Properties and skin compatibility of films based on poly(lactic acid) (PLA) bionanocomposites incorporating chitin nanofibrils (CN)

Nanobiocomposites suitable for preparing skin compatible films by flat die extrusion were prepared by using plasticized poly(lactic acid) (PLA), poly(butylene succinate-co-adipate) (PBSA), and Chitin nanofibrils as functional filler. Chitin nanofibrils (CNs) were dispersed in the blends thanks to the preparation of pre-nanocomposites containing poly(ethylene glycol). Thanks to the use of a melt strength enhancer (Plastistrength) and calcium carbonate, the processability and thermal properties of bionanocomposites films containing CNs could be tuned in a wide range. Moreover, the resultant films were flexible and highly resistant. The addition of CNs in the presence of starch proved not advantageous because of an extensive chain scission resulting in low values of melt viscosity. The films containing CNs or CNs and calcium carbonate resulted biocompatible and enabled the production of cells defensins, acting as indirect anti-microbial. Nevertheless, tests made with Staphylococcus aureus and Enterobacter spp. (Gram positive and negative respectively) by the qualitative agar diffusion test did not show any direct anti-microbial activity of the films. The results are explained considering the morphology of the film and the different mechanisms of direct and indirect anti-microbial action generated by the nanobiocomposite based films

Mutational screening of splicing factor genes in cases with autosomal dominant retinitis pigmentosa.

PURPOSE: Mutations in genes encoding proteins from the tri-snRNP complex of the spliceosome account for more than 12% of cases of autosomal dominant retinitis pigmentosa (adRP). Although the exact mechanism by which splicing factor defects trigger photoreceptor death is not completely clear, their role in retinitis pigmentosa has been demonstrated by several genetic and functional studies. To test for possible novel associations between splicing factors and adRP, we screened four tri-snRNP splicing factor genes (EFTUD2, PRPF4, NHP2L1, and AAR2) as candidate disease genes. METHODS: We screened up to 303 patients with adRP from Europe and North America who did not carry known RP mutations. Exon-PCR and Sanger methods were used to sequence the NHP2L1 and AAR2 genes, while the sequences of EFTUD2 and PRPF4 were obtained by using long-range PCRs spanning coding and non-coding regions followed by next-generation sequencing. RESULTS: We detected novel missense changes in individual patients in the sequence of the genes PRPF4 and EFTUD2, but the role of these changes in relationship to disease could not be verified. In one other patient we identified a novel nucleotide substitution in the 5' untranslated region (UTR) of NHP2L1, which did not segregate with the disease in the family. CONCLUSIONS: The absence of clearly pathogenic mutations in the candidate genes screened in our cohort suggests that EFTUD2, PRPF4, NHP2L1, and AAR2 are either not involved in adRP or are associated with the disease in rare instances, at least as observed in this study in patients of European and North American origin

Epigenetic remodelling in human hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is the most frequent primary liver cancer, being the sixth most commonly diagnosed cancer and the fourth leading cause of cancer-related death. As other heterogeneous solid tumours, HCC results from a unique synergistic combination of genetic alterations mixed with epigenetic modifications. In HCC the patterns and frequencies of somatic variations change depending on the nearby chromatin. On the other hand, epigenetic alterations often induce genomic instability prone to mutations. Epigenetics refers to heritable states of gene expression without alteration to the DNA sequence itself and, unlike genetic changes, the epigenetic modifications are reversible and affect gene expression more extensively than genetic changes. Thus, studies of epigenetic regulation and the involved molecular machinery are greatly contributing to the understanding of the mechanisms that underline HCC onset and heterogeneity. Moreover, this knowledge may help to identify biomarkers for HCC diagnosis and prognosis, as well as future new targets for more efficacious therapeutic approaches. In this comprehensive review we will discuss the state-of-the-art knowledge about the epigenetic landscape in hepatocarcinogenesis, including evidence on the diagnostic and prognostic role of non-coding RNAs, modifications occurring at the chromatin level, and their role in the era of precision medicine. Apart from other better-known risk factors that predispose to the development of HCC, characterization of the epigenetic remodelling that occurs during hepatocarcinogenesis could open the way to the identification of personalized biomarkers. It may also enable a more accurate diagnosis and stratification of patients, and the discovery of new targets for more efficient therapeutic approaches

