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PubMed1995

Myofibrillogenesis in rodent skeletal muscle in vitro: two pathways involving thick filament aggregates.

Thick filament aggregates play an important role in myofibrillogenesis in rodent skeletal muscle in vitro. This ultrastructural study describes these aggregates, shows their involvement in the process of myofibril formation, and correlates their appearance and function with current models of myofibrillogenesis. Initially, following myoblast fusion in normal mouse skeletal muscle in vitro, abundant stress fiber-like structures (SFLS) are found near the periphery of early myotubes. These undergo internal rearrangements, forming subcortical sarcomeres and early myofibrils. However, additional thick filaments are synthesized, and some join appositionally to the nascent myofibrils, increasing their diameter. More interiorly, this thick filament synthesis accelerates, with filaments aligning into aggregates resembling discrete A-bands, usually with M-lines and M-regions. The ends of these 'A-band' aggregates are infiltrated with ribosomes and capped by flocculent material. Ultimately, aggregates are incorporated into preexisting myofibrils or associate end-to-end to form new, parallel myofibrils, the flocculent material forming putative I-bands with diminished Z-lines and few thin filaments. As differentiation continues, Z-lines and thin filaments appear, forming true myofibrils. Dysgenic mouse skeletal muscle develops similarly, but when this non-contractile cell matures (i.e., generates action potentials), filaments and their organization break down. Cloned myogenic rat L5/A10 cells also follow this developmental pattern, but in mature, contracting myotubes, Z-lines remain irregular and thin filaments are reduced. In all three types of muscle developing in vitro, thick filament aggregates are a common and predominant feature and as such appear to constitute an additional or alternate pathway to previously described models of myofibrillogenesis.

Animals↗

PubMed2001

Interaction between Cryptosporidium oocysts and water treatment coagulants.

The electrokinetic properties of gamma-irradiated Cryptosporidium oocysts in the presence of coagulants (ferric chloride and alum) and coagulant aids (DADMAC based cationic polyelectrolytes) have been studied. The zeta potential of the oocysts was unaffected by the addition of ferric chloride at all pH values (3-10) studied. Addition of alum resulted in reversal of the oocysts charge, which suggests that the initial stage in the coagulation process leading to floc formation proceeds via the adsorption of hydrolysed aluminium species. The cationic polyelectrolyte Magnafloc LT35 was adsorbed onto iron flocs at doses of 0.1 mg/L even against an electrostatic barrier. The cationic polyelectrolyte only adsorbed and caused charge reversal at the oocyst surface at around 0.4 mg/L, suggesting a lower affinity for this surface. These results indicate that the oocysts, unlike inorganic colloidal materials such as metal oxides, appear to possess a lower surface density of active or charged sites. The lower density of sites, combined with the rapid precipitation of iron salts, may be responsible for the lack of specific adsorption of either hydroxylated ferric species or primary iron hydroxide particles on the oocysts. Further, this suggests that a process of sweep flocculation, where oocysts are engulfed in flocs during coagulation and floc formation, is the more likely mechanism involved. By comparison, it is likely that the specific interaction of hydrolysed aluminium species with the oocysts surface would result in a stronger link at the oocyst-floc interface and that the flocculation process may initially proceed via charge neutralisation.

Alum Compounds↗

PubMed1975

Cirrhosis associated with partial deficiency of alpha-1-antitrypsin: a clinical and autopsy study.

A 63 year old woman with cryptogenic cirrhosis, ascites, portal hypertension, and intermediate levels of alpha-1-antitrypsin of protease inhibitor SZ phenotype who died of esophageal variceal hemorrhage is described. The partial deficiency of alpha-1-antitrypsin and the diagnosis of cirrhosis were suspected one year prior to death because a needle biopsy of liver showed PAS positive, diastase resistant cytoplasmic bodies within hepatocytes. This report illustrates three previously undescribed features: (1) Heterozygous protease inhibitor SZ phenotype may be associated with coarsely nodular cirrhosis in the older adult. (2) The large intracytoplasmic glycoprotein droplets that are distinctive by light microscopy are probably formalin induced aggregates of submicroscopic flocculent material. (3) In the older patients with aberrant alpha-1-antitrypsin the flocculent material is present not only in the granular endoplasmic reticulum but also in smooth endoplasmic reticulum vesicles and cytolysosomes.

