Nanoparticle drug delivery systems for inner ear therapy: An overview

open7noembargoed_20180701Valente, Filippo; Astolfi, Laura; Simoni, Edi; Danti, Serena; Franceschini, Valeria; Chicca, Milvia; Martini, AlessandroValente, Filippo; Astolfi, Laura; Simoni, Edi; Danti, Serena; Franceschini, Valeria; Chicca, Milvia; Martini, Alessandr

Transcription Pattern of Neurotrophic Factors and Their Receptors in Adult Zebrafish Spinal Cord

In vertebrates, neurotrophins and their receptors play a fundamental role in the central and peripheral nervous systems. Several studies reported that each neurotrophin/receptor signalling pathway can perform various functions during axon development, neuronal growth, and plasticity. Previous investigations in some fish species have identified neurotrophins and their receptors in the spinal cord under physiological conditions and after injuries, highlighting their potential role during regeneration. In our study, for the first time, we used an excellent animal model, the zebrafish (Danio rerio), to compare the mRNA localization patterns of neurotrophins and receptors in the spinal cord. We quantified the levels of mRNA using qPCR, and identified the transcription pattern of each neurotrophin/receptor pathway via in situ hybridization. Our data show that ngf/trka are the most transcribed members in the adult zebrafish spinal cord

Analysis of clasp2 Transcription Pattern in Male Germ Cells during Spermatogenesis: A Comparative Study in Zebrafish (Danio rerio) and Guppy (Poecilia reticulata)

Cytoplasmic linker-associated protein-2 (CLASP2) is a member of the CLIP-associating proteins (CLASPs) family involved in the structure and function of microtubules and Golgi apparatus. Several studies performed using different mammalian and non-mammalian model organisms reported that CLASP2 controls microtubule dynamics and the organization of microtubule networks. In Drosophila and mice, an important role of CLASP2 during the development of germ cell lines has been uncovered. However, no study has clearly defined its role during fish germ cell differentiation. In the present study, we used two excellent aquatic animal models among teleost fish: zebrafish (Danio rerio) and guppy (Poecilia reticulata). Using qPCR, we found that the clasp2 transcript level is significantly high in the testis of both fish. Then, by in situ hybridization, we localized the clasp2 transcript in the spermatozoa of zebrafish and the spermatozeugmata of guppy. Our data suggest a potential role for this gene in the last stage of spermiogenesis in fis

Stroke Gait Rehabilitation: A Comparison of End-Effector, Overground Exoskeleton, and Conventional Gait Training

Gait recovery is one of the main goals of post-stroke rehabilitation and Robot-Assisted Gait Training (RAGT) has shown positive outcomes. However, there is a lack of studies in the literature comparing the effects of different devices. This paper aims to study the effects, in terms of clinical and gait outcomes, of treadmill-based and overground RAGT, compared to conventional gait training in stroke subjects. The results showed a significant improvement of clinical outcomes in both robotic treatments and in conventional therapy. The performance of locomotor tasks was clinically significant in the robotic groups only. The spatio-temporal gait parameters did not reveal any significant difference. Results suggest future multicentre studies on a larger number of subjects

VSV-G-Enveloped Vesicles for Traceless Delivery of CRISPR-Cas9

The method of delivery of CRISPR-Cas9 into target cells is a strong determinant of efficacy and specificity in genome editing. Even though high efficiency of Cas9 delivery is necessary for optimal editing, its long-term and high levels of expression correlate with increased off-target activity. We developed vesicles (VEsiCas) carrying CRISPR-SpCas9 ribonucleoprotein complexes (RNPs) that are efficiently delivered into target cells through the fusogenic glycoprotein of the vesicular stomatitis virus (VSV-G). A crucial step for VEsiCas production is the synthesis of the single guide RNA (sgRNA) mediated by the T7 RNA polymerase in the cytoplasm of producing cells as opposed to canonical U6-driven Pol III nuclear transcription. In VEsiCas, the absence of DNA encoding SpCas9 and sgRNA allows rapid clearance of the nuclease components in target cells, which correlates with reduced genome-wide off-target cleavages. Compared with SpCas9 RNPs electroporation, which is currently the method of choice to obtain transient SpCas9 activity, VEsiCas deliver the nuclease with higher efficiency and lower toxicity. We show that a wide variety of cells can be edited through VEsiCas, including a variety of transformed cells, induced pluripotent stem cells (iPSCs), and cardiomyocytes, in vivo. VEsiCas is a traceless CRISPR-Cas9 delivery tool for efficient and safe genome editing that represents a further advancement toward the therapeutic use of the CRISPR-Cas9 technology

