A Review on Composite Liposomal Technologies for Specialized Drug Delivery

The combination of liposomes with polymeric scaffolds could revolutionize the current state of drug delivery technology. Although liposomes have been extensively studied as a promising drug delivery model for bioactive compounds, there still remain major drawbacks for widespread pharmaceutical application. Two approaches for overcoming the factors related to the suboptimal efficacy of liposomes in drug delivery have been suggested. The first entails modifying the liposome surface with functional moieties, while the second involves integration of pre-encapsulated drug-loaded liposomes within depot polymeric scaffolds. This attempts to provide ingenious solutions to the limitations of conventional liposomes such as short plasma half-lives, toxicity, stability, and poor control of drug release over prolonged periods. This review delineates the key advances in composite technologies that merge the concepts of depot polymeric scaffolds with liposome technology to overcome the limitations of conventional liposomes for pharmaceutical applications

Caractérisation et conservation de la diversité bactérienne d’un lait fermenté traditionnel breton, le Gwell en lien avec la préservation d’une race locale de vache, la Bretonne Pie Noir

Le Gwell est un lait fermenté traditionnel spécifique de la Bretagne. Il est obtenu à partir de lait de vaches de race Bretonne Pie Noir, inoculé avec une portion de la fabrication précédente (appelé ferment) sans aucun recours à des levains commerciaux. Les productions de Gwell partagent une texture ferme et onctueuse et un gout frais et acidulé, avec des caractéristiques organoleptiques propres à chaque producteur. Les producteurs sont malheureusement parfois confrontés à la perte de leur ferment et doivent alors avoir recours à la solidarité d’autres producteurs pour réacquérir un ferment opérationnel. Ces pertes de ferments sont un frein au développement de la production de Gwell et donc à la valorisation de lait issu de vaches Bretonne Pie Noir. Cette race emblématique de la Bretagne, caractérisée par une rusticité hors du commun et un lait très riche en matière grasse totalisait au milieu du 19ème siècle près de 900 000 têtes. La modernisation des pratiques agricoles alliée à une orientation productiviste forte a conduit à une quasi extinction de l’espèce, ce qui a conduit à initier en 1976 un programme de sauvegarde de l’espèce. Le nombre de vaches s’élève ainsi aujourd’hui à près de 2500 femelles. La transformation du lait en Gwell est, pour les éleveurs, un moyen de valoriser la qualité du lait de Bretonne Pie Noir en conservant sa valeur ajoutée. Les éleveurs qui transforment le lait en Gwell œuvrent ainsi à la sauvegarde de l’espèce Bretonne Pie Noir, mais aussi à la préservation de la diversité microbienne, du patrimoine et des savoir-faire paysans associés. La caractérisation de l’écosystème microbien du ferment Gwell, pour mieux maitriser sa conservation et sécuriser ainsi la production de Gwell, participe de ce fait au maintien de la race Bretonne Pie Noir. Dans ce contexte notre étude visait à caractériser l’écosystème microbien du Gwell pour sécuriser les souches à l’origine de la typicité du produit. Nous avons ainsi montré que toutes les productions de Gwell avaient une flore bactérienne dominante similaire, composée de deux sous-espèces de la bactérie lactique Lactococcus lactis (subsp. lactis et subsp. cremoris). En fonction des producteurs, le nombre de souches de chaque sous-espèce peut varier avec dans certain cas la présence de Streptococcus thermophilus. De plus, nous avons identifié et caractérisé des souches spécifiques à chaque producteur et montré une forte résilience de l’écosystème pouvant expliquer en partie les différences organoleptiques observées entre les Gwell de différents producteurs

Mutations in TUBG1, DYNC1H1, KIF5C and KIF2A cause malformations of cortical development and microcephaly

The genetic causes of malformations of cortical development (MCD) remain largely unknown. Here we report the discovery of multiple pathogenic missense mutations in TUBG1, DYNC1H1 and KIF2A, as well as a single germline mosaic mutation in KIF5C, in subjects with MCD. We found a frequent recurrence of mutations in DYNC1H1, implying that this gene is a major locus for unexplained MCD. We further show that the mutations in KIF5C, KIF2A and DYNC1H1 affect ATP hydrolysis, productive protein folding and microtubule binding, respectively. In addition, we show that suppression of mouse Tubg1 expression in vivo interferes with proper neuronal migration, whereas expression of altered gamma-tubulin proteins in Saccharomyces cerevisiae disrupts normal microtubule behavior. Our data reinforce the importance of centrosomal and microtubule-related proteins in cortical development and strongly suggest that microtubule-dependent mitotic and postmitotic processes are major contributors to the pathogenesis of MCD

