Organotypic spinal cord cultures: An in vitro 3D model to preliminary screen treatments for spinal muscular atrophy

Spinal muscular atrophy (SMA) is a severe neuromuscular disease affecting children, due to mutation/deletion of survival motor neuron 1 (SMN1) gene. The lack of functional protein SMN determines motor neuron (MN) degeneration and skeletal muscle atrophy, leading to premature death due to respiratory failure. Nowadays, the Food and Drug Administration approved the administration of three drugs, aiming at increasing the SMN production: although assuring noteworthy results, all these therapies show some non-negligible limitations, making essential the identification of alternative/synergistic therapeutic strategies. To offer a valuable in vitro experimental model for easily performing preliminary screenings of alternative promising treatments, we optimized an organotypic spinal cord culture (derived from murine spinal cord slices), which well recapitulates the pathogenetic features of SMA. Then, to validate the model, we tested the effects of human Mesenchymal Stem Cells (hMSCs) or murine C2C12 cells (a mouse skeletal myoblast cell line) conditioned media: 1/3 of conditioned medium (obtained from either hMSCs or C2C12 cells) was added to the conventional medium of the organotypic culture and maintained for 7 days. Then the slices were fixed and immunoreacted to evaluate the MN survival. In particular we observed that the C2C12 and hMSCs conditioned media positively influenced the MN soma size and the axonal length respectively, without modulating the glial activation. These data suggest that trophic factors released by MSCs or muscular cells can exert beneficial effects, by acting on different targets, and confirm the reliability of the model. Overall, we propose the organotypic spinal cord culture as an excellent tool to preliminarily screen molecules and drugs before moving to in vivo models, in this way partly reducing the use of animals and the costs

Frontal lobe metabolic alterations in autism spectrum disorder: a 1H-magnetic resonance spectroscopy study.

Recently, neuroimaging studies were performed using 1H-magnetic resonance spectroscopy (1H-MRS), revealing a quantitative alteration of neurochemicals (such as neurotransmitters and metabolites) in several brain regions of patients with autism spectrum disorder (ASD). The involvement of the frontal lobe in the neurobiology of ASD has long been documented in the literature. Therefore, the aim of this study was to analyze the alterations of N-acetylaspartate/creatine (NAA/Cr) and choline/Cr (Cho/Cr) ratios in the frontal lobe subcortical white matter (WM) in ASD patients, in order to reveal any alteration of metabolites that might be the expression of specific clinical features of the disorder

Development of a High Oleic Cardoon Cell Culture Platform by SAD Overexpression and RNAi-Mediated FAD2.2 Silencing

The development of effective tools for the sustainable supply of phyto-ingredients and natural substances with reduced environmental footprints can help mitigate the dramatic scenario of climate change. Plant cell cultures-based biorefineries can be a technological advancement to face this challenge and offer a potentially unlimited availability of natural substances, in a standardized composition and devoid of the seasonal variability of cultivated plants. Monounsaturated (MUFA) fatty acids are attracting considerable attention as supplements for biodegradable plastics, bio-additives for the cosmetic industry, and bio-lubricants. Cardoon (Cynara cardunculus L. var. altilis) callus cultures accumulate fatty acids and polyphenols and are therefore suitable for large-scale production of biochemicals and valuable compounds, as well as biofuel precursors. With the aim of boosting their potential uses, we designed a biotechnological approach to increase oleic acid content through Agrobacterium tumefaciens-mediated metabolic engineering. Bioinformatic data mining in the C. cardunculus transcriptome allowed the selection and molecular characterization of SAD (stearic acid desaturase) and FAD2.2 (fatty acid desaturase) genes, coding for key enzymes in oleic and linoleic acid formation, as targets for metabolic engineering. A total of 22 and 27 fast-growing independent CcSAD overexpressing (OE) and CcFAD2.2 RNAi knocked out (KO) transgenic lines were obtained. Further characterization of five independent transgenic lines for each construct demonstrated that, successfully, SAD overexpression increased linoleic acid content, e.g., to 42.5%, of the relative fatty acid content, in the CcSADOE6 line compared with 30.4% in the wild type (WT), whereas FAD2.2 silencing reduced linoleic acid in favor of the accumulation of its precursor, oleic acid, e.g., to almost 57% of the relative fatty acid content in the CcFAD2.2KO2 line with respect to 17.7% in the WT. Moreover, CcSADOE6 and CcFAD2.2KO2 were also characterized by a significant increase in total polyphenolic content up to about 4.7 and 4.1 mg/g DW as compared with 2.7 mg/g DW in the WT, mainly due to the accumulation of dicaffeoyl quinic and feruloyl quinic acids. These results pose the basis for the effective creation of an engineered cardoon cells-based biorefinery accumulating high levels of valuable compounds from primary and specialized metabolism to meet the industrial demand for renewable and sustainable sources of innovative bioproducts

