Role of carboxyl pendant groups of medium chain length poly(3‐hydroxyalkanoate)s in biomedical temporary applications

The poly(3‐hydroxyoctanoate) (PHO) is a biodegradable polyester containing hydrophobic side chains. One way to obtain more hydrophilic polyester consisted in the introduction of polar groups in the side chains. Carboxyl groups (PHO₇₅COOH₂₅) were introduced by chemical modifications. The role of carboxyl groups was investigated in the first part as potential support for cell seeding by studying the cell adhesion and proliferation, and in the second part as potential drug carrier by comparing the abilities of PHO and PHO₇₅COOH₂₅ to form degradable particles. Measurements of human bladder RT112 cells adhesion were done with or without collagen IV. Adhesive RT112 cells were counted by a colorimetric MTT test. The results showed that the COOH pendant groups of PHO₇₅COOH₂₅ films promoted cell adhesion after 4 h of incubation. The proliferation of cells is not improved after 4 days of incubation because of a reorganization of macromolecular chains and reorientation of COOH groups. This surface restructuration when the film was in contact with water was showed by contact angle measurements. We showed that the presence of COOH groups modified the hydrophobic/hydrophilic balance and enhanced the formation of particles. Stable lyophilisable particles were then obtained with diblock copolymer P(HO₇₅COOH₂₅‐b‐CL); the caprolactone block (CL) was necessary to improve particles stability. The results showed that the release of doxorubicin from the particles is enhanced in presence of hydrophilic and degradable block (PHO₇₅COOH₂₅). It was possible to obtain a degradable functional polyester based on PHO with carboxyl pendant groups to improve degradation rate by simple hydrolysis required for drug delivery systems. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2010     

Sequencing analysis of a 40·2 kb fragment of yeast chromosome X reveals 19 open reading frames including URA2 (5′ end), TRK1, PBS2, SPT10, GCD14, RPE1, PHO86, NCA3, ASF1, CCT7, GZF3, two tRNA genes,three remnant delta elements and a Ty4 transposon

The complete sequence of a 40 247 bp DNA segment located on the left arm of chromosome X of Saccharomyces cerevisiae has been determined and analysed. The sequence encodes the 5′ coding region of the URA2 gene and 18 open reading frames of at least 100 amino acids. Ten of these correspond to known genes, whereas eight correspond to new genes. In addition, the sequence contains a tRNA-Ala gene, a tRNA-Asp gene, a Ty4 transposable element and three delta elements. The sequence has been deposited in the EMBL databank under Accession Numbers: Z49405, Z49404, Z49403, Z49402, Z49401, Z49400, Z49399, Z49398, Z49397, Z49396, Z49394, Z49392, Z49391, Z49390, Z49389, Z49387, Z49386, Z49385.     

Regulatory Changes Affecting the Production and Use of Fats and Oils: Focus on Partially Hydrogenated Oils

Partially hydrogenated oils (PHO), the products of incomplete catalytic hydrogenation of food oils, have been widely employed by the food industry for more than a century. Their exceptional stability and technologic characteristics made them the preferred choice for the production of several food products including margarines, bakery goods, and frying oils. Some of these highly prized characteristics were provided by the high content in trans fatty acids (TFA), defined as fatty acids with one or more isolated double bond in trans configuration. The discovery of negative health effects associated with dietary intake of TFA triggered world-wide a wave of regulatory actions aimed to curb their consumption. PHO became the main target of most campaigns aimed to reduce consumption of TFA, and their fortune in the food industry progressively faded. At the 2017, AOCS Annual Meeting in Orlando, a group of experts from regulatory agencies and industry from North America and Europe met to discuss the current status of government regulations and industry adaptations regarding the productions and use of PHO. The discussion was enriched by including the impact on fats and oils production of the 2016 amendment of the Toxic Substances Control Act of 1976 (TSCA). The present publication may not include all regulatory changes that took place after this symposium, in 2017.     

The PHO80/TUP7 locus in Saccharomyces cerevisiae is on the left arm of chromosome XV: mapping by chromosome engineering.