Nonthrombogenic, Biodegradable Elastomeric Polyurethanes with Variable Sulfobetaine Content

For applications where degradable polymers are likely to have extended blood contact, it is often important for these materials to exhibit high levels of thromboresistance. This can be achieved with surface modification approaches, but such modifications may be transient with degradation. Alternatively, polymer design can be altered such that the bulk polymer is thromboresistant and this is maintained with degradation. Toward this end a series of biodegradable, elastic polyurethanes (PESBUUs) containing different zwitterionic sulfobetaine (SB) content were synthesized from a polycaprolactone-diol (PCL-diol):SB-diol mixture (100:0, 75:25, 50:50, 25:75 and 0:100) reacted with diisocyanatobutane and chain extended with putrescine. The chemical structure, tensile mechanical properties, thermal properties, hydrophilicity, biodegradability, fibrinogen adsorption and thrombogenicity of the resulting polymers was characterized. With increased SB content some weakening in tensile properties occurred in wet conditions and enzymatic degradation also decreased. However, at higher zwitterionic molar ratios (50% and 75%) wet tensile strength exceeded 15 MPa and breaking strain was >500%. Markedly reduced thrombotic deposition was observed both before and after substantial degradation for both of these PESBUUs and they could be processed by electrospinning into a vascular conduit format with appropriate compliance properties. The mechanical and degradation properties as well as the acute in vitro thrombogenicity assessment suggest that these tunable polyurethanes could provide options appropriate for use in blood contacting applications where a degradable, elastomeric component with enduring thromboresistance is desired

Stretch-activated ion channel TMEM63B associates with developmental and epileptic encephalopathies and progressive neurodegeneration

By converting physical forces into electrical signals or triggering intracellular cascades, stretch-activated ion channels allow the cell to respond to osmotic and mechanical stress. Knowledge of the pathophysiological mechanisms underlying associations of stretch-activated ion channels with human disease is limited. Here, we describe 17 unrelated individuals with severe early-onset developmental and epileptic encephalopathy (DEE), intellectual disability, and severe motor and cortical visual impairment associated with progressive neurodegenerative brain changes carrying ten distinct heterozygous variants of TMEM63B, encoding for a highly conserved stretch-activated ion channel. The variants occurred de novo in 16/17 individuals for whom parental DNA was available and either missense, including the recurrent p.Val44Met in 7/17 individuals, or in-frame, all affecting conserved residues located in transmembrane regions of the protein. In 12 individuals, hematological abnormalities co-occurred, such as macrocytosis and hemolysis, requiring blood transfusions in some. We modeled six variants (p.Val44Met, p.Arg433His, p.Thr481Asn, p.Gly580Ser, p.Arg660Thr, and p.Phe697Leu), each affecting a distinct transmembrane domain of the channel, in transfected Neuro2a cells and demonstrated inward leak cation currents across the mutated channel even in isotonic conditions, while the response to hypo-osmotic challenge was impaired, as were the Ca2+ transients generated under hypo-osmotic stimulation. Ectopic expression of the p.Val44Met and p.Gly580Cys variants in Drosophila resulted in early death. TMEM63B-associated DEE represents a recognizable clinicopathological entity in which altered cation conductivity results in a severe neurological phenotype with progressive brain damage and early-onset epilepsy associated with hematological abnormalities in most individuals. Genetics of disease, diagnosis and treatmen

AMPA receptor GluA2 subunit defects are a cause of neurodevelopmental disorders.

