Agarose–gelatin conjugate for adherent cell-enclosing capsules

Spherical capsules were prepared by extruding aqueous agarose–gelation conjugate solution into co-flowing liquid paraffin at 38°C and cooling the resultant emulsion. Capsule diameter was controlled between 40 and 250 μm by changing the velocity of the liquid paraffin. Adherent Crandall–Reese feline kidney cells enclosed in conjugate capsules of 141 ± 23 μm diam. had a higher degree of proliferation than those in unmodified agarose capsules. Mitochondrial activity, detected for cell-enclosing conjugate capsules normalized against unit volume of gel, was about double that of unmodified agarose capsules over 28 days. These results demonstrated the feasibility of agarose–gelatin conjugate as a material of cell-enclosing capsules.     

Oriented Embedding of Biological Materials and Accurate Localization for Ultrathin Sectioning

Gelatin capsules with rounded ends clipped off and open ends moistened, affixed to a glass slide and sealed with a 15% gelatin solution are used to embed blocks of tissue in plastic. The surface of the slide serves as an orientation plane for structures of the tissue. The plane end of capsules of polymerized plastic containing no tissue is used in embedding frozen tissue sections. The plastic-infiltrated section is flattened against the capsule end under the weight of a 3/4 inch square of plate glass so that larger sections may be cut and surveyed. Embedding cultured cell monolayers grown on coverslips is accomplished in a comparable manner, but the square of plate glass is not needed as a weight. Block-face localization methods depend on the type of material embedded. With blocks of tissue it is achieved by moistening the face with xylene to develop relief. Thin tissue sections are examined by transmitted light, while cell monolayers are stained on the capsule end with methylene blue.     

Immunohistochemical reactivity of polyclonal antibodies against Sphaerospora testicularis and Ceratomyxa labracis (Myxosporea: Bivalvulida), with other myxosporean parasites.

Immunological staining with rabbit antibodies raised against Sphaerospora testicularis and Ceratomyxa labracis was used to characterise their specificity and their reactivity towards other fish parasites. Polar capsules and valves of S. testicularis and C. labracis were labelled with their homologous antibody and cross reaction was observed with all the myxosporean parasites assayed from marine and freshwater fish hosts. All polar capsules were stained with both antibodies, except those of Zschokkella mugilis, which were not labelled with anti-S. testicularis serum. These observations suggest that polar capsules may be very conserved structures in myxosporean parasites from different hosts.     

Reference material for the evaluation of a standard method for the detection of salmonellas in foods and feeding stuffs

To evaluate a standard salmonella isolation method a reference material consisting of 0.2 g spray-dried milk inoculated with Salmonella typhimurium and contained in gelatin capsules was prepared. The organisms were distributed homogeneously between capsules, and their numbers were stable for 120 d when the capsules were stored in dry conditions at 4 degrees C. Addition of these capsules with or without food samples to pre-enrichment broth gave low and reproducible levels of Salm. typhimurium contamination without altering the pre-enrichment and without influencing the other bacterial flora present. As a result of an interlaboratory trial, the reference material indicated that the food and/or its competitive flora may have a negative influence on the detection of salmonellas.     

Reference material for the evaluation of a standard method for the detection of salmonellas in foods and feeding stuffs

To evaluate a standard salmonella isolation method a reference material consisting of 0.2 g spray-dried milk inoculated with Salmonella typhimurium and contained in gelatin capsules was prepared. The organisms were distributed homogeneously between capsules, and their numbers were stable for 120 d when the capsules were stored in dry conditions at 4C. Addition of these capsules with or without food samples to pre-enrichment broth gave low and reproducible levels of Salm. typhimurium contamination without altering the pre-enrichment and without influencing the other bacterial flora present. As a result of an interlaboratory trial, the reference material indicated that the food and/or its competitive flora may have a negative influence on the detection of salmonellas.     

Mechanical characterization of an unusual elastic biomaterial from the egg capsules of marine snails (Busycon spp.)

