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At least 1,243 records · Page 69Linked to original sources
PubMed2000

One step purification of peanut phospholipase D by precipitation with alginate.

A simple titrimetric assay with soybean lecithin has been used for screening phospholipase D activity from some plant sources, viz. peanut, wheat germ, cabbage and carrot. The enzyme from peanut has been purified by binding to alginate which is a water soluble polymer. The purification consisted of co-precipitation of enzyme with alginate upon addition of 0.06 M Ca++. The enzyme was eluted from the polymer using 0.2 M sodium chloride. The activity recovery was 61% with 34 fold purification.

Alginates↗
PubMed1992

Calcium alginate beads as core carriers of 5-aminosalicylic acid.

The utilization of calcium alginate beads as core carriers for delayed dissolution followed by burst release as a potential method of intestinal site specific drug delivery was investigated. 5-Aminosalicylic acid was spray-coated on dried calcium alginate beads and then coated with different percentages of enteric coating polymer and/or sustained-release polymer. Beads coated with more than 6% (w/w) methacrylic copolymer plastisized with dibutyl sebacate and triethyl citrate resisted release in 2-hr acid fluid challenge and allowed immediate dissolution upon transfer to simulated intestinal fluid. With 6% (w/w) methacrylic copolymer on top of 4% (w/w) ethylcellulose polymer, the major portion of drug did not release in 2 hr of acid treatment or the next 3 hr of simulated intestinal fluid treatment. This dosage form provides the possibility to deliver drug to the lower intestinal tract with minimal early release, followed by sustained release in the colon.

Alginates↗
PubMed2002

Encapsulation of recombinant adenovirus into alginate microspheres circumvents vector-specific immune response.

Pre-existing immunity against adenoviruses may compromise the efficacy of adenoviral vectors for vaccination and gene therapy. The purpose of this study was to determine whether encapsulation of adenovirus recombinants into biodegradable alginate microparticles could circumvent the vector-specific immune response. Mice were immunized either intranasally (i.n.) or intraperitoneally (i.p.) with human adenovirus type 5 (HAd5), resulting in the development of virus-specific antibodies. Immunized and nai;ve mice were inoculated with AdCA36lacZ (an E1-deleted HAd5 recombinant containing the bacterial beta-galactosidase (LacZ) gene), encapsulated (E) into alginate microparticles, or nonencapsulated (NE) ie, as a virus suspension. LacZ expression in animals immunized once (1x) or twice (2x) with HAd5 and subsequently inoculated with NE-AdCA36lacZ (NE-Z) was significantly (P<0.001) reduced compared to those levels observed in NE-Z inoculated nai;ve mice, suggesting that the immune response against the vector adversely affected transgene expression. In contrast, there was only slight reduction (P>0.05) in LacZ expression in mice immunized 1x or 2x with HAd5 that were subsequently inoculated with E-AdCA36lacZ (E-Z) compared to those levels obtained in E-Z inoculated nai;ve animals. Similar results were obtained with i.n. or i.p. inoculated animals. These results indicate that microencapsulation of recombinant adenovirus effectively circumvented the vector-specific immune response.

Adenoviridae↗
PubMed2003

A strategic study using mutant-strain entrapment in calcium alginate for the production of Saccharomyces cerevisiae cells with high invertase activity.

Entrapped cells and entrapped cells grown inside of a calcium alginate matrix as well as free cells have been investigated using Saccharomyces cerevisiae mutant strains with regard to their pattern of growth and invertase activity. The repression of invertase by glucose and glucose-consumption ability were considered in the selection process of the mutants. Efficient sucrose bioconversion due to high invertase activity was obtained when entrapped mutant strain Q6R2 cells were grown within calcium alginate gel beads using sucrose plus glucose as the carbon source. Under these conditions, 1 mg (dry weight) of entrapped cells is able to produce 20 micromol of inverted sugar in 3 min (the maximum activity obtained was 20 units x mg(-1)). The experiments were carried out for 6 months without appreciable loss of either bead integrity or invertase activity. The biocatalyst was also stored at 4 degrees C for 6 months without appreciable loss of the invertase activity. This work shows that entrapped yeast cells with a weak ability to consume sugar may be used to produce inverted sugar.

