Spontaneous Responses Research Articles

Intraganglionic reactive oxygen species mediate inflammatory pain and hyperalgesia through TRPA1 in the rat.

Reactive oxygen species (ROS) are generated in nociceptive pathways in response to inflammation and injury. ROS are accumulated within the sensory ganglia following peripheral inflammation, but the functional role of intraganlionic ROS in inflammatory pain is not clearly understood. The aims of this study were to investigate whether peripheral inflammation leads to prolonged ROS accumulation within the trigeminal ganglia (TG), whether intraganglionic ROS mediate pain hypersensitivity via activation of TRPA1, and whether TRPA1 expression is upregulated in TG during inflammatory conditions by ROS. We demonstrated that peripheral inflammation causes excess ROS production within TG during the period when inflammatory mechanical hyperalgesia is most prominent. Additionally, scavenging intraganglionic ROS attenuated inflammatory mechanical hyperalgesia and a pharmacological blockade of TRPA1 localized within TG also mitigated inflammatory mechanical hyperalgesia. Interestingly, exogenous administration of ROS into TG elicited mechanical hyperalgesia and spontaneous pain-like responses via TRPA1, and intraganglionic ROS induced TRPA1 upregulation in TG. These results collectively suggest that ROS accumulation in TG during peripheral inflammation contributes to pain and hyperalgesia in a TRPA1 dependent manner, and that ROS further exacerbate pathological pain responses by upregulating TRPA1 expression. Therefore, any conditions that exacerbate ROS accumulation within somatic sensory ganglia can aggravate pain responses and treatments reducing ganglionic ROS may help alleviate inflammatory pain.

Inhibition of Tumor Metastasis by Liquid-Nitrogen-Shocked Tumor Cells with Oncolytic Viruses Infection.

Despite the superior tumor lytic efficacy of oncolytic viruses (OVs), their systemic delivery still faces the challenges of limited circulating periods, poor tumor tropism, and spontaneous antiviral immune responses. Herein, a virus-concealed tumor-targeting strategy enabling OVs' delivery toward lung metastasis via systemic administration is described. The OVs can actively infect, be internalized, and cloak into tumor cells. Then the tumor cells are subsequently treated with liquid-nitrogen-shocking to eliminate the pathogenicity. Such a Trojan Horse-like vehicle avoids virus neutralization and clearance in the bloodstream and facilitates tumor-targeted delivery for more than 110-fold virus enrichment in the tumor metastasis. In addition, this strategy can serve as a tumor vaccine and initiate endogenous adaptive antitumor effects through increasing the memory T cells and modulating the tumor immune microenvironment, including reducing the M2 macrophage, downregulating Treg cells, and priming T cells.

(094) Synergy Between Orally Administered Phosphodiesterase-type-5-inhibitors and Topically Delivered Nitric-oxide in Eliciting Erectile Responses