The PHO80/TUP7 locus in Saccharomyces cerevisiae is reported to be located on the right arm of chromosome XV close to its centromere. In the present study, the locus has been reassigned to the left arm of the same chromosome by reciprocal recombination between chromosomes V and XV at URA3 (on chromosome V) and PHO80/TUP7 loci by using the site-specific recombination system of the yeast plasmid pSR1.     

Vimentin Cytoskeleton Architecture Analysis on Polylactide and Polyhydroxyoctanoate Substrates for Cell Culturing

Polylactide (PLA), widely used in bioengineering and medicine, gained popularity due to its biocompatibility and biodegradability. Natural origin and eco-friendly background encourage the search of novel materials with such features, such as polyhydroxyoctanoate (P(3HO)), a polyester of bacterial origin. Physicochemical features of both P(3HO) and PLA have an impact on cellular response 32, i.e., adhesion, migration, and cell morphology, based on the signaling and changes in the architecture of the three cytoskeletal networks: microfilaments (F-actin), microtubules, and intermediate filaments (IF). To investigate the role of IF in the cellular response to the substrate, we focused on vimentin intermediate filaments (VIFs), present in mouse embryonic fibroblast cells (MEF). VIFs maintain cell integrity and protect it from external mechanical stress, and also take part in the transmission of signals from the exterior of the cell to its inner organelles, which is under constant investigation. Physiochemical properties of a substrate have an impact on cells’ morphology, and thus on cytoskeleton network signaling and assembly. In this work, we show how PLA and P(3HO) crystallinity and hydrophilicity influence VIFs, and we identify that two different types of vimentin cytoskeleton architecture: network “classic” and “nutshell-like” are expressed by MEFs in different numbers of cells depending on substrate features.     

缺失PHO13基因与进化工程构建高效利用木糖酵母

过表达了木糖还原酶、木糖醇脱氢酶和木酮糖激酶基因的重组工业酿酒酵母菌株KAM-6X内缺失了编码对硝基苯磷酸盐磷酸化酶的PHO13基因,接着通过EMS诱变和进化工程筛选,获得了一株高效利用木糖的菌株,命名为PE。有氧条件下,在含50 g/L木糖和100 g/L木糖的YPX中最大比生长速率分别为0.299和0.282 h^-1,分别比出发菌提高了95.43%和102.87%,同时PE菌株能在前24 h内耗掉36.12 g/L木糖,48 h内耗掉70.25 g/L木糖。微好氧条件下副产物产量降低,糖醇转化率最高达到0.382 g/g,证明PE是一株高效利用木糖发酵的工业酵母菌株。     

Grafting of poly(3‐hydroxyalkanoate) and linoleic acid onto chitosan

Poly(3‐hydroxy octanoate) (PHO), poly(3‐hydroxy butyrate‐co‐3‐hydroxyvalerate) (PHBV), and linoleic acid were grafted onto chitosan via condensation reactions between carboxylic acids and amine groups. Unreacted PHAs and linoleic acid were eliminated via chloroform extraction and for elimination of unreacted chitosan were used 2 wt % of HOAc solution. The pure chitosan graft copolymers were isolated and then characterized by FTIR, ¹³C‐NMR (in solid state), DSC, and TGA. Microbial polyester percentage grafted onto chitosan backbone was varying from 7 to 52 wt % as a function of molecular weight of PHAs, namely as a function of steric effect. Solubility tests were also performed. Graft copolymers were soluble, partially soluble or insoluble in 2 wt % of HOAc depending on the amount of free primary amine groups on chitosan backbone or degree of grafting percent. Thermal analysis of PHO‐g‐Chitosan graft copolymers indicated that the plastizer effect of PHO by means that they showed melting transitions T_ms at 80, 100, and 113°C or a broad T_ms between 60.5–124.5°C and 75–125°C while pure chitosan showed a sharp T_m at 123°C. In comparison of the solubility and thermal properties of graft copolymers, linoleic acid derivatives of chitosan were used. Thus, the grafting of poly(3‐hydroxyalkanoate) and linoleic acid onto chitosan decrease the thermal stability of chitosan backbone. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103:81–89, 2007