Egg capsule material serves as a putative protection mechanism for developing snail embryos facing the perils of the marine environment. We conducted the first quantitative study of this acellular structural protein with the goals of characterizing its chemical and mechanical properties and the relationship of these properties to its biological protective function. We have found that this protein polymer exhibits long-range elasticity with an interesting recoverable yield evidenced by an order of magnitude decrease in elastic modulus (apparent failure) that begins at 3%-5% strain. This material differs significantly from other common structural proteins such as collagen and elastin in mechanical response to strain. Qualitative similarities in stress/strain behavior to keratin, another common structural protein, are more than coincidental when composition and detailed mechanical quantification are considered. This suggests the possibility of alpha-helical structure and matrix organization that might be similar in these two proteins. Indeed, the egg capsule protein may be closely related to vertebrate keratins such as intermediate filaments. We conclude that while this material's bimodal tensile properties may serve as useful protection against the impact loading egg capsules encounter in the intertidal zone, the full biological importance of these capsules is not known.     

Short-chain basement membrane collagen. Further characterization and its biosynthesis by F-9 embryonal carcinoma cells

The paper describes further characterization of the 55-kDa short-chain collagen from lens capsule. Lens capsules were extracted with 5.5 M guanidine.HCl and the extracted material was fractionated on agarose A-5M followed by high-pressure liquid chromatography (HPLC). By amino acid composition, the major fraction obtained from HPLC was found to be different than type-IV collagen fragments. The 55-kDa short-chain collagen on pepsin digestion produced a 45-kDa pepsin-resistant fragment. The undifferentiated embryonal carcinoma (F-9) cells were found to synthesize increased amounts of 55-kDa short-chain collagen. The identity of this biosynthesized molecule with 55-kDa short-chain collagen from lens capsules was established by immunoprecipitation experiments. The results indicated a close similarity or identical nature of the short-chain collagens from these two sources.     

The polysaccharide capsule of the pathogenic fungus Cryptococcus neoformans enlarges by distal growth and is rearranged during budding

The capsule of Cryptococcus neoformans can undergo dramatic enlargement, a phenomenon associated with virulence. A prior study that used Ab to the capsule as a marker for older capsular material concluded that capsule growth involved the intermixing of new and old capsular material with displacement of older capsular polysaccharide towards the surface. Here we have revisited that question using complement (C), which binds to capsular polysaccharide covalently, and cannot redistribute by dissociation and binding at different sites. The experimental approach involved binding of C to cells with small capsules, inducing capsule growth, and following the location of C relative to the cell wall as the capsule enlarged. C remained close to the cell wall during capsule growth, indicating that capsule enlargement occurred by addition of new polysaccharide near the capsule edge. This conclusion was confirmed by an independent method that employed radioactive metabolic labelling of newly synthesized capsule with 3H-mannose followed by gradual capsular stripping with gamma-radiation. Capsule growth proceeded to a certain size, which was a function of cell size, and was not degraded when the cells were transferred to a non-inducing medium. During budding, an opening appeared in the capsule of the mother cell that permitted the nascent bud to separate. Scanning EM suggested that a physical separation formed between the capsules of the mother and daughter cells during budding, which may avoid mixture between both capsules. Our results indicate that C. neoformans capsular enlargement also occurs by apical growth and that budding results in capsular rearrangements.     

Demonstration by light and electron microscopy of capsules on gonococci recently grown in vivo.

A study by light microscopy, using Leishman's stain alone or Leishman's stain followed by nigrosin, showed the presence of capsules on gonococci of two strains subcultured from subcutaneous chambers in guinea pigs. With the Alcian blue method of preparation for electron microscopy, gonococci of both strains recently grown in vivo showed densely stained capsules on some cells, while others in the same preparation showed only irregular masses of dense material on their surfaces with strands connecting adjacent bacteria. Treatment with antiserum, complement and conglutinin revealed irregular masses and strands of extracellular material with fixatives that did not contain Alcian blue.     

Preparation and Thermal Energy Storage of Carboxymethyl Cellulose-Modified Nanocapsules

A series of nanocapsules with carboxymethyl cellulose (CMC)-modified melamine-formaldehyde as the shell material and phase change paraffin as the core material were prepared by in situ polymerization. The modified capsules were examined using Fourier transform infrared spectra, scanning electronic microscope, differential scanning calorimeter, and optical microscopy, and two factors that influence paraffin emulsion preparation (emulsifier type and stirring rate) were investigated. The effects of the synthesis conditions used for the prepolymer on the surface morphology of the capsules were also studied. We found that phase change capsules prepared with both anionic and nonionic emulsifiers were superior to those prepared with a simple emulsifier. The best performance of the paraffin emulsion was obtained when the emulsion was stirred at 8,000 rpm during preparation. The optimal prepolymer reaction conditions to give smooth capsules with good dispersibility and complete morphology were reaction temperature 72.5 °C, reaction time 75 min, and pH?8.5. The CMC-modified nanocapsules have a phase change enthalpy of 83.46 J/g, are fully encased, and are uniform, with an average particle size of 50 nm.     