Abstract Introduction Phosphodiesterase-type-5-inhibitors (PDE5i) represent first-line FDA-approved oral treatments for erectile dysfunction (ED). However, because their mode of action is dependent on the production of nitric oxide (NO) from nerve endings, patients that suffer neurogenic ED (for example, following the procedures for radical prostatectomy (RP) or resulting from diabetes) are commonly refractory to PDE5i treatment. We recently demonstrated proof-of-principle for increased efficacy of topically applied NO-releasing microparticles (NO-MP) in eliciting an erectile response when used in combination with the orally administered PDE5i, sildenafil. However, there are several other FDA approved PDE5i used to treat ED, which, although acting through the common mechanism of PDE5 inhibition, exhibit several characteristics that potentially could impact clinical and commercial translation of combination therapy. Objective The goal of the present studies was to determine if other commercially available PDE5i besides sildenafil (avanafil, tadalafil and vardenafil) also act cooperatively when used in combination with NO-MP to elicit an erectile response. Methods Sprague-Dawley rats underwent bi-lateral transection of the cavernous nerve (CN), as a model of RP, one-week prior to determining erectile response to treatment. We investigated the effect of oral administration of 0.05mg sildenafil/kg, 0.005mg tadalafil/kg, 0.01mg vardenafil/kg and 0.1mg avanafil/kg (approximately equivalent to clinical doses of 2.5mg, 0.25mg, 0.5mg and 5mg, respectively, administered to an 80 kg patient) 30 minutes prior to being anesthetized for the determination of erectile response by intracorporal pressure (ICP) and systemic blood pressure (BP). Baseline ICP and BP were recorded for 30 minutes, at which time 100 mg of NO-MP (or control, blank-MP) were applied to the rat penile shaft. The ICP and systemic BP were used to determine the time to the first erectile response, duration of the erectile responses, the maximal and average ICP/BP responses and the number of spontaneous erections per hour. Results Although oral treatment by PDE5i alone resulted in no observable erectile response, topical application of NO-MP resulted in spontaneous erectile responses. When NO-MP were applied in combination with oral PDE5i treatment, all PDE5i resulted in faster times to the first erectile response (on average 11 minutes compared to 22 minutes when NO-MP were used alone; P-value=0.041) and greater numbers of spontaneous erections (on average 4.6 erections per hour compared to 2.0 when NO-MP were used alone, P-value=0.043). Although there were no significant differences between any of the PDE5i when used in combination with NO-MP with respect to the average erectile responses per hour, duration of erectile response, and maximal ICP/BP, there was a trend in the vardenafil and avanafil treated animals for a more rapid onset of the erectile responses than observed with sildenafil and tadalafil. Conclusions Although knowledge of the molecular pathways that lead to an erection provide a rationale for a synergistic effect between PDE5i and NO in eliciting an erectile response, our results provide the first evidence of such synergy. Our results suggest a novel treatment strategy for patients with ED refractory to PDE5i, namely any of the FDA-approved oral PDE5 inhibitors used in combination with formulations that topically deliver NO to corporal tissue. Disclosure Yes, this is sponsored by industry/sponsor: Zylo Therapeutics, Inc Clarification Industry funding only - investigator initiated and executed study Any of the authors act as a consultant, employee or shareholder of an industry for: Zylo Therapeutics, Inc

Development of abnormalities at the neuromuscular junction in the SOD1-G93A mouse model of ALS: dysfunction then disruption of postsynaptic structure precede overt motor symptoms.

The ultimate deficit in amyotrophic lateral sclerosis (ALS) is neuromuscular junction (NMJ) loss, producing permanent paralysis, ultimately in respiratory muscles. However, understanding the functional and structural deficits at NMJs prior to this loss is crucial for therapeutic strategy design. Should early interventions focus on reversing denervation, or supporting largely intact NMJs that are functionally impaired? We therefore determined when functional and structural deficits appeared in diaphragmatic NMJs relative to the onset of hindlimb tremor (the first overt motor symptoms) in vivo in the SOD1-G93A mouse model of ALS. We employed electrophysiological recording of NMJ postsynaptic potentials for spontaneous and nerve stimulation-evoked responses. This was correlated with fluorescent imaging microscopy of the postsynaptic acetylcholine receptor (AChR) distribution throughout the postnatal developmental timecourse from 2 weeks to early symptomatic ages. Significant reduction in the amplitudes of spontaneous miniature endplate potentials (mEPPs) and evoked EPPs emerged only at early symptomatic ages (in our colony, 18-22 weeks). Reductions in mEPP frequency, number of vesicles per EPP, and EPP rise time were seen earlier, at 16weeks, but this reversed by early symptomatic ages. However, the earliest and most striking impairment was an inability to maintain EPP amplitude during a 20 Hz stimulus train, which appeared 6 weeks before overt in vivo motor symptoms. Despite this, fluorescent α-bungarotoxin labelling revealed no systematic, progressive changes in 11 comprehensive NMJ morphological parameters (area, shape, compactness, number of acetylcholine receptor, AChR, regions, etc.) with disease progression. Rather, while NMJs were largely normally-shaped, from 16 weeks there was a progressive and substantial disruption in AChR concentration and distribution within the NMJ footprint. Thus, NMJ functional deficits appear at least 6 weeks before motor symptoms in vivo, while structural deficits occur 4 weeks later, and predominantly within NMJs. These data suggest initial therapies focused on rectifying suboptimal NMJ function could produce effective relief of symptoms of weakness.

