poly(vinylpyrrolidone)-poly(vinylacetate-co-crotonic acid) (PVP: PVAc-CA) interpolymer complex formed in scCO₂. Research indicated that this method improves the stability of encapsulated bacteria in simulated gastrointestinal fluids in vitro. However, further analysis indicated release of lower numbers of encapsulated bacteria from the encapsulating matrix. The aims of this study were to determine a method that would release high numbers of bacteria from the PVP: PVAc-CA interpolymer complex matrix microparticles, and furthermore, to determine the effects of milling on the morphological properties of the microparticles. Three release methods, namely sonication, homogenization in a stomacher and incubation in simulated intestinal fluid were compared. Released viable bacteria were assayed using plate counts. Viable bacteria released using a stomacher were three orders of magnitude higher than those released by incubation and an order of magnitude higher than those released using sonication. SEM indicated no negative effects such as exposure of encapsulated bacteria on the matrix due to milling of product. Homogenization in a stomacher is the most efficient method for releasing bacteria from the PVP: PVAc-CA interpolymer complex matrix. Particle size of the PVP: PVAc-CA microparticles encapsulating bacteria can be reduced further by grinding, without exposing the enclosed bacteria.     

Kinetic study of Lactococcus lactis strains (SLT6 and SLT10) growth on papain-hydrolysed whey

The effect of controlled whey hydrolysis by papain on growth of two lactic acid bacteria isolated from artisanal Leben: Lactococcus lactis var. diacetylactis (SLT6 and SLT10) was investigated. The higher biomass and maximum specific growth rate (μ _max) were obtained after 30 min of hydrolysis. HPLC analysis of peptides showed that whey hydrolysis reduced the amount of peptides of MW > 400 Da and increased those peptides of MW < 400 Da. The two studied strains exhibited different peptide requirements. The pH-controlled batch cultures in 30 min hydrolysed whey followed the Monod kinetic for growth and for lactate production. The values of the key kinetic constants were: maximum specific growth rate (μ _max), 1.08 and 0.56 h^?1; yield biomass on lactose (Y _x/s), 0.20 and 0.18 g g^?1 and saturation constant K _s, 4.2 and 2.8 g L^?1 for SLT6 and SLT10, respectively. When compared with batch experimental data, the model provided good predictions for growth, lactose utilisation and lactate production profiles.     

Dujardinascaris mormyropsis n. sp. (Nematoda: Anisakidae) from the osteoglossiform fish Mormyrops anguilloides (Linnaeus) (Mormyridae) in Central Africa

A new nematode species, Dujardinascaris mormyropsis n. sp. (Anisakidae), is described from specimens found in the stomach and intestine of the cornish jack Mormyrops anguilloides (Linnaeus) (Osteoglossiformes; Mormyridae) in the Sangha River (Congo River basin) in Dzanga-Sangha Protected Areas, Central African Republic. Based on light and scanning electron microscopical examination, the new species differs from the only other congeneric species parasitising fishes in Africa, D. malapteruri (Baylis, 1923), mainly in the presence of dentigerous ridges on lips, absence of lateral caudal alae in the cloacal region, in the anteriorly curved, non-bifid distal tip of the gubernaculum and larger eggs (60–90 × 52–78 vs 45 × 30 μm); males of D. mormyropsis are characterised by the presence of a ventral precloacal cuticular ornamentation not observed in other Dujardinascaris spp. Dujardinascaris mormyropsis and D. malapteruri also differ in the order of their fish hosts (Osteoglossiformes vs Siluriformes). Specimens previously reported as Dujardinascaris graberi Troncy, 1969, a junior synonym of D. malapteruri, from Mormyrops engystoma Boulenger in Chad belong to the new species D. mormyropsis.     

