Conditioning of hiPSC-derived cardiomyocytes using surface topography obtained with high throughput technology

Surface functionalization of polymers aims to introduce novel properties that favor bioactive responses. We have investigated the possibility of surface functionalization of polyethylene terephthalate (PET) sheets by the combination of laser ablation with hot embossing and the application of such techniques in the field of stem cell research. We investigated the response of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to topography in the low micrometer range. HiPSC-CMs are expected to offer new therapeutic tools for myocardial replacement or regeneration after an infarct or other causes of cardiac tissue loss. However, hiPSC-CMs are phenotypically immature compared to myocytes in the adult myocardium, hampering their clinical application. We aimed to develop and test a high-throughput technique for surface structuring that would improve hiPSC-CMs structural maturation. We used laser ablation with a ps-laser source in combination with nanoimprint lithography to fabricate large areas of homogeneous micron- to submicron line-like pattern with a spatial period of 3 µm on the PET surface. We evaluated cell morphology, alignment, sarcomeric myofibrils assembly, and calcium transients to evaluate phenotypic changes associated with culturing hiPSC-CMs on functionalized PET. Surface functionalization through hot embossing was able to generate, at low cost, low micrometer features on the PET surface that influenced the hiPSC-CMs phenotype, suggesting improved structural and functional maturation. This technique may be relevant for high-throughput technologies that require conditioning of hiPSC-CMs and may be useful for the production of these cells for drug screening and disease modeling applications with lower costs.Fil: Cortella, Lucas R. X.. Universidade de Sao Paulo; BrasilFil: Cestari, Idágene A.. Universidade de Sao Paulo; BrasilFil: Lahuerta, Ricardo D.. Universidade de Sao Paulo; BrasilFil: Arana, Matheus C.. Universidade de Sao Paulo; BrasilFil: Soldera, Marcos Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigación y Desarrollo en Ingeniería de Procesos, Biotecnología y Energías Alternativas. Universidad Nacional del Comahue. Instituto de Investigación y Desarrollo en Ingeniería de Procesos, Biotecnología y Energías Alternativas; ArgentinaFil: Rank, Andreas. Technische Universität Dresden; AlemaniaFil: Lasagni, Andrés F.. Technische Universität Dresden; AlemaniaFil: Cestari, Ismar N.. Universidade de Sao Paulo; Brasi

Long-lasting positive effects of collaborative remembering on false assents to misleading questions

Previous studies showed that collaborative remembering can reduce false memories through a process of mutual error checking, although conclusions were limited by the nature of the memory tasks (very few errors). The present experiments extend these findings to eyewitness memory by using a paradigm designed to increase the frequency of memory errors. Collaborative and nominal pairs viewed a video-clip illustrating a bank robbery, provided an immediate free recall, were forced to confabulate answers to false-event questions, and, after a short- (1 h: Experiment 1) or a long-term delay (1 week: Experiment 2), were administered a yes/no recognition task in which the misleading statements either matched the questions presented in the confabulation phase (answered questions) or not (control questions). Collaborative pairs recalled fewer correct details in the immediate free recall task, replicating the negative effects of collaborative inhibition. Most importantly, in the final recognition test, collaborative pairs were less likely to provide false assents to misleading statements, regardless of whether they had provided a response to the related false-event questions 1 h or 1 week earlier. Our results suggest that collaboration can increase the eyewitnesses' tendency to check the accuracy of others' responses and reject false memories through discussion

Current status of local penile therapy

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Ivar, an interpretation‐oriented tool to manage the update and revision of variant annotation and classification

The rapid evolution of Next Generation Sequencing in clinical settings, and the resulting challenge of variant reinterpretation given the constantly updated information, require robust data management systems and organized approaches. In this paper, we present iVar: a freely available and highly customizable tool with a user‐friendly web interface. It represents a platform for the unified management of variants identified by different sequencing technologies. iVar accepts variant call format (VCF) files and text annotation files and elaborates them, optimizing data organization and avoiding redundancies. Updated annotations can be periodically re‐uploaded and associated with variants as historically tracked attributes, i.e., modifications can be recorded whenever an updated value is imported, thus keeping track of all changes. Data can be visualized through variant‐centered and sample‐centered interfaces. A customizable search function can be exploited to periodically check if pathogenicity‐related data of a variant has changed over time. Patient recontacting ensuing from variant reinterpretation is made easier by iVar through the effective identification of all patients present in the database carrying a specific variant. We tested iVar by uploading 4171 VCF files and 1463 annotation files, obtaining a database of 4166 samples and 22,569 unique variants. iVar has proven to be a useful tool with good performance in terms of collecting and managing data from a medium‐throughput laboratory