ANK2 loss-of-function variants are associated with epilepsy, and lead to impaired axon initial segment plasticity and hyperactive network activity in hiPSC-derived neuronal networks.

To characterize a novel neurodevelopmental syndrome due to loss-of-function (LoF) variants in Ankyrin 2 (ANK2), and to explore the effects on neuronal network dynamics and homeostatic plasticity in human induced pluripotent stem cell-derived neurons. We collected clinical and molecular data of twelve individuals with heterozygous de novo LoF variants in ANK2. We generated a heterozygous LoF allele of ANK2 using CRISPR/Cas9 in human induced pluripotent stem cells (hiPSCs). HiPSCs were differentiated towards excitatory neurons of which we measured spontaneous electrophysiological responses using micro-electrode arrays (MEAs), characterized somatodendritic morphology and axon initial segment (AIS) structure and plasticity. We found a broad neurodevelopmental disorder, comprising intellectual disability, autism spectrum disorders, and early onset epilepsy. Using MEAs, we found that hiPSC-derived neurons with heterozygous LoF of ANK2 show a hyperactive and desynchronized neuronal network. ANK2 deficient neurons also showed increased somatodendritic structures and altered AIS structure of which its plasticity is impaired upon activity-dependent modulation. Phenotypic characterization of patients with de novo ANK2 LoF variants define a novel NDD with early onset epilepsy. Our functional in vitro data of ANK2-deficient human neurons show a specific neuronal phenotype in which reduced ANKB expression leads to hyperactive and desynchronized neuronal network activity, increased somatodendritic complexity and AIS structure and impaired activity-dependent plasticity of the AIS.

Tax Revolts in Yorubaland, 1925-1955

Most researchers have generally regarded taxation as the main reason for the revolt in colonial Nigeria. There are good reasons to believe that there were other significant tax-related considerations, other than taxation, that were responsible for the revolts that were thrown up in late colonial southwestern Nigeria. Some scholars have also noted a comparative process that is currently unfolding; that it was the need to raise money that forced the hands of the colonial authority into democratization and that the despondency of the indigenous population was a major reason for the crises that enveloped Yorubaland from 1925 up to 1955. These claims have been fully tested. In the article ‘Tax Revolts in Yoruba land, 1925-1955’, questions are asked and arguments explored using archival and oral evidence. The study found that the high-handedness of the Native Authority and other social and economic considerations were genuine reasons for the discontent of the indigenous population. These were significant constraints that morphed into spontaneous violent responses across Yorubaland against British colonialism.

Chronic Intermittent Hypoxia Facilitates Action Potential Firing in PVN-projecting MnPO Neurons

