Control of cell penetration enhancer shielding and endosomal escape-kinetics crucial for efficient and biocompatible siRNA delivery.

Although cationic liposomes are efficient carriers for nucleic acid delivery, their toxicity often hampers the clinical translation. Polyethylene glycol (PEG) coating has been largely used to improve their stability and reduce toxicity. Nevertheless, it has been found to decrease the transfection process. In order to exploit the advantages of cationic liposomes and PEG decoration for nucleic acid delivery, liposomes decorated with tetraArg-[G-1]-distearoyl glycerol (Arg4-DAG) dendronic oligo-cationic lipid enhancer (OCE) and PEG-lipid have been investigated. Non decorated or OCE-decorated lipoplexes (OCEfree-LPX and OCE-LPX, respectively) were obtained by lipid film hydration using oligonucleotide (ON) solutions. PEG and OCE/PEG decorated lipoplexes (PEG-OCEfree-LPX and PEG-OCE-LPX, respectively) were obtained by post-insertion of 2 or 5 kDa PEG-DSPE on preformed lipoplexes. The OCE decoration yielded lipoplexes with size of about 240 nm, 84% loading efficiency at 10 N/P ratio, ten times higher than OCEfree-LPX, and prevented the ON release when incubated with physiological heparin concentration or with plasma. The PEG decoration reduced the zeta potential, enhanced the lipoplex stability in serum and decreased both hemolysis and cytotoxicity, while it did not affect the lipoplex size and ON loading. With respect to OCEfree-LPX, the OCE-LPX remarkably associated with cells and were taken up by different cancer cell lines (HeLa and MDA-MB-231). Interestingly, 2 or 5 kDa PEG decoration did not reduce either the cell interaction or the cell up-take of the cationic lipoplexes. With siRNA as a payload, OCE enabled efficient internalization, but endosomal release was hampered. Post-transfection treatment with the lysosomotropic drug chloroquine allowed to identify the optimal time point for endosomal escape. Chloroquine treatment after 12 to 20 h of LPX pre-incubation enabled siRNA mediated target knockdown indicating that this is the time window of endo-lysosomal processing. This indicates that OCE can protect siRNA from lysosomal degradation for up to 20 h, as shown by these rescue experiments.

Influence of Folate-Targeted Gold Nanoparticles on Subcellular Localization and Distribution into Lysosomes.

The cell interaction, mechanism of cell entry and intracellular fate of surface decorated nanoparticles are known to be affected by the surface density of targeting agents. However, the correlation between nanoparticles multivalency and kinetics of the cell uptake process and disposition of intracellular compartments is complicated and dependent on a number of physicochemical and biological parameters, including the ligand, nanoparticle composition and colloidal properties, features of targeted cells, etc. Here, we have carried out an in-depth investigation on the impact of increasing folic acid density on the kinetic uptake process and endocytic route of folate (FA)-targeted fluorescently labelled gold nanoparticles (AuNPs). A set of AuNPs (15 nm mean size) produced by the Turkevich method was decorated with 0-100 FA-PEG3.5kDa-SH molecules/particle, and the surface was saturated with about 500 rhodamine-PEG2kDa-SH fluorescent probes. In vitro studies carried out using folate receptor overexpressing KB cells (KBFR-high) showed that the cell internalization progressively increased with the ligand surface density, reaching a plateau at 50:1 FA-PEG3.5kDa-SH/particle ratio. Pulse-chase experiments showed that higher FA density (50 FA-PEG3.5kDa-SH molecules/particle) induces more efficient particle internalization and trafficking to lysosomes, reaching the maximum concentration in lysosomes at 2 h, than the lower FA density of 10 FA-PEG3.5kDa-SH molecules/particle. Pharmacological inhibition of endocytic pathways and TEM analysis showed that particles with high folate density are internalized predominantly by a clathrin-independent process.

Design and in vitro/in vivo Evaluation of Polyelectrolyte Complex Nanoparticles Filled in Enteric-Coated Capsules for Oral Delivery of Insulin.

