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PubMed2001

Physical characteristics of poly(vinyl alcohol) and calcium alginate hydrogels for the immobilization of activated sludge.

Hydrogels based on poly(vinyl alcohol), PVA, and calcium alginate were prepared by a freezing and thawing cycle process and characterized, in terms of the role of the polymer mixture percentage and the number of treatment cycles, on their weight swelling ratio, WSR, gel fraction, and activated sludge entrapment and immobilization. The results show that the morphology of these hydrogels is highly dependent on the PVA-Ca alginate ratio of 5 wt % total polymer content in the initial aqueous solution and that the number of entrapped microorganisms which survive the freezing-thawing procedure is independent of this ratio. For 80/20 PVA-Ca alginate hydrogels, results also show that for up to three freezing and thawing cycles, the WSR, which is in average 24, is not severely affected by the number of the cycles. For the hydrogels with three cycles, the calculated gel fraction for the composite hydrogel is 0.99. Immobilized microorganisms from sedimented activated sludge, constituted by bacteria and fungi, die in high numbers during the freezing and thawing treatment. However, with a proper time of incubation with glucose as carbon source, the population of bacteria is recovered and mainly proliferate inside the hydrogel, attached on top of the fibril network formed by the polymers, while fungi are recovered predominantly on the surface of the spheres.

Alginates↗
PubMed2006

Characterization of the hydrolysis mechanism of polyalternating alginate in weak acid and assignment of the resulting MG-oligosaccharides by NMR spectroscopy and ESI-mass spectrometry.

Alginate with long strictly alternating sequences of mannuronic (M) and guluronic (G) acid residues, F(G) = 0.47 and F(GG) = 0.0, was prepared by incubating mannuronan with the recombinant C-5 epimerase AlgE4. By partial acid hydrolysis of this PolyMG alginate at pH values from 2.8 to 4.5 at 95 degrees C, alpha-L-GulpA-(1-->4)-beta-D-ManpA (G-M) linkages were hydrolyzed far faster than beta-D-ManpA-(1-->4)-alpha-L-GulpA (M-G) linkages in the polymer chain. The ratio of the rates (kG-M/kM-G) decreased with increasing pH. The dominant mechanism for hydrolysis of (1-->4)-linked PolyMG in weak acid was thus proved to be an intramolecular catalysis of glycosidic cleavage of the linkages at C-4 by the undissociated carboxyl groups at C-5 in the respective units. The higher degradation rate of G-M than M-G glycosidic linkages in the polymer chain of MG-alginate at pH 3.5 and 95 degrees C was exploited to make oligomers mainly consisting of M on the nonreducing and G on the reducing end and, thus, a majority of oligomers with an even number of residues. The ratio of the rate constants kG-M/kM-G at this pH was 10.7. The MG-hydrolysate was separated by size exclusion chromatography and the MG oligosaccharide fractions analyzed by electrospray ionization-mass spectrometry together with 1H and 13C NMR spectroscopy. Chemical shifts of MG-oligomers (DP2-DP5) were elucidated by 2D 1H and 13C NMR.

Acids↗
PubMed2006

Removable colored coatings based on calcium alginate hydrogels.

This article describes the creation of a nontoxic, biodegradable coating using calcium alginate and FD&C approved dyes. The coating is robust but is rapidly removed upon treatment with disodium ethylenediamine tetraacetate (EDTA). Dye leaching from calcium alginate films was studied, and it was determined that the efficiency of dye retention is proportional to the degree of cross-linking. Degradation rates were studied on calcium alginate beads serving as a model for a coating. We determined that degradation rates depend on the gel's cross-linking and on the amount of EDTA used. Bead size also influenced the degradation rates; smaller beads degraded faster than larger beads. We show that the coating can be used as an easily removable and environmentally friendly logotype on an artificial turf surface. Applications of these coatings can be extended to food, cosmetic, medicinal, and textile uses and to wherever nontoxic, easily removable colored coating is desired.

Alginates↗
PubMedDate not supplied

A smart bioconjugate of alginate and pectinase with unusual biological activity toward chitosan.

The commercial preparation of pectinase (Pectinex Ultra SP-L) was conjugated to alginate by noncovalent interactions by employing 1% alginate during the conjugation protocol. The optimum "immobilization efficiency" was 0.76. The pH optimum and the thermal stability of the enzyme remained unchanged upon conjugation with alginate. The soluble bioconjugate showed a 3-fold increase in V(max)/K(m) as compared to the free enzyme when the smart biocatalyst was used for chitosan hydrolysis. Time course hydrolysis of chitosan thus showed higher conversion of chitosan into reducing oligosaccharides/sugars. The smart bioconjugate could be reused five times without any detectable loss of chitosanase activity.