Our previous results indicate that Chronic intermittent hypoxia (CIH) enhances glutamatergic neurotransmitter release in PVN-projecting MnPO neurons. Very little information exists regarding how CIH affects the excitability of PVN-projecting MnPO neurons. MnPO neurons exhibit a wide diversity of spontaneous and evoked firing patterns. Initial studies indicate CIH modulates resting membrane properties and AP firing patterns which may be associated with alterations in hyperpolarization-activated cyclic nucleotide-gated currents (Ih). The Ih current is an inward current activated by hyperpolarization from the resting potential and is an important modulator of action potential firing frequency. Data were collected in PVN-projecting MnPO neurons which play an important role in CIH-induced hypertension. Adult male Sprague Dawley rats (250-350g) received bilateral PVN injections of 200 nL retrograde adeno-associated virus pAAV-CAG-tdTomato. Three weeks following injections, rats were either exposed to a 7-day intermittent hypoxia protocol consisting of 6 min cycles (3 min 21% O2, 3 min 10% O2) repeated 10x/hr for 8 hours (during the normal inactive/sleep phase) or room air (Norm). Following CIH/Norm treatment, rats were deeply anesthetized and sagittal brain slices of MnPO were prepared using standard slice procedures. Spontaneous activity and responses to current injections were recorded from PVN projecting MnPO neurons using whole cell current-clamp. Data was analyzed off-line using Clampfit. There was a trend for spontaneous activity to be higher in PVN-projecting MnPO neurons from rats exposed to 7 days CIH when compared to normoxic controls (Norm: 2.29 ± 0.51 vs CIH: 3.3 ± 0.84) that was not due to a change in resting membrane potential. The threshold to evoke AP potentials was significantly lower in CIH neurons (5 pA injecting current, 17/19 CIH neurons vs 9/13 Norm neurons, CIH 8.6 ± 0.79 mv vs Norm 13.06±1.53 mv, p<0.01; 10 pA injecting current, 19 CIH neurons vs 12/13 Norm neurons CIH 14.74±0.92 mv vs Norm 19.72 ±1.64 mv, p<0.01). However, evoked action potentials were less sensitive to increasing current injection amplitudes in CIH vs Norm treated rats. Following bath application of the specific Ih blocker ZD7288 (ZD, 30 μm) spontaneous action potential activity was reduced by 24% in Norm neurons and 47% in CIH neurons (p=0.0313). These data demonstrate that CIH facilitates action potential generation in PVN projecting MnPO neurons and changes in Ih currents may contribute to this increase in excitability. R01 HL155977 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Effects of neck proprioceptive modulation on pallidal network connectivity in dystonia.

Cervical dystonia (CD) is the third most common movement disorder affecting 1 million people worldwide. Proprioceptive modulation is the hallmark of contemporary therapies for dystonia, but the mechanism for this intervention is unclear. We studied proprioceptive influence on CD by measuring the spontaneous single-neuron responses and local field potentials (LFP) from the globus pallidus interna (GPi) in 17 CD patients (9 isolated CD and 8 with CD as a feature of generalized dystonia). The goal was to examine how high-frequency neck vibration, a putative modulator of neck proprioception changes pallidal physiology. We found that the neck vibration instantaneously alters the pallidal single neuron activity. We also found that neck vibration modulates pallido-cerebellar connectivity by changing alpha band in LFP recordings. The effects were more robust in those with isolated CD. The vibration also affects pallido-hippocampal connectivity by modulating theta-band power. These effects were more robust in CD with generalized dystonia. Vibration changed LFP only in select pallidal regions. Regions where LFP power was substantially modulated had a prominent proportion of burst subtypes of neurons, compared to pause or tonic subtypes. Such disparity in subtype was absent in regions where the LFP power was not modulated or subtly reduced with neck vibration. When changes in the theta, alpha and beta bands of the LFP recordings were compared against each other in response to vibration, high correlation was observed.

Celecoxib alleviates nociceptor sensitization mediated by interleukin-1beta-primed annulus fibrosus cells