Autopsy↗

PubMed1999

Submicroscopic immunodetection of PrP in the brain of a patient with a new-variant of Creutzfeldt-Jakob disease.

We analyzed the distribution and organization of the pathological prion protein isoform (PrPsc) in the brain of new variant Creutzfeldt-Jakob disease using a sensitive post-embedding immunogold electron microscopy method. On methacrylate semithin sections, silver-PrP staining showed florid plaques, containing microvacuoles. It also revealed scattered granular and perivacuolar deposits. At the electron microscope level, plaque PrP-gold labeling was associated with filaments and flocculent amorphous material sometimes observed inside microvacuoles, considered as degenerative neurites. Outside the plaques, PrP-gold labeling was predominantly found over flocculent amorphous material and the presynaptic domain of synapses. Some lysosome-like organelles seen in the neuron perikaryon, were also found to be PrP-immunoreactive. These results are consistent with the hypothesis that the synapse is a privileged target in prion disease.

Adult↗

PubMed2000

Strategies for maintaining the particle size of peptide DNA condensates following freeze-drying.

The particle size of peptide DNA condensates were studied after freeze-drying and rehydration as a function of sugar excipient, concentration, pH, DNA concentration, and peptide condensing agent. In the absence of an excipient, freeze-dried 50 microg/ml AlkCWK(18) (iodoacetic acid alkylated Cys-Typ-Lys(18)) DNA condensates formed large fibrous flocculates on rehydration. Of the sugars tested as lyoprotectants, sucrose proved most effective at preserving particle size during rehydration. The addition of 5 wt/vol% sucrose preserved a mean particle diameter of less than 50 nm during rehydration of AlkCWK(18) DNA condensates prepared at DNA concentrations up to 200 microg/ml; however, higher DNA concentrations led to the formation of insoluble fibrous flocculates. Substitution of polyethylene glycol (PEG)-CWK(18) as a DNA condensing peptide eliminated the need for sucrose, resulting in peptide DNA condensates that retained particle size when rehydrated in water or normal saline at concentrations up to 5 mg/ml. The results suggest that sucrose functions primarily as a bulking agent during freeze-drying that only preserves the particle size of AlkCWK(18) DNA condensates up to a maximum concentration of 200 microg/ml. Alternatively, the steric layer created on the surface of PEG-CWK(18) DNA condensates provides far more efficient lyoprotection, preserving their particle size at a concentration of 5 mg/ml without a bulking agent.

DNA↗

PubMed2003

Chitosan-coated liposomes: characterization and interaction with leuprolide.

The objective of the present work was to investigate the effect of chitosan concentration and lipid type on the characteristics of chitosan-coated liposomes and their interactions with leuprolide. Liposomes from lipid of high purity and low purity were prepared and coated by chitosan. Physical properties, drug entrapment efficiency, and stability upon dilution were respectively compared. Results showed that the particle size increment of liposomes from low purity lipid was larger than that from high purity lipid, indicating a thicker coating layer. The high zeta potential of particles from low purity lipid was thought to play an important role in the resistance to flocculation. As to particles from high purity lipid, polymer bridging caused flocculation at low polymer concentration while at high concentration, the adsorbed chitosan molecule led to steric stabilization. Drug entrapment efficiency decreased as chitosan was added to liposomes, showing the disturbance of bilayers. Upon dilution, the leakage of leuprolide from low purity liposomes was larger than that from high purity liposomes. In conclusion, low purity lipid possessed more negative charge and formed thicker adsorptive layer by stronger electrostatic attraction with chitosan. The interaction between chitosan and the polar head groups on the surface of phospholipid bilayers may interfere with leuprolide entrapped in liposomes and result in the leakage of leuprolide.

Chitin↗

PubMed2001

The fractal nature of Escherichia coli biological flocs.

