First report of molecular diagnosis of Tunisian hemophiliacs A: identification of 8 novel causative mutations

ABSTRACT: INTRODUCTION: Hemophilia A is an X linked recessive hemorrhagic disorder caused by mutations in the F8 gene that lead to qualitative and/or quantitative deficiencies of coagulation factor VIII (FVIII). Molecular diagnosis of hemophilia A is challenging because of the high number of different causative mutations that are distributed throughout the large F8 gene. Molecular studies of these mutations are essential in order to reinforce our understanding of their pathogenic effect responsible for the disorder. Aim In this study we have performed molecular analysis of 28 Tunisian hemophilia A patients and analyzed the F8 mutation spectrum. METHODS: We screened the presence of intron 22 and intron 1 inversion in severe hemophilia A patients by southern blotting and polymerase chain reaction (PCR). Detection of point mutations was performed by dHPLC/sequencing of the coding F8 gene region. We predict the potential functional consequences of novel missense mutations with bioinformatics approaches and mapping of their spatial positions on the available FVIII 3D structure. RESULTS: We identified 23 different mutations in 28 Tunisian hemophilia A patients belonging to 22 unrelated families. The identified mutations included 5 intron 22 inversions, 7 insertions, 4 deletions and 7 substitutions. In total 18 point mutations were identified, of which 9 are located in exon 14, the most mutated exonic sequence in the F8 gene. Among the 23 mutations, 8 are novel and not deposited in the HAMSTeRS database nor described in recently published articles. CONCLUSION: The mutation spectrum of Tunisian hemophilia A patients is heterogeneous with the presence of some characteristic features. Virtual slides The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1693269827490715.     

Exposure of Vicia faba and Pisum sativum to copper-induced genotoxicity

The potential genotoxicity of Cu(2+) was investigated in Vicia faba and Pisum sativum seedlings in hydroponic culture conditions. Cu(2+) caused a dose-dependent increase in micronuclei frequencies in both plant models. Cytological analysis of root tips cells showed clastogenic and aneugenic effects of this heavy metal on V. faba root meristems. Cu(2+) induced chromosomal alterations at the lowest concentration used (2.5 mM) when incubated for 42 h, indicating the potent mutagenic effect of this ion. A spectrum of chromosomal abnormalities was observed in V. faba root meristems, illustrating the genotoxic events leading to micronuclei formation.     

Chemical and physicochemical investigation of an aminoalkylalkoxysilane as strengthening agent for cellulosic materials

AMDES (aminopropylmethyldiethoxysilane) was used to investigate the physicochemical and chemical events related to the introduction of aminoalkylalkoxysilanes in cellulosic materials. Using (29)Si CP-MAS and (1)H NMR to study the reactivity and structural modification of AMDES in the paper it was shown that polymerization occurs in situ. The distribution of the active compound on the surface of the fibers and throughout the fibers' thickness was visualized by SEM-EDS. A relation between moisture content, fiber swelling, and uptake of AMDES was found. To better represent old and brittle documents, the paper was predegraded by oxidation with sodium hypochlorite. XRD confirmed the advanced destruction of the amorphous areas of cellulose. Adding AMDES in the oxidized paper resulted in improved mechanical properties, a roughly unmodified degree of polymerization of cellulose, but a slight increase in the yellowing, probably due to several possible reaction products such as imines, amine, amides, and Maillard reactions products. The deacidification efficacy was established and the strengthening effect was shown to arise from the interaction of AMDES with the cellulose fibers.     

L’adoption en Tunisie comme réponse a la stérilité masculine

The sterility problem opens to that of procreating desire. Artificial insemination using donor sperm, being assimilated to adultery in Islam rules, is forbidden in Tunisia. The law n°58-27 (March 4th, 1958) introducing adoption protects abandoned children, offers an answer to male infertility and gives to the couple the possibility of realizing their procreating desire. The statistic analysis of 1988 gives a real approach about the importance of adoption in Tunisia.     

Dipeptidyl peptidase III as a DNA marker to investigate epidemiology and taxonomy of Old World Leishmania species