Myxobolus goensis n. sp. (Myxozoa, Myxosporea, Myxobolidae), a parasite of the gills of Mugil cephalus (Osteichthyes, Mugilidae) from Goa, India

Two species of Myxobolus are reported from the gills of Mugil cephalus collected at Goa, India: M. goensis n. sp. and M. parvus Shulman, 1962. Myxobolus goensis n. sp. forms digitiform or rounded plasmodia between the gill rakers. Their spores are oval in frontal view, with tapered anterior extremity, and lemon-shaped in lateral view, measuring 9.7 (9.5-10.5) microm in length, 6.6 (6-7.5) microm in width, and 5.2 (5-6) microm in thickness. The polar capsules are pyriform and unequal in size. The larger ones are 5.3 (4.5-6) microm long and 2.4 (2-3) microm wide; the smaller ones are 2.4 (2-3) microm long and 1.8 (1.5-2) microm wide. The polar filament forms five turns aligned perpendicularly to the longitudinal axis of the larger polar capsules. Within the smaller polar capsules the polar filament is difficult to observe and, apparently, forms three coils. The spores are distinctly different from other Myxobolus species infecting M. cephalus and other Mugil spp. Furthermore, the present material is also different from 204 Myxobolus species presenting differently sized polar capsules, representing nearly all the known species with this characteristic. The fact that only the M. cephalus specimens were infected among a sample of 206 fish specimens, comprising 27 different species, strongly suggests that this parasite is specific to M. cephalus.     

Detection of light signals by microencapsulated cis-cinnamoyl chymotrypsin

For the production of a new photographic material, microencapsulation of light-sensitive cis-cinnamoyl chymotrypsin into semipermeable nylon membranes is proposed. It has been shown that microencapsulation of a light-sensitive preparation precludes lateral diffusion of the light-induced enzyme in the photosensitive layer. Indoxylacetate that diffuses inside the capsules to be hydrolysed there by the enzyme to indoxyl is suggested for use as a substrate for chymotrypsin (EC 3.4.21). Indoxyl is then oxidized with oxygen to form insoluble bright-blue indigo which precipitates inside the capsules and stains them. It has been shown that the use of such substrates prevents the product of the enzymatic reaction from diffusing into the photosensitive layer. As a result, in such a photographic material the enzyme is regenerated only in exposed capsules in which a stained product is formed and fixed.     

Encapsulation of Trichoderma harzianum conidia as a method of conidia preservation at room temperature and propagation in submerged culture

The objective of the present work was to develop a method for the preservation of T. harzianum conidia at room temperature and the immobilised conidia propagation in submerged culture. This was accomplished by immobilising the strain in sodium alginate capsules (white capsules) and subsequently propagating them in a column bubble reactor (green capsules). Three capsule diameters were tested (micro, medium and large capsules), which were produced by emulsion internal gelation and dripping methods. Tested variables were the immobilised conidia propagation in submerged culture for free conidia production, the immobilised conidia viability throughout the time (two years), the resistance of the encapsulated conidia to the UV irradiation of short and long wavelength, and the antagonistic effect of the encapsulated T. harzianum against four phytopathogenic fungi. It was found that the medium capsules (1.5?±?0.3?mm) favoured the massive production of released conidia in submerged culture and that the higher the density of conidia per capsule, the greater the protection against the ultraviolet irradiation. Regarding the conidia preservation in calcium alginate, a viability loss of around 30% was observed two years after storage at environmental temperature in both white and green capsules; along the two years that the viability of conidia was analyzed, the purity of the formulation was corroborated. The results presented here show the efficacy of the green and white capsules for T. harzianum preservation at room temperature for a long period of time.     