Insulin is one of the most important drugs in the treatment of diabetes. There is an increasing interest in the oral administration of insulin as it mimics the physiological pathway and potentially reduces the side effects associated with subcutaneous injection. Therefore, insulin-loaded polyelectrolyte complex (PEC) nanoparticles were prepared by the ionic cross-linking method using protamine sulfate as the polycationic and sodium alginate as the anionic polymer. Taguchi experimental design was used for the optimization of nanoparticles by varying the concentration of sodium alginate, the mass ratio of sodium alginate to protamine, and the amount of insulin. The optimized nanoparticle formulation was used for further in vitro characterization. Then, insulin-loaded PEC nanoparticles were placed in hard gelatin capsules and the capsules were enteric-coated by Eudragit L100-55 (PEC-eCAPs). Hypoglycemic effects PEC-eCAPs were determined in vivo by oral administration to diabetic rats. Furthermore, in vivo distribution of PEC nanoparticles was evaluated by fluorescein isothiocyanate (FITC) labelled nanoparticles. The experimental design led to nanoparticles with a size of 194.4 nm and a polydispersity index (PDI) of 0.31. The encapsulation efficiency (EE) was calculated as 95.96%. In vivo studies showed that PEC-eCAPs significantly reduced the blood glucose level of rats at the 8th hour compared to oral insulin solution. It was concluded that PEC nanoparticles loaded into enteric-coated hard gelatin capsules provide a promising delivery system for the oral administration of insulin.

Outcomes of bladder preservation therapy on survival in patients with muscle-invasive bladder cancer / Resultados de preservación vesical en pacientes con tumor vesical musculoinfiltrante

INTRODUCCIÓN: El estándar de tratamiento en el CVMI es la cistectomía radical, aunque la RTUv + RTP+ quimioterapia sistémica demuestra resultados comparables a la cistectomía radical en términos de control local y supervivencia global. OBJETIVOS: Evaluar nuestros resultados en terapia trimodal en cáncer de vejiga músculo-invasivo que rechazan la cistectomía radical MÉTODOS: Análisis retrospectivo de 23 pacientes con estadio T3 TVMI tratados con preservación vesical (RTUv + 3 ciclos de gemcitabina, cisplatino+ 64Gy RTP adyuvante). KM estimados y log Rank fueron calculados. RESULTADOS: La mediana de seguimiento fue de 58 meses (15-158). El intervalo libre de enfermedad y la supervivencia global a los 5 años fue de 56% y 64%, respectivamente. La Supervivencia cáncer especifica fue de 67%. No se objetivaron complicaciones grado 3 o más. CONCLUSIONES: Nuestra serie de tratamiento preservación vesical demuestra que el uso de este tratamiento en pacientes debidamente seleccionados que no quieren cistectomía radical es apropiado

INTRODUCCIÓN: El estándar de tratamiento en el CVMI es la cistectomía radical, aunque la RTUv + RTP+ quimioterapia sistémica demuestra resultados comparables a la cistectomía radical en términos de control local y supervivencia global. OBJETIVOS: Evaluar nuestros resultados en terapia trimodal en cáncer de vejiga músculo-invasivo que rechazan la cistectomía radical. MÉTODOS: Análisis retrospectivo de 23 pacientes con estadio T3 TVMI tratados con preservación vesical (RTUv +3 ciclos de gemcitabina, cisplatino+ 64Gy RTP adyuvante). KM estimados y log Rank fueron calculados. RESULTADOS: La mediana de seguimiento fue de 58 meses (15-158). El intervalo libre de enfermedad y la supervivencia global a los 5 años fue de 56% y 64%, respectivamente. La Supervivencia cáncer especifica fue de 67%. No se objetivaron complicaciones grado 3 o más. CONCLUSIONES: Nuestra serie de tratamiento preservación vesical demuestra que el uso de este tratamiento en pacientes debidamente seleccionados que no quieren cistectomía radical es apropiado

Upper Zone of Growth Plate and Cartilage Matrix (UCMA) Levels in Patients with Chronic Kidney Disease

