Rosuvastatin effects on the HDL proteome in hyperlipidemic patients

The advancements in proteomics have provided a better understanding of the functionality of apolipoproteins and lipoprotein-associated proteins, with the HDL lipoprotein fraction being the most studied. The focus of this study was to evaluate the HDL proteome in dyslipidemic subjects without an established cardiovascular disease, as well as to test whether rosuvastatin treatment alters the HDL proteome. Patients with primary hypercholesterolemia or mixed dyslipidemia were assigned to 20 mg/day rosuvastatin and blood samples were drawn at study entry and after 12 weeks of treatment. A label-free LC-MS/MS protein profiling was conducted, coupled with bioinformatics analysis. Sixty-nine HDL proteins were identified, belonging to four main biological function clusters: lipid transport and metabolism; platelet activation, degranulation, and aggregation, wound response and wound healing; immune response; inflammatory and acute phase response. Five HDL proteins showed statistically significant differences in the abundance (Anova ≤ 0.05), before and after rosuvastatin treatment. Platelet factor 4 variant (PF4V1), Pregnancy-specific beta-1-glycoprotein 2 (PSG2), Profilin-1 (PFN1) and Keratin type II cytoskeletal 2 epidermal (KRT2) showed decreased expressions, while Integrin alpha-IIb (ITGA2B) showed an increased expression after treatment with rosuvastatin. The ELISA validation of PFN1 segregated the subjects into responders and non-responders, as PFN1 levels after rosuvastatin were shown to mostly depend on the subjects\u27 inflammatory phenotype. Findings from this study introduce novel insights into the HDL proteome and statin pleiotropism

Poli(laktid-ko-glikolid) mikročestice kao sustav za kontrolirano oslobađanje proteina: priprava i karakterizacija

Poly(DL-lactide-co-glycolide) (PDLLGA) and poly(L-lactide-co-glycolide) (PLLGA) copolymers were prepared by bulk ring opening polymerization of lactide and glycolide and characterized by GPC, FTIR, 1H NMR and DSC. Copolymers with different molar masses at a constant lactide/glycolide ratio were used for preparation of bovine serum albumin (BSA)-loaded microparticles by the double emulsion w/o/w method. The influence of the copolymer molar mass and composition on the microparticle morphology, size, yield, degradation rate, BSA-loading efficiency and BSA release profile were studied. For microparticles prepared from PDLLGA copolymers, a biphasic profile for BSA release was found and for those made from PLLGA copolymers the release profile was typically triphasic; both of them were characterized by high initial burst release. Possible reasons for such behavior are discussed.Poli(DL-laktid-ko-glikolid) (PDLLGA) i poli(L-laktid-ko-glikolid) (PLLGA) kopolimeri priređeni su polimerizacijom laktida i glikolida uz otvaranje prstenova i karakterizirani pomoću GPC, FTIR, 1H NMR i DSC. Kopolimeri različitih molarnih masa i stalnog omjera laktida i glikolida upotrebljeni su za pripravu mikročestica s goveđim serumskim albuminom (BSA) metodom dvostruke emulzije tipa voda/ulje/voda. Proučavan je utjecaj molarne mase i sastava kopolimera na oblik, veličinu, iskorištenje i stupanj razgradnje mikročestica, uklapanje i oslobađanje BSA. Za mikročestice pripravljene s PDLLGA kopolimerom utvrđen je bifazični profil oslobađanja BSA, a za mikročestice s PLLGA kopolimerom trifazičan profil. Za obje vrste karakteristično je brzo početno oslobađanje. Razmatrani su mogući uzroci takvog ponašanja

Poli(laktid-ko-glikolid) mikročestice kao sustav za kontrolirano oslobađanje proteina: priprava i karakterizacija

