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PubMed1993

Protection of islets of Langerhans from antibodies by microencapsulation with alginate-poly-L-lysine membranes.

Microencapsulation of islets has been proposed to prevent their immune destruction following transplantation. An indirect immunofluorescence technique has been developed and used to study the permeability of the alginate-poly-L-lysine microcapsules to antibodies. Wistar rat islets were incubated with the R2D6 monoclonal mouse IgM antibody against rat islets, microencapsulated, and incubated with fluorescein-labeled goat IgG antibodies against mouse IgG and IgM. For the negative controls, the first antibody was omitted or both antibodies were omitted. The positive controls included islets incubated with both antibodies before they were encapsulated. Our study demonstrated that the alginate-poly-L-lysine membranes are not permeable to IgG when poly-L-lysine of molecular weights ranging from 21,000 to 390,000 are used. This simple immunofluorescence technique demonstrated the nonpermeability of the microcapsules to IgG, and could be useful for the initial evaluation of new types of membranes.

Alginates↗

PubMed1993

Macrophage-generated nitric oxide as cytotoxic factor in destruction of alginate-encapsulated islets. Protection by arginine analogs and/or coencapsulated erythrocytes.

Isolated rat islets were microencapsulated in alginate beads of about 1.5 mm in diameter. These were cocultured with activated or resident peritoneal macrophages of syngeneic rats for 24 hr. Examination of the encapsulated islets by transmission electron microscopy showed that the islets were lysed by activated (80.0 +/- 12.8% of islets lysed), but not by resident, macrophages (17.5 +/- 12.2% lysis) despite encapsulation. Islet lysis was inhibited in a concentration-dependent manner by a specific nitric oxide-synthase inhibitor (0.5 mM NG-methyl-L-arginine: 5.9 +/- 3.9% lysis) in an L-arginine-reversible manner (0.5 mM NG-methyl-L-arginine + 10 mM L-arginine: 55.1 +/- 16.6% lysis). Incubation of encapsulated islets with 3 different nitric oxide-generating compounds also resulted in a concentration-dependent islet lysis. Coencapsulation of autologous erythrocytes was found to be an effective and easy way of protection from macrophage-mediated lysis. Protection was dependent upon the number of erythrocytes coencapsulated. This in vitro study demonstrates that nitric oxide secreted by activated macrophages is able to destroy islets despite encapsulation in alginate, and that both, inhibition of nitric oxide formation using enzyme inhibitors and scavenging of nitric oxide once formed exploiting the hemoglobin of autologous erythrocytes, protect encapsulated islets from destruction.

Alginates↗

PubMed2002

Synthesis of a Kunitz-type serine proteinase inhibitory protein that shares homology with bovine pancreatic trypsin inhibitor by ovine intervertebral disc cells in serum-free alginate bead culture.

The objective of this study was to determine whether disc cells could be cultured under serum-free conditions and whether they synthesized bovine pancreatic trypsin inhibitor (BPTI)-like serine proteinase inhibitory proteins (SPIs) previously demonstrated for ovine chondrocytes. Intervertebral discs from 1- to 2-year-old merino wether sheep were dissected into the annulus fibrosus and nucleus pulposus, and the cells isolated by sequential enzymatic digestion. The cells were grown encapsulated in calcium alginate microspheres under serum-free conditions for 10 days. They remained more than 92% viable as assessed using the vital fluorescent dyes chloromethyl fluorescein diacetate and ethidium homodimer-1 to delineate live/dead cells, respectively. Western and affinity blotting identified a 12-16-kDa media SPI and an additional 34-36-kDa BPTI-like species in solubilized bead samples. This study has demonstrated ovine disc cells synthesized BPTI-like SPIs in serum-free alginate bead culture similar to chondrocyte SPIs; however, the 58-kDa precursor SPI form was not detected suggesting differences in the endogenous processing of these SPIs.

Alginates↗

PubMedDate not supplied

Calcium alginate dressing-related hypercalcemia.