BackgroundDipeptidyl peptidase III (DPPIII) member of M49 peptidase family is a zinc-dependent metallopeptidase that cleaves dipeptides sequentially from the N-terminus of its substrates. In Leishmania, DPPIII, was reported with other peptidases to play a significant role in parasites’ growth and survival. In a previous study, we used a coding sequence annotated as DPPIII to develop and evaluate a PCR assay that is specific to dermotropic Old World (OW) Leishmania species. Thus, our objective was to further assess use of this gene for Leishmania species identification and for phylogeny, and thus for diagnostic and molecular epidemiology studies of Old World Leishmania species.MethodologyOrthologous DDPIII genes were searched in all Leishmania genomes and aligned to design PCR primers and identify relevant restriction enzymes. A PCR assays was developed and seventy-two Leishmania fragment sequences were analyzed using MEGA X genetics software to infer evolution and phylogenetic relationships of studied species and strains. A PCR-RFLP scheme was also designed and tested on 58 OW Leishmania strains belonging to 8 Leishmania species and evaluated on 75 human clinical skin samples.FindingsSequence analysis showed 478 variable sites (302 being parsimony informative). Test of natural selection (dN-dS) (-0.164, SE = 0.013) inferred a negative selection, characteristic of essential genes, corroborating the DPPIII importance for parasite survival. Inter- and intra-specific genetic diversity was used to develop universal amplification of a 662bp fragment. Sequence analyses and phylogenies confirmed occurrence of 6 clusters congruent to L. major, L. tropica, L. aethiopica, L. arabica, L. turanica, L. tarentolae species, and one to the L. infantum and L. donovani species complex.A PCR-RFLP algorithm for Leishmania species identification was designed using double digestions with HaeIII and KpnI and with SacI and PvuII endonucleases. Overall, this PCR-RFLP yielded distinct profiles for each of the species L. major, L. tropica, L. aethiopica, L. arabica and L. turanica and the L. (Sauroleishmania) L. tarentolae. The species L. donovani, and L. infantum shared the same profile except for strains of Indian origin. When tested on clinical samples, the DPPIII PCR showed sensitivities of 82.22% when compared to direct examination and was able to identify 84.78% of the positive samples.ConclusionThe study demonstrates that DPPIII gene is suitable to detect and identify Leishmania species and to complement other molecular methods for leishmaniases diagnosis and epidemiology. Thus, it can contribute to evidence-based disease control and surveillance.     

Formation of brown lines in paper: characterization of cellulose degradation at the wet-dry interface

Brown lines were generated at the wet-dry interface on Whatman paper No. 1 by suspending the sheet vertically in deionized water. Formic acid and acetic acid were quantified in three areas of the paper defined by the wet-dry boundary (above, below, and at the tideline) using capillary zone electrophoresis with indirect UV detection. Their concentration increased upon accelerated aging of the paper and was highest in the tideline. The hydroperoxides have been quantified using reverse phase high performance liquid chromatography with UV detection based on the determination of triphenylphosphine oxide produced from the reaction with triphenylphosphine, and their highest concentration was found in the tideline as well. For the first time, it was shown that various types of hydroperoxides were present, water-soluble and non-water-soluble, most probably in part hydroperoxide functionalized cellulose. After accelerated aging, a significant increase in hydroperoxide concentration was found in all the paper areas. The molar masses of cellulose determined using size-exclusion chromatography with multiangle light scattering detection showed that, upon aging, cellulose degraded significantly more in the tideline area than in the other areas of the paper. The area below the tideline was more degraded than the area above. A kinetic study of the degradation of cellulose allowed determining the constants for glycosidic bond breaking in each of the areas of the paper.     

Colloidal properties of biodegradable nanoparticles influence interaction with amyloid-β peptide

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by the extracellular deposition of amyloid-β peptides (Aβ). During the past few years, promising approaches based on nanotechnologies have emerged to alter Aβ aggregation and its related toxicity. This study aims to investigate the influence of the nanoparticle colloidal properties over the interaction with Aβ peptide 1–42 (Aβ_1–42). Using capillary electrophoresis with laser-induced fluorescence detection, it was shown that biodegradable poly(ethylene glycol)-block-polylactide (PEG-b-PLA) nanoparticles were able to interact with Aβ_1–42 peptide leading to its uptake in rather short time periods. In addition, we highlighted the crucial role of the nanocarrier colloidal properties on the uptake kinetics. Whereas nanoparticles stabilized by sodium cholate (lower size and higher negative surface charge) gave optimum uptake kinetics, nanoparticles stabilized with others surfactants presented lower interactions. In contrast, PEG density seemed to have no influence on the interaction when sodium cholate was used for the preparation. This study intends to give new insights into Aβ_1–42 peptide interaction with nanoparticulate systems by helping to determine suitable nanoparticle characteristics regarding forthcoming therapeutic strategies against AD.     

Impuissance sexuelle iatrogène et responsabilité médicale

Medical responsibility means that the Doctor must assume his acts. It enables not only to repair prejudices met but also discourage prejudiciable acts to society by sanctioning them. Its founding is based on the principle of error, of the damage caused and the establishing of a cause link between error and damage. The authors report a clinical case observed during a medical expertise of an iatrogenic accident (priapism) secondary to the treatment of a psychogenic impotence with papaverine injection in intracavernous. It’s a 27 year old patient, single, teacher, without pathological antecedents, who, after a psycho-affective shock (break with his girl-friend), showed a diminished sewal desire and erectile power. These complaints led him to consult an urologist, who prescribed a hormonal check-up and a medical treatment based on Yohimbine^* for two weeks. Seeing no improvement of his state, he consulted another urologist who gave him an intracavernous Papaverine chlorydrate injection (40 mg). The immediate main consequen?s were a priapism necessitating a surgical intervention (caverno-spongious anastomosis), practised one day after the injection. Following the intervention, the patient complains of a persistant impoten?, which led him to sue his Doctor for a professional error having led to a permanent sexual prejudice. The error retained against the Doctor consists in a bad control of his patient, which led to delay in taking the latter in charge, a classical complication resulting from the treatment prescribed. Because of the existence of an error, a damage caused and a cause link, the responsibility of the Doctor is in question. From this case, the authors insist on the necessity of a psychologic evaluation prior to every medicinal treatment of a psychogenic impotence.