In Vitro Methods for Specific IgE Detection on Cow’s Milk Allergy

Background: A new method for determining serum specific IgE (IMMULITE“ 2000 3gAllergy) has recently become available. Objective: To evaluate the clinical performance of IMMULITE 2000 in the diagnosis of cow’s milk allergy compared with that of UniCAP“. Additionally, we verified the behavior of both methods at two diagnostic decision points proposed by other authors. Methods: The study population consisted of 31 children with cow’s milk allergy (group A) and a control group of 19 atopic children without food allergy(group B). A blood sample from each child was tested using both methods and the results were compared. Results: In group A, the values for cow’s milk IgE ranged from 0.35 kU/L (the lowest common detection limit) to above 100 kU/L. In group B, the values were less than 1.1 kU/L for IMMULITE 2000 and less than 1.6 kU/L for UniCAP. An agreement of 90 % in IgE classes was obtained. Both methods demonstrated exactly the same diagnostic performance(sensitivity: 100 %; specificity: 78.9 %; negative predictive value: 100%; positive predictive value: 84.6%;efficiency: 90.2 %). The evaluation of the two methods at the two different decision points proposed in the literature showed a better positive predictive value with UniCAP, but we obtained equivalent performance with IMMULITE 2000 by choosing higher cutoff values. Conclusions: We conclude that IMMULITE 2000 is as effective as UniCAP in the diagnosis of cow’s milk allergy. Both methods can be used to obtain site-specific decision points that are population, age and disease dependent.

Differential Internalization of Amphotericin B - Conjugated Nanoparticles in Human Cells and the Expression of Heat Shock Protein 70

Although a variety of nanoparticles (NPs) functionalized with amphotericin B, an antifungal agent widely used in the clinic, have been studied in the last years their cytotoxicity profile remains elusive. Here we show that human endothelial cells take up high amounts of silica nanoparticles (SNPs) conjugated with amphotericin B (AmB) (SNP-AmB) (65.4 12.4 pg of Si per cell) through macropinocytosis while human fibroblasts internalize relatively low amounts (2.3 0.4 pg of Si per cell) because of their low capacity for macropinocytosis. We further show that concentrations of SNP-AmB and SNP up to 400 mg/mL do not substantially affect fibroblasts. In contrast, endothelial cells are sensitive to low concentrations of NPs (above 10 mg/mL), in particular to SNP-AmB. This is because of their capacity to internalize high concentration of NPs and high sensitivity of their membrane to the effects of AmB. Low-moderate concentrations of SNP-AmB (up to 100 mg/mL) induce the production of reactive oxygen species (ROS), LDH release, high expression of pro-inflammatory cytokines and chemokines (IL-8, IL-6, G-CSF, CCL4, IL-1b and CSF2) and high expression of heat shock proteins (HSPs) at gene and protein levels. High concentrations of SNP-AmB (above 100 ug/mL) disturb membrane integrity and kill rapidly human cells(60% after 5 h). This effect is higher in SNP-AmB than in SNP.

The Photoreceptor Cell-Specific Nuclear Receptor Gene (PNR ) Accounts for Retinitis Pigmentosa in the Crypto-Jews from Portugal (Marranos), Survivors from the Spanish Inquisition

The last Crypto-Jews (Marranos) are the survivors of Spanish Jews who were persecuted in the late fifteenth century, escaped to Portugal and were forced to convert to save their lives. Isolated groups still exist in mountainous areas such as Belmonte in the Beira-Baixa province of Portugal. We report here the genetic study of a highly consanguineous endogamic population of Crypto-Jews of Belmonte affected with autosomal recessive retinitis pigmentosa (RP). A genome-wide search for homozygosity allowed us to localize the disease gene to chromosome 15q22-q24 (Zmax=2.95 at θ=0 at the D15S131 locus). Interestingly, the photoreceptor cell-specific nuclear receptor (PNR) gene, the expression of which is restricted to the outer nuclear layer of retinal photoreceptor cells, was found to map to the YAC contig encompassing the disease locus. A search for mutations allowed us to ascribe the RP of Crypto-Jews of Belmonte to a homozygous missense mutation in the PNR gene. Preliminary haplotype studies support the view that this mutation is relatively ancient but probably occurred after the population settled in Belmonte.

Validação da Versão Portuguesa da "Activities-specific Balance Confidence Scale"

Introdução: Muitos dos instrumentos disponíveis para a avaliação do equilíbrio, risco de queda e medo de cair medem apenas actividades simples no domicílio e apresentam tendência para um “efeito de tecto” em idosos residentes na comunidade. A “Activities-specific Balance Confidence (ABC) Scale” foi concebida para avaliar o equilíbrio de forma abrangente, num conjunto de actividades de vida diária associadas a um largo espectro de dificuldade. Objectivos: Traduzir para português e adaptar culturalmente para Portugal a “Activities-specific Balance Confidence (ABS) Scale” e avaliar a sua fiabilidade. População e Métodos: Tradução e adaptação cultural do instrumento e subsequente aplicação a uma população idosa portuguesa, para determinação da sua fiabilidade inter-observador, fiabilidade intra-observador e consistência interna. Resultados: Os resultados foram muito homogéneos na grande maioria das comparações realizadas, quer intra-observador, quer inter-observador. A avaliação da consistência interna revelou valores muito elevados. Estes níveis de fiabilidade mantiveram-se mesmo removendo qualquer uma das 16 questões que compõem o questionário, mantendo-se praticamente constantes os valores registados com o questionário completo. Conclusões: A versão portuguesa da escala ABC demonstrou boa fiabilidade intra-observador, fiabilidade inter-observador e consistência interna na avaliação da auto-percepção do equilíbrio para diversas actividades de vida diária numa população idosa portuguesa. Outros trabalhos serão necessários para avaliar a utilidade desta escala na avaliação do risco de queda e do efeito de intervenções terapêuticas nesta população.

Genetic Characterization of an Insect-Specific Flavivirus Isolated from Culex Theileri Mosquitoes Collected in Southern Portugal

We describe the full genetic characterization of an insect-specific flavivirus (ISF) from Culex theileri (Theobald) mosquitoes collected in Portugal. This represents the first isolation and full characterization of an ISF from Portuguese mosquitoes. The virus, designated CTFV, for Culex theileri flavivirus, was isolated in the C6/36 Stegomyia albopicta (=Aedes albopictus) cell line, and failed to replicate in vertebrate (Vero) cells in common with other ISFs. The CTFV genome encodes a single polyprotein with 3357 residues showing all the features expected for those of flaviviruses. Phylogenetic analyses based on all ISF sequences available to date, place CTFV among Culex-associated flaviviruses, grouping with recently published NS5 partial sequences documented from mosquitoes collected in the Iberian Peninsula, and with Quang Binh virus (isolated in Vietnam) as a close relative. No CTFV sequences were found integrated in their host’s genome using a range of specific PCR primers designed to the prM/E, NS3, and NS5 region.