Alginates↗
PubMed2004

Modified alginate matrices for the immobilization of bioactive agents.

Bioactive agents (catalase - an enzyme, and nisin - a bacteriocin) were covalently immobilized on alginate activated with sodium periodate (oxidatively converting 2,3-dihydroxy groups into dialdehyde residues), followed or preceded by ionotropic gelation. For the same protein coupling yield, the retained enzyme activity of the immobilized enzyme (ImE) can be markedly increased by diminishing the bead diameter, a phenomenon that illustrates the role of substrate/product diffusion through the bead gel layer. When the amount of enzyme introduced for coupling was about 15 mg/100 mg of support and the bead diameter was about 100 microm, a high retained specific activity (95-98%) was obtained. Diffusion phenomena can be markedly decreased by enzyme immobilization on the surface of microbeads (obtained by gelation of activated alginate prior to immobilization). In this case, the retained activity was approx. 75% of that of the free enzyme. A slightly higher K (m) value of ImE suggested that the enzyme-substrate affinity was almost maintained. The profiles of ImE activities at various pH values, at various temperatures and when undergoing proteolysis showed a overall higher stability for the immbolized than that for the free enzyme. Nisin immobilized on the microbead surface, when submitted to proteolysis, conserved its bacteriocin activity, strongly inhibiting the growth of Lactobacillus sake when subjected to an agar spot test, whereas free nisin totally lost its activity.

Adsorption↗
PubMed2004

Overall volumetric oxygen transfer coefficient in an aerated bench-top stirred fermenter in aqueous dispersions of sodium alginate.

The dynamic gassing-out method was used to assess the effect of stirrer speed, air flow rate and alginate concentration on the overall volumetric oxygen mass transfer coefficient (K(L)a) in a bench-top stirred fermenter equipped with two Rushton-type turbines. As the alginate concentration in the medium was increased from 0 to 2% (w/v), the liquid film time constant (tau(F)) increased from approx. 30 to 300% of the electrode time constant (tau(E)), clearly showing that the liquid-probe diffusion film affected the oxygen-electrode response. By accounting for delayed probe response, liquid film time constant and gas residence time, the corrected values of K(L)a were empirically correlated with the above independent variables as converted into gassed power input and aeration number per unit liquid volume and the effective liquid/water shear viscosity ratio. Despite an average error of 19%, such a correlation might be useful to adjust the oxygen transfer in bench-top stirred reactors during viscous aerobic fermentations.

Alginates↗
PubMed2006

A simple non-destructive method for the fixation and immunostaining of cultured cells encapsulated in alginate.

In the present paper we report a simple non-destructive method for the analysis of cells and their proteins encapsulated in alginate. Investigations with a co-culture of the human hepatoma cell line HepG2 and primary human fibroblasts are reported. We studied the expression of three proteins, namely intracellular alpha-foetoprotein and extracellular albumin, expressed by HepG2, and membrane-bound CD90 (Thy-1), expressed by the fibroblasts. Fluorimetric and colorimetric staining methods were evaluated and compared. We optimized the method by investigating a range of concentrations of the washing buffers. The technique protected the capsule and cell structures and enabled the investigated proteins to be clearly and reliably visualized within the encapsulated co-culture in their respective intracellular, membrane-bound and extracellular domains. These results demonstrate that this simple method is suitable for the non-destructive analysis of protein expression by cells encapsulated in alginate.

Albumins↗
PubMed2004

Biochemical analysis of the processive mechanism for epimerization of alginate by mannuronan C-5 epimerase AlgE4.

The enzymes mannuronan C-5 epimerases catalyse the in-chain epimerisation of beta-D-mannuronic acid to alpha-L-guluronic acid in the last step of alginate biosynthesis. The recombinant C-5 epimerase AlgE4, encoded by the soil bacteria Azotobacter vinelandii and expressed in Escherichia coli, exhibits a non-random mode of action when acting on mannuronan and alginates of various monomeric compositions. The observed residue sequence has been suggested previously to be due to either a preferred attack or a processive mode of action. Based on methodologies involving specific degrading enzymes, NMR, electrospray ionisation mass spectrometry and capillary electrophoresis we show here that on average 10 residues are epimerised for each enzyme-substrate encounter. A subsite model for the enzyme is analysed by the same methodology using native and 13C-labelled mannuronan oligomers as substrate for the AlgE4 epimerase. A hexameric oligomer is the minimum size to accommodate activity. For hexa-, hepta- and octameric substrates the third M residue from the non-reducing end is epimerised first.

