Enhancement of a biotechnological platform for the purification and delivery of a human papillomavirus supercoiled plasmid DNA vaccine.

New biotechnological strategies are being explored, aimed at rapid and economic manufacture of large quantities of DNA vaccines with the required purity for therapeutic applications, as well as their correct delivery as biopharmaceuticals to target cells. This report describes the purification of supercoiled (sc) HPV-16 E6/E7 plasmid DNA (pDNA) vaccine from a bacterial lysate, using an arginine-based monolith, presenting a spacer arm in its configuration. To enhance the performance of the purification process, monolith modification with the spacer arm can improve accessibility of the arginine ligand. By using a low NaCl concentration at pH 7.0, a condition to eliminate the RNA impurity directly in the flow through was established. The pH increase to 7.5 allowed the elimination of non-functional pDNA isoforms, the sc pDNA being recovered by increasing the ionic strength. As well as a binding capacity of 2.53 mg/mL obtained with a pre-purified sc pDNA sample, the column also purified sc pDNA from high lysate loading, with capacities above 1 mg/mL. Due to the sample displacement phenomena, non-functional pDNA isoforms were eliminated throughout column loading, favoring the degree of purity of final sc pDNA of 93.3%-98.5%. Thereafter, purified sc pDNA was successfully encapsulated into CaCO3-gelatin nano-complexes. Delivery of the pDNA-carriers to THP-1 cells was assessed through pDNA cellular uptake evaluation and correct E6 expression was verified by mRNA and protein detection. A biotechnological platform was established for sc pDNA purification and delivery to dendritic cells, stimulating further in vivo studies.

Ultrasonographic methods for evaluation of testicles in cats

The testicles are the primary sexual organs of male and their function is to produce sperm and sexual hormones. Disorders of the testicles are common in domestic cats. Therefore, detailed assessment of the testes is of great importance in veterinary medicine. Considering the recent advances in diagnostic imaging in companion animals, this review aims to describe the applicability elastography (qualitative and quantitative), Doppler, contrast-enhanced ultrasonography and B-mode ultrasonography in testes evaluation in cats. B-mode ultrasonography of the testicles combined with haemodynamic analysis in real time by Doppler and contrast enhanced ultrasonography can assist as diagnostic tool in evaluating testicular abnormalities in sick cats. Furthermore, ARFI elastography is a new ultrasound method that evaluates tissue elasticity by elastogram and shear weave. Ultrasonographic study of the testes is a common diagnostic imaging procedure.

Los testículos son los principales órganos sexuales de los machos y su función es producir espermatozoides y hormonas sexuales. Los trastornos de estos órganos son comunes en gatos domésticos. La ultrasonografía convencional, combinada con el análisis hemodinámico en tiempo real por Doppler y ultrasonido contrastado son herramientas diagnósticas importantes para la evaluación de enfermedades testiculares en gatos. Además, La elastografía-ARFI es un método ecográfico actual que evalúa la elasticidad tisular por medio del elastograma y de la velocidad de propagación de las ondas de cizallamiento. El estudio ecográfico de los testículos es un procedimiento de diagnóstico por imágenes común.

Ultrasonography of the canine pancreas / La ecografía del páncreas canino

ABSTRACT This study describes the ultrasonographic techniques currently used in the evaluation of the canine pancreas. Ultrasonography was the first method to enable direct visualization of the pancreas in humans and it has been subsequently applied to animals. Currently, it is the method of choice for pancreatic evaluation and is essential as a diagnostic tool in the detection of abnormalities, especially tumors. Innovative equipment technology has led to the emergence of techniques complementary to B-mode ultrasound; such as Doppler, elastography, and contrast-enhanced ultrasonography, which have enabled more accurate diagnosis. Doppler provides information on vascular architecture and the hemodynamic aspect of blood vessels in multiple organs. ARFI elastography provides detailed images of the alterations detected by conventional examination (qualitative method) and assists in differentiating between benign and malignant processes (quantitative method). Microbubble contrast agents determine parameters related to homogeneous and heterogeneous filling of organs with microbubbles, mainly nodular areas, thus defining high and low intensity patterns.