The fractal structures of Escherichia coli biological flocs were characterised in terms of fractal dimension, which is a measurement of how the bacteria in the flocs occupy space. The dimensional analysis methods, based on power law correlations between floc perimeter, projected area and maximum length, were used to determine the one- and two-dimensional fractal dimensions (D(1) and D(2)) of E. coli flocs formed by flocculation in chitosan solution with a concentration of 10.0 mg chitosan per g dry cell weight (DCW), giving D(1)=1.07+/-0.06 and D(2)=1.70+/-0.08 (+/-S.D.). The three-dimensional fractal dimension (D(3)) of the E. coli flocs was determined by the two-slopes method, using cumulative size distributions of floc length and solid volume, to be 1.99+/-0.08 (+/-S.D.), which is close to the value of D(3)=2.14+/-0.04 (+/-S.D.) measured by the small angle light scattering method. The results demonstrate that E. coli flocs flocculated with chitosan have a fractal nature, as their fractal dimensions D(1), D(2) and D(3) differ from the values of 1, 2 and 3 expected for the spherical Euclidean object, respectively.

Journal Article↗

PubMed1999

Application of co-grinding to formulate a model pMDI suspension.

The objective of this study was to investigate the effect of co-grinding the model drug, triamcinolone acetonide (TAA), with a polymeric surfactant on the in vitro performance of a model pMDI suspension system. The physicochemical properties of TAA after co-grinding with the surfactant, Pluronic F77, were determined by laser light diffraction, helium pycnometry and equilibrium solubility measurements. TAA-surfactant interaction was investigated by differential scanning calorimetry and Fourier transform infrared spectroscopy (FTIR). The suspension characteristics of pMDI formulations prepared with co-ground TAA and surfactant were investigated by determining their in situ sedimentation, rheological profiles and vapor pressure. The performance characteristics of the pMDI formulations were determined by cascade impaction and dose delivery through-the-valve (DDV) measurements. It was found that the presence of Pluronic F77 decreased the solubility of TAA in the propellant medium. Co-grinding TAA particles with Pluronic F77 influenced the particle size distribution, sedimentation and flocculation characteristics of the pMDI suspension formulation. The addition of Pluronic F77 decreased the viscosity of the pMDI formulation. Formulating the suspension pMDI system with co-ground TAA and Pluronic F77 decreased the mass median aerodynamic diameter (MMAD) of the emitted aerosol and increased the percent respirable fraction (%RF). The co-ground TAA and Pluronic F77 pMDI suspension formulation exhibited greater physical stability which was due to the influence of the co-grinding technique on the physicochemical properties of the TAA particle surface and the propellant dispersion medium. The changes induced by co-grinding with Pluronic F77 improved the performance characteristics of a pMDI suspension formulation by stabilizing the suspension and influencing the flocculation characteristics. Co-grinding is a process which may be useful when developing new pMDI systems containing HFA propellants.

Administration, Inhalation↗

PubMed2000

Effective onion vinegar production by a two-step fermentation system.

A two-step fermentation system combining a repeated batch process using a flocculating yeast with a charcoal pellet bioreactor was developed for onion vinegar production. Juice from the red onion R-3, which contained 67.3 g/l total sugar, was smoothly converted to onion alcohol containing 30.6 g/l ethanol by repeated batch operation using the flocculating yeast Saccharomyces cerevisiae strain IR-2. Stable operation was possible and the maximum productivity was about 8.0 g/l/h. A packed bed bioreactor containing charcoal pellets produced from waste mushroom medium was then applied to continuous onion vinegar production from the onion alcohol. Onion vinegar was successfully produced, with a maximum productivity and acetic acid concentration of about 3.3 g/l/h and 37.9 g/l, respectively. The total acetic acid yield calculated from the amount of sugar consumed was 0.86. The two-step system was operated for 50 d and proved to be competitive with other systems in terms of its high productivity, high acetic acid yield, operational stability and low production costs.

Journal Article↗

PubMed1985

Ultrastructural observations on the interaction in vitro between bovine eosinophils and juvenile Fasciola hepatica.

