Pectin Conformation in Solution.

The interplay of degree of methylesterification (DM), pH, temperature, and concentration on the macromolecular interactions of pectin in solution has been explored. Small-angle X-ray scattering complemented by atomic force microscopy and molecular dynamics was employed to probe chain dimensions and solution structure. Two length scales have been observed with the first level of structure characterising chain clusters with sizes ranging between 100-200 nm. The second level of structure arises from single biopolymer chains with a radius of gyration between ∼6 and 42 nm. The development of a range of macromolecular dimensions in vitro and in silico shows that the chain flexibility increases with DM and at acidic pH, whereas hydrogen bonding is the responsible thermodynamic driving force for cluster formation. High methyl pectins create structures of lower fractal dimension with less efficient packing. This work unveils pectin conformations covering most of its industrially and biologically relevant environments, enabling rational design of advanced biomaterials based on pectin.

In vivo degradation and release kinetics of chloramphenicol-loaded poly(D,L)-lactide sponges.

Poly(d,l)-lactide (PDLLA) homopolymer, with an average molecular weight of 20,000 daltons, was produced by the ring-opening polymerization of d,l-lactide in the presence of SnCl(2).2H(2)O as the catalyst. The PDLLA sponges loaded with chloramphenicol were prepared by a solvent evaporation technique. The drug loadings achieved were 14.84 and 25.23 mg for the PDLLA sponges with 35 and 70 mg total weights, respectively. These sponges were implanted in Wistar rats, and in vivo degradation, drug release, and tissue reactions were followed. The PDLLA sponges carrying no drug degraded with time linearly. Almost 80% of the sponges were degraded in about 180 days. While the drug-loaded PDLLA sponges were degraded much faster in 4 weeks (about 35% of the matrix was degraded), then the degradation slowed down significantly. Drug release from the sponges was parallel to the degradation. Almost 60% of the loaded drug released in 4 weeks. There were no acute inflammatory reactions in the initial period, either for the plain or for the drug-loaded PDLLA sponges. Macrophages and multinuclear giant cells start to appear after 7 days of implantation. The fibroblastic activity also started after the same period. After that, there were decreases in the number of some cells (neutrophils, lymphocytes, and macrophages), while multinuclear giant cells and fibroblastic activities gradually increased. Granulation tissue started at about 1 month, and new connective tissue was gradually formed until 180 days of implantation. There were significant numbers of inflammatory cells after 60 days, which were replaced by fibroblasts after 180 days. There was almost no significant neovascularization after 180 days, but implant fragmentation gradually increased (which slows the degradation) with time. It was concluded that this novel drug release sponge may be safely and effectively used as an active soft tissue-filling material.

Observations on the crystallization of oligogalacturonates.

Oligogalacturonates were produced by the limited enzymic hydrolysis of polygalacturonic acid and purified by ion-exchange chromatography. The fractions obtained were of limited polydispersity, determined by analytical ion-exchange chromatography. Oligomers with an average degree of polymerization of 10-15 were readily crystallized from aqueous salt solutions at neutral pH as single crystals. Crystal morphology of the salts examined, Na+, K+ and Ca2+ were characteristic of the salt. The wide-angle X-ray diffraction patterns obtained for the sodium salt were consistent with published fibre diffraction data of this salt form.

Factors affecting the release of flavor encapsulated in carbohydrate matrixes.

The effects of water content and temperature variation on the release of flavor components into the headspace over flavors, encapsulated by an extrusion process, in low water content carbohydrate matrixes is studied. The largest amounts of release occurred when the matrix was above its glass transition temperature, whether this was due to increased water content or elevated temperature. Under these conditions up to 70% of the sucrose in the matrix crystallized over a period of 10 days, as quantified using Fourier transform Raman spectroscopy. Smaller amounts of headspace release occurred when the water content of the encapsulated flavor system was decreased from 3. 5 to 3.1% w/w. Small amounts of release occurred from the "as prepared" materials, which were associated with the presence of small amounts of unencapsulated flavor oil with direct access to the headspace. It was concluded that release due to matrix permeability was relatively slow as compared with the above mechanisms.

Growth of food-borne pathogenic bacteria in oil-in-water emulsions: I--Methods for investigating the form of growth.

Methods are presented for investigating the site and form of growth of bacteria in model oil-in-water emulsions and in dairy cream. Following growth of the bacteria, the continuous aqueous phase is gelled using agarose and the oil phase removed using a mixture of chloroform and methanol. Using this method, the authors have found that Listeria monocytogenes, Salmonella typhimurium and Yersinia enterocolitica grow in the form of colonies in concentrated oil-in-water emulsions. Colonies of L. monocytogenes and Y. enterocolitica also form in artificially-inoculated fresh and tinned dairy cream. If information about the precise site of growth is not required, the authors have discovered that intact colonies can be liberated from the model emulsions by dissolving away the oil phase with chloroform:methanol.

Growth of food-borne pathogenic bacteria in oil-in-water emulsions: II--Effect of emulsion structure on growth parameters and form of growth.

The growth rates and yields of Listeria monocytogenes and Yersinia enterocolitica were determined in liquid culture media, and in model oil-in-water emulsions that contained 30, 70 or 83% (v/v) hexadecane. In emulsions with a mean droplet size of 2 microns containing 83% (v/v) hexadecane, the growth of both organisms resulted in decreased yields. Additionally, in these emulsions adjusted to pH 5.0 or 4.4 the growth rate of L. monocytogenes was significantly less than in other model systems which had an aqueous phase of equivalent chemical composition. Microscopic examination of the 83% (v/v) emulsion showed that its microstructure immobilized the bacteria, which were constrained to grow as colonies. Bacteria behaved similarly in model emulsions of either hexadecane or sunflower oil. Manipulation of the droplet size distribution of the emulsions changed the form and rate of growth of bacteria within them.

The microstructure and distribution of micro-organisms within mature Serra cheese.

The distribution of micro-organisms in mature Serra, a traditional Portuguese cheese made from unpasteurised ewes' milk without added starter culture, was examined by light microscopy and electron microscopy. Four populations of micro-organisms were recognized according to their position within the cheese: (i) those present as apparently axenic colonies within the curd matrix; (ii) bacteria growing along curd junctions; (iii) yeasts and bacteria present in the smear on the surface of the cheese and (iv) bacteria found in cracks which penetrated the outer part of the cheese from the rind. Two types of crystals were observed, together with contaminants of vegetable origin and somatic cells originating from the milk.

Fracture mechanics of the cell wall of Chara corallina.

Previous mechanical studies using algae have concentrated on cell extension and growth using creep-type experiments, but there appears to be no published study of their failure properties. The mechanical strength of single large internode cell walls (up to 2 mm diameter and 100 mm in length) of the charophyte (giant alga) Chara corallina was determined by dissecting cells to give sheets of cell wall, which were then notched and fractured under tension. Tensile tests, using a range of notch sizes, were conducted on cell walls of varying age and maturity to establish their notch sensitivity and to investigate the propagation of cracks in plant cell walls. The thickness and stiffness of the walls increased with age whereas their strength was little affected. The strength of unnotched walls was estimated as 47+/-13 MPa, comparable to that of some grasses but an order of magnitude higher than that published for model bacterial cellulose composite walls. The strength was notch-sensitive and the critical stress intensity factor K1c was estimated to be 0.63+/-0.19 MNm(-3/2), comparable to published values for grasses.