Il minore vittima di abusi sessuali e le garanzie del giusto processo penale

Criminal proceedings concerning the assessment of sexual abusive behaviour affecting minors includes, among its most delicate stages, the collection of the young victim’s statements, especially if the victims are in their earliest years.The collection of a minor’s testimony must follow rules and procedures whose main goal is to safeguard the minor.Up to now, Italian Criminal Justice System does not provide an organic system for the Young victim’s protection; on the part of the legislator, there is a lack of specific rules and regolations for the minor’s testimony.According to Article 196, paragraph 1 and 2 of the Italian Code of Criminal Procedures, “every person is competent to testify,” but “in order to evaluate the witness’ statement, the judge can order appropriate verifications using all lawful means, even ex officio.”More effectively, Article 10 of the Carta of Noto recommend, for children under 12 and apart from exceptional cases, “that a valuation must always occur in order to verify the victim’s fitness to testify about the matters,” unless the judge can evaluate the reliability of the testimony.Therefore, is it possible for the judge to establish the truthfulness of the minor’s testimony apart from a technical advice of an expert, that is without making a psychological evaluation of the child first?” The prevailing jurisprudential orientation in recent years confirms the prevalence of the so-called principle of the all-encompassing evaluation, whereby “on the subject of sexual offenses against young children, it is illegal, for violation of the principle of the formation of evidence in contradictory, the refusal of judge to arrange a psychological examination, in order to ascertain the adherence to the reality or not of the narration of the facts, depending on possible fanciful elaborations of the age or of the personological structure of the minor2”.Nei procedimenti penali aventi ad oggetto l’accertamento della condotta di abuso sessuale a danno di minore, fra le fasi più delicate, vi è quella inerente l’acquisizione delle dichiarazioni della giovane vittima specie se in tenera età.E’ pacifico che l’assunzione della testimonianza di un minore, perché tale, deve avvenire con criteri e modalità che abbiano quale obiettivo principale, quello della tutela del minore stesso. Il sistema penale italiano a tutt’oggi rileva l’assenza di un organico sistema di protezione del minore vittima di abusi; manca una specifica disciplina da parte del legislatore, della testimonianza del soggetto minore di età.Secondo quanto disposto dall’art. 196, comma 1 e 2 c.p.p.“Ogni persona ha la capacità di testimoniare” tuttavia “qualora, al fine di valutare le dichiarazioni del testimone, sia necessario verificarne l’idoneità física o mentale a rendere testimonianza, il giudice anche di ufficio può ordinare gli accertamenti opportuni con i mezzi consentiti dalla legge”.Più efficacemente la Carta di Noto all’art. 10 raccomanda per i minori di anni dodici e salvo casi eccezionali “che sia sempre disposta perizia al fine di verificarne la idoneità a testimoniare sui fatti oggetto d’indagine”, salvo al giudice valutare l’attendibilità della testimonianza resa.E’ possibile allora per il decidente, accertare la veridicità della condotta abusante dal racconto del minore, prescindendo dall’ausilio del parere tecnico di un perito, ovvero senza aver prima disposto una perizia psicologica sul bambino?L’orientamento giurisprudenziale prevalente negli ultimi anni, conferma il prevalere del principio cosiddetto della valutazione onnicomprensiva, per cui “in tema di reati sessuali su minori in tenera età, è illegittimo, per violazione del principio della formazione della prova in contraddittorio, il rifiu to dei giudice di disporre una perizia psicologica, al fine di accertare l’aderenza alla realtà o meno della narrazione dei fatti, in dipendenza di eventuali elaborazioni fantasiose proprie dell’età o della struttura personologica del minore”1

A New Environmentally Safe Formulation and of Low Cost for Prolonged Release System of Atrazine and Diuron

Diuron and atrazine were incorporated in new formulations developed with the purpose to improve herbicides action through release systems, as well as to reduce the environmental toxicity. A low cost formulation (ALG/ESC) was obtained by combining sodium alginate (ALG) with fish scales of the Piau fish (ESC) from the Leporinus elongatus species. From the crosslinking of ALG/ESC with CaCl2, the formulation ALG/ESC-CaCl2 was obtained. For ALG/ESC-CaCl2, the results are successful, showing a prolonged release of 3.5 and 4.5 days for atrazine and diuron, respectively. Based on parameters of an empirical equation used to fit the herbicide release data, it appears that the release systems of diuron and atrazine from ALG/ESC-CaCl2 are by diffusion processes due to anomalous transport, which did not follow Fick’s laws of diffusion. DOI: http://dx.doi.org/10.17807/orbital.v9i3.99