This ultrastructural study has shown that there is a layer of dense flocculent material on the surface of juvenile Fasciola hepatica incubated in vitro with specific bovine antiserum. This material corresponds to the complexes of secreted glycocalyx and bovine antibody previously characterized by fluorescence microscopy. Bovine eosinophils attach closely to those regions of the parasite's surface that are free of flocculent precipitates. This close attachment is followed by degranulation of the eosinophils into the narrow zone between the cells and the parasite. Only in these regions is damage, in the form of vacuolation of the tegument, seen within the juvenile F. hepatica. It is concluded that the inability of bovine eosinophils to kill juvenile F. hepatica in the presence of specific antiserum results from the presence of a protective layer, consisting of antigen/antibody complexes, on the parasite's surface.

Animals↗

PubMedDate not supplied

Versatile and efficient formation of colloids of biopolymer-based polyelectrolyte complexes.

The formation of colloids based on polyelectrolyte complexes (PECs) of biopolymers was investigated through the complexation between two charged polysaccharides, chitosan as polycation, and dextran sulfate as polyanion. The slow dropwise addition of components, generally used for the formation of PECs, allowed to elaborate both cationic or anionic particles with an excess of chitosan or dextran sulfate, respectively. The PEC particles featured a core/shell structure, the hydrophobic core resulting from the segregation of complexed segments whereas excess component in the outer shell ensured the colloidal stabilization against further coagulation. Considering the host/guest concept for the formation of PECs, the influence of the molecular weight of components on particles sizes could be well explained by the chain length ratios of the two polymers. As an irreversible flocculation occurred with a dropwise approach for both cationic and anionic PEC particles when the mixing ratio was close to unity, a more versatile, and simpler to setup, method was designed: the one-shot addition of one solution to the other. Because process of addition is faster than the flocculation, cationic or anionic particles could be elaborated irrespective of the order of addition of the reactant. Characterization of these particles by quasielastic light scattering, electrophoresis, and scanning electron microscopy revealed very similar properties to those obtained by a slow dropwise approach. Critical coagulation concentrations of 0.12 and 0.09 M (with sodium chloride) for cationic and anionic particles evidenced a mostly electrostatic stabilization.

Biopolymers↗

PubMed2005

Copper-glyphosate sorption to microcrystalline gibbsite in the presence of soluble Keggin Al13 polymers.

Among the most reactive yet largely neglected adsorbents of toxicant species occurring in acidic aquatic environments are the epsilon-Keggin Al13 polyoxocations [AlO4Al12(OH)24(H2O)12(7+)], known generally as Al13 polymers. Here, we report on the sorption of Cu(II), a common ingredient of pesticides, and glyphosate {N-[phosphonomethyl]glycine (PMG)}, a widely applied herbicide, to microcrystalline gibbsite [gamma-Al(OH)3] in the presence of soluble Al13 polymers over the pH range 4-7. In the presence of gibbsite and soluble Al13 polymers, dissolved Cu(II) decreased gradually with pH, achieving a minimum at pH 5.5. Between pH 5.5 and 6.0, however, soluble Cu increased markedly, with approximately 80% of the added metal remaining in solution at pH 5.86. At pH > 6.0, soluble Cu once again decreased, becoming undetectable at pH 7. The anomalous Cu solubilization was attributed to a concomitant deprotonation of soluble Al13 polymers, yielding surface OH groups possessing high affinity for Cu(II). Removal of Cu from solution at pH > 6.0 is facilitated by flocculation of the Al13 polymers to which Cu had sorbed. The sorption behavior of the zwitterionic PMG in the presence of gibbsite and Al13 polymers was consistent with this interpretation, there being a dramatic increase in sorbed PMG at pH > 6.0 as the Al13 polymers deprotonated and flocculated. Copper and PMG loss from solution with increasing pH when both adsorptives were added to the gibbsite-Al13 polymer system was broadly similar to what was observed in the PMG-free systems, although small differences were detected in response to varying the order of adsorptive addition. The inclusion of soluble Al polymers in our experiments exposes a fundamental limitation of models based on but a single inorganic adsorbent as a means to predict the behavior of trace metals and xenobiotic organic compounds in natural systems.

Adsorption↗

PubMed1998

Mechanism of the Topotactic Formation of gamma-Zirconium Phosphate Covalently Pillared with Diphosphonate Groups.