Abstract Chronic kidney disease (CKD) is an important health problem across the world affecting the adult population with an enormous social and economic burden. Calcium regulation is also affected in patients with CKD, and related to several disorders including vascular calcifications, mineral bone disorders, and cardiovascular diseases (CVD). Upper zone of growth plate and cartilage matrix (UCMA) is vitamin K-dependent protein (VKDP) and acts as a calcification inhibitor in the cardiovascular system. The molecular mechanism of UCMA action remains unclear in CKD. In the current study, we aimed to investigate serum total UCMA levels and its association with calcium metabolism parameters in CKD patients including hemodialysis (HD) patients. Thirty-seven patients with CKD stage 3-5, 41 HD patients, and 34 healthy individuals were enrolled in this cross-sectional study. Serum UCMA and calcification related protein levels (Matrix Gla Protein (MGP), Osteocalcin (OC), and Fetuin-A) were analyzed with enzyme-linked immunosorbent assay (ELISA). Calcium mineral disorder parameters (Serum Ca, P, iPTH) were quantified with routine techniques. We, for the first time, report the potential biomarker role of UCMA in CKD including HD. Serum total UCMA levels were significantly higher in patients with CKD including HD patients than the healthy controls. Also, serum UCMA levels showed negative correlations with serum calcium, and eGFR, while showed positive relationships with P, iPTH, MGP, OC. Increased total UCMA levels may have a role in the Ca metabolism disorder and related to the pathogenesis of Vascular Calcification in patients with CKD.

Thermally and photocatalytically active multifunctional porous coatings for high temperature applications.

In this study, the thermal catalytic powder mixtures of Mn, Ce and Co oxides supported on Al2O3 were synthesized. In order to obtain thermally active porous coating on the steel substrate, a binder solution produced via sol-gel method was used to bind the synthesized powder. Additionally by incorporation of TiO2 nanoparticles to previously described thermally active coating, both thermally and photocatalytically active multifunctional surfaces were obtained. The self-cleaning property and the rate of volatile organic compounds (VOC) degradation of the coatings were studied, in which pyrazine, thiazole, maltol, vanillin and toluene were applied as VOC. In TiO2 applications UV illuminations were applied. In the self-cleaning performance tests using soybean oil, the thermally active coating was slightly better than the TiO2 containing coatings. Under UV illumination, the high rates of VOC (photo) degradation was achieved with the thermally and photocatalytically active coating.

Hairy Polyp of the Supratonsillar Fossa Causing Intermittent Airway Obstruction

Introduction Dermoids frequently called "hairy polyps" and their nature have not been completely clarified. Objectives To discuss the unusual presentation, symptoms, incidence, histology, and perioperative management of hairy polyps in the light of a case and current literature. Resumed Report A 3-year-old boy presented with intermittent respiratory distress since birth. Oropharyngeal examination revealed a nasopharyngeal mass originating from the supratonsillar fossa. The mass was so mobile that it moved between the oropharynx and the nasopharynx during swallowing. The radiologic and pathologic examinations confirmed the mass as a hairy polyp. Conclusion In a pediatric age group with airway obstruction, hairy polyps of the oropharyngeal region must also be included in the differential diagnosis. .

Photocatalytic antibacterial performance of Sn(4+)-doped TiO(2) thin films on glass substrate.

Pure anatase, nanosized and Sn(4+) ion doped titanium dioxide (TiO(2)) particulates (TiO(2)-Sn(4+)) were synthesized by hydrothermal process. TiO(2)-Sn(4+) was used to coat glass surfaces to investigate the photocatalytic antibacterial effect of Sn(4+) doping to TiO(2) against gram negative Escherichia coli (E. coli) and gram positive Staphylococcus aureus (S. aureus). Relationship between solid ratio of TiO(2)-Sn(4+) in coatings and antibacterial activity was reported. The particulates and the films were characterized using particle size analyzer, zeta potential analyzer, Brunauer-Emmett-Teller (BET), X-ray diffractometer (XRD), SEM, AAS and UV/VIS/NIR techniques. The results showed that TiO(2)-Sn(4+) is fully anatase crystalline form and easily dispersed in water. Increasing the solid ratio of TiO(2)-Sn(4+) from 10 to 50% in the coating solution increased antibacterial effect.

Preparation of re-usable photocatalytic filter for degradation of Malachite Green dye under UV and vis-irradiation.

Sn(4+) doped and undoped nano-TiO(2) particles easily dispersed in water were synthesized without using organic solvent by hydrothermal process. Nanostructure-TiO(2) based thin films were prepared on flyswatter substrate, made with stainless steel, by dip-coating technique. The structure, surface and optical properties of the particles and thin films were characterized by element analysis and XRD, BET, SEM and UV/vis/NIR techniques. The photocatalytic performance of the films were tested for degradation of Malachite Green dye in solution under UV and vis-lights. The results showed that the coated flyswatter has a very high photocatalytic performance for the photodegradation of Malachite Green irradiated with UV and vis-lights. The results also proved that the hydrothermally synthesized nano-TiO(2) particles are fully anatase crystalline form and are easily dispersed in water, the coated surfaces are hydrophilic, and the doping of transition metal ion efficiently improved the degradation performance of TiO(2)-coated flyswatter. The photocatalytic performances determined at both irradiation conditions were very good and were almost similar to each other for Sn(4+) doped TiO(2)-coated flyswatter and it can be repeatedly used with increasing photocatalytic activity compared to undoped TiO(2)-coated flyswatter.