Lactobacillus acidophilus La5 and Bifidobacterium lactis Bb12 cell surface hydrophobicity and survival of the cells under adverse environmental conditions

Changes in the cell surface hydrophobicity (CSH) of probiotic bacteria Lactobacillus acidophilus La5 and Bifidobacterium lactis Bb12 and the survival of these cells were examined in response to varied cultivation conditions and adverse environmental conditions. An inverse linear relationship (P < 0.01) was detected between the CSH of intact L. acidophilus La5 and B. lactis Bb12 and survival of cells subjected to subsequent freezing/thawing, long-term storage or exposure to mineral and bile acids. The observed relationships were supported by significant correlations between the CSH and changes in composition of the cell envelopes (proteins, lipids and carbohydrates) of L. acidophilus La5 and B. lactis Bb12 examined using FT-IR spectroscopy and conventional biochemical analysis methods. The results also suggest that the estimates of hydrophobicity, being a generalized characteristic of cell surfaces, are important parameters to predict the ability of intact probiotic bacteria to endure extreme environments and therefore should be monitored during cultivation. A defined balance of cell components, which can be characterized by the reduced CSH values, apparently helps to ensure the resistance, improved viability and hence the overall probiotic properties of bacteria.     

Yeast Cell Microcapsules as a Novel Carrier for Cholecalciferol Encapsulation: Development,Characterization and Release Properties

In this study Saccharomyces cerevisiae yeast cells was used as a novel vehicle for encapsulation of vitamin D₃. The effects of initial cholecalciferol concentration (100,000 and 500,000 IU/g yeast), yeast cell pretreatment (plasmolysis with NaCl) and drying method (spray or freeze drying) on microcapsules properties were investigated. It was found that the vitamin concentration and drying method had significant influence on encapsulation efficiency (EE) and size of yeast microcapsules. Furthermore, EE values were more increased by the plasmolysis treatment. The highest EE was obtained for plasmolysed and spray dried yeast cells prepared using initial cholecalciferol concentration of 2.5 mg per gram of yeast cells (76.10?±?6.92%). The values of mean particle size were 3.43–7.91 μm. The presence of cholecalciferol in yeast microcapsules was confirmed by X-ray diffraction (XRD) and Fourier transform-infrared (FT-IR) analyses. The in vitro cholecalciferol release from yeast microcapsules in phosphate buffer saline solution (PBS) followed a controlled release manner consistent with a Fickian diffusion mechanism. In addition, the release studies in simulated gastrointestinal tract showed sustained release of cholecalciferol in the stomach condition and significant release in intestinal medium.     

Casein Microparticles from Blend Films Forming Casein/α-Tocopherol Emulsion Droplets in Solution

Microparticles continue to receive widespread attention from food professionals because of their potential use as carriers of bioactive substances. This study demonstrates a novel method to prepare casein microparticles, which co-assemble with α-tocopherol (αT) into emulsion droplets. When the particles were extracted from the pectin matrix via enzymatic degradation, they remained stable in a buffer solution for at least 3 weeks. Optical microscopy showed that the size distribution of the microparticles is between 5 μm and 50 μm, which is in accordance with previous observations in blend films. High-performance liquid chromatography revealed that the amounts of α_S1- and κ-casein in the microparticles are significantly higher than those in native casein micelles. Confocal Raman microscopy showed that in the presence of α-tocopherol, the microparticles assemble into emulsion droplets, with phenol in the core and casein in the shell. Herein, we demonstrate a new method to form casein-based emulsion droplets for potential use as carriers of bioactive substances.     

A new species of Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) from Carlia spp. (Sauria: Scincidae) from Papua New Guinea

A new species of Eimeria Schneider, 1875 from rainbow skinks, Carlia ailanpalai Zug and Carlia eothen Zug is described from specimens collected in Papua New Guinea (PNG). Oöcysts of Eimeria zugi n. sp. from one of one (100%) C. eothen are ellipsoidal to cylindroidal, with a smooth, colourless, bi-layered wall, measure 25.1 × 15.5 μm and have a length/width ratio of 1.6. The micropyle and the oöcyst residuum are absent, but a polar granule is present. The sporocysts are ovoidal to ellipsoidal and 10.3 × 7.1 μm in size and do not contain Stieda, sub-Stieda or para-Stieda bodies; and the sporocyst residuum is composed of a compact mass of large globules. The sporozoites are elongate, 12.8 × 2.9 μm in size, and contain anterior and posterior refractile bodies with a nucleus between them. This is the ninth species of coccidium described from skinks from PNG, and the new species described herein is apparently endemic to the skink genus Carlia (Gray).     