Alginates↗
PubMedDate not supplied

Evaluation of microbeads of calcium alginate as a fluidized bed medium for affinity chromatography of Aspergillus niger Pectinase.

Calcium alginate microbeads (212-425 microm) were prepared by spraying 2% (w/v) alginate solution into 1 M CaCl2 solution. The fluidization behavior of these beads was studied, and the bed expansion index and terminal velocity were found to be 4.3 and 1808 cm h(-1), respectively. Residence time distribution curves showed that the dispersion of the protein was much less with these microbeads than with conventionally prepared calcium alginate macrobeads when both kinds of beads were used for chromatography in a fluidized bed format. The fluidized bed of these beads was used for the purification of pectinase from a commercial preparation. The media performed well even with diluted feedstock; 90% activity recovery with 211-fold purification was observed.

Alginates↗
PubMedDate not supplied

Alginate coating of Xenopus laevis embryos.

Xenopus laevis eggs were coated, immediately after squeeze-stripping and fertilization, with a thin layer ( approximately 50 microm) of film based on one of three different types of alginates which varied in their mannuronic/guluronic acid ratio. The alginate was cross-linked with either Ca or Ba ions at three different concentrations. The developmental, survival, and hatching of these embryos and the swelling of their natural jelly coats or hydrocolloid coatings were studied over 7 days, while embryos were maintained in flowing aerated water at a ratio of 85 mL per embryo or at a very diminished ratio of 1.6 mL of sterile or nonsterile MMR solution per embryo. All experiments were conducted in triplicate at 20+/-1 degrees C. Oxygen was monitored continuously. Mineral content was determined in the alginate-jelly coat and within the embryos over time. The coating conferred major advantages when the ratio between the embryos and the surrounding medium was at a minimum under nonsterile conditions, perhaps as a result of the film's resistance to diffusion. In the studied systems, the coating seemed to postpone embryo hatching to a more developed stage. In addition, the coating served as a barrier to microbial contamination and thus improved survival prospects.

Alginates↗
PubMed2006

Controlled release of isoproturon, imidacloprid, and cyromazine from alginate-bentonite-activated carbon formulations.

Different alginate-based systems of isoproturon, imidacloprid, and cyromazine have been investigated in order to obtain controlled release (CR) properties. The basic formulation [sodium alginate (1.50%), pesticide (0.30%), and water] was modified using different amounts of bentonite and activated carbon. The higher values of encapsulation efficiency corresponded to those formulations prepared with higher percentages of activated carbon, showing higher encapsulation efficiency values for isoproturon and imidacloprid than for cyromazine, which has a higher water solubility. The kinetic experiments of imidacloprid/isoproturon release in water have shown us that the release rate is higher in imidacloprid systems than in those prepared with isoproturon. Moreover, it can be deduced that the use of bentonite and/or activated carbon sorbents reduces the release rate of the isoproturon and imidacloprid in comparison with the technical product and with alginate formulation without modifying agents. The highest decrease in release rate corresponds to the formulations prepared with the highest percentage of activated carbon. The water uptake, permeability, and time taken for 50% of the active ingredient to be released into water, T50, were calculated to compare the formulations. On the basis of a parameter of an empirical equation used to fit the pesticide release data, the release of isoproturon and imidacloprid from the various formulations into water is controlled by a diffusion mechanism. The sorption capacity of the sorbents and the permeability of the formulations were the most important factors modulating pesticide release. Finally, a linear correlation of the T50 values and the content of activated carbon in formulations were obtained.

Alginates↗
PubMed2000

Preparation of neoglycoprotein from carp myofibrillar protein and alginate oligosaccharide: improved solubility in low ionic strength medium.

Alginate oligosaccharide (AO) was conjugated with carp myofibrillar protein (Mf) by using the controlled Maillard reaction, and the change in the solubility of Mf in low ionic strength media as affected by the glycosylation was investigated. AO was prepared by degrading sodium alginate using alginate lyase, which was purified from the culture supernatant of Pseudoalteromonas elyakovii. When a lyophilized Mf and AO mixture was incubated at 40 degrees C and 65% relative humidity, the conjugation of AO was confirmed at myosin heavy chain, actin, and tropomyosin. When >30 microg/mg of AO was conjugated to Mf, the protein solubility in a low ionic strength medium was greatly improved without significant loss of available lysine. These results indicate that the conjugation with AO is a superior manner for improving the water solubility of Mf in view of its nutritional aspect.