Photocatalytic performance of Sn-doped and undoped TiO2 nanostructured thin films under UV and vis-lights.

Sn-doped and undoped nano-TiO(2) particles have been synthesized by hydrotermal process without solvent at 200 degrees C in 1h. Nanostructure-TiO(2) based thin films have been prepared on glass substrate by spin-coating technique. The structure, surface morphology and optical properties of the thin films and the particles have been investigated by element analysis and XRD, SEM, BET and UV-vis-NIR techniques. The photocatalytic performance of the films were tested for degradation of Malachite Green dye in solution under UV and vis-lights. The results showed that (a) hydrothermally synthesized nano-TiO(2) particles are fully anatase crystalline form and are easily dispersed in water, (b) the coated surfaces have nearly super-hydrophilic properties and, (c) the doping of transition metal ion efficiently improved the photocatalytic performance of the TiO(2) thin film.

Photocatalytic performance of Sn-doped TiO2 nanostructured mono and double layer thin films for Malachite Green dye degradation under UV and vis-lights.

Nanostructure Sn(4+)-doped TiO(2) based mono and double layer thin films, contain 50% solid ratio of TiO(2) in coating have been prepared on glass surfaces by spin-coating technique. Their photocatalytic performances were tested for degradation of Malachite Green dye in solution under UV and vis irradiation. Sn(4+)-doped nano-TiO(2) particle a doping ratio of about 5[Sn(4+)/Ti(OBu(n))(4); mol/mol%] has been synthesized by hydrotermal process at 225 degrees C. The structure, surface and optical properties of the thin films and/or the particles have been investigated by XRD, BET and UV/vis/NIR techniques. The results showed that the double layer coated glass surfaces have a very high photocatalytic performance than the other one under UV and vis lights. The results also proved that the hydrothermally synthesized nano-TiO(2) particles are fully anatase crystalline form and are easily dispersed in water. The results also reveal that the coated surfaces have hydrophilic property.

Characterization of the hydrothermally synthesized nano-TiO2 crystallite and the photocatalytic degradation of Rhodamine B.

Pure anatase-TiO2 nanoparticles with 8 nm average crystallite size was synthesized hydrothermally at 200 degrees C in 2 h. The structural and physico-chemical properties of nano-TiO2 were determined by powder XRD, FT-IR, BET and SEM analyses. The behavior of anatase nano-TiO2 in catalytic degradation of Rhodamine B (RB) dye in transparent nano-TiO2 sol under UV-light was examined as a function of irradiation power of UV-light, irradiation time, amount of nano-TiO2 and initial RB concentration in the sol. Rhodamine B was fully degraded with the catalysis of the nano-TiO2 in a short time as low as 60 min. Photocatalytic activity of the nano-TiO2 for degradation of RB was compared with Degussa P-25 at optimum catalysis conditions determined for the nano-TiO2. It was found that, when compared to Degussa P-25, the nano-TiO2 could be repeatedly used with increasing photocatalytic activity. It was found that the photodegradation obeys the pseudo first-order reaction kinetics with the rate constant of 0.0658 min(-1), and the half period t1/2 was 10.53 min.

The production and application of a regenerable filter system for adsorption of some atmospheric contaminants.

In this work, a new filter, used to prevent atmospherical pollution, has been developed. Spherical amorphous shaped silicates (such as KC-Siliperl AF 125 and Aluminium silicate 596 FA) were coated with different materials which were prepared from the hydrolysis-condensation products of organically modified silanes and metal alkoxides. The adsorption capacities of such silicates for different solvents; ethylacetate, toluene, n-hexane and cyclohexanone were investigated. It was found that KC-Siliperl AF 125 coated with IMEO (a commercial silane) adsorbed all the solvents better than Aluminium silicate 596 FA, and ethylacetate was adsorbed more than the other solvents.