Bioavailability and metabolism of fucoxanthin in rats: structural characterization of metabolites by LC-MS (APCI)

This study reports bioavailability and metabolism of fucoxanthin (FUCO) from brown algae Padina tetrastromatica in rats. Rats were divided into two groups (n = 25/group). Group one was fed basal diet (control) while the group two received retinol deficient diet (RD group) for 8 weeks. After confirmed RD in blood (0.53 μmol/l), rats were further sub-grouped (n = 5/sub group), intubated a dose of FUCO (0.83 μmol) and killed after 0, 2, 4, 6 and 8 h. The plasma levels (area under curve/8 h) of FUCO (fucoxanthinol (FUOH) + amarouciaxanthin (AAx)) was 2.93 (RD group) and 2.74 pmol/dl (control), respectively. No newly formed retinol was detected in RD rats intubated with FUCO. Besides FUOH (m/z 617 (M+H)⁺) and AAx (m/z 617 (M+H^?)⁺), other deacetylated, hydrolyzed and demethylated metabolites of bearing molecular mass at m/z 600.6 (FUOH–H₂O), m/z 597 (AAx–H₂O), m/z 579 (AAx–2H₂O+1), m/z 551 (AAx–2H₂O–2CH₃+2) and m/z 523 (AAx–2H₂O–4CH₃+4) were also detected in plasma and liver by LC-MS (APCI). Although biological functions of FUCO metabolites need thorough investigation, this is the first detailed report on FUCO metabolites in rats.     

Tuning the Optical and Rheological Properties of Fullerene C<Subscript>60</Subscript>/Poly (Vinyl Pyrrolidone) Nanofluids via Inclusion of Nanogold

In this article, development of nanogold (NG) containing fullerene C₆₀ nanofluids (NFs) with poly (vinyl pyrrolidone) PVP polymer in water via a wet chemical route is reported and studied their rheo-optical properties. An inclusion of NG into a C₆₀:PVP nanofluid thus results in a gold (Au) surface plasmon resonance (SPR) enhanced π?→?π* C₆₀ (sp²) electron transition over 250–450 nm of absorption spectrum in water. Evolution of a broad Au-SPR band in the 450–750 nm region signifies that a controlled Au³⁺?→?Au reduction reaction had occurred on it and was carried in C₆₀:PVP NFs. The average maximum wavelength value has thus varied along with a molar extinction coefficient in a function of the Au-uploads and shape and size of the flocculates in the resulting Au:C₆₀-PVP NFs. A systematic rheological study performed on the Au:C₆₀-PVP NFs in water by varying the NG content up to 85.0 μM reveals a non-Newtonian behavior with an enhanced yield stress is a signature of Bingham flow. NG of different shapes serves as filler in C₆₀:PVP NFs so as it adapts tailored shear viscosity. The shear viscosity relaxes slowly to the base value on increasing the shear rate from 10 to 100 s^?1 as it leads to breaking up of soft Au:C₆₀-PVP assemblies into a fine structures. Synthesis of Au:C₆₀-PVP NFs with various Au-contents could be potential nanostructure hybrid composite materials for development of photovoltaic nanodevices.     

Synthesis of nylon 4 from gamma-aminobutyrate (GABA) produced by recombinant Escherichia coli

In this study, we developed recombinant Escherichia coli strains expressing Lactococcus lactis subsp. lactis Il1403 glutamate decarboxylase (GadB) for the production of GABA from glutamate monosodium salt (MSG). Syntheses of GABA from MSG were examined by employing recombinant E. coli XL1-Blue as a whole cell biocatalyst in buffer solution. By increasing the concentration of E. coli XL1-Blue expressing GadB from the OD₆₀₀ of 2–10, the concentration and conversion yield of GABA produced from 10 g/L of MSG could be increased from 4.3 to 4.8 g/L and from 70 to 78 %, respectively. Furthermore, E. coli XL1-Blue expressing GadB highly concentrated to the OD₆₀₀ of 100 produced 76.2 g/L of GABA from 200 g/L of MSG with 62.4 % of GABA yield. Finally, nylon 4 could be synthesized by the bulk polymerization using 2-pyrrolidone that was prepared from microbially synthesized GABA by the reaction with Al₂O₃ as catalyst in toluene with the yield of 96 %.     