PurposeThis study aims to analyze the effect of pro-inflammatory cytokine-stimulated human annulus fibrosus cells (hAFCs) on the sensitization of dorsal root ganglion (DRG) cells. We further hypothesized that celecoxib (cxb) could inhibit hAFCs-induced DRG sensitization.MethodshAFCs from spinal trauma patients were stimulated with TNF-α or IL-1β. Cxb was added on day 2. On day 4, the expression of pro-inflammatory and neurotrophic genes was evaluated using RT-qPCR. Levels of prostaglandin E2 (PGE-2), IL-8, and IL-6 were measured in the conditioned medium (CM) using ELISA. hAFCs CM was then applied to stimulate the DRG cell line (ND7/23) for 6 days. Then, calcium imaging (Fluo4) was performed to evaluate DRG cell sensitization. Both spontaneous and bradykinin-stimulated (0.5 μM) calcium responses were analyzed. The effects on primary bovine DRG cell culture were performed in parallel to the DRG cell line model.ResultsIL-1ß stimulation significantly enhanced the release of PGE-2 in hAFCs CM, while this increase was completely suppressed by 10 µM cxb. hAFCs revealed elevated IL-6 and IL-8 release following TNF-α and IL-1β treatment, though cxb did not alter this. The effect of hAFCs CM on DRG cell sensitization was influenced by adding cxb to hAFCs; both the DRG cell line and primary bovine DRG nociceptors showed a lower sensitivity to bradykinin stimulation.ConclusionCxb can inhibit PGE-2 production in hAFCs in an IL-1β-induced pro-inflammatory in vitro environment. The cxb applied to the hAFCs also reduces the sensitization of DRG nociceptors that are stimulated by the hAFCs CM.

The Personality Predictors of Art Preferences in Children

Art has been considered as a means of enriching oneself with values, transferring socio-cultural experience and traditions. Children articulate their wonder and excitement while engaging with an artwork and it is evident through their spontaneous responses. Aesthetic experience is regarded as fundamental to human existence. Experiencing and enjoying artworks help children to encounter diversities and prepare them for a facing challenge in life. The objective of the present quantitative study is to explore the relation between personality traits and art appreciation of children. The sample constituted sixty children of grade IX from the state of Telangana. The number of boys and girls participated in the study from rural and urban backgrounds are equal and children are in the age group of 13-15 years. Children’s artistic preferences are measured by showing 48 images of artworks of two dimensions: ‘abstractedness’ and ‘complexity’. The ‘abstractedness’ dimension include representational/realistic, semi-representational and highly abstract artworks and the ‘complexity’ dimension encompass low complexity, medium complexity, and high complexity artworks. Each group has four types of paintings: landscape, portrait, still life and animal representations. The results of analysis of the relation between children’s personality traits and their art preferences using Pearson product-moment correlation coefficient (r) reveals that Agreeableness and Conscientiousness is moderately positively correlated with the perceived attractiveness of abstract paintings which is statistically significant. The results do not show any statistically significant correlation between Openness to Experience and perceived attractiveness for abstract artworks. Further, there exists a low negative correlation between Neuroticism and perceived attractiveness for abstract artworks which is statistically significant. There are no significant correlations between children’s personality traits and perceived attractiveness for Realistic, High Complexity and Low Complexity artworks. The research findings are an indication for teachers to better design their classroom activities for enriching the lives of children.

Gone with the sound: critical perspectives on studying imaginability of American cinematic experience in the “sound” era, 1927-1935

This article explores the human-technology relationship during the transition from silent films to sound films in American history and analyzes how this specific technology has influenced film audiences’ experiences. The author revisits the importance of human-centered technology and uses the term “imaginability” to describe the likelihood for both spontaneous bodily responses and technologically cultivated thinking activities inspired by cinematic engagement. The article challenges the discourse of technology as progress while focusing on the audiences’ experience during their cinematic engagement. The author's hypothesis is that sound film provides a linear channel of synchronization and hence produces restrictions on the audience’s imagination by limiting their temporal and spatial sensibilities. This synchronized sound technology offers a streamlined audiovisual reality through matching sounds and images in real-time, leading to a shortened processing of images, sounds, and their interconnections. The transitions between filmmaking and cinematic presenting have also revolutionized theater design, leading to the liminality that describes the state of transitioning under this circumstance led by sound technology, which generates metrics, measuring how fast and how thoroughly technology conquers an outdated society by enforcing innovations. The article focuses on the interplay between the spectatorship that addresses the condition of viewing films and the sound consciousness led by the synchronized sound system as applied to filmmaking. The analysis of imaginability becomes measurable, descriptive, and referential. The author suggests that the preservation and scrutiny of silent films are urgent and necessary to recognize the values of silent film production and the audience’s cinematic experience during the silent film era.