To date, no adverse effects or contraindications have been observed in association with the use of calcium alginate dressing on an exuding wound. This is the first case in the literature in which it is suggested that using a large quantity of calcium alginate dressing could be harmful. The scenario leading to the complication and suggestions to prevent similar conditions in the future are described.

Aged, 80 and over↗

PubMed1980

Alginate synthesis in mucoid Pseudomonas aeruginosa: a chromosomal locus involved in control.

Mucoid variants of Pseudomonas aeruginosa isolated in vitro or in vivo could be classified into two phenotypic groups based on whether alginate was produced on a chemically defined medium. Mucoid strains yielded lowe recombination frequencies than the non-mucoid parent when used as donors in FP2-mediated plate matings. The mucoid characteristic (muc) was co-inherited by a proportion of recombinants selected for the inheritance of chromosomal markers his-5075+ or cys-5605+. The results of further experiments using either a mucoid recipient or a mucoid donor carrying plasmid R68.45 suggested that the control of alginate production in P. aeruginosa involves at least one chromosomal locus.

Alginates↗

PubMed1981

Isolation of alginate-producing mutants of Pseudomonas fluorescens, Pseudomonas putida and Pseudomonas mendocina.

Spontaneous alginate-producing (muc) variants were isolated from strains of Pseudomonas fluorescens, P. putida and P. mendocina at a frequency of 1 in 10(8) by selecting for carbenicillin resistance. The infrared spectrum of the bacterial exopolysaccharide was typical of an acetylated alginate similar to that previously described in Azotobacter vinelandii and in mucoid variants of P. aeruginosa. Mucoid variants were not isolated from P. stutzeri, P. pseudoalcaligenes, P. testosteroni, P. diminuta, P. acidovorans, P. cepacia or P. maltophilia.

Alginates↗

PubMed1997

A novel gene, algK, from the alginate biosynthesis cluster of Pseudomonas aeruginosa.

Colonization of the cystic fibrosis lung by Pseudomonas aeruginosa is greatly facilitated by the production of an exopolysaccharide called alginate. Many of the enzymes involved in alginate biosynthesis are clustered in an operon at 34 min on the P. aeruginosa chromosome. This paper reports the nucleotide sequence of a previously uncharacterized gene, algK, which lies between the alg44 and algE genes of the operon. DNA sequencing data for algK predicted a protein product of approximately 52.5 kDa which contains a putative 27 amino acid N-terminal signal sequence and a consensus cleavage and lipid attachment site for signal peptidase II. Expression of algK using either T7 or tac promoter expression systems, in vivo labelling studies with [35S]methionine, indicated that algK encodes a polypeptide of approximately 53 kDa which is processed to a mature protein of approximately 50 kDa when expressed in Escherichia coli or P. aeruginosa, in agreement with the nucleotide sequence analysis. Results from an algK-beta-lactamase fusion survey support this interpretation and also provide evidence that mature AlgK is entirely periplasmic and is probably membrane-anchored.

Alginates↗

PubMed2002

Lysophosphatidic acid inhibition of the accumulation of Pseudomonas aeruginosa PAO1 alginate, pyoverdin, elastase and LasA.

The pathogenesis of Pseudomonas aeruginosa is at least partially attributable to its ability to synthesize and secrete the siderophore pyoverdin and the two zinc metalloproteases elastase and LasA, and its ability to form biofilms in which bacterial cells are embedded in an alginate matrix. In the present study, a lysophospholipid, 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphate [also called monopalmitoylphosphatidic acid (MPPA)], which accumulates in inflammatory exudates, was shown to inhibit the extracellular accumulation of P. aeruginosa PAO1 alginate, elastase, LasA protease and the siderophore pyoverdin. MPPA also inhibited biofilm formation. The inhibitory effects of MPPA occur independently of rpoS expression and without affecting the accumulation of the autoinducers N-(3-oxododecanoyl) homoserine lactone and N-butyryl-L-homoserine lactone, and may be due, at least in part, to the ability of MPPA to bind divalent cations.