Description of Choleoeimeria duszynskii n. sp. (Apicomplexa: Eimeriidae) from the gallbladder of the Middle Eastern short-fingered gecko Stenodactylus doriae (Blanford) (Sauria: Gekkonidae) in Saudi Arabia

Choleoeimeria duszynskii n. sp. is described from the gallbladder of the Middle Eastern short-fingered gecko Stenodactylus doriae (Blanford) from Salasel, Central region, Saudi Arabia. Oöcysts are ellipsoidal (rarely ovoidal), 23–25 × 16–18 (24 × 17) μm, with mean length/width ratio 1.4. Oöcyst wall is smooth, bi-layered, c.1.0 μm thick. Micropyle, oöcyst residuum and polar granule are absent. Sporocysts are ellipsoidal, 8–10 × 4–6 (9 × 5) μm, with a smooth, colourless sporocyst wall and mean length/width ratio 1.7. Sporozoites are sausage-shaped, with one end slightly pointed, arranged head to tail around sporocyst residuum. Refractile bodies and nuclei are not discernible. The endogenous development is restricted to the epithelium of the gallbladder and bile duct. Meronts are rounded, 12 μm in diameter, containing up to c.15 merozoites. Microgamonts are irregular in shape, 22 × 17 μm, containing a large number of microgametes. Macrogamonts are spherical, 17 μm in diameter, with centrally located nucleus and wall-forming bodies at the periphery.     

Fabricating PLGA microparticles with high loads of the small molecule antioxidant N‐acetylcysteine that rescue oligodendrocyte progenitor cells from oxidative stress

Reactive oxygen species (ROS), encompassing all oxygen radical or non‐radical oxidizing agents, play key roles in disease progression. Controlled delivery of antioxidants is therapeutically relevant in such oxidant‐stressed environments. Encapsulating small hydrophilic molecules into hydrophobic polymer microparticles via traditional emulsion methods has long been a challenge due to rapid mass transport of small molecules out of particle pores. We have developed a simple alteration to the existing water‐in‐oil‐in‐water (W/O/W) drug encapsulation method that dramatically improves loading efficiency: doping external water phases with drug to mitigate drug diffusion out of the particle during fabrication. PLGA microparticles with diameters ranging from 0.6 to 0.9 micrometers were fabricated, encapsulating high loads of 0.6–0.9 µm diameter PLGA microparticles were fabricated, encapsulating high loads of the antioxidant N‐acetylcysteine (NAC), and released active, ROS‐scavenging NAC for up to 5 weeks. Encapsulation efficiencies, normalized to the theoretical load of traditional encapsulation without doping, ranged from 96% to 400%, indicating that NAC‐loaded external water phases not only prevented drug loss due to diffusion, but also doped the particles with additional drug. Antioxidant‐doped particles positively affected the metabolism of oligodendrocyte progenitor cells (OPCs) under H₂O₂‐mediated oxidative stress when administered both before (protection) or after (rescue) injury. Antioxidant doped particles improved outcomes of OPCs experiencing multiple doses of H₂O₂ by increasing the intracellular glutathione content and preserving cellular viability relative to the injury control. Furthermore, antioxidant‐doped particles preserve cell number, number of process extensions, cytoskeletal morphology, and nuclear size of H₂O₂‐stressed OPCs relative to the injury control. These NAC‐doped particles have the potential to provide temporally‐controlled antioxidant therapy in neurodegenerative disorders such as multiple sclerosis (MS) that are characterized by continuous oxidative stress.     

Nutrient uptake rates by the alien alga Undaria pinnatifida (Phaeophyta) (Nuevo Gulf, Patagonia, Argentina) when exposed to diluted sewage effluent

In the early nineties, Undaria pinnatifida has been accidentally introduced to Nuevo Gulf (Patagonia, Argentina) where the environmental conditions would have favored its expansion. The effect of the secondary treated sewage discharge from Puerto Madryn city into Nueva Bay (located in the western extreme of Nuevo Gulf) is one of the probable factors to be taken into account. Laboratory cultures of this macroalgae were conducted in seawater enriched with the effluent. The nutrients (ammonium, nitrate and phosphate) uptake kinetics was studied at constant temperature and radiation (16?°C and 50 μE m^?2 s^?1 respectively). Uptake kinetics of both inorganic forms of nitrogen were described by the Michaelis–Menten model during the surge phase (ammonium: V _max ^sur: 218.1 μmol h^?1 g^?1, K _s ^sur: 476.5 μM and nitrate V _max ^sur: 10.7 μmol h^?1 g^?1, K _s ^sur: 6.1 μM) and during the assimilation phase (ammonium: V _max ^ass: 135.6 μmol h^?1 g^?1, K _s ^ass: 407.2 μM and nitrate V _max ^ass: 1.9 μmol h^?1 g^?1, K _s ^ass: 2.2 μM), with ammonium rates always higher than those of nitrate. Even though a net phosphate disappearance was observed in all treatments, uptake kinetics of this ion could not be properly estimated by the employed methodology.     

