Ipsilateral and Contralateral Interactions in Spinal Locomotor Circuits Mediated by V1 Neurons: Insights from Computational Modeling.

We describe and analyze a computational model of neural circuits in the mammalian spinal cord responsible for generating and shaping locomotor-like oscillations. The model represents interacting populations of spinal neurons, including the neurons that were genetically identified and characterized in a series of previous experimental studies. Here, we specifically focus on the ipsilaterally projecting V1 interneurons, their possible role in the spinal locomotor circuitry, and their involvement in the generation of locomotor oscillations. The proposed connections of these neurons and their involvement in different neuronal pathways in the spinal cord allow the model to reproduce the results of optogenetic manipulations of these neurons under different experimental conditions. We suggest the existence of two distinct populations of V1 interneurons mediating different ipsilateral and contralateral interactions within the spinal cord. The model proposes explanations for multiple experimental data concerning the effects of optogenetic silencing and activation of V1 interneurons on the frequency of locomotor oscillations in the intact cord and hemicord under different experimental conditions. Our simulations provide an important insight into the organization of locomotor circuitry in the mammalian spinal cord.

Genomic Epidemiology of SARS-CoV-2 Infection During the Initial Pandemic Wave and Association With Disease Severity.

Importance: Understanding of SARS-CoV-2 variants that alter disease outcomes are important for clinical risk stratification and may provide important clues to the complex virus-host relationship. Objective: To examine the association of identified SARS-CoV-2 variants, virus clades, and clade groups with disease severity and patient outcomes. Design, Setting, and Participants: In this cross-sectional study, viral genome analysis of clinical specimens obtained from patients at the Cleveland Clinic infected with SARS-CoV-2 during the initial wave of infection (March 11 to April 22, 2020) was performed. Identified variants were matched with clinical outcomes. Data analysis was performed from April to July 2020. Main Outcomes and Measures: Hospitalization, intensive care unit (ICU) admission, mortality, and laboratory outcomes were matched with SARS-CoV-2 variants. Results: Specimens sent for viral genome sequencing originated from 302 patients with SARS-CoV-2 infection (median [interquartile range] age, 52.6 [22.8 to 82.5] years), of whom 126 (41.7%) were male, 195 (64.6%) were White, 91 (30.1%) required hospitalization, 35 (11.6%) needed ICU admission, and 17 (5.6%) died. From these specimens, 2531 variants (484 of which were unique) were identified. Six different SARS-CoV-2 clades initially circulated followed by a rapid reduction in clade diversity. Several variants were associated with lower hospitalization rate, and those containing 23403A>G (D614G Spike) were associated with increased survival when the patient was hospitalized (64 of 74 patients [86.5%] vs 10 of 17 patients [58.8%]; χ21 = 6.907; P = .009). Hospitalization and ICU admission were similar regardless of clade. Infection with Clade V variants demonstrated higher creatinine levels (median [interquartile range], 2.6 [-0.4 to 5.5] mg/dL vs 1.0 [0.2 to 2.2] mg/dL; mean creatinine difference, 2.9 mg/dL [95% CI, 0.8 to 5.0 mg/dL]; Kruskal-Wallis P = .005) and higher overall mortality rates (3 of 14 patients [21.4%] vs 17 of 302 patients [5.6%]; χ21 = 5.640; P = .02) compared with other variants. Infection by strains lacking the 23403A>G variant showed higher mortality in multivariable analysis (odds ratio [OR], 22.4; 95% CI, 0.6 to 5.6; P = .01). Increased variants of open reading frame (ORF) 3a were associated with decreased hospitalization frequency (OR, 0.4; 95% CI, 0.2 to 0.96; P = .04), whereas increased variants of Spike (OR, 0.01; 95% CI, <0.01 to 0.3; P = .01) and ORF8 (OR, 0.03; 95% CI, <0.01 to 0.6; P = .03) were associated with increased survival. Conclusions and Relevance: Within weeks of SARS-CoV-2 circulation, a profound shift toward 23403A>G (D614G) specific genotypes occurred. Replaced clades were associated with worse clinical outcomes, including mortality. These findings help explain persistent hospitalization yet decreasing mortality as the pandemic progresses. SARS-CoV-2 clade assignment is an important factor that may aid in estimating patient outcomes.

Flexor and Extensor Ankle Afferents Broadly Innervate Locomotor Spinal Shox2 Neurons and Induce Similar Effects in Neonatal Mice.

Central pattern generators (CPGs) in the thoracolumbar spinal cord generate the basic hindlimb locomotor pattern. The locomotor CPG integrates descending commands and sensory information from the periphery to activate, modulate and halt the rhythmic program. General CPG function and response to sensory perturbations are well described in cat and rat models. In mouse, roles for many genetically identified spinal interneurons have been inferred from locomotor alterations following population deletion or modulation. However, the organization of afferent input to specific genetically identified populations of spinal CPG interneurons in mouse remains comparatively less resolved. Here, we focused on a population of CPG neurons marked by the transcription factor Shox2. To directly test integration of afferent signaling by Shox2 neurons, sensory afferents were stimulated during patch clamp recordings of Shox2 neurons in isolated spinal cord preparations from neonatal mice. Shox2 neurons broadly displayed afferent-evoked currents at multiple segmental levels, particularly from caudal dorsal roots innervating distal hindlimb joints. As dorsal root stimulation may activate both flexor- and extensor-related afferents, preparations preserving peripheral nerves were used to provide more specific activation of ankle afferents. We found that both flexor- and extensor-related afferent stimulation were likely to evoke similar currents in a given Shox2 neuron, as assessed by response polarity, latency, duration and amplitude. It has been proposed that Shox2 neurons can be divided into neurons which contribute to rhythm generation and neurons that are premotor by the absence and presence of the V2a marker Chx10, respectively. Response to afferent stimulation did not differ based on Chx10 expression. Although currents evoked in response to flexor and extensor afferent activation did not follow expected functional antagonism, they were consistent with the observation that stimulation of flexor- and extensor-related afferents both reset the phase of ongoing fictive locomotion to flexion in neonatal mice. Together, the data suggest that Shox2 neurons are interposed in multiple sensory pathways and low threshold proprioceptive input reinforces sensory perturbation of ongoing locomotion by similarly activating or inhibiting both the rhythm and patterning layers of the CPG.

