Cholesterol improves the transfection efficiency of polyallylamine as a non-viral gene delivery vector

ABSTRACT Cationic polymers such as polyallylamine (PAA) having primary amino groups are poor transfection agents and possess a high cytotoxicity index when used without any chemical modification. In this study, PAA was modified with cholesterol in order to improve transfection efficiency and to reduce cytotoxicity. PAA polymers with molecular weights of 15 and 65 kDa were selected and grafted with cholesterol at percentages of 5, 10, 15, 30, and 50. After purification, the efficacy of the synthetic vectors was evaluated in terms of DNA condensation using the ethidium bromide test, buffering capacity, particle size, zeta potential, transfection efficiency, and cytotoxicity assay in Neuro2A cell lines. According to the ethidium bromide test, these vectors can condense DNA at moderate and high carrier to plasmid (C/P) ratios. The buffering capacity of the prepared vector in both molecular weights was less than unmodified PAA. Particle size measurements demonstrated that modified PAAs were able to form nanoparticles ranging in size from 125 to 530 nm. The vectors based on PAA 15 kDa demonstrated a better transfection efficiency than the vectors made of PAA 65 kDa. Cytotoxicity studies showed that toxicity of all vectors was less than PAA. Some cholesterol modified polymers composed of PAA (15 kDa) were suitable vectors for gene delivery with low cytotoxicity.

Treatment of patients with aortic atherosclerotic disease with paclitaxel-associated lipid nanoparticles

OBJECTIVE: The toxicity of anti-cancer chemotherapeutic agents can be reduced by associating these compounds, such as the anti-proliferative agent paclitaxel, with a cholesterol-rich nanoemulsion (LDE) that mimics the lipid composition of low-density lipoprotein (LDL). When injected into circulation, the LDE concentrates the carried drugs in neoplastic tissues and atherosclerotic lesions. In rabbits, atherosclerotic lesion size was reduced by 65% following LDE-paclitaxel treatment. The current study aimed to test the effectiveness of LDE-paclitaxel on inpatients with aortic atherosclerosis. METHODS: This study tested a 175 mg/m2 body surface area dose of LDE-paclitaxel (intravenous administration, 3/3 weeks for 6 cycles) in patients with aortic atherosclerosis who were aged between 69 and 86 yrs. A control group of 9 untreated patients with aortic atherosclerosis (72-83 yrs) was also observed. RESULTS: The LDE-paclitaxel treatment elicited no important clinical or laboratory toxicities. Images were acquired via multiple detector computer tomography angiography (64-slice scanner) before treatment and at 1-2 months after treatment. The images showed that the mean plaque volume in the aortic artery wall was reduced in 4 of the 8 patients, while in 3 patients it remained unchanged and in one patient it increased. In the control group, images were acquired twice with an interval of 6-8 months. None of the patients in this group exhibited a reduction in plaque volume; in contrast, the plaque volume increased in three patients and remained stable in four patients. During the study period, one death unrelated to the treatment occurred in the LDE-paclitaxel group and one death occurred in the control group. CONCLUSION: Treatment with LDE-paclitaxel was tolerated by patients with cardiovascular disease and showed the potential to reduce atherosclerotic lesion size.

Clinical use of a ceramide-based moisturizer for treating dogs with atopic dermatitis

In humans, skin barrier dysfunction is thought to be responsible for enhanced penetration of allergens. Similar to conditions seen in humans, canine atopic dermatitis (CAD) is characterized by derangement of corneocytes and disorganization of intercellular lipids in the stratum corenum (SC) with decreased ceramide levels. This study was designed to evaluate the effects of a moisturizer containing ceramide on dogs with CAD. Dogs (n = 20, 3~8 years old) with mild to moderate clinical signs were recruited and applied a moisturizer containing ceramide for 4 weeks. Transepidermal water loss (TEWL), skin hydration, pruritus index for canine atopic dermatitis (PICAD) scores, and canine atopic dermatitis extent and severity index (CADESI) scores of all dogs were evaluated. Skin samples from five dogs were also examined with transmission electron microscopy (TEM) using ruthenium tetroxide. TEWL, PICAD, and CADESI values decreased (p < 0.05) and skin hydration increased dramatically over time (p < 0.05). Electron micrographs showed that the skin barrier of all five dogs was partially restored (p < 0.05). In conclusion, these results demonstrated that moisturizer containing ceramide was effective for treating skin barrier dysfunction and CAD symptoms.

