Reduced transcription of TCOF1 in adult cells of Treacher Collins syndrome patients

<p>Abstract</p> <p>Background</p> <p>Treacher Collins syndrome (TCS) is an autosomal dominant craniofacial disorder caused by frameshift deletions or duplications in the <it>TCOF1 </it>gene. These mutations cause premature termination codons, which are predicted to lead to mRNA degradation by nonsense mediated mRNA decay (NMD). Haploinsufficiency of the gene product (treacle) during embryonic development is the proposed molecular mechanism underlying TCS. However, it is still unknown if <it>TCOF1 </it>expression levels are decreased in post-embryonic human cells.</p> <p>Methods</p> <p>We have estimated <it>TCOF1 </it>transcript levels through real time PCR in mRNA obtained from leucocytes and mesenchymal cells of TCS patients (n = 23) and controls (n = 18). Mutational screening and analysis of NMD were performed by direct sequencing of gDNA and cDNA, respectively.</p> <p>Results</p> <p>All the 23 patients had typical clinical features of the syndrome and pathogenic mutations were detected in 19 of them. We demonstrated that the expression level of <it>TCOF1 </it>is 18-31% lower in patients than in controls (<it>p < 0.05</it>), even if we exclude the patients in whom we did not detect the pathogenic mutation. We also observed that the mutant allele is usually less abundant than the wild type one in mesenchymal cells.</p> <p>Conclusions</p> <p>This is the first study to report decreased expression levels of <it>TCOF1 </it>in TCS adult human cells, but it is still unknown if this finding is associated to any phenotype in adulthood. In addition, as we demonstrated that alleles harboring the pathogenic mutations have lower expression, we herein corroborate the current hypothesis of NMD of the mutant transcript as the explanation for diminished levels of <it>TCOF1 </it>expression. Further, considering that <it>TCOF1 </it>deficiency in adult cells could be associated to pathologic clinical findings, it will be important to verify if TCS patients have an impairment in adult stem cell properties, as this can reduce the efficiency of plastic surgery results during rehabilitation of these patients.</p

Administration of M. leprae Hsp65 Interferes with the Murine Lupus Progression

The heat shock protein [Hsp] family guides several steps during protein synthesis, are abundant in prokaryotic and eukaryotic cells, and are highly conserved during evolution. The Hsp60 family is involved in assembly and transport of proteins, and is expressed at very high levels during autoimmunity or autoinflammatory phenomena. Here, the pathophysiological role of the wild type [WT] and the point mutated K409A recombinant Hsp65 of M. leprae in an animal model of Systemic Lupus Erythematosus [SLE] was evaluated in vivo using the genetically homogeneous [NZBxNZW]F1 mice. Anti-DNA and anti-Hsp65 antibodies responsiveness was individually measured during the animal's life span, and the mean survival time [MST] was determined. The treatment with WT abbreviates the MST in 46%, when compared to non-treated mice [p<0.001]. An increase in the IgG2a/IgG1 anti-DNA antibodies ratio was also observed in animals injected with the WT Hsp65. Incubation of BALB/c macrophages with F1 serum from WT treated mice resulted in acute cell necrosis; treatment of these cells with serum from K409A treated mice did not cause any toxic effect. Moreover, the involvement of WT correlates with age and is dose-dependent. Our data suggest that Hsp65 may be a central molecule intervening in the progression of the SLE, and that the point mutated K409A recombinant immunogenic molecule, that counteracts the deleterious effect of WT, may act mitigating and delaying the development of SLE in treated mice. This study gives new insights into the general biological role of Hsp and the significant impact of environmental factors during the pathogenesis of this autoimmune process

2-Hydroxylethyl methacrylate (HEMA), a tooth restoration component, exerts its genotoxic effects in human gingival fibroblasts trough methacrylic acid, an immediate product of its degradation

HEMA (2-hydroxyethyl methacrylate), a methacrylate commonly used in dentistry, was reported to induce genotoxic effects, but their mechanism is not fully understood. HEMA may be degraded by the oral cavity esterases or through mechanical stress following the chewing process. Methacrylic acid (MAA) is the primary product of HEMA degradation. In the present work we compared cytotoxic and genotoxic effects induced by HEMA and MAA in human gingival fibroblasts (HGFs). A 6-h exposure to HEMA or MAA induced a weak decrease in the viability of HGFs. Neither HEMA nor MAA induced strand breaks in the isolated plasmid DNA, but both compounds evoked DNA damage in HGFs, as evaluated by the alkaline comet assay. Oxidative modifications to the DNA bases were monitored by the DNA repair enzymes Endo III and Fpg. DNA damage induced by HEMA and MAA was not persistent and was removed during a 120 min repair incubation. Results from the neutral comet assay indicated that both compounds induced DNA double strand breaks (DSBs) and they were confirmed by the γ-H2AX assay. Both compounds induced apoptosis and perturbed the cell cycle. Therefore, methacrylic acid, a product of HEMA degradation, may be involved in its cytotoxic and genotoxic action

Fauna used in popular medicine in Northeast Brazil

<p>Abstract</p> <p>Background</p> <p>Animal-based remedies constitute an integral part of Brazilian Traditional Medicine. Due to its long history, zootherapy has in fact become an integral part of folk medicine both in rural and urban areas of the country. In this paper we summarize current knowledge on zootherapeutic practices in Northeast of Brazil, based on information compiled from ethnobiological scientific literature.</p> <p>Methods</p> <p>In order to examine the diversity of animals used in traditional medicine in Northeast of Brazil, all available references or reports of folk remedies based on animals sources were examined. 34 sources were analyzed. Only taxa that could be identified to species level were included in assessment of medicinal animal species. Scientific names provided in publications were updated.</p> <p>Results</p> <p>The review revealed that at least 250 animal species (178 vertebrates and 72 invertebrates) are used for medicinal purposes in Northeast of Brazil. The inventoried species comprise 10 taxonomic categories and belong to 141 Families. The groups with the greatest number of species were fishes (n = 58), mammals (n = 47) and reptiles (n = 37). The zootherapeutical products are used for the treatment of different illnesses. The most widely treated condition were asthma, rheumatism and sore throat, conditions, which had a wide variety of animals to treat them with. Many animals were used for the treatment of multiple ailments. Beyond the use for treating human diseases, zootherapeutical resources are also used in ethnoveterinary medicine</p> <p>Conclusion</p> <p>The number of medicinal species catalogued was quite expressive and demonstrate the importance of zootherapy as alternative therapeutic in Northeast of Brazil. Although widely diffused throughout Brazil, zootherapeutic practices remain virtually unstudied. There is an urgent need to examine the ecological, cultural, social, and public health implications associated with fauna usage, including a full inventory of the animal species used for medicinal purposes and the socio-cultural context associated with their consumption.</p