Ação moduladora da microbiota de portadores de síndrome de Down sobre bactérias cariogênicas

Carriers of Down Syndrome have low prevalence of tooth decay, despite the fact that they show precarious oral hygiene. This research was carried out with the intent of evaluating the possible action of strains that produce bacteriocins as regulators of the carrier of Down Syndrome cariogenic microbiota. Streptococcus mutans and Streptococcus sobrinus are the main acid producers in the oral cavity.Consequently, they were tested antagonistically with strains isolated from the individuals with Down Syndrome in order to verify if cariogenic bacteria were being inhibited by the by the carrier's strains. Streptococcus mutans and Streptococcus sobrinus were isolated with the utilization of agar saccharose bacitracin medium and identified through standard biochemical tests. Samples of carriers'supragingival plaques were collected, spread through dissemination in Agar Mitis Salivarius and after isolation were cultivated in BHI liquid medium.10 cultures from syndromics were seeded in the Muller Hinton medium. After the growth, the colonies in the Petri dishes were treated by chloroform. Then, one portion of the cariogenic bacterium cultures was mixed with soft brain-heart infusion medium and immediately transferred to the Petri dish with the test cultures. Two of the ten tested strains presented a zone of inhibition against Streptococcus mutans. The discovery of strains that produce antibacterial substances may be responsible for the low prevalence of tooth decay in individuals with Down Syndrome. These findings may help to understand the relationship between oral microbiota and pathological tooth decay.Os portadores da Síndrome de Down possuem baixa prevalência de cárie, apesar de apresentarem uma higiene oral deficiente. O estudo foi realizado para detectar cepas produtoras de bacteriocinas, com o intuito de avaliar a sua possível ação como moduladoras da microbiota cariogênica dos sindrômicos. Streptococcus mutans e Streptococcus sobrinus são os principais produtores de ácidos na cavidade oral. Em função disso, esses estreptococos foram testados antagonicamente com cepas isoladas dos indivíduos com Síndrome de Down, para que fosse avaliado se havia inibição das bactérias cariogênicas pelas cepas dos sindrômicos. Streptococcus mutans e Streptococcus sobrinus foram isolados utilizando o meio ágar sacarose bacitracina e identificados através de provas bioquímicas. Uma amostra de placa supragengival dos sindrômicos foi coletada e semeada por disseminação em Ágar Mitis-Salivarius, após o isolamento foram cultivados em caldo BHI. Dez culturas dos sindrômicos foram semeadas no Ágar Muller Hinton. Após o crescimento, as placas foram tratadas com clorofórmio. Sucessivamente, uma porção das culturas de bactérias cariogênicas foi transferida para meio semi-sólido BHI e vertidas nas placas com as culturas testes. Duas das dez cepas testadas apresentaram halo de inibição de crescimento contra cepas de Streptococcus mutans. A descoberta de cepas que produzem substâncias antimicrobianas pode ser responsável pela baixa prevalência de cárie em indivíduos com Síndrome de Down. Esse achado pode ajudar a entender a relação entre microbiota oral e patologia da cárie dentária

Increase of poly(ADP-ribose) polymerase mRNA levels during TPA-induced differentiation of human lymphocytes

AbstractThe non-mitogenic stimulation of human peripheral blood mononuclear cells (PBMC) with low concentrations of the phorbol ester 12-O-tetradecanoylphorbol 13-acatate (TPA) caused a progressive increase in the percent fraction of the cells that were positive for the early activating antigen CD69. At the same time, it caused a progressive increase in the steady-state levels of poly(ADP-ribose) polymerase (pADPRP) transcripts. A further increase in TPA concentration, while inducing the maximal expression of the levels of CD69 activating surface antigen, both in the presence or in the absence of proliferative activity, did not evoke any additional hightening of pADPRP mRNA levels. Time course of PBMC stimulation with a non-mitogenic dose or TPA showed an early increase in the accumulation of pADPRP mRNA, which changed at 8-16 h. and remained high for several days thereafter. On the basis of these data, we suggest flat the increase in pADPRP mRNA may be associated with the commitment of human lymphocytes from a quiescent (G0) to an activated (G1) state

