Comparison of different methods for release of Bifidobacterium longum Bb46 from the poly(vinylpyrrolidone)-poly(vinylacetate-co-crotonic acid) interpolymer complex matrix, and the effect of grinding on the microparticles

Bifidobacteria have been efficiently encapsulated in poly(vinylpyrrolidone)-poly(vinylacetate-co-crotonic acid) (PVP:PVAc-CA) interpolymer complex formed in scCO2. Research indicated that this method improves the stability of encapsulated bacteria in simulated gastrointestinal fluids in vitro. However, further analysis indicated release of lower numbers of encapsulated bacteria from the encapsulating matrix. The aims of this study were to determine a method that would release high numbers of bacteria from the PVP:PVAc-CA interpolymer complex matrix microparticles, and furthermore, to determine the effects of milling on the morphological properties of the microparticles. Three release methods, namely sonication, homogenization in a stomacher and incubation in simulated intestinal fluid (SIF) were compared. Released viable bacteria were assayed using plate counts. Viable bacteria released using a stomacher were three orders of magnitude higher than those released by incubation and an order of magnitude higher than those released using sonication. SEM indicated no negative effects such as exposure of encapsulated bacteria on the matrix due to milling of product. Homogenization in a stomacher is the most efficient method for releasing bacteria from the PVP:PVAc-CA interpolymer complex matrix. Particle size of the PVP:PVAc-CA microparticles encapsulating bacteria can be reduced further by grinding, without exposing the enclosed bacteria.The authors would like to thank the National Research Foundation of South Africa for funding of the project.http://www.springer.com/chemistry/biotech/journal/1127

Molecular autopsy in maternal-fetal medicine

Purpose: The application of genomic sequencing to investigate unexplained death during early human development, a form of lethality likely enriched for severe Mendelian disorders, has been limited.& para;& para;Methods: In this study, we employed exome sequencing as a molecular autopsy tool in a cohort of 44 families with at least one death or lethal fetal malformation at any stage of in utero development. Where no DNA was available from the fetus, we performed molecular autopsy by proxy, i.e., through parental testing.& para;& para;Results: Pathogenic or likely pathogenic variants were identified in 22 families (50%), and variants of unknown significance were identified in further 15 families (34%). These variants were in genes known to cause embryonic or perinatal lethality (ALPL, GUSB, SLC17A5, MRPS16, THSD1, PIEZO1, and CTSA), genes known to cause Mendelian phenotypes that do not typically include embryonic lethality (INVS, FKTN, MYBPC3, COL11A2, KRIT1, ASCC1, NEB, LZTR1, TTC21B, AGT, KLHL41, GFPT1, and WDR81) and genes with no established links to human disease that we propose as novel candidates supported by embryonic lethality of their orthologs or other lines of evidence (MS4A7, SERPINA11, FCRL4, MYBPHL, PRPF19, VPS13D, KIAA1109, MOCS3, SVOPL, FENI, HSPB11, KIF19, and EXOC3L2).& para;& para;Conclusion: Our results suggest that molecular autopsy in pregnancy losses is a practical and high-yield alternative to traditional autopsy, and an opportunity for bringing precision medicine to the clinical practice of perinatology

Supercritical CO2 interpolymer complex encapsulation improves heat stability of probiotic bifidobacteria

The probiotic industry faces the challenge of retention of probiotic culture viability as numbers of these cells within their products inevitably decrease over time. In order to retain probiotic viability levels above the therapeutic minimum over the duration of the product’s shelf life, various methods have been employed, among which encapsulation has received much interest. In line with exploitation of encapsulation for protection of probiotics against adverse conditions, we have previously encapsulated bifidobacteria in poly-(vinylpyrrolidone)-poly-(vinylacetate-co-crotonic acid) (PVP:PVAc-CA) interpolymer complex microparticles under supercritical conditions. The microparticles produced had suitable characteristics for food applications and also protected the bacteria in simulated gastrointestinal fluids. The current study reports on accelerated shelf life studies of PVP:PVAc-CA encapsulated Bifidobacterium lactis Bb12 and Bifidobacterium longum Bb46. Samples were stored as free powders in glass vials at 30 °C for 12 weeks and then analysed for viable counts and water activity levels weekly or fortnightly. Water activities of the samples were within the range of 0.25–0.43, with an average a w = 0.34, throughout the storage period. PVP:PVAc-CA interpolymer complex encapsulation retained viable levels above the recommended minimum for 10 and 12 weeks, for B. longum Bb46 and B. lactis Bb12, respectively, thereby extending their shelf lives under high storage temperature by between 4 and 7 weeks. These results reveal the possibility for manufacture of encapsulated probiotic powders with increased stability at ambient temperatures. This would potentially allow the supply of a stable probiotic formulation to impoverished communities without proper storage facilities recommended for most of the currently available commercial probiotic products.University of Pretoria, National Research Foundation (NRF), South Africa and The Council for Scientific and Industrial Research (CSIR), Pretoria.http://www.springer.com/chemistry/biotech/journal/11274hb201