RESUMEN Este estudio describe las técnicas ecográficas frecuentemente utilizadas para evaluar el pancreas del perro. La ecografía fue el primer método que permitió la visualización directa del páncreas en seres humanos y que luego se aplicó en animales. Es actualmente el método de elección para la evaluación del páncreas y es esencial como herramienta diagnóstica en la detección de anomalías, especialmente tumores. La tecnología innovadora de los equipos, llevó a la aparición de técnicas complementarias al modo B, tales como Doppler, elastografía, ecografía de contraste, que han permitido realizar diagnósticos más precisos. El Doppler proporciona información sobre la arquitectura vascular y aspectos hemodinámicos de los vasos sanguíneos en múltiples órganos. La elastografía ARFI ofrece imágenes detalladas de las alteraciones detectadas en exámenes convencionales (método cualitativo) y ayuda a diferenciar entre procesos benignos y malignos (método cuantitativo). Los agentes de contraste con microburbujas permiten determinar parámetros relacionados con el llenado homogéneo o heterogéneo de los órganos, principalmente áreas nodulares, definiendo, por tanto, patrones de alta o baja intensidad.

[Non steroidal anti-inflammatory drugs prescription in General Practice in the centre of Portugal from 2007 to 2009]. / Prescrição de anti-inflamatórios não esteróides em medicina geral e familiar no centro de Portugal entre 2007 e 2009.

BACKGROUND: Non steroidal anti-inflammatory drugs computer-assisted prescription in General Practice/Family Medicine (GP/FM) ambience can give information about the prescription profile during a period of time and understand its trend. Such prescription profile could vary according to the urban (city) vs non urban (village) setting of the GP/FM practice. OBJECTIVES: To characterize the prescription trends of non steroidal inflammatory drugs (NSAIDs) in General Practice, from 2007 to 2009, by public sales price per inscript, daily defined doses (DDD) per inscript and price of DDD. To verify differences of the three fractions according to the urban vs non urban health centre. MATERIAL AND METHODS: Retrospective observational study conducted in February 2010, by the analysis of prescription informatic files automatically generated when the prescription was made. Population calculated at the middle point of each period of study. The differences in volume prescription in Daily Defined Dose (DDD) per registered patient in the middle of 2006 and 2007 two semesters, as well as the value of prescription measured by the price per registered patient were calculated and medicines were studied by the third level of the Portuguese medicines classification very similar to the ATC. The price per DDD was calculated as well. RESULTS: The studied population was of 315.377 subjects in 2007, 321.784 in 2008 and 331.106 in 2009. In price per inscript urban health centres showed a growth from 2007 to 2009 in "Indol and Inden" derivatives (+51.6%), in "Propionic acid derivatives" (+39.4%) and in "Selective cox-2 inhibitors" (+33.6%). For "Sulfanilamidic derivatives, a reduction of 3.8% was verified. For non urban Health Centres prescription a generalised reduction in price per inscript was observed. For DDD/per inscript from 2007 to 2009 we found a generalised increase for urban health centres, except for "Oxicans". For non urban Health Centres there is a generalised decrease of DDD/per inscript from 2007 to 2009, with a special decrease for non-acidic compounds. As for Price of DDD from 2007 to 2009 there is a generalised reduction of such ratio with the only exception of "non acidic compounds" (+9,1%) for urban Health Centres. In view of the calculated ratios, the differences by urban vs non urban Health Centre in each of the studied years there is a significant higher value in non urban health centre, with the exception of Price of DDD for 2008 and 2009 where such ratio is non significantly higher in urban heath centre. CONCLUSIONS: During the study period, demographic growth was higher than the prescription growth in volume and in value. Prescription was more frequent and expensive in non-urban Health Centres. Price of DDD decreases from 2007 to 2009 signifying a much cheaper NSAIDs therapeutics.