Pharmacokinetic conversion study of a new cyclosporine formulation in stable adult renal transplant recipients

Cyclosporine A (CyA) is a standard component of immunosuppressive regimens. It is a critical-dose drug for which a minor change in absorption can have important clinical consequences. The aim of the study was to compare the pharmacokinetics and safety of the new generic CyA formulation, Equoral capsules, after a switch from original formulation, Neoral capsules, in seventy stable adult renal transplant recipients. The extent and rate of pharmacokinetic parameters for bioequivalence were compared in a non-randomized, steady-state clinical study with fixed non-replicate study design. Pharmacokinetic analysis of CyA have shown that both the rate and extent of absorption of Equoral does not differ significantly from that of Neoral. At identical dosing, the new formulation was found to have geometric means of C(max) 717 ng/ml and AUCtau 3108 ng/ml.h, while corresponding results of comparator were 725 ng/ml and AUCtau 3039 ng/ml.h, respectively. The 90 % confidence intervals of C(max) and AUCtau were within 80- 125 % interval of the mean values. The results suggest that Equoral capsules can be used as an alternative treatment to Neoral capsules in CyA regimen.     

Selection of different genotype larvae and adult worms for anthelmintic resistance by persistent and short-acting avermectin/milbemycins

To understand the factors that influence selection for anthelmintic resistance, it is necessary to examine the impact of drug treatment, particularly persistent drugs, on all phases of the worm life cycle. The efficacy of various avermectin/milbemycin anthelmintics was determined against resident worms, incoming larvae (L3) and development of eggs in faecal culture. Homozygote-resistant and maternal and paternal F1-heterozygote genotypes of Haemonchus contortus were used to infect sheep before or after treatment with ivermectin (IVM) oral, IVM capsule, moxidectin (MOX) oral or MOX injectable. Total worm count and quantitative larval culture were used to determine efficacy against parasitic and free-living stages, respectively. Selection for resistance by IVM capsules occurred at the adult and L3 stages because of poor efficacy against these stages for all resistant genotypes. However, the selective advantage of these surviving worms was reduced due to the low development of their eggs to L3 in faecal culture. For MOX, selection for resistance predominantly occurred after treatment because of high efficacy against resident adult worms of all resistant genotypes but poor efficacy against resistant L3 ingested after drug administration. The results indicated no evidence of sex-linked inheritance for IVM resistance. Mean IVM efficacies against homozygous and heterozygous resistant adult worms were not different, and IVM capsule efficacy against incoming L3 was approximately 70% for all resistant genotypes, consistent with a dominant trait. MOX was highly effective against adults of all resistant genotypes and approximately 76% effective against incoming L3 regardless of resistance genotype, also consistent with a dominant trait. These results will enable the impact of persistent drugs on worm control and anthelmintic resistance to be estimated. The results indicate that IVM capsules should not be used in populations where avermectin/milbemycin resistance is present.     

Zonal rate model for axial and radial flow membrane chromatography. Part I: Knowledge transfer across operating conditions and scales

The zonal rate model (ZRM) has previously been applied for analyzing the performance of axial flow membrane chromatography capsules by independently determining the impacts of flow and binding related non‐idealities on measured breakthrough curves. In the present study, the ZRM is extended to radial flow configurations, which are commonly used at larger scales. The axial flow XT5 capsule and the radial flow XT140 capsule from Pall are rigorously analyzed under binding and non‐binding conditions with bovine serum albumin (BSA) as test molecule. The binding data of this molecule is much better reproduced by the spreading model, which hypothesizes different binding orientations, than by the well‐known Langmuir model. Moreover, a revised cleaning protocol with NaCl instead of NaOH and minimizing the storage time has been identified as most critical for quantitatively reproducing the measured breakthrough curves. The internal geometry of both capsules is visualized by magnetic resonance imaging (MRI). The flow in the external hold‐up volumes of the XT140 capsule was found to be more homogeneous as in the previously studied XT5 capsule. An attempt for model‐based scale‐up was apparently impeded by irregular pleat structures in the used XT140 capsule, which might lead to local variations in the linear velocity through the membrane stack. However, the presented approach is universal and can be applied to different capsules. The ZRM is shown to potentially help save valuable material and time, as the experiments required for model calibration are much cheaper than the predicted large‐scale experiment at binding conditions. Biotechnol. Bioeng. 2013; 110: 1129–1141. © 2012 Wiley Periodicals, Inc.     