Alginates↗
PubMedDate not supplied

Radiopaque alginate microcapsules for X-ray visualization and immunoprotection of cellular therapeutics.

Alginate-poly-L-lysine-alginate (APA) microcapsules have been explored as vehicles for therapeutic drug and cell delivery. The permselectivity of these capsules provides a unique means of controlled drug release and immunoisolation of encapsulated cells. Immunoisolation is especially attractive as it abrogates the need for chronic immunosuppressive therapy and opens up the possibility for the delivery of numerous cell sources including xenogeneic grafts. APA microcapsules containing cellular therapeutics have proven effective in the short-term treatment of a wide range of diseases requiring enzyme or endocrine replacement therapy, including type I diabetes. If these microcapsules could be noninvasively monitored with X-ray imaging modalities (i.e., fluoroscopy, CT, and digital subtraction angiography), questions such as the ideal transplantation site, the best means of delivery, and the long-term survival of grafts could be better addressed. We have developed two novel alginate-based radiopaque microcapsule formulations containing either barium sulfate (Ba X-Caps) or bismuth sulfate (Bi X-Caps). As compared to conventional, nonradiopaque APA capsules, Ba X-Caps and Bi X-Caps containing human cadaveric islets resulted in a decrease in cellular viability of less than 5% up to 14 days after encapsulation. Both radiopaque capsules were found to be permeable to lectins < or =75 kDa, but were impermeable to lectins > or =120 kDa, thus ensuring the blockage of the penetration of antibodies while allowing free diffusion of insulin and nutrients. The glucose-responsive insulin secretion of the radiopaque encapsulated human islets was found to be unaltered compared to that of unlabeled controls, with human C-peptide levels ranging from 3.21 to 2.87 (Ba X-Caps) and 3.23 to 2.87 (Bi X-Caps) ng/islet at 7 and 14 days postencapsulation, respectively. Using fluoroscopy, both Ba X-Caps and Bi X-Caps could be readily visualized as single radiopaque entities in vitro. Furthermore, following transplantation in vivo in mice and rabbits, single capsules could be identified with no significant change in contrast for at least 2 weeks. This study represents the first attempt at making radiopaque microcapsules for X-ray guided delivery and imaging of cellular therapeutics. While human cadaveric islets were used as a proof-of-principle, these radiopaque capsules may have wide ranging therapeutic applications for a variety of cell types.

Alginates↗
PubMed2001

Survival and proliferation of alginate encapsulated Trichoderma spp. in Egyptian soil in comparison with allyl alcohol soil fumigation.

Conidia of Trichoderma harzianum and T. pseudokoningii (Rifai) were formulated to make alginate pellets with or without 10% cellulose as a food-base material. The formulations were compared for their ability for survival and proliferation of Trichoderma spp. in clay-loamy soil (50% moisture content) with allyl alcohol fumigation (0.05, 0.1 and 0.2 ml/1,000 ml space). Trichoderma medium E (TME) containing 100 microg/ml pentachloronitrobenzene (PCNB) was valuable for isolation and counting of Trichoderma spp. from the tested soil than the Glucose-Czapek's agar medium containing 1:15,000 Rose-bengal. The promotive effect of Trichoderma by different doses of allyl alcohol fumigation still enhanced after two-month incubation period. Conidia entrapped in alginate with or without cellulose and introduced into the soil survived better than conidia added directly to the same soil after three months incubation period. Sterile soil provided a more favorable environment for the proliferation and survival of immobilized conidia than the non-sterile soil, and the addition of 10% cellulose increased the survival of the entrapped conidia more than those prepared without cellulose. Soil fumigation inhibited the occurrence of other fungal species; however, inoculation of the soil with alginate immobilized conidia or conidial suspension had such inhibitory effect but in a less extent.

Alginates↗
PubMed2003

Components in the inoculum determine the kinetics of Azotobacter vinelandii cultures and the molecular weight of its alginate.

In cultures of Azotobacter vinelandii inoculated using washed cells (avoiding exhausted broth components) alginates of a higher molecular weight (1200 kDa) than those obtained in cultures conventionally inoculated (350 kDa), were produced. Also, when comparing conventionally inoculated cultures with those inoculated with washed-cells, the alginate lyase activity was delayed and the final polymer concentration decreased from 4.8 to 3.5 g l(-1). This suggests that components in the exhausted inoculum broth play important regulatory roles in alginate biosynthesis and needs to be taken into account when describing polymer biosynthesis.