Poly(DL-lactide-co-glycolide) (PDLLGA) and poly(L-lactide-co-glycolide) (PLLGA) copolymers were prepared by bulk ring opening polymerization of lactide and glycolide and characterized by GPC, FTIR, 1H NMR and DSC. Copolymers with different molar masses at a constant lactide/glycolide ratio were used for preparation of bovine serum albumin (BSA)-loaded microparticles by the double emulsion w/o/w method. The influence of the copolymer molar mass and composition on the microparticle morphology, size, yield, degradation rate, BSA-loading efficiency and BSA release profile were studied. For microparticles prepared from PDLLGA copolymers, a biphasic profile for BSA release was found and for those made from PLLGA copolymers the release profile was typically triphasic; both of them were characterized by high initial burst release. Possible reasons for such behavior are discussed.Poli(DL-laktid-ko-glikolid) (PDLLGA) i poli(L-laktid-ko-glikolid) (PLLGA) kopolimeri priređeni su polimerizacijom laktida i glikolida uz otvaranje prstenova i karakterizirani pomoću GPC, FTIR, 1H NMR i DSC. Kopolimeri različitih molarnih masa i stalnog omjera laktida i glikolida upotrebljeni su za pripravu mikročestica s goveđim serumskim albuminom (BSA) metodom dvostruke emulzije tipa voda/ulje/voda. Proučavan je utjecaj molarne mase i sastava kopolimera na oblik, veličinu, iskorištenje i stupanj razgradnje mikročestica, uklapanje i oslobađanje BSA. Za mikročestice pripravljene s PDLLGA kopolimerom utvrđen je bifazični profil oslobađanja BSA, a za mikročestice s PLLGA kopolimerom trifazičan profil. Za obje vrste karakteristično je brzo početno oslobađanje. Razmatrani su mogući uzroci takvog ponašanja

The impact of molecular tumor profiling on the design strategies for targeting myeloid leukemia and EGFR/CD44-positive solid tumors

Nanomedicine has emerged as a novel cancer treatment and diagnostic modality, whose design constantly evolves towards increasing the safety and efficacy of the chemotherapeutic and diagnostic protocols. Molecular diagnostics, which create a great amount of data related to the unique molecular signatures of each tumor subtype, have emerged as an important tool for detailed profiling of tumors. They provide an opportunity to develop targeting agents for early detection and diagnosis, and to select the most effective combinatorial treatment options. Alongside, the design of the nanoscale carriers needs to cope with novel trends of molecular screening. Also, multiple targeting ligands needed for robust and specific interactions with the targeted cell populations have to be introduced, which should result in substantial improvements in safety and efficacy of the cancer treatment. This article will focus on novel design strategies for nanoscale drug delivery systems, based on the unique molecular signatures of myeloid leukemia and EGFR/CD44-positive solid tumors, and the impact of novel discoveries in molecular tumor profiles on future chemotherapeutic protocols

Mikroinkapsuliranje bakterije Lactobacillus casei u izolatu sojinog proteina i alginata pomoću sušenja raspršivanjem