Tree growth and stem carbon accumulation in human-modified Amazonian forests following drought and fire

Human-modified forests are an ever-increasing feature across the Amazon Basin, but little is known about how stem growth is influenced by extreme climatic events and the resulting wildfires. Here we assess for the first time the impacts of human-driven disturbance in combination with El Niño–mediated droughts and fires on tree growth and carbon accumulation. We found that after 2.5 years of continuous measurements, there was no difference in stem carbon accumulation between undisturbed and human-modified forests. Furthermore, the extreme drought caused by the El Niño did not affect carbon accumulation rates in surviving trees. In recently burned forests, trees grew significantly more than in unburned ones, regardless of their history of previous human disturbance. Wood density was the only significant factor that helped explain the difference in growth between trees in burned and unburned forests, with low wood–density trees growing significantly more in burned sites. Our results suggest stem carbon accumulation is resistant to human disturbance and one-off extreme drought events, and it is stimulated immediately after wildfires. However, these results should be seen with caution—without accounting for carbon losses, recruitment and longer-term changes in species composition, we cannot fully understand the impacts of drought and fire in the carbon balance of human-modified forests. This article is part of a discussion meeting issue ‘The impact of the 2015/2016 El Nino on the terrestrial tropical carbon cycle: patterns, mechanisms and implications’

Religious experience and aesthetic experience in plastic artists : perspectives from the psychology of religion

Pode a experiência religiosa ser substituída pela experiência estética? Os estudiosos se dividem na resposta. Situando a pergunta no quadro de referência conceitual e epistemológico que distingue entre religião substantiva e religião funcional, levantamos a hipótese de que não há substituição substantiva, mas pode haver substituição funcional entre essas experiências. Levantamos a hipótese ulterior de que a experiência estética pode ser uma experiência do sagrado. As hipóteses foram examinadas com os dados obtidos em entrevistas com oito renomados artistas plásticos, por ocasião da 23ª Bienal de São Paulo, em 1996. Os dados revelaram diferenças, continuidades e analogias entre arte e religião assim como entre arte e sagrado, que permitiram reconhecer em alguns casos substituição funcional da religião pela arte, principalmente se dotada da densidade do sagrado.ABSTRACT Can aesthetic experience replace religious experience? Researchers are quite divided about this issue. From a conceptual and epistemological framework that distinguishes between substantive and functional religion, we formulated the hypothesis that there is not a substantive replacement, but there can be a functional replacement of religious experience by an aesthetic experience. We also considered the additional hypothesis that aesthetic experience can be an experience of the sacred. The hypotheses were examined with the help of interviews with eight well known Brazilian artists, by the time of the 23rd Biennial International Exhibition of Arts of São Paulo, 1996. The data revealed differences, continuities, and analogies between art and religion and between art and the sacred, that allowed to identify in some cases a functional replacement of religion by art, especially if art was endowed with the density of the sacred

The clinical effectiveness of an integrated multidisciplinary evidence-based program to prevent intraoperative pressure injuries in high-risk children undergoing long-duration surgical procedures: a quality improvement study

The prevention of hospital-acquired pressure injuries (HAPIs) in children undergoing long-duration surgical procedures is of critical importance due to the potential for catastrophic sequelae of these generally preventable injuries for the child and their family. Long-duration surgical procedures in children have the potential to result in high rates of HAPI due to physiological factors and the difficulty or impossibility of repositioning these patients intraoperatively. We developed and implemented a multi-modal, multi-disciplinary translational HAPI prevention quality improvement program at a large European Paediatric University Teaching Hospital. The intervention comprised the establishment of wound prevention teams, modified HAPI risk assessment tools, specific education, and the use of prophylactic dressings and fluidized positioners during long-duration surgical procedures. As part of the evaluation of the effectiveness of the program in reducing intraoperative HAPI, we conducted a prospective cohort study of 200 children undergoing long-duration surgical procedures and compared their outcomes with a matched historical cohort of 200 children who had undergone similar surgery the previous year. The findings demonstrated a reduction in HAPI in the intervention cohort of 80% (p < 0.01) compared to the comparator group when controlling for age, pathology, comorbidity, and surgical duration. We believe that the findings demonstrate that it is possible to significantly decrease HAPI incidence in these highly vulnerable children by using an evidence-based, multi-modal, multidisciplinary HAPI prevention strategy