An Association in the Aquila Star-Forming Region: High Resolution Infrared Spectroscopy of T Tauri Stars

We present the properties of a group of young stars associated with the well-studied T Tauri star system AS 353, located in the Aquila star-forming region. The association is identified using radial velocity measurements of sample objects selected from the Herbig and Bell Catalog based on their spatial proximity to AS 353. Radial velocities of nine objects were measured from multi-epoch high-resolution (R~30,000) H-band spectra obtained with NIRSPEC on Keck II. High-resolution K-band spectra were also obtained for most of the sample objects. Spectral types and rotational velocities are determined for all objects in the sample. The multi-epoch H-band spectra were examined for radial velocity variations in order to detect possible spectroscopic binaries. Eight of the nine objects have radial velocities that are consistent within the 1-sigma scatter of the sample. From their mean of -8.6 km/s these eight objects have a standard deviation of 2 km/s, which suggests that the sample stars are related. The ninth object shows significant radial velocity variations between epochs, characteristic of a spectroscopic binary. The overall multiplicity of the sample is high; we observed 13 stars in seven systems, identifying three new candidate binary components in this project. Many of the spectra reveal hydrogen emission lines typical of strong accretion processes, indicating that most of these objects harbor circumstellar disks and are less than a few million years old. We discuss possible interpretations of the enigmatic pure emission line spectrum of HBC 684. This work represents the highest spectral resolution infrared observations to date of these intriguing, nearby young stars.Comment: 7 figures, accepted to Astrophysical Journa

Identification of two novel mutations in CDHR1 in consanguineous Spanish families with autosomal recessive retinal dystrophy.

Inherited retinal dystrophies present extensive phenotypic and genetic heterogeneity, posing a challenge for patients' molecular and clinical diagnoses. In this study, we wanted to clinically characterize and investigate the molecular etiology of an atypical form of autosomal recessive retinal dystrophy in two consanguineous Spanish families. Affected members of the respective families exhibited an array of clinical features including reduced visual acuity, photophobia, defective color vision, reduced or absent ERG responses, macular atrophy and pigmentary deposits in the peripheral retina. Genetic investigation included autozygosity mapping coupled with exome sequencing in the first family, whereas autozygome-guided candidate gene screening was performed by means of Sanger DNA sequencing in the second family. Our approach revealed nucleotide changes in CDHR1; a homozygous missense variant (c.1720C > G, p.P574A) and a homozygous single base transition (c.1485 + 2T > C) affecting the canonical 5' splice site of intron 13, respectively. Both changes co-segregated with the disease and were absent among cohorts of unrelated control individuals. To date, only five mutations in CDHR1 have been identified, all resulting in premature stop codons leading to mRNA nonsense mediated decay. Our work reports two previously unidentified homozygous mutations in CDHR1 further expanding the mutational spectrum of this gene

P07-05. HIV and STI prevalence among men who have sex with men (MSM) recruited through respondent driven sampling (RDS) in Buenos Aires, Argentina