AMPA receptors (AMPARs) are tetrameric ligand-gated channels made up of combinations of GluA1-4 subunits encoded by GRIA1-4 genes. GluA2 has an especially important role because, following post-transcriptional editing at the Q607 site, it renders heteromultimeric AMPARs Ca2+-impermeable, with a linear relationship between current and trans-membrane voltage. Here, we report heterozygous de novo GRIA2 mutations in 28 unrelated patients with intellectual disability (ID) and neurodevelopmental abnormalities including autism spectrum disorder (ASD), Rett syndrome-like features, and seizures or developmental epileptic encephalopathy (DEE). In functional expression studies, mutations lead to a decrease in agonist-evoked current mediated by mutant subunits compared to wild-type channels. When GluA2 subunits are co-expressed with GluA1, most GRIA2 mutations cause a decreased current amplitude and some also affect voltage rectification. Our results show that de-novo variants in GRIA2 can cause neurodevelopmental disorders, complementing evidence that other genetic causes of ID, ASD and DEE also disrupt glutamatergic synaptic transmission

Zymogram and Preliminary Characterization of Lactobacillus helveticus Autolysins

The autolysins of Lactobacillus helveticus ISLC5 were detected and partially characterized by renaturing sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis with substrate-containing gels (zymogram). By using lyophilized Micrococcus luteus cells or heated whole cells of L. helveticus ISLC5 (0.2% [wt/vol]) as a substrate, several lytic activities were detected in the whole-cell SDS extract of strain ISLC5 (i) one activity at 42.4 kDa, which was named autolysin A, and (ii) six other activities having very similar molecular weights (29.1, 29.6, 30, 30.8, 31.7, and 32.8 kDa), which were named autolysins B (B1 through B6, respectively). As regards the temporal distribution of the enzymes, autolysins A and B were detected in the cells harvested from the beginning of the exponential growth phase. Autolysin A appeared to be associated only with viable cells, whereas the autolysins B remained associated with the cell envelope several days after the complete loss of culture viability. When SDS-treated walls of L. helveticus ISLC5 were used as a substrate, a supplementary lytic activity appeared at 37.5 kDa; it was considered a peptidoglycan hydrolase, since it was not able to induce lysis of whole-cell substrate. The autolysins of 30 other strains of L. helveticus from various geographical origins were also analyzed by zymogram; all the activity profiles obtained were similar to that of strain ISLC5 in terms of the number of lytic bands and their apparent molecular weights. Only the relative intensities of the lytic bands corresponding to autolysins A and B were variable depending on the strains. This observation suggested that autolysins are highly conserved enzymes. A concentrated crude lysate of the virulent bacteriophage 832-B1 infecting L. helveticus was also analyzed by zymogram; one lytic activity with an apparent molecular weight of 31.7 kDa, very close to the weights of the autolysins B, was observed. Finally, the autolysins of L. helveticus ISLC5 were successfully extracted from whole cells by using a 1 M lithium chloride solution; they were partially purified by precipitation, selective resolubilization, and gel filtration chromatography, which led to a 20-fold increase in specific activity

Evaluation of Antibacterial Activities of Tanzanian Moringa oleifera Extracts against Escherichia coli and Klebsiella pneumonia Clinical Isolates

In vitro antibacterial activities of methanolic leaf and seed extracts of Moringa oleifera grown in Dodoma, Tanzania were evaluated using standard microdilution and disc diffusion methods against extended spectrum beta lactamase producing Gram negative bacteria (Escherichia coli and Klebsiella pneumoniae). Microdilution method showed no activity at concentration of 20 mg/mL for all the extracts except leaf extract which exhibited minimum inhibition at concentration of  2.5 µg/mL for E. Coli but when high concentrations of the extracts were used in the disc diffusion method then the results exhibited the highest killing susceptibility at concentration of 0.4 g/mL with zone of inhibition 31 mm for leaf extract and 26 mm for seed extract against E. coli, and 27 mm for leaf extract and 29 mm for seed extract against K. pneumonia. Additionally, when both extracts were screened qualitatively for phytoconstituents using standard methods, leaf extract confirmed the presence of carbohydrates, cardiac glycosides,  tannins and quinones, and seed extract confirmed the presence of proteins, carbohydrates, cardiac glycosides, alkaloids, terpenoids and quinones. These phytoconstituents can be new sources of future antibiotics that potentially combat the existing problem of antimicrobial resistance and thus, creating an awareness in the community regarding the usage of M. oleifera growing widely but neglected in Tanzania in spite of its nutritional values and traditional uses. Keywords: Moringa oleifera; Phytoconstituents; Antibacterial activity; Escherichia coli and Klebsiella pneumonia