Alginates↗

PubMed1993

Relapse rate of patients after healing of oesophagitis--a prospective study of alginate as self-care treatment for 6 months. French Co-operative Study Group.

In order to estimate the rate of symptomatic relapse and to identify factors associated with this relapse, 1030 patients were included in a multicentre study. All patients included had clinically and endoscopically healed oesophagitis (grade I (erosion, 57%), grade II (confluent erosion, 33%) or grade III (circular erosion, 9%)) after treatment with an H2-blocker or omeprazole. Patients were given conservative advice and were told to take sodium alginate only in response to pain. Relapse was defined as pain justifying more than eight sodium alginate sachets over 48 hours or treatment with another drug, or the need for a new endoscopy. Forty risk factors were recorded initially and patients were seen every 2 months; they filled out diary-cards describing symptoms and drug consumption. Data were available for 883 patients during follow up. The cumulative percentage of patients without relapse at 6 months estimated by Kaplan Meier method was 76 +/- 2% (mean +/- S.E.M.). Among these patients without relapse, 95% took less than two sachets per day. Only two factors were identified as associated with relapse--the grade of oesophagitis (P < 0.007), and the occurrence of stress during follow-up (P < 0.05). The percentage of patients free from relapse at six months was 85 +/- 2% in patients with grade I oesophagitis, 69 +/- 3% in patients with grade II, and 56 +/- 6% in patients with grade III oesophagitis (P < 0.0001).

Adolescent↗

PubMed2005

An evaluation of the antireflux properties of sodium alginate by means of combined multichannel intraluminal impedance and pH-metry.

BACKGROUND: Alginate-based preparations act as mechanical antireflux barrier, which can reduce both acid and non-acid reflux events and limit the proximal migration of oesophageal refluxate. AIM: To evaluate all the above features with a novel technique, multichannel electrical impedance and pH-metry. METHODS: Ten reflux patients underwent stationary impedancemetry and pH-metry after eating a refluxogenic meal. They were studied 1 h in basal conditions and 1 h after taking 10 mL of Gaviscon Advance. In both sessions, measurements were obtained in right lateral and supine decubitus. RESULTS: Alginate preparation was able to decrease significantly (P < 0.05) the number of acid reflux events and the percentage time pH < 4.0 compared with baseline. There was no effect of the drug on non-acid refluxes. Gaviscon Advance was also significantly effective (P < 0.05) in reducing the height of proximal migration of reflux events compared with baseline in both decubitus positions. CONCLUSIONS: Our findings explain how Gaviscon Advance controls acid reflux episodes, limits the proximal migration of refluxed material and thereby decreases symptoms in clinical practice.

Adult↗

PubMed1992

The role of calcium alginate swabs in adenotonsillectomy.

A randomized controlled study using swabs of calcium alginate versus gauze was undertaken to evaluate the haemostatic capabilities and the shortening of operative time. Haemostasis in adenoidectomy was shown to be more effective and the operating time shortened (P = 0.05) by the use of calcium alginate. Haemostasis and operating time for tonsillectomy were not affected.

Adenoidectomy↗

PubMed1993

Identification and characterization of a siderophore regulatory gene (pfrA) of Pseudomonas putida WCS358: homology to the alginate regulatory gene algQ of Pseudomonas aeruginosa.