Alginates↗
PubMed2000

Drug utilization evaluation of nonprescription H2-receptor antagonists and alginate-containing preparations for dyspepsia.

AIMS: To evaluate the use, efficacy and adverse effects of nonprescription H2-receptor antagonists and alginate-containing preparations obtained from community pharmacies. METHODS: Questionnaires were distributed to customers from 39 pharmacies in Scotland and Wales. RESULTS: Of 767 customers recruited, 608 (79.3%) returned an initial questionnaire and 472 (61.5%) customers a second questionnaire. The vast majority of respondents (424, 69.7%) had suffered their symptoms on three or more occasions and 369 (60.7%) had previously tried medicines to relieve their symptoms. Referrals to a doctor were less frequent than recommended in guidelines and few of those who were referred actually saw a doctor. Over a quarter of those returning the second questionnaire claimed to be taking more than one product simultaneously for symptom control. Eight customers who were taking prescribed ulcer-healing drugs obtained H2-receptor antagonists. The majority of respondents (355/472, 75. 2%) obtained some or complete symptom relief using the product obtained and 369/472 (78.2%) were completely satisfied with their product. H2-receptor antagonists were more likely to produce complete relief of symptoms than alginate-containing preparations (P < 0.05). Only 14 respondents (3.0%) reported side-effects from the product used which were mostly gastro-intestinal. CONCLUSIONS: The study demonstrated that drug utilization studies are feasible to carry out in a community pharmacy setting. While the results support published evidence of the efficacy and minimal toxicity of these products, they also highlight the possibility of H2-receptor antagonists being used outwith their licenced indications.

Adult↗
PubMed1999

Pseudomonas aeruginosa AlgZ, a ribbon-helix-helix DNA-binding protein, is essential for alginate synthesis and algD transcriptional activation.

The Pseudomonas aeruginosa algD gene is the first gene of an operon encoding most of the enzymes necessary for biosynthesis of the exopolysaccharide alginate. Transcriptional activation of algD results in the high-level synthesis of alginate, an important P. aeruginosa virulence factor with antiphagocytic and adherence properties. Previously, we have identified a protein(s), AlgZ, expressed in mucoid P. aeruginosa CF isolates that specifically bound to sequences located 280 bp upstream of the algD promoter. Mutagenesis of the AlgZ DNA binding site and transcription assays were used to show that AlgZ was an activator of algD transcription. In the current study, the monomeric size of AlgZ was estimated to be between 6 kDa and 15 kDa by electroelution of a protein preparation from an SDS-PAGE gel and analysis of the fractions via protein staining and electrophoretic mobility shift assays. A biochemical enrichment procedure, resulting in a 130-fold enrichment for AlgZ, was devised, the protein identified and a partial amino-terminal sequence obtained. Using the P. aeruginosa Genome Project database, a complete sequence was obtained, and algZ was cloned and expressed in Escherichia coli. Expression of algZ was sufficient for the observed AlgZ DNA binding previously observed from extracts of P. aeruginosa. A protein database search revealed that AlgZ is homologous to the Mnt and Arc repressors of the ribbon-helix-helix family of DNA-binding proteins. An algZ deletion mutant was constructed in the mucoid CF isolate FRD1. The resulting strain was non-mucoid and exhibited no detectable algD transcription. As an indirect role in transcription would probably result in some residual algD transcription, these data suggest that AlgZ is an integral activator of algD and support the hypothesis that both AlgZ and the response regulator AlgR are involved in direct contact with RNA polymerase containing the alternative sigma factor, AlgT. The cloning of algZ is a crucial step in determining the mechanism of algD activation.

Alginates↗
PubMed2001

The roles of mucD and alginate in the virulence of Pseudomonas aeruginosa in plants, nematodes and mice.