This article presents a novel formulation for preparation of Lactobacillus casei 01 encapsulated in soy protein isolate and alginate microparticles using spray drying method. A response surface methodology was used to optimise the formulation and the central composite face-centered design was applied to study the effects of critical material attributes and process parameters on viability of the probiotic after microencapsulation and in simulated gastrointestinal conditions. Spherical microparticles were produced in high yield (64 %), narrow size distribution (d50=9.7 µm, span=0.47) and favourable mucoadhesive properties, with viability of the probiotic of 11.67, 10.05, 9.47 and 9.20 log CFU/g after microencapsulation, 3 h in simulated gastric and intestinal conditions and four-month cold storage, respectively. Fourier-transform infrared spectroscopy confirmed the probiotic stability after microencapsulation, while differential scanning calorimetry and thermogravimetry pointed to high thermal stability of the soy protein isolate-alginate microparticles with encapsulated probiotic. These favourable properties of the probiotic microparticles make them suitable for incorporation into functional food or pharmaceutical products.U radu je prikazan novi način pripreme mikrokapsula bakterije Lactobacillus casei 01 u izolatu sojinog proteina i alginata pomoću sušenja raspršivanjem. Metodologijom odzivnih površina optimiran je sastav mikrokapsula, a korištenjem plošno centriranog plana ispitan je utjecaj glavnih sastojaka kapsula te parametara procesa na preživljavanje mikroinkapsuliranog probiotika u simuliranim uvjetima gastrointestinalnog trakta. Najviše je proizvedeno kuglastih mikročestica (64 %), s raspodjelom veličina od d50=9,7 μm (raspon od 0,47) i povoljnim mukoadhezivnim svojstvima. Nakon mikroenkapsulacije preživjelo je 11,67 log CFU/g probiotika, nakon 3 sata u simuliranom želučanom soku 10,50 log CFU/g, u simuliranom soku tankog crijeva 9,47 log CFU/g, a nakon četiri mjeseca hladnog skladištenja 9,20 log CFU/g. Stabilnost probiotika nakon mikroinkapsulacije potvrđena je infracrvenom spektroskopijom s Fourierovom transformacijom, a toplinska stabilnost mikrokapsula probiotika u izolatu proteina soje i alginatu diferencijalnom pretražnom kalorimetrijom i termogravimetrijom. Zaključeno je da se zbog njihovih povoljnih svojstava mikrokapsule probiotika mogu upotrijebiti u proizvodnji funkcionalne hrane i farmaceutskih proizvoda

Design of Experiments (DoE)-based approach for improvement of dry mixing processes in the production of low-dose Alprazolam tablets using Raman spectroscopy for content uniformity monitoring

A low-dose tablet formulation, containing a potent Benzodiazepine derivative Alprazolam was developed, considering the achievement of appropriate content uniformity of the active substance in powder blends and tablets as a major challenge. Two different types of lactose monohydrate (Tablettose 80 and Granulac 200) and two different types of dry mixing processes (high-shear mixing and "in bulk" mixing) were employed. To evaluate the influence of the variables (mixing speed, mixing time, filling level of the high-shear and cube mixer, lactose monohydrate type) and their interactions upon the response (content uniformity of Alprazolam in the powder blends), a Factorial 2 4 design (with 4 factors at 2 levels in 1 block) was generated for each type of mixer. For high-shear dry mixing the Response Surface, D-optimal Factorial 2 4 design (with 2 replications and 31 experiments) was used, while for the "in bulk" dry mixing the Response Surface, Central Composite Factorial 2 4 design (with 34 experiments) was used. The process parameters for the high-shear mixer were varied within the following ranges: filling level of 70-100%, impeller mixing speed of 50-300 rpm and mixing time of 2-10 minutes. For the cube mixer the following process parameter ranges were employed: filling level of 30-60%, mixing speed of 20-390 rpm and mixing time of 2-10 minutes. Raman spectroscopy in conjunction with a validated Partial Least Square (PLS) regression model was used as a Process Analytical Technology (PAT) tool for Alprazolam content determination and content uniformity monitoring. The DoE model was further employed to optimize the powder blending process in regard to the achievement of appropriate Alprazolam content uniformity using high-shear mixing and Tabletosse 80 as filler. The desirability function revealed that the following process parameters: a mixing time of 2 minutes, a mixing speed of 300 rpm and a 70% filling level of the mixer would produce powder blends with the lowest variability in Alprazolam content. The three independent lab batches of low-dose Alprazolam tablets, produced with high-shear mixing using these process parameters, conformed to the requirements of the European Pharmacopoeia for content (assay) of Alprazolam and uniformity of the dosage units

(Benzoylamino)methyl 4-Hydroxybenzoate

(Benzoylamino)methyl 4-hydroxybenzoate (“Benzamidomethylparaben”) (3) was obtained from a reaction of 4-hydroxybenzoic acid (2) with a dioxane suspension of (benzamidomethyl)triethylammonium chloride (1). The phenolic group in 2 cannot be benzamidomethylated with 1 in aqueous media