Genes encoding biosynthesis of pseudobactin 358 (a microbial iron transport agent) and its cognate outer membrane receptor protein, PupA, are transcribed only under iron limitation in plant growth-promoting Pseudomonas putida WCS358. Two cosmid clones were identified from a gene bank of WCS358 DNA which could independently and in an iron-dependent manner activate transcription from a WCS358 siderophore gene promoter in heterologous Pseudomonas strain A225. The functional region of one of the clones was localized by subcloning, transposon Tn3Gus mutagenesis, and DNA sequencing. Genomic transposon insertion mutants in the functional region lost the capacity to activate a siderophore gene promoter fusion transcriptionally; furthermore, these mutants no longer produced pseudobactin 358. The activating region consisted of a single gene designated pfrA (Pseudomonas ferric regulator). The pfrA gene codes for a single polypeptide, PfrA, of approximately 18 kDa, which has 58% identity to AlgQ (also known as AlgR2), a positive regulator involved in transcriptionally regulating alginate biosynthesis in Pseudomonas aeruginosa. Cross-complementation studies between the pfrA gene of P. putida and the algQ gene of P. aeruginosa revealed that pfrA can restore mucoidy (alginate production) in an algQ mutant and that algQ could poorly complement a pfrA genomic mutant. It is concluded that PfrA is involved in the positive regulation of siderophore biosynthetic genes in response to iron limitation; furthermore, pfrA and algQ appeared to be interchangeable between P. putida and P. aeruginosa.

Alginates↗

PubMed2006

Cell wall-inhibitory antibiotics activate the alginate biosynthesis operon in Pseudomonas aeruginosa: Roles of sigma (AlgT) and the AlgW and Prc proteases.

A bioassay was developed to identify stimuli that promote the transcriptional induction of the algD operon for alginate biosynthesis in Pseudomonas aeruginosa. Strain PAO1 carried the algD promoter fused to a chloramphenicol acetyl-transferase cartridge (PalgD-cat), and > 50 compounds were tested for promoting chloramphenicol resistance. Most compounds showing PalgD-cat induction were cell wall-active antibiotics that blocked peptidoglycan synthesis. PalgD-cat induction was blocked by mutations in the genes for sigma22 (algT/algU) or regulators AlgB and AlgR. Anti-sigma factor MucA was the primary regulator of sigma22 activity. A transcriptome analysis using microarrays verified that the algD operon undergoes high induction by D-cycloserine. A similar sigma(E)-RseAB complex in Escherichia coli responds to envelope stress, which requires DegS protease in a regulated intramembrane proteolysis (RIP) cascade to derepress the sigma. Mutant phenotypic studies in P. aeruginosa showed that AlgW (PA4446) is likely to be the DegS functional homologue. A mutation in algW resulted in a complete lack of PalgD-cat induction by D-cycloserine. Overexpression of algW in PAO1 resulted in a mucoid phenotype and alginate production, even in the absence of cell wall stress, suggesting that AlgW protease plays a role in sigma22 activation. In addition, a mutation in gene PA3257 (prc), encoding a Prc-like protease, resulted in poor induction of PalgD-cat by D-cycloserine, suggesting that it also plays a role in the response to cell wall stress.

Alginates↗

PubMed1996

Reduction of Brochothrix thermosphacta on beef surfaces following immobilization of nisin in calcium alginate gels.

Lean and adipose beef carcass tissues inoculated with Brochothrix thermosphacta (BT) (approx. 4.50 log10 cfu cm-2) were left untreated (U) or treated with 100 micrograms ml-1 nisin (N), calcium alginate (A) or 100 micrograms ml-1 nisin immobilized in a calcium alginate gel (AN). Tissue samples were refrigerated after treatments and bacterial populations and nisin activity were determined at 0, 1, 2 and 7 d. U, A and N treatments of lean and adipose tissues did not suppress bacterial growth ( > 6 log10 cfu cm-2 by day 7) while treatments of lean and adipose tissues with AN suppressed bacteria ( > 2.42 log10 cfu cm-2 by day 7). Bacteriocin titres from both tissues were higher in AN vs N samples after the 7 d incubation. This study demonstrates that immobilization of nisin in a gel may be a more effective delivery system of a bacteriocin to the carcass surface than direct application.

Alginates↗

PubMed1986

Synthesis and secretion of protein by hepatocytes entrapped within calcium alginate.