Association of protein function-altering variants with cardiometabolic traits:the strong heart study

Clinical and biomarker phenotypic associations for carriers of protein function-altering variants may help to elucidate gene function and health effects in populations. We genotyped 1127 Strong Heart Family Study participants for protein function-altering single nucleotide variants (SNV) and indels selected from a low coverage whole exome sequencing of American Indians. We tested the association of each SNV/indel with 35 cardiometabolic traits. Among 1206 variants (average minor allele count = 20, range of 1 to 1064), similar to 43% were not present in publicly available repositories. We identified seven SNV-trait significant associations including a missense SNV at ABCA10 (rs779392624, p= 8 x 10(-9)) associated with fasting triglycerides, which gene product is involved in macrophage lipid homeostasis. Among non-diabetic individuals, missense SNVs at four genes were associated with fasting insulin adjusted for BMI (PHIL, chr6:79,650,711, p= 2.1 x 10(-6); TRPM3, rs760461668, p= 5 x10(-8); SPTY2D1, rs756851199, p= 1.6 x 10(-8); and TSPO, rs566547284, p= 2.4 x 10(-6)). PHIL encoded protein is involved in pancreatic beta-cell proliferation and survival, and TRPM3 protein mediates calcium signaling in pancreatic beta-cells in response to glucose. A genetic risk score combining increasing insulin risk alleles of these four genes was associated with 53% (95% confidence interval 1.09, 2.15) increased odds of incident diabetes and 83% (95% confidence interval 1.35, 2.48) increased odds of impaired fasting glucose at follow-up. Our study uncovered novel gene-trait associations through the study of protein-coding variants and demonstrates the advantages of association screenings targeting diverse and high-risk populations to study variants absent in publicly available repositories

Pandemic nightmares: effects on dream activity of the COVID-19 lockdown in Italy

COVID-19 has critically impacted the world. Recent works have found substantial changes in sleep and mental health during the COVID-19 pandemic. Dreams could give us crucial information about people's well-being, so here we have directly investigated the consequences of lockdown on the oneiric activity in a large Italian sample: 5,988 adults completed a web-survey during lockdown. We investigated sociodemographic and COVID-19-related information, sleep quality (by the Medical Outcomes Study-Sleep Scale), mental health (by the Depression, Anxiety, and Stress Scales), dream and nightmare frequency, and related emotional aspects (by the Mannheim Dream Questionnaire). Comparisons between our sample and a population-based sample revealed that Italians are having more frequent nightmares and dreams during the pandemic. A multiple logistic regression model showed the predictors of high dream recall (young age, female gender, not having children, sleep duration) and high nightmare frequency (young age, female gender, modification of napping, sleep duration, intrasleep wakefulness, sleep problem index, anxiety, depression). Moreover, we found higher emotional features of dream activity in workers who have stopped working, in people who have relatives/friends infected by or who have died from COVID-19 and in subjects who have changed their sleep habits. Our findings point to the fact that the predictors of high dream recall and nightmares are consistent with the continuity between sleep mentation and daily experiences. According to the arousal-retrieval model, we found that poor sleep predicts a high nightmare frequency. We suggest monitoring dream changes during the epidemic, and also considering the implications for clinical treatment and prevention of mental and sleep disorders