Alginates↗
PubMed2004

Alginate coated loofa sponge discs for the removal of cadmium from aqueous solutions.

A biosorbent was prepared by coating the fibrous network of loofa sponge (Luffa cylindrica) with a thin film of calcium alginate. Alginate-coated loofa sponge removed Cd(II) rapidly, reaching equilibrium loading of 124 mg g(-1) in 30 min. Seventy % of equilibrium uptake was achieved in 10 min. In contrast, it took 240 min for alginate beads to reach a loading equilibrium of 88 mg g(-1) under identical conditions. The biosorption behaviour followed the Langmuir adsorption isotherm and the ACLS biosorbent was shown to be highly effective in removing Cd(II) from a 10 mg l(-1) solution in a continuous flow fixed-bed column bioreactor.

Adsorption↗
PubMed2003

Macrophage depletion improves survival of porcine neonatal pancreatic cell clusters contained in alginate macrocapsules transplanted into rats.

BACKGROUND: Macrophages can accumulate on the surface of empty and islet-containing alginate capsules, leading to loss of functional tissue. In this study, the effect of peritoneal macrophage depletion on the biocompatibility of alginate macrocapsules and function of macroencapsulated porcine neonatal pancreatic cell clusters (NPCCs) was investigated. METHODS: Clodronate liposomes were injected into the peritoneal cavities of normoglycemic Lewis rats 5 and 2 days before the transplantation. Empty or NPCC-containing Ca-alginate poly L-lysine (PLL)-coated macrocapsules were transplanted into the peritoneal cavities of rats injected with either clodronate liposomes or saline. On days 7, 14 and 21, samples were evaluated by immunohistochemistry for cellular immune responses on the surface of the macrocapsules and for macrophage populations in omental tissue. To assess the function of macroencapsulated NPCCs, insulin secretory responses to glucose and theophylline were measured after capsule retrieval. RESULTS: In saline-injected control groups, all of the empty and NPCC-containing macrocapsules were overgrown with macrophages, this being especially severe on NPCC-containing macrocapsules. In the clodronate liposomes-injected group, the majority of the empty macrocapsules were free of macrophage accumulation and the NPCC-containing macrocapsules were less overgrown than in control animals. Higher insulin responses to glucose and theophylline were observed in NPCCs retrieved from rats injected with clodronate liposomes. CONCLUSION: We conclude that depletion of peritoneal macrophages with clodronate liposomes improve the survival of macroencapsulated NPCCs.

Alginates↗
PubMed1998

A randomised trial of calcium alginate swabs to control blood loss in 3-5-year-old children.

OBJECTIVE: To compare the effectiveness of calcium alginate swabs and traditional cotton swabs in the control of blood loss after extraction of deciduous teeth from healthy children, aged 3-5 years, under general anaesthetic. DESIGN: Prospective, randomised clinical trial. SETTING: General anaesthetic suite in a dental teaching hospital. SUBJECTS: 101 patients were recruited to the study, comprising 50 randomised to be treated with cotton swabs and 51 with alginate swabs. INTERVENTIONS: Teeth were extracted under general anaesthesia and blood collected for measurement in order to compare blood loss using the two systems. MAIN OUTCOME MEASURES: The number of teeth extracted ranged from 1-14. The total blood loss ranged from 0.53-78.13 ml with a median of 12.9 ml. CONCLUSION: Calcium alginate swabs were not found to produce any clinical or statistical advantage over traditional cotton swabs in this surgical setting.

Alginates↗
PubMed2006

The use of acupuncture in controlling the gag reflex in patients requiring an upper alginate impression: an audit.