We are exploiting the broad host range of the human opportunistic pathogen Pseudomonas aeruginosa strain PA14 to elucidate the molecular basis of bacterial virulence in plants, nematodes, insects and mice. In this report, we characterize the role that two PA14 gene products, MucD and AlgD, play in virulence. MucD is orthologous to the Escherichia coli periplasmic protease and chaperone DegP. DegP homologues are known virulence factors that play a protective role in stress responses in various species. AlgD is an enzyme involved in the biosynthesis of the exopolysaccharide alginate, which is hyperinduced in mucD mutants. A PA14 mucD mutant was significantly impaired in its ability to cause disease in Arabidopsis thaliana and mice and to kill the nematode Caenorhabditis elegans. Moreover, MucD was found to be required for the production of an extracellular toxin involved in C. elegans killing. In contrast, a PA14 algD mutant was not impaired in virulence in plants, nematodes or mice. A mucDalgD double mutant had the same phenotype as the mucD single mutant in the plant and nematode pathogenesis models. However, the mucDalgD double mutant was synergistically reduced in virulence in mice, suggesting that alginate can partially compensate for the loss of MucD function in mouse pathogenesis.

Alginates↗
PubMed1997

Incorporation of nisin in micro-particles of calcium alginate.

Nisin was successfully incorporated into a matrix of calcium alginate and ground into micro-particles smaller than 150 microns. Formation of micro-particles and incorporation of nisin was verified by scanning electron microscopy and by the reduction in the inactivation of nisin activity with proteolytic enzymes. Incorporation efficiency was 87-93% and the nisin in the alginate-incorporated form was 100% active against an indicator culture of Lactobacillus curvatus both in MRS broth and reconstituted skim milk.

Alginates↗
PubMed1994

Large-scale production of Ba(2+)-alginate-coated islets of Langerhans for immunoisolation.

Islet xenografts immunisolated in alginate capsules have been proposed by many groups for clinical islet transplantation. However, diffusion limitations and the total volume of microcapsules required for transplantation are, among other things, factors which have so far prevented successful clinical application. In this study, these problems have been overcome by immobilisation of rat and porcine islets in a Ba(2+)-cross-linked alignate matrix using an air jet droplet generator technique in combination with subsequent density-gradient purification. This procedure leads to high yields of islets coated with a tailored, thin layer of cross-linked alginate which protects the islets against cytotoxic compounds present in human plasma. The recovery of encapsulated rat and porcine islets was about 70%. Empty capsules were nearly completely removed. Using this technique, the total volume of the resulting microcapsules increased only by a factor of about 1.5 compared to the volume of free islets. The technique can be used for large-scale production of coated islets.

Alginates↗
PubMed1993

Long-term function of porcine islets and single cells embedded in barium-alginate matrix.

The influence of alginate-embedding on the maintenance of functioning and morphological integrity in long-term culture of isolated porcine islets and islet cells was studied. Free-floating islets and islet cells served for control. Function was tested after the 1st, 2nd and 4th week. Basal and glucose-stimulated insulin secretion of embedded islets decreased slightly, but significantly after the first week (from 4.39 +/- 0.64 to 2.87 +/- 0.47 at normal and from 11.96 +/- 1.44 to 4.76 +/- 0.78 microU pro 24 h pro islet at elevated glucose concentration, p < 0.05 and < 0.01, resp.) and remained unchanged thereafter. Glucose-stimulation resulted in significant increases in insulin secretion at all three testings (p < 0.001, < 0.01 and < 0.01). Single cells in alginate matrix had even more stable insulin secretion throughout the whole cultivation with significant increases to glucose challenge (p < 0.01, < 0.01 and < 0.05). In contrast, insulin secretion of free-floating islet cells decreased from 5.70 +/- 1.19 to 2.04 +/- 0.64 and to 1.05 +/- 0.33 at basal conditions (p < 0.01 and < 0.05) and from 11.39 +/- 1.87 to 2.76 +/- 0.76 and to 2.15 +/- 0.71 microU pro 24 h pro islet under stimulation (p < 0.01 and not sign). In addition, the secretory response to glucose challenge was significant only at the first testing (p < 0.05). Non-embedded islets could be tested only at the first week since after this time they dissociated to single cells. Embedded islets and single cells showed intact morphology after four weeks with trypan blue (TPB) positivity of less than 5%.(ABSTRACT TRUNCATED AT 250 WORDS)

Alginates↗
PubMedDate not supplied

Lipoinsulin encapsulated alginate-chitosan capsules: intestinal delivery in diabetic rats.