The topotactic reaction of gamma-ZrPO(4)[O(2)P(OH)(2)].2H(2)O (gamma-ZrP) with benzenediphosphonic acid was examined in water and in acetone-water mixtures. This reaction was found to take place in water only on the external surface of the microcrystals, and pillared compounds were never obtained, even after very long reaction times. On the contrary, covalently pillared compounds were quickly obtained in acetone-water mixtures. The mechanism of the latter topotactic reaction was investigated by determining the rate of the phosphate groups released and the rate of the benzenediphosphonates taken up by gamma-ZrP over a long time (50 days). These data showed that pillared derivatives of gamma-ZrP can be obtained because colloidal dispersions of exfoliated lamellae are formed in acetone-water mixtures. The diphosphonate group acts initially as a monovalent species, replacing only one dihydrogen phosphate group on the surface of the exfoliated gamma-lamellae. The colloidal and partially derivatized lamellae thus formed can interact with each other by forming polylamellar pillared systems. When the number of pillared lamellae exceeds a given value (usually 5-6), flocculation of the colloidal gamma-ZrP takes place. Topotactic reactions between packets of pillared lamellae may also continue in the flocculated system. Therefore, the average number of the pillared lamellae slowly increases over time.

Journal Article↗

PubMed2005

Separation of single-walled carbon nanotubes on silica gel. Materials morphology and Raman excitation wavelength affect data interpretation.

In this report, procedures are discussed for the enrichment of single-walled carbon nanotube (SWNT) types by simple filtration of the functionalized SWNTs through silica gel. This separation uses nanotube sidewall functionalization employing two different strategies. In the first approach, a crude mixture of metallic and semiconducting SWNTs was heavily functionalized with 4-tert-butylphenyl addends to impart solubility to the entire sample of SWNTs. Two major polarity fractions were rapidly filtered through silica gel, with the solvent being removed in vacuo, heated to 700 degrees C to remove the addends, and analyzed spectroscopically. The second approach uses two different aryldiazonium salts (one with a polar grafting group and one nonpolar), appended selectively onto the different SWNTs by means of titration and monitoring by UV analysis throughout the functionalization process. The different addends accentuate the polarity differences between the band-gap-based types permitting their partial separation on silica gel. Thermal treatment regenerated pristine SWNTs in enriched fractions. The processed samples were analyzed and characterized by Raman spectroscopy. A controlled functionalization method using 4-fluorophenyl and 4-iodophenyl addends was performed, and XPS analyses yielded data on the degree of functionalization needed to affect the van Hove singularities in the UV/vis/NIR spectra. Finally, we demonstrate that relative peak intensity changes in Raman spectra can be caused by morphological changes in SWNT bundling based on differing flocculation or deposition methods. Therefore a misleading impression of separations can result, underscoring the care needed in assessing efficacies in SWNT enrichment and the prerequisite use of multiple excitation wavelengths and similar flocculation or deposition methods in comparative analyses.

Journal Article↗

PubMed2004

Influence of polysaccharides on the rate of coalescence in oil-in-water emulsions formed with highly hydrolyzed whey proteins.

The objective of this study was to examine the effects of added xanthan gum, guar gum, or kappa-carrageenan on the formation and properties of emulsions (4 wt % corn oil) formed with an extensively hydrolyzed commercial whey protein (WPH) product under a range of conditions. The rate of coalescence was calculated on the basis of the changes in the droplet size of emulsions during storage of the emulsions at 20 degrees C. Compared with the emulsion made without the addition of polysaccharides, the rate of creaming and coalescence in emulsions containing xanthan gum, guar gum, or kappa-carrageenan was markedly enhanced with increasing concentration of polysaccharides during storage for up to 7 days. At a given concentration, the rate of coalescence was highest in the emulsions containing guar gum, whereas it was lowest in the emulsions containing kappa-carrageenan. All emulsions containing xanthan gum, guar gum, or kappa-carrageenan showed flocculation of oil droplets by a depletion mechanism. This flocculation was considered to enhance the coalescence of oil droplets. The different rates of coalescence could be explained on the basis of the strength of the depletion potential, which was dependent on the molecular weight and the radius of gyration of the polysaccharides.

Carrageenan↗

PubMed2004

Stabilization of oil-in-water emulsions by cod protein extracts.