Patterns of sudanophilic lesions at ostia of descending thoracic and abdominal aorta in cholesterol-fed adult rabbits / Patrones de lesiones sudanofílicas en el ostio de las partes torácica y abdominal de la aorta en conejos adultos alimentados con colesterol

Sudan-positive atherosclerotic lesions preferentially occur at downstream and lateral margins of arterial branch ostia in human neonates and weanling rabbits but tend to develop at lateral and upstream margins in old subjects. We investigated (i) the pattern of sudanophilic lesions at aortic ostia of cholesterol-fed adult rabbits and (ii) determined any differences in lesion distribution between descending thoracic and abdominal aorta. Ten adult males, New Zealand white rabbits were fed 2 percent high cholesterol diet. After six weeks, aortas were excised, opened longitudinally and stained with Sudan-IV for gross examination of atherosclerotic lesions. A total of 156 descending thoracic and 90 abdominal ostia were examined. Mean lesion frequencies upstream, downstream and at lateral margins of the affected ostia were calculated and compared. Sudanophilic lesions were detected around 32 percent ostia of descending thoracic aorta and 25 percent those of abdominal aorta. At ostia of descending thoracic aorta, lesion frequencies were significantly higher (P<0.001) downstream (95 percent) and at lateral margins (92 percent) than upstream (2 percent). In abdominal aorta, lateral (100 percent) and upstream (43 percent) margins were significantly (P<0.05) more affected while minimal lesion frequencies were seen at downstream branch points (9 percent). Comparison between descending thoracic and abdominal aorta showed an insignificant difference of lesion frequencies at lateral margins (P>0.05) but a highly significant difference at upstream versus downstream of ostia (P<0.001). We concluded that in cholesterol-fed adult rabbits, juvenile pattern of downstream lipid deposition persists at ostia of descending thoracic aorta while a switch towards the upstream pattern of old subjects occurs at ostia of abdominal aorta.

Las lesiones ateroscleróticas Sudán-positivas se producen preferentemente en los márgenes posteriores y laterales del ostio de las ramas arteriales en neonatos humanos y en conejos destetados, pero tienden a desarrollarse en los márgenes laterales y superiores en sujetos de edad avanzada. Investigamos el patrón de lesiones sudanofílicas en el ostio aórtico de conejos adultos alimentados con colesterol y determinamos las diferencias en la distribución de lesiones entre la aorta torácica descendente y abdominal. Diez conejos machos blancos adultos New Zealand fueron alimentados con una dieta alta en colesterol al 2 por ciento. Después de seis semanas, fueron extraídas las aortas, se disecaron longitudinalmente y se tiñeron con Sudan-IV para el examen macroscópico y se observaron las lesiones ateroscleróticas. Fueron examinados 156 ostios de aortas torácicas descendentes y 90 de aortas abdominales. Fueron calculadas y comparadas las frecuencias medias de lesiones superiores, inferiores y de los márgenes laterales de los ostios afectados. Las lesiones sudanofílicas se detectaron en alrededor del 32 por ciento de ostios de la aorta torácica descendente y en el 25 por ciento de las aortas abdominales. En el ostio de la aorta torácica descendente, las frecuencias de lesiones fueron significativamente mayores (p <0,001) por superior (95 por ciento) y en los márgenes laterales (92 por ciento) que por inferior (2 por ciento). En la aorta abdominal, los márgenes laterales (100 por ciento) y superiores (43 por ciento) fueron significativamente más afectados (p <0,05), mientras que las frecuencias mínimas de lesiones se observaron en los puntos de la rama descendente (9 por ciento). La comparación entre la aorta torácica descendente y abdominal mostró una diferencia no significativa de las frecuencias de lesiones en los márgenes laterales (p> 0,05), pero muy significativa al comparar el margen superior con el inferior (p <0,001). Llegamos a la conclusión que en los conej...

¿Será útil la litotripsia en el Hospital ABC? / Is lithotripsy useful at the ABC Hospital?

La lotrotripsia por ondas de choque extracorporeas (LOCE) ha recibido atención creciente como una alternativa viable a la colecistectomía. No existen datos sobre que porciento de los pacientes actualmente tratados por patología biliar por año en el hospital ABC pueden ser elegibles para este procedimiento. Estudiamos 47 pacientes consecutivos operados de colecistectomía y aplicamos los criterios de exclusión actuales para determinar la posibilidad de ser candidatos a LOCE. En cada paciente se obtuvo historia clínica preoperatoria y hallazgos quirúrgicos. El tamaño, número y características macroscópicas de loscálculos fue evaluado. En 36 por ciento los cálculos fueron de colesterol, en 34 por ciento mixtos, en 15 por ciento de pigmento y en 15 por ciento no hubo cálculos. De los 47 pacientes únicamente 3(6.3 por ciento) no tuvieron criterios de exclusión, y por lo tanto, hubieran sido elegibles. Estos datos sugieren que aproximadamente 18 entre 300 pacientes por año podrían ser candidatos a LOCE en nuestro hospital.