Fil: Pando, María A. Centro Nacional de Referencia para el SIDA; Argentina.Fil: Marone, Rubén. Nexo Asociación Civil; Argentina.Fil: Balán, Iván C. Columbia University. HIV Center for Clinical and Behavioral Studies; Estados Unidos.Fil: Dolezal, Curtis. Columbia University. HIV Center for Clinical and Behavioral Studies; Estados Unidos.Fil: Squiquera, Luis. Nexo Asociación Civil; Argentina. Fil: Balan, Iván C. Columbia University. HIV Center for Clinical and Behavioral Studies; Estados Unidos.Fil: Picconi, María Alejandra. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Servicio Virus Oncogénicos; Argentina.Fil: Gonzales, J. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Servicio Virus Oncogénicos; Argentina.Fil: Rey, Jorge. Hospital de Clínicas José de San Martín; Argentina.Fil: Fernandez Toscano, M. Hospital de Clínicas José de San Martín; Argentina.Fil: Rodriguez Fermepín, Marcelo. UBA. Laboratorio de Inmunología Clínica; Argentina.Fil: Gallo Vaulet, Lucia. UBA. Laboratorio de Inmunología Clínica; Argentina.Fil: Carballo Dieguez, Alex. Columbia University. HIV Center for Clinical and Behavioral Studies; Estados Unidos.Fil: Avila, María M. Centro Nacional de Referencia para el SIDA; Argentina.Background MSM constitute one of the populations most affected by HIV and other STIs in Argentina. Previous prevalence studies were based on convenience samples. RDS, a methodology designed to access hidden populations, is being used for the first time in Argentina to recruit MSM. Methods RDS recruitment started in November 2007 with 16 first generation participants (seeds) who were selected for their potential to tap in MSM networks. Recruitment is ongoing. Men must be 18 years of age or older, be residents of Buenos Aires, self-report having sex with men at least 10 times in their lives and at least once in the past six months. They must have a coupon indicating they have been referred by a study participant. Specimens are being collected for HIV and STI diagnosis. All data are weighted using the RDS Analysis Tool (RDSAT). Results To date, 333 MSM were recruited through RDS showing a prevalence of 11.3, 16.6, 7.9, 17.6, 4.0 and 88.3% for HIV, HBV, HCV, T. pallidum, Chlamydia and HPV, respectively. Chlamydia and HPV diagnoses were only performed in 73 and 79 participants, respectively. Among HPV positive individuals, 47.8% had almost one of these high risk types (16, 58, 33, 45, 18 and 31) and 32% had multiple infections with 2 or more types. Conclusion Compared with previous studies, these results show similar HIV, HBV and T. pallidum prevalences but higher HCV prevalence. For the first time, an elevated prevalence of HPV was detected on MSM, with high frequency of types associated with ano-genital cancer. Preliminary analyses of socio demographic data show that RDS helps recruit a diversity of MSM, particularly of lower socio-economic level usually missed with other recruitment methods. The high prevalence of coinfections in this population should be taken into account if MSM are part of vaccine trials given that STI may increase HIV sexual transmission

Apoptotic microtubules delimit an active caspase free area in the cellular cortex during the execution phase of apoptosis

Apoptotic microtubule network (AMN) is organized during apoptosis, forming a cortical structure beneath plasma membrane, which has an important role in preserving cell morphology and plasma membrane permeability. The aim of this study was to examine the role of AMN in maintaining plasma membrane integrity during the execution phase of apoptosis. We demonstrated in camptothecin-induced apoptosis in H460 cells that AMN delimits an active caspase free area beneath plasma membrane that permits the preservation of cellular cortex and transmembrane proteins. AMN depolymerization in apoptotic cells by a short exposure to colchicine allowed active caspases to reach the cellular cortex and cleave many key proteins involved in plasma membrane structural support, cell adhesion and ionic homeostasis. Cleavage of cellular cortex and plasma membrane proteins, such as a-spectrin, paxilin, focal adhesion kinase (FAK), E-cadherin and integrin subunit b4 was associated with cell collapse and cell detachment. Otherwise, cleavage-mediated inactivation of calcium ATPase pump (PMCA-4) and Naþ/Ca2þ exchanger (NCX) involved in cell calcium extrusion resulted in calcium overload. Furthermore, cleavage of Naþ/Kþ pump subunit b was associated with altered sodium homeostasis. Cleavage of cell cortex and plasma membrane proteins in apoptotic cells after AMN depolymerization increased plasma permeability, ionic imbalance and bioenergetic collapse, leading apoptotic cells to secondary necrosis. The essential role of caspase-mediated cleavage in this process was demonstrated because the concomitant addition of colchicine that induces AMN depolymerization and the pan-caspase inhibitor z-VAD avoided the cleavage of cortical and plasma membrane proteins and prevented apoptotic cells to undergo secondary necrosis. Furthermore, the presence of AMN was also critical for proper phosphatidylserine externalization and apoptotic cell clearance by macrophages. These results indicate that AMN is essential to preserve an active caspase free area in the cellular cortex of apoptotic cells that allows plasma membrane integrity during the execution phase of apoptosis