Neuropilin 1 Receptor Is Up-Regulated in Dysplastic Epithelium and Oral Squamous Cell Carcinoma.

Neuropilins are receptors for disparate ligands, including proangiogenic factors such as vascular endothelial growth factor and inhibitory class 3 semaphorin (SEMA3) family members. Differentiated cells in skin epithelium and cutaneous squamous cell carcinoma highly express the neuropilin-1 (NRP1) receptor. We examined the expression of NRP1 in human and mouse oral mucosa. NRP1 was significantly up-regulated in oral epithelial dysplasia and oral squamous cell carcinoma (OSCC). NRP1 receptor localized to the outer suprabasal epithelial layers in normal tongue, an expression pattern similar to the normal skin epidermis. However, dysplastic tongue epithelium and OSCC up-regulated NRP1 in basal and proliferating epithelial layers, a profile unseen in cutaneous squamous cell carcinoma. NRP1 up-regulation is observed in a mouse carcinogen-induced OSCC model and in human tongue OSCC biopsies. Human OSCC cell lines express NRP1 protein in vitro and in mouse tongue xenografts. Sites of capillary infiltration into orthotopic OSCC tumors correlate with high NRP1 expression. HSC3 xenografts, which express the highest NRP1 levels of the cell lines examined, showed massive intratumoral lymphangiogenesis. SEMA3A inhibited OSCC cell migration, suggesting that the NRP1 receptor was bioactive in OSCC. In conclusion, NRP1 is regulated in the oral epithelium and is selectively up-regulated during epithelial dysplasia. NRP1 may function as a reservoir to sequester proangiogenic ligands within the neoplastic compartment, thereby recruiting neovessels toward tumor cells.

Fabrication and characterization of porous EH scaffolds and EH-PEG bilayers.

Biomaterials made from synthetic polymers are becoming more pervasive in the medical field. Synthetic polymers are particularly advantageous as their chemical and mechanical properties can be easily tailored to a specific application. This work characterizes polymer scaffolds derived from the cyclic acetal monomer 5-ethyl-5-(hydroxymethyl)-ß,ß-dimethyl-1,3-dioxane-2-ethanol diacrylate (EHD). Both porous scaffolds and bilayer scaffolds based upon the EHD monomer were fabricated, and the resulting scaffolds' degradation and mechanical properties were studied. The results showed that by modifying the architecture of an EH scaffold, either by adding a porous network or a poly(ethylene glycol) (PEG) coating, the degradation and Young's modulus of the biomaterial can be significant altered. However, results also indicated that these architectural modifications can be accomplished without a significant loss in the flexural strength of the scaffold. Therefore, we suggest that porous EH scaffolds, and particularly porous EH-PEG bilayers, may be especially useful in dynamic tissue environments due to their advantageous architectural and mechanical properties.

Porous EH and EH-PEG scaffolds as gene delivery vehicles to skeletal muscle.

PURPOSE: Synthetic biomaterials are widely used in an attempt to control the cellular behavior of regenerative tissues. This can be done by altering the chemical and physical properties of the polymeric scaffold to guide tissue repair. This paper addresses the use of a polymeric scaffold (EH network) made from the cyclic acetal monomer, 5-ethyl-5-(hydroxymethyl)-ß,ß-dimethyl-1,3-dioxane-2-ethanol diacrylate (EHD), as a release device for a therapeutic plasmid encoding for an insulin-like growth factor-1 green fluorescent protein fusion protein (IGF-1 GFP). METHODS: Scaffolds were designed to have different porous architectures, and the impact of these architectures on plasmid release was determined. We hypothesized that IGF-1 could be delivered more effectively using a porous scaffold to allow for the release of IGF-1. RESULTS: We showed that by altering the number of pores exposed to the surface of the network, faster plasmid loading and release were achieved. In addition, the IGF-1 GFP plasmids were found to be effective in producing IGF-1 and GFP within human skeletal muscle myoblast cell cultures. CONCLUSIONS: This work aims to show the utility of EH biomaterials for plasmid delivery for potentially localized skeletal muscle regeneration.

Solanum erianthum intoxication mimicking an acute cerebrovascular disease.

We describe a case of Solanum erianthum poisoning that happened to a 75-year-old man. He ate the S erianthum that he gathered from the countryside, believing that it would be helpful in improving the numbness of his distal limbs. S erianthum is a solanaceous plant that contains a variable concentration of solanum alkaloids, causing gastrointestinal irritation, and tropane alkaloids that have anticholinergic properties producing typical and sometimes severe atropinelike symptoms. The diagnosis of S erianthum poisoning was made based on clinical symptoms and signs of anticholinergic syndrome associated with a history of vegetable meal from countryside and, moreover, on identification of the vegetable obtained from his family. Gastrointestinal decontamination and symptomatic treatment were useful in treatment of acute poisoning.