An oral formulation based on liposome encapsulated alginate-chitosan gel capsules was developed for insulin delivery for the treatment of diabetes. Liposome encapsulation helped to increase the encapsulation efficiency of insulin in alginate-chitosan capsules. This formulation delivers insulin in the neutral environment of the intestine, by-passing the acidic media in the stomach, with increased drug absorption and bioavailability. Oral administration of this formulation was found to reduce blood glucose levels when tested in diabetic rats.

Administration, Oral↗
PubMed2004

Alginate microparticles for Polymyxin B Peyer's patches uptake: microparticles for antibiotic oral administration.

Microparticles with size less than 3 microm, able to be taken up by M cell of Peyer's patches for the drug delivery to the Gut Associated Limphoid Tissue (GALT), were developed in order to improve oral bioavailability of Polymyxin B (PMB). Less than 3 microm alginate microparticles resistant to gastro-intestinal media were prepared by spray-drying technique and cross-linking by calcium ions and chitosan. The cross-linked microparticles were evaluated for PMB content by spectrophotometric method, alginate/PMB interaction by rheological study, cross-linking degree by EDS analysis and PMB activity by microbiological assay. By modulating the polymer cross-linking degree, cationic PMB interacted on alginate chains leading to a proper PMB loading as well as antibiotic retention in gastric environment and sustained delivery in intestinal fluid. Moreover, the procedure resulted suitable for PMB biological activity preservation.

Administration, Oral↗
PubMed2006

Formulation and statistical optimization of novel double-incorporated PLA-PLGA microparticles within an alginate-pectinate platform for the delivery of nicotine.

The application of nicotine in the research phase of Alzheimer's disease (AD) treatment has shown promise. The present study was aimed at understanding the incorporation of nicotine into poly-lactic acid (PLA) and poly(D,L-lactic-co-glycolic) acid (PLGA) microparticles which were then re-incorporated into a cross-linked zinc-alginate-pectinate polyspheric multi-particulate system to be employed as a possible brain implant for the treatment of AD. The Box-Behnken design was employed to prepare 15 PLA-PLGA formulations, which were tested for their drug incorporation efficiency and release potential and subsequently optimized using multiple regression and an artificial neural network generalized feed-forward model. Based on the rapid burst effect from the microparticles, further incorporation was conducted in a zinc-alginate-pectinate system using ionotropic gelation. Although double incorporation continued to provide a burst effect, this was followed by a lag period for 7 days and a second phase of drug release.

Alginates↗
PubMed2006

Preparation of stable alginate microcapsules coated with chitosan or polyethyleneimine for extraction of heavy metal ions.

Stable alginate microcapsules in dried form containing bis(2,4,4-trimethylpentyl) monothiophosphinic acid (HA) were prepared by coating of fresh alginate microcapsules with chitosan or polyethyleneimine (PEI). The thickness of coatings was estimated by electron probe microanalysis (EPMA), along with electron microscopy (SEM), as well as comparison of uptake percentage of coated and uncoated hollow capsules. Characterization of microcapsules was carried out by Ag(+) uptake experiments, destructive chemical analyses and thermogravimetric methods (TG and DTA). Chemical stability tests in HNO(3) and NaNO(3) media indicated that the coating with 4-double layer chitosan or mono-layer PEI led to an appreciable enhancement of impermeability in the range of pH > 1 or [Na(+)] < 1 M (mol dm(-3)). Unfortunately, multiple coating causes some extractant losses due to effect of physical stress during the coating; however, PEI-coated microcapsules nearly completely hold their extractant content. Stable extractive microcapsules have an appreciable potential for the selective removal of heavy metal ions.

Alginates↗