The ability of entrapped hepatocytes to secrete plasma proteins was examined for the purpose of developing a biological artificial liver. Hepatocytes were isolated from adult rat liver by perfusion with collagenase. Isolated hepatocytes were entrapped within calcium alginate. The entrapped cells induced tyrosine aminotransferase (TAT) in the presence of dexamethasone and dibutyryl-cyclic AMP and retained the ability to induce TAT for 7 days. Moreover, entrapped cells could synthesize and secrete a biologically active form of coagulation Factor II, prothrombin. Two plasma proteins, lecithin: cholesterol acyltransferase and cholinesterase, were also secreted into the medium. Thus, hepatocytes within calcium alginate showed liver-specific characteristics, and these activities were almost comparable with those of monolayer-cultured cells.

Alginates↗

PubMed2004

Microencapsulation of parathyroid tissue with photosensitive poly(L-lysine) and short chain alginate-co-MPEG.

Human parathyroid glands were encapsulated using the alginate-PLL system in this study. In order to improve the mechanical strength and the biocompatibility, the microcapsules were fabricated with a three-layer structure that consisted of alginate/photosensitive poly(L-lysine)/short chain alginate-co-MPEG. These modified microcapsules were used for encapsulating human parathyroid tissue. In vitro experiments revealed that microencapsulated parathyroid glands maintained differentiative properties in culture, and the capsular membrane was freely permeable to the human parathyroid hormone. For in vivo experiments, these capsules were transplanted into parathyroidectomized SD-rats. After parathyroidectomy, serum calcium decreased from 2.25 to 1.68 mmol/L and remained in a constantly low concentration until transplantation. Parathyroidectomized SD-rats were normocalcemic after transplant of encapsulated parathyroid tissue. The microcapsules were then explanted at 12 weeks for examination. Histological evaluations of excised transplants revealed that the microcapsules remained intact structurally and were free of cell adhesions. The results demonstrated that human parathyroid tissue microencapsulated by this system retains stability and is functional both in vitro and in vivo. This encapsulating system will have valuable application for endocrine surgery in the future.

Alginates↗

PubMed2005

Proliferation rates of HepG2 cells encapsulated in alginate are increased in a microgravity environment compared with static cultures.

This study investigates the effect of rotary culture compared with static culture on the proliferation, cell viability, synthetic function and detoxificatory capacity of HepG2 cells encapsulated in 1% alginate. Cell viability and alginate bead morphology were maintained in the rotary culture system at day 10, while cell number showed a 4.5-fold increase compared with static culture. Protein production was increased in rotary cultures with a 4.1-fold increase in total albumin and a 4.4-fold increase in alpha1 antitrypsin levels in rotary compared with static culture at day 10. CYP4501A1/2 activity was maintained between the two culture systems. In conclusion, rotary culture increases proliferation rates leading to improved bead packing and a concomitant increase in total protein synthesis, along with maintenance of detoxificatory capacity. This allows a greater level of hepatic function to be expressed in a given volume, offering clear advantages for the design of liver support systems.

Alginates↗

PubMed2006

Initiation of mesenchymal condensation in alginate hollow spheres--a useful model for understanding cartilage repair?

A promising strategy for the regeneration of degenerated cartilage tissue structure in osteoarthritic joints is the use of mesenchymal precursor cells. These cells can be triggered to undergo differentiation into functional active chondrocytes resulting in newly synthesized cartilage. Because chondrogenic differentiation is initiated by the step of mesenchymal condensation in vitro, it is of great interest to fully characterize the first lineage specific step in vitro. Therefore, a modified culture system was developed which mimics the process in vitro and may finally help to identify the key factors that are essential for the induction of chondrogenic differentiation in vivo. Compared to other established 3D culture systems like alginate beads and micromass cultures, the use of alginate hollow spheres bears the advantage to analyze different phases of cell aggregation starting from a single cell suspension of previously isolated and expanded human primary cells of mesenchymal origin.

Alginates↗