<Emphasis Type="Italic">Isospora bocamontensis</Emphasis> n. sp. (Apicomplexa: Eimeriidae) from the yellow cardinal <Emphasis Type="Italic">Gubernatrix cristata</Emphasis> (Vieillot) (Passeriformes: Emberizidae) in South America

A new coccidian species (Protozoa: Apicomplexa: Eimeriidae) is reported from the endangered yellow cardinal Gubernatrix cristata (Vieillot) in southern Brazil. Isospora bocamontensis n. sp. has oöcysts which are subspheroidal, measure 32.1 × 28.9 μm and have a smooth, bilayered wall c.1.5 μm thick. The micropyle and the oöcyst residuum are absent, but a polar granule is sometimes present. Its sporocysts are ellipsoidal and 17.3 × 12.2 μm in size and contain a half-moon-shaped Stieda body, a prominent, homogeneous substieda body; and a sporocyst residuum composed of a compact mass of granules. The sporozoites have one refractile body and a nucleus.     

Karyotype analysis and DNA content in some species of Chrysolaena (Vernonieae,Asteraceae)

Cytogenetic characterization by karyotyping and determination of DNA content by flow cytometry of five species of Chrysolaena (Vernonieae, Asteraceae) was performed. This is the first study of nuclear DNA content realized in the genus. The 2C-values were compared with the ploidy level and the total karyotype length (TKL) of each species. Mitotic analysis revealed a base chromosome number x = 10 for all entities and different ploidy levels, from diploid (2n = 2x = 20) to octoploid (2n = 8x = 80). All species showed bimodal karyotypes composed of metacentric and submetacentric chromosomes. The average chromosome size (ML) varied from 1.86 μm to 2.70 μm, while the TKL ranged from 18.65 μm to 80.55 μm. The intrachromosomal asymmetry index (A₁) varied from 0.27 to 0.38, while the interchromosomal asymmetry index (A₂) ranged from 0.19 to 0.25. A new cytotype is reported for the first time for C. propinqua. Accessory chromosomes found in C. verbascifolia, C. cognata, C. flexuosa, and C. propinqua are also reported as new.     

Choleoeimeria ghaffari n. sp. (Apicomplexa: Eimeriidae) from the gallbladder of Eryx jayakari Boulenger (Serpentes: Boidae) in Saudi Arabia

Choleoeimeria ghaffari n. sp. is described from the gallbladder of Eryx jayakari Boulenger in Saudi Arabia. Oöcysts are tetrasporocystic, cylindroidal, 23 × 14 μm, with a smooth bi-layered wall and length/width ratio of 1.5, without micropyle, oöcyst residuum and polar granule. Sporocysts are subspheroidal to ellipsoidal, 8 × 6 μm, with length/width ratio of 1.4, without Stieda, sub-Stieda and para-Stieda bodies but with sporocyst residuum. Sporozoites are banana-shaped measuring 10 × 1.5 μm. The endogenous development was found to occur in the gallbladder epithelium and the extrahepatic bile ducts. Mature meronts are spheroidal, c.10 μm wide, and suspected to produce 12–16 merozoites. Microgamonts are irregular in shape, 13 × 10 μm, whereas macrogamonts are mostly subspheroidal, c.12 μm wide, with a prominent centrally-located nucleus. Based on oöcyst morphology and the site of endogenous development (epithelium of the gallbladder and bile ducts) the new eimeriid coccidian was placed in the genus Choleoeimeria Paperna & Landsberg, 1989.     