The ability of two protein fractions extracted from cod to form and stabilize oil-in-water emulsions was examined: a high salt extracted fraction (HSE protein) and a pH 3 acid extracted fraction (AE protein). Both fractions consisted of a complex mixture of different proteins, with the predominant one being myosin (200 kDa). The two protein fractions were used to prepare 5 wt % corn oil-in-water emulsions at ambient temperature (pH 3.0, 10 mM citrate-imidazole buffer). Emulsions with relatively small mean droplet diameters (d(3,2) < 1 microm) and good creaming stability (> 9 days) could be produced at protein concentrations > or =0.2 wt % for both fractions. The isoelectric point of droplets stabilized by both protein fractions was pH approximately 5. The emulsions were stable to droplet flocculation and creaming at relatively low pH (< or =4) and NaCl concentrations (< or =150 mM) when stored at room temperature. In the absence of salt, the emulsions were also stable to thermal treatment (30-90 degrees C for 30 min), but in the presence of 100 mM NaCl droplet flocculation and creaming were observed in some of the emulsions, particularly those stabilized by the AE fraction. The results suggest that protein fractions extracted from cod can be used as emulsifiers to form and stabilize food emulsions.

Animals↗

PubMed2004

Influence of pH and iota-carrageenan concentration on physicochemical properties and stability of beta-lactoglobulin-stabilized oil-in-water emulsions.

The influence of pH and iota-carrageenan concentration on the properties of beta-lactoglobulin (beta-Lg)-stabilized oil-in-water emulsions was investigated by measuring the particle charge, particle size distribution, and creaming stability. Emulsions containing droplets stabilized by beta-Lg were produced by homogenization, and then, iota-carrageenan was added. At pH 3, the droplet charge did not change for iota-carrageenan concentrations <or=0.1 wt % but decreased rapidly at high concentrations, while the mean particle diameter increased slightly as the iota-carrageenan concentration was increased. These results suggest that the interaction between iota-carrageenan and beta-Lg was weak at pH 3 probably because some sulfate groups were protonated (pK(a) = 2). At pH 4 and pH 5, the droplet charge decreased dramatically as the iota-carrageenan concentration was increased from 0 to 0.15 wt %, but droplet aggregation and creaming occurred in the emulsions, indicating that interfacial complexes between iota-carrageenan and beta-Lg could not stabilize the emulsions, probably due to bridging flocculation. At pH 6, the droplet charge in the primary emulsions was negative and became more negative as the iota-carrageenan concentration was increased. The mean particle diameter was relatively small at all iota-carrageenan concentrations, and emulsions were stable to creaming after 1 week of storage. We propose that carrageenan adsorbed to the droplet surfaces and increased the electrostatic repulsion between droplets. At pH 7 and pH 8, the droplet charge did not change as the iota-carrageenan concentration was increased, but these emulsions became unstable to creaming above a critical carrageenan concentration, which was attributed to depletion flocculation.

Carrageenan↗

PubMed2005

Interactions of high methoxyl pectin with whey proteins at oil/water interfaces at acid pH.

The interactions between whey protein isolate (WPI) and high methoxyl pectin (HMP) at pH 3.5 were investigated in situ using ultrasound (US) and diffusing wave spectroscopy (DWS). HMP was added to 10% oil-in-water emulsions containing 1% WPI. At neutral pH, no protein-pectin interactions were observed as both molecules are negatively charged, while at pH 3.5 bridging flocculation occurred via electrostatic interactions. Four different stages were distinguished during the addition of HMP in WPI-stabilized emulsions at pH 3.5. At a concentration below a critical value, no interactions were observed. At concentrations >0.02% HMP, a change in the l factor indicated a change in the ordering of the emulsion droplets, influenced by long-range interactions. At higher concentrations (in the range between 0.04 and 0.06% HMP), attenuation showed significant changes in the surface of the oil droplets, changes which affected the droplet-droplet interactions. At pectin concentrations >0.05%, attenuation of sound and 1/l* decreased, while velocity of sound and particle size increased, as a result of bridging flocculation. These results demonstrated for the first time that methods such as US and DWS combined permit the observation of the early stages of the interactions between two biopolymers at the interface. This is significant in light of increasing efforts in engineering complex interfacial layers.

Chemical Phenomena↗