Tissue reactions to breast implants coated with polyurethane.

The histological features noted in the capsules from 7 polyurethane coated silicone breast prostheses are described. The polyurethane provoked a definite foreign body reaction and was slowly degraded, with some particles ejected from the capsule into the surrounding tissues. Separation of the polyurethane coating from the silicone prosthesis and the degradation of the polyurethane took about two years. Another much more resistant foreign material was found to occur in conjunction with the polyurethane in the capsules. It may be an adhesive or flakes off the silicone shell. Vacuolated spaces were noted in the inner layers of 3 capsules; it was assumed that they contained liquid silicone.     

Compression of biocompatible liquid-filled HSA-alginate capsules: determination of the membrane mechanical properties

Compression experiments between two parallel plates are performed on a series of biocompatible HSA-alginate capsules with two different membrane thicknesses. The capsule geometry and size as well as the average membrane thickness are first measured. The compression set-up is fitted with a sensitive force transducer that allows measurement of the compression force as a function of plate separation. The response of the capsule is analyzed by assuming different constitutive models for the membrane, where the shear and surface dilatation effects are accounted. An apparent area dilatation modulus is then computed for different values of the plate separation and required to remain constant as the capsule deformation increases. This allows identification of plausible constitutive laws for the membrane material.     

Investigation of the impact of feeding Lactobacillus plantarum CRL 1815 encapsulated in microbially derived polymers on the rat faecal microbiota

Aims

The aim of this study was to evaluate the impact of the administration of microencapsulated Lactobacillus plantarum CRL 1815 with two combinations of microbially derived polysaccharides, xanthan : gellan gum (1%:0·75%) and jamilan : gellan gum (1%:1%), on the rat faecal microbiota.

Methods and Results

A 10‐day feeding study was performed for each polymer combination in groups of 16 rats fed either with placebo capsules, free or encapsulated Lact. plantarum or water. The composition of the faecal microbiota was analysed by fluorescence in situ hybridization and temporal temperature gradient gel electrophoresis. Degradation of placebo capsules was detected, with increased levels of polysaccharide‐degrading bacteria. Xanthan : gellan gum capsules were shown to reduce the Bifidobacterium population and increase the Clostridium histolyticum group levels, but not jamilan : gellan gum capsules. Only after administration of jamilan : gellan gum‐probiotic capsules was detected a significant increase in Lactobacillus‐Enterococcus group levels compared to controls (capsules and probiotic) as well as two bands were identified as Lact. plantarum in two profiles of ileum samples.

Conclusions

Exopolysaccharides constitute an interesting approach for colon‐targeted delivery of probiotics, where jamilan : gellan gum capsules present better biocompatibility and promising results as a probiotic carrier.

Significance and Impact of Study

This study introduces and highlights the importance of biological compatibility in the encapsulating material election, as they can modulate the gut microbiota by themselves, and the use of bacterial exopolysaccharides as a powerful source of new targeted‐delivery coating material.     

potamolejeunea Holtii (Spruce) nov.comb.

Abstract

Bryophytes usually have anti-feeding properties to defend against microbial and herbivore attack; however, the consumption of Haplocladium microphyllum (Hedw.) Broth. capsules by Agrotis sp. larvae is rather common in Shanghai in the spring. To test whether H. microphyllum is the only moss eaten, and why the gametophytes of H. microphyllum are not eaten, a series of quantitative experiments were carried out to understand the feeding habits of Agrotis larvae on the given moss materials at three growth stages of larval life. The results show that the larvae can feed on the capsules of six moss species to different degrees: Funaria hygrometrica Hedw., H. microphyllum, Physcomitrium sphaericum (C.F.Ludw. ex Schkuhr) Brid., Trematodon longicollis Michx., Ditrichum pallidum (Hedw.) Hampe, and Pogonatum inflexum (Lindb.) Sande Lac. The capsules of the first four species were grazed heavily by the larvae, compared with limited consumption of the latter two (D. pallidum and P. inflexum), which even induced a high mortality rate among the larvae. With the growth of the larval instar, the daily demand for moss capsules by the larvae increases gradually. The lipid content of the capsules may play an important role when the larva selects its feeding target.