Reduced pancreatic β-cell mass is associated with decreased FoxO1 and Erk1/2 protein phosphorylation in low-protein malnourished rats

A low-protein diet leads to functional and structural pancreatic islet alterations, including islet hypotrophy. Insulin-signaling pathways are involved in several adaptive responses by pancreatic islets. We determined the levels of some insulin-signaling proteins related to pancreatic islet function and growth in malnourished rats. Adult male Wistar rats (N = 20 per group) were fed a 17% protein (normal-protein diet; NP) or 6% protein (low-protein diet; LP), for 8 weeks. At the end of this period, blood glucose and serum insulin and albumin levels were measured. The morphometric parameters of the endocrine pancreas and the content of some proteins in islet lysates were determined. The β-cell mass was significantly reduced (≅65%) in normoglycemic but hypoinsulinemic LP rats compared to NP rats. Associated with these alterations, a significant 30% reduction in insulin receptor substrate-1 and a 70% increase in insulin receptor substrate-2 protein content were observed in LP islets compared to NP islets. The phosphorylated serine-threonine protein kinase (pAkt)/Akt protein ratio was similar in LP and NP islets. The phosphorylated forkhead-O1 (pFoxO1)/FoxO1 protein ratio was decreased by 43% in LP islets compared to NP islets (P < 0.05). Finally, the ratio of phosphorylated-extracellular signal-related kinase 1/2 (pErk1/2) to total Erk1/2 protein levels was decreased by 71% in LP islets compared to NP islets (P < 0.05). Therefore, the reduced β-cell mass observed in LP rats is associated with the reduction of phosphorylation in mitogenic-related signals, FoxO1 and Erk proteins. The cause/effect basis of this association remains to be determined4210935941FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2/04310-4; 04/11684-9; 03/10829-

First report about saxitoxins in freshwater fish Hoplias malabaricus through trophic exposure.

Cyanobacterial waterblooms, such as the saxitoxin (STX) producer Cylindrospermopsis raciborskii, have been a worldwide concern in environmental health. However, the bioaccumulation of this neurotoxin in the trophic chain is not completely known. The aim of the present work was to evaluate STX bioaccumulation through chemical analyses and the toxic and trophic effects using biomarkers in the tropical freshwater fish Hoplias malabaricus. They were fed once every five days with Astyanax sp. before being subjected to intraperitoneal inoculation with STX extract (0.08 mg/100 g) obtained by lysis of toxic C. raciborskii strain (T3). After 20 days the brain was collected for acetylcholinesterase (AChE), superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase (GST), glutathione peroxidase (GPx), glutathione (GSH), lipoperoxidation (LPO), protein carbonylation (PCO), and comet assay analysis. The muscle was collected for STX chemical analysis. The activities of SOD and concentrations of PCO and LPO increased. The CAT, GST, and GPx activities decreased. Genotoxicity was observed in the experimental group. STX was not detected in muscle samples. Thus, an oxidative stress was observed in the brain, leading to the damage of lipids, proteins, and DNA. The mechanism of action of the neurotoxin in this subchronic exposure suggests an apoptotic cellular process

Impact of N-tau on adult hippocampal neurogenesis, anxiety, and memory.

Different pathological tau species are involved in memory loss in Alzheimer’s disease, the most common cause of dementia among older people. However, little is known about how tau pathology directly affects adult hippocampal neurogenesis, a unique form of structural plasticity implicated in hippocampusdependent spatial learning and mood-related behavior. To this aim, we generated a transgenic mouse model conditionally expressing a pathological tau fragment (26e230 aa of the longest human tau isoform, or N-tau) in nestin-positive stem/progenitor cells. We found that N-tau reduced the proliferation of progenitor cells in the adult dentate gyrus, reduced cell survival and increased cell death by a caspase- 3eindependent mechanism, and recruited microglia. Although the number of terminally differentiated neurons was reduced, these showed an increased dendritic arborization and spine density. This resulted in an increase of anxiety-related behavior and an impairment of episodic-like memory, whereas less complex forms of spatial learning remained unaltered. Understanding how pathological tau species directly affect neurogenesis is important for developing potential therapeutic strategies to direct neurogenic instructive cues for hippocampal function repair