BACKGROUND: A pronounced gag reflex (GR) can be a problem to both the acceptance and delivery of dental treatment. Despite a range of management strategies, some patients cannot accept even simple dental treatment. The aim of this study was to evaluate the use of acupuncture point CV-24 in controlling a profound gag reflex during dental treatment requiring an upper alginate impression. METHOD: Members of the British Dental Acupuncture Society were invited to take part in an audit of the role of acupuncture point CV-24 in controlling the gag reflex. They were issued with patient inclusion criteria, a standardised procedure instruction sheet and a recording form. All patients fulfilling the inclusion criteria had an upper dental alginate impression taken (or an attempt made at it) before acupuncture, and a second upper alginate impression taken immediately after acupuncture of point CV-24. The GR assessment was undertaken prior to insertion of the acupuncture needle using the Gagging Severity Index (GSI); and after the acupuncture and impression taking using the Gagging Prevention Index (GPI). Both the GSI and GPI were recorded at three stages of the dental impression taking procedure, ie, when the empty impression tray was tried in the mouth, when the loaded tray was inserted into the mouth, and on completion of the impression taking. RESULTS: Twenty-one dentists submitted 64 case reports of which 37 fulfilled the inclusion criteria. Prior to acupuncture all 37 patients (20 females and 17 males with a mean age of 46.8 years) were unable to accept the impression taking. After acupuncture of point CV-24, an improvement of between 51-55% (mean 53%) for the three stages of impression taking was noticed. Thirty patients (81%) were able to accept the impression taking, whereas seven (19%) remained unable to tolerate the procedure. Assessed by the GSI and GPI, there was a significant decrease in GR scores at all three stages of the impression taking procedure (median 3 vs 1; 4 vs 2; 4 vs 2; p < 0.0001). Thus before acupuncture, the patients had moderate to severe GR and after acupuncture the GR had reduced to a level which only complicated dental treatment slightly. CONCLUSION: Our results indicate that acupuncture of point CV-24 is an effective method of controlling severe GR during dental treatment including impression taking. However, the results of the current audit need to be tested in a randomised controlled study in order to substantiate the effectiveness of this method.

Acupuncture Points↗
PubMed2001

Survival, root colonisation and biocontrol capacities of Pseudomonas fluorescens F113 LacZY in dry alginate microbeads.

Cells of Pseudomonas fluorescens F113 LacZY were encapsulated in alginate and their survival and ability to colonise sugar beet were evaluated. To assess survival, the formulation, composed of dry alginate microbeads of 300- to 700-microm diameter, was stored 1 year at 28+/-2 and 4+/-2 degrees C and then tested against pathogenic fungi Pythium ultimum and Rhizoctonia solani in in vitro inhibition experiments. The same material was also used as inoculant for protection of sugar beet against Py. ultimum in microcosm experiments. The results obtained indicated that, although drying alginate beads resulted in a significant reduction of bacterial viability, the use of microbeads enabled a satisfactory level of root colonisation and protection, at least under microcosm conditions. The capability of the encapsulated cells to produce the antifungal metabolite 2,4-diacetylphloroglucinol (Phl) was not significantly affected by 12 months storage.

Alginates↗
PubMed2005

Gentle cell trapping and release on a microfluidic chip by in situ alginate hydrogel formation.

Microfluidic devices are increasingly used to perform biological experiments on a single-cell basis. However, long-term stability of cell positions is still an issue. A novel biocompatible method for cell entrapment and release on a microchip is presented. It is based on the controlled formation of an alginate hydrogel by bringing two laminar flows of alginate and calcium ions in the range of 2 mM to 40 mM into contact. The resulting growth of a gel bar is used to enclose and immobilize yeast cells. Adding ethylenediaminetetraacetic acid (EDTA) to the alginate solution allows for control of the hydrogel growth, and by varying the ratio of Ca(2+) to EDTA concentrations gel growth or gel shrinkage can be induced at will. Trapped cells are released during shrinkage of the gel. The trapping efficiency for different cell speeds is investigated and the properties of gel growth are discussed using a diffusion model. Precise positioning of a single cell is demonstrated. The technique presented allows not only the reversible immobilization of cells under gentle conditions but also offers the potential of long-term cell cultures as shown by on-chip incubation of yeast cells. The procedure may provide a simple and fully biocompatible technique for a multitude of innovative experiments on cells in microsystems.

Alginates↗
PubMed2006

Manipulating the generation of Ca-alginate microspheres using microfluidic channels as a carrier of gold nanoparticles.

In this paper the manipulation of Ca-alginate microspheres, using a microfluidic chip, for the encapsulation of gold nanoparticles is presented. Our strategy is based on hydrodynamic-focusing on the forming of a series of self-assembling sphere structures, the so-called water-in-oil (w/o) emulsions, in the cross-junction microchannel. These fine emulsions, consisting of aqueous Na-alginates, are then dripped into a solution of 20% calcium salt to accomplish Ca-alginate microspheres in an efficient manner. Experimental data show that microspheres with diameters ranging from 50 microm to 2000 microm with a variation less than 5% were precisely generated. The size and gap of the droplets are tunable by adjusting the relative sheath/sample flow rate ratio. Furthermore, we applied them to encapsulated gold nanoparticles, and this one shot operation performs the 'Lab on a Chip'.

Alginates↗