The human dopamine (DA) transporter (hDAT) mediates clearance of DA. Genetic variants in hDAT have been associated with DA dysfunction, a complication associated with several brain disorders, including autism spectrum disorder (ASD). Here, we investigated the structural and behavioral bases of an ASD-associated in-frame deletion in hDAT at N336 (∆N336). We uncovered that the deletion promoted a previously unobserved conformation of the intracellular gate of the transporter, likely representing the rate-limiting step of the transport process. It is defined by a “half-open and inward-facing” state (HOIF) of the intracellular gate that is stabilized by a network of interactions conserved phylogenetically, as we demonstrated in hDAT by Rosetta molecular modeling and fine-grained simulations, as well as in its bacterial homolog leucine transporter by electron paramagnetic resonance analysis and X-ray crystallography. The stabilization of the HOIF state is associated both with DA dysfunctions demonstrated in isolated brains of Drosophila melanogaster expressing hDAT ∆N336 and with abnormal behaviors observed at high-time resolution. These flies display increased fear, impaired social interactions, and locomotion traits we associate with DA dysfunction and the HOIF state. Together, our results describe how a genetic variation causes DA dysfunction and abnormal behaviors by stabilizing a HOIF state of the transporter.

In this study we describe the site and moment of histospecific differentiation in developmental stages of the annelid Platynereis dumerilii by use of biochemical markers. The monoclonal antibody (mab) OI7 and uncloned hybridoma supernatants (pAb's) OI8, OI10, OI46 and OI69 recognize neural antigens that appear asynchronously during development. By an enzymatic test, acetylcholinesterase (AChE) was found specific for nervous tissue as well. The patterns of neural structures labelled by antibodies differ, however, from those revealed by AChE staining. Experimental inhibition of transcription (with actinomycin D) and of translation (using puromycin) demonstrate that the expression of histospecific neural markers depends on both zygotic transcription and subsequent translation. The mAb OI64 labels epidermal (and neural) gland cells. The antibody 4D9, raised against the engrailed protein of Drosophila, labels single rows of ciliated cells at the posterior border of segments.

Parylene deposition on 3D prints creates biocompatible microdevices and facilitates fabrication of master molds.

In vitro models of the human central nervous system (CNS), particularly those derived from induced pluripotent stem cells (iPSCs), are becoming increasingly recognized as useful complements to animal models for studying neurological diseases and developing therapeutic strategies. However, many current 3D CNS models suffer from deficits that limit their research utility. In this work, we focused on improving the interactions between the extracellular matrix (ECM) and iPSC-derived neurons to support model development. The most common ECMs used to fabricate 3D CNS models often lack the necessary bioinstructive cues to drive iPSC-derived neurons to a mature and synaptically connected state. These ECMs are also typically difficult to pattern into complex structures due to their mechanical properties. To address these issues, we functionalized gelatin methacrylate (GelMA) with an N-cadherin extracellular peptide epitope to create a biomaterial termed GelMA-Cad. After photopolymerization, GelMA-Cad forms soft hydrogels (on the order of 2 kPa) that can maintain patterned architectures. The N-cadherin functionality promotes survival and maturation of single-cell suspensions of iPSC-derived glutamatergic neurons into synaptically connected networks as determined by viral tracing and electrophysiology. Immunostaining reveals a pronounced increase in presynaptic and postsynaptic marker expression in GelMA-Cad relative to Matrigel, as well as extensive co-localization of these

Robust data on hepatitis C virus (HCV) population prevalence are essential to inform national HCV services. In 2016, we undertook a survey to estimate HCV prevalence among the adult population in Ireland. We used anonymised residual sera available at the National Virus Reference Laboratory. We selected a random sample comprising persons ≥ 18 years with probability proportional to the general population age-sex distribution. Anti-HCV and HCV Ag were determined using the Architect anti-HCV and HCV Ag assays. Fifty-three of 3,795 specimens were seropositive (age-sex-area weighted seroprevalence 0.98% (95% confidence interval (CI): 0.73–1.3%)). Thirty-three specimens were HCV-antigen and antibody-positive (age-sex-area weighted prevalence of chronic infection 0.57% (95% CI: 0.40–0.81%)). The prevalence of chronic infection was higher in men (0.91%; 95% CI: 0.61–1.4%), in specimens from the east of the country (1.4%; 95%CI: 0.99–2.0%), and among persons aged 30–39 years and 40–49 years (1.1% (95% CI: 0.59–2.0%) and 1.1% (95% CI: 0.64–1.9%) respectively). Ireland ranks at the lower end of the spectrum of prevalence of chronic HCV infection internationally. Men born between 1965 and 1984 from the east of the country have the highest rate of chronic HCV infection.