Mutational and genetic origin of LDL receptor gene mutations detected in both Belgian and Dutch familial hypercholesterolemics

DNA samples from 100 unrelated Belgian patients with familial hypercholesterolemia (FH) were screened for the presence of specific low-density lipoprotein receptor (LDLR) gene mutations, previously shown to be prevalent in related populations. Two point mutations, viz., P664L and a G to A splicing defect at position 1359-1, were detected in single Flemish-speaking families. A long-distance polymerase chain reaction (PCR) assay, used to screen for the 4-kb and 2.5-kb deletions previously identified by Southern blot analyses in different parts of The Netherlands, revealed a 3-kb deletion in two Belgian patients. Comparison of PCR product length showed that both Dutch deletions of exons 7-8 are identical to that found in Belgians, but different from the 2.5-kb deletion previously described in South Africans of mixed ancestry. The Belgian patients probably share a common ancestor, for each mutation identified, with FH patients from The Netherlands, since all three mutations were associated with the same LDLR gene haplotype as described for the Dutch population. Analysis of the deletion junctions demonstrated the role of a 31-bp repetitive sequence in the generation of large rearrangements involving exons 7 and 8 of the LDLR gene. The finding that only 4 out of 100 analyzed Belgian hypercholesterolemics carry a known LDLR mutation that is prevalent in related populations suggests that the Belgian FH population has its own spectrum of mutations.Articl

Effect of Cysticercus cellulosae fractions on the respiratory burst of pig neutrophils Efeito de frações de Cysticercus cellulosae sobre a explosão respiratória de neutrófilos de suínos

Neutrophils, eosinophils and macrophages are cells that interact with invading parasites and naive hosts have been shown to have anti-parasitic activity. The initial reaction of these leukocytes is the generation of reactive oxygen species (ROS) to play in parasite expulsion. The present work was carried out to study the effect of total extract, scolex and membrane fractions from Cysticercus cellulosae on respiratory burst by pig neutrophils. Hydrogen peroxide (H2O2) production by neutrophils incubated with metacestode fractions from C. cellulosae showed an increase of: 190% (total extract), 120% (scolex) and 44% (membrane). High antioxidant catalatic activity (33%, 28%, 28% by total extract, scolex and membrane, respectively) was observed in neutrophils incubated with metacestode fractions, which could be an attempt at self-protection. Scolex and membrane fractions increased the phagocytic capacity of neutrophils (44% and 28%, respectively). On the other hand, total cysticerci did not alter the phagocytosis, possibly due to modifications in membrane function, caused by high ROS production from neutrophils in the presence of total cysticerci. Total fraction from C. cellulosae is toxic for neutrophils as shown by the decrease in phagocytic capacity, probably caused by high levels of ROS formation. The difference in toxicity of total extract, scolex and membrane fractions on neutrophils can be explained by the presence of an antigenic effect of the vesicular fluid in the total extract of C. cellulosae.<br>Neutrófilos, eosinófilos e macrófagos são células que interagem com os parasitas no corpo do hospedeiro desenvolvendo atividade antiparasitária. A reação inicial destes leucócitos é a geração de espécies reativas de oxigênio (ERO) a fim de expulsar os parasitas. No presente trabalho estudou-se o efeito da fração total, de escolex e de membrana de Cysticercus cellulosae sobre a explosão respiratória de neutrófilos de suínos. A produção de peróxido de hidrogênio (H2O2) pelos neutrófilos incubados com as frações de C. cellulosae apresentou acréscimo de 190% (extrato total), 120% (escolex) e 44% (membrana). Alta atividade de catalase (33%, 28% e 28% para extrato total, escolex e membrana respectivamente) foi observada nos neutrófilos incubados com as frações de metacestodeo, podendo representar a própria proteção celular do neutrófilo. Frações de escolex e de membrana aumentaram a capacidade fagocitária dos neutrófilos (44% e 28%, respectivamente). Por outro lado, a fração total do cisticerco não alterou a capacidade fagocitária dos neutrófilos, o que pode estar relacionada com modificações na função da membrana celular causadas pela alta produção de ERO na presença da fração total. O extrato total de C. cellulosae é tóxico para os neutrófilos, indicada pela diminuição da capacidade fagocitária, provavelmente pela indução de alto nível de ERO. A diferença de toxicidade do extrato total, de escolex e de membrana para os neutrófilos pode ocorrer pelo efeito antigênico presente no fluido vesicular no extrato total de C. cellulosae