Method of testing insecticide susceptibility of Stenotus rubrovittatus (Hemiptera: Miridae) by topical application

Appropriate droplet size and the preferred sex and age of insects for insecticides used in topical application were investigated to standardize the method of testing the susceptibility of Stenotus rubrovittatus (Matsumura). The overflow of acetone droplets applied on the notum of the thorax of S. rubrovittatus adults occurred when the droplet size exceeded 0.20 μl per insect, and the overflow occurred more frequently in males than in females. No effect of acetone solution on mortality was observed at less than 0.20 μl droplet size per insect within 72 h after treatment. The LD₅₀ values and relative potencies of fenitrothion 24 and 48 h after treatment were almost the same in each sex. The fenitrothion susceptibility of males was twice higher than that of females. The fenitrothion susceptibility increased with age after emergence. However, there were small differences in LD₅₀ values and relative potencies between 4–7 and 8–11 days after emergence because the females 4–11 days after emergence had a small difference in body weight. These results led us to conclude that the recommended size of acetone droplets per insect is less than 0.20 μl and that it is suitable to use S. rubrovittatus females 4–11 days after emergence for topical application.     

The comparative pollen morphology of genera Matricaria L. and Tripleurospemum Sch. Bip. (Asteraceae) in Turkey

Pollen morphology of four Matricaria species and 28 Tripleurospermum species was investigated with light microscopies (LM) and scanning electron microscopies (SEM). Pollen slides were prepared using Wodehouse technique. Measurements were based on 20 or more pollen grains per specimen. For SEM studies, dried pollen grains were transferred on aluminum stubs and coated with gold for 4 min in a sputter-coater. The pollen grains of Matricaria and Tripleurospermum are radially symmetric and isopolar. The pollen grains of the Matricaria are oblate-spheroidal with the polar axes 16.6–31.2 μm and the equatorial axes 18.7–23.9 μm. Tripleurospermum is oblate-spheroidal, suboblate and prolate-spheroidal with the polar axes 15.6–32.2 μm and the equatorial axes 17.7-38.5 μm. The pollen grains of Tripleurospermum are operculate and tricolporate. Matricaria is operculate and usually tricolporate or rarely syncolporate, tricolpate and tetracolporate. The pollen grain of both taxa shows echinate ornamentation. The spines are commonly conical with a broadened base and a tapered apical portion. The spine length varies between 1.8–4 μm in Tripleurospermum and 2.3–3.3 μm in Matricaria. The width of spines varies between 2.8–4.6 μm in Tripleurospermum and 2.4–3.6 μm in Matricaria. Inter-spinal area shows granulate–perforate, reticulate–perforate, rugulate–perforate ornamentations and the tectum surrounding the spine base is micro perforate. Overall exine thickness ranges from 2.8 to 4.8 μm in Tripleurospermum, 3.6 to 5.2 μm in Matricaria. Intine is thicker under pores in Tripleurospermum (0.3–0.62 μm) than in Matricaria (0.6–0.8 μm). Inter-spinal ornamentations, pollen shape and the numbers of perforations at the spin base have been observed as important morphological characters.     

Physicochemical and Preclinical Evaluation of a Novel Buccal Measles Vaccine

The aim of this study is to develop an orally disintegrating film (ODF) containing a microparticulate measles vaccine formulation for buccal delivery. The measles vaccine microparticles were made with biocompatible and biodegradable bovine serum albumin (BSA) and processed by spray drying. These vaccine microparticles were incorporated in the ODF, consisting of Lycoat RS720®, Neosorb P60W® and Tween 80. The yield of the microparticles was approximately 85–95%, w/w. The mean size of the vaccine microparticles was 3.65?±?1.89 μm and had a slightly negative surface charge of 32.65?±?2.4 mV. The vaccine particles were nontoxic to normal cells at high concentrations (500 μg/2.5?×?10⁵ cells) of vaccine particles. There was a significant induction of innate immune response by vaccine microparticles which was observed in vitro when compared to blank microparticles (P?<?0.05). The vaccine microparticles also significantly increased the antigen presentation and co-stimulatory molecules expression on antigen presenting cells, which is a prerequisite for Th1 and Th2 immune responses. When the ODF vaccine formulation was dosed in juvenile pigs, significantly higher antibody titers were observed after week 2, with a significant increase at week 4 and plateauing through week 6 comparative to naïve predose titers. The results suggest that the ODF measles vaccine formulation is a viable dosage form alternative to noninvasive immunization that may increase patient compliance and commercial distribution.