Astrocytic purinergic signalling contributes to the development and maintenance of neuropathic pain via modulation of glutamate release.

Although activation of astrocytes is critical in developing neuropathic pain (NP) following nerve injury, the underlying mechanisms of NP and therapeutic management for NP are still vague. Importantly, the decreases in the levels of astrocytic glutamate transporter-1 (GLT-1) in the spinal dorsal horn result in enhanced excitatory transmission and cause persistent pain. P2Y1 purinergic receptor (P2Y1R) has been shown to enhance many inflammatory processes. The up-regulated expression of astrocytic P2Y1R is crucial to participate in pain transduction under conditions of nerve injury and peripheral inflammation considering that P2Y1R is potentially involved in glutamate release and synaptic transmission. This study indicates that the expression of P2Y1R in the spinal cord was increased accompanied by the activation of A1 phenotype astrocytes in the rat model of spinal nerve ligation (SNL). Astrocyte-specific knockdown of P2Y1R alleviated SNL-induced nociceptive responses and mitigated A1 reactive astrocytes, which subsequently increased GLT-1 expression. Conversely, in naïve rats, P2Y1R over-expression induced a canonical NP-like phenotype and spontaneous hypernociceptive responses and increased the concentration of glutamate in the spinal dorsal horn. Besides, our in vitro data showed that the proinflammatory cytokine tumour necrosis factor-alpha contributes to A1/A2 astrocyte reactivity and Ca2+-dependent release of glutamate. Conclusively, our results provide novel insights that as a significant regulator of astrocytic A1/A2 polarization and neuroinflammation, P2Y1R may represent a potential target for the treatment of SNL-induced NP.

Caregiver Implemented Dialogic Reading: Impact on Children with and without Autism in Brazil

There is evidence about the effectiveness of using caregiver-implemented, shared book reading protocols for mitigating language and literacy deficits in children with autism spectrum disorders (ASD). In Brazil, studies of this nature are scarce but essential for identifying successful early intervention practices for a population that has significantly grown in the last three decades. The aim of this investigation was to evaluate the effects of Reading to Engage Children with Autism in Language and Learning (RECALL), an intervention protocol that employs dialogic reading principles on the language and literacy skills of a 3-year-old child with ASD and a 4-year-old typically developing peer. Using a single-subject A-B research design, the effects of RECALL, employed by mothers, on their children's responses were evaluated. The results revealed that: (a) average effect size for strategy use following caregiver training was large for the mother of the child with ASD (Tau-U = 0,7) and moderate for the mother of the typically developing child (Tau-U = 0,5) ; (b) both children became more responsive, significantly increasing the frequency of communication interactions following caregiver intervention (Tau-U = 1); (d) the child with typical development showed an increase in the frequency of correct and spontaneous responses, and (c) the intervention was positively evaluated by the caregivers. Adaptations of the protocol, as well as its limitations, are discussed.

Caregiver Implemented Dialogic Reading: impact on children with and without autism in Brazil

There is evidence about the effectiveness of using caregiver-implemented, shared book reading protocols for mitigating language and literacy deficits in children with autism spectrum disorders (ASD). In Brazil, studies of this nature are scarce but essential for identifying successful early intervention practices for a population that has significantly grown in the last three decades. The aim of this investigation was to evaluate the effects of Reading to Engage Children with Autism in Language and Learning (RECALL), an intervention protocol that employs dialogic reading principles on the language and literacy skills of a 3-year-old child with ASD and a 4-year-old typically developing peer. Using a single-subject A-B research design, the effects of RECALL, employed by mothers, on their children's responses were evaluated. The results revealed that: (a) average effect size for strategy use following caregiver training was large for the mother of the child with ASD (Tau-U = 0,7) and moderate for the mother of the typically developing child (Tau-U = 0,5) ; (b) both children became more responsive, significantly increasing the frequency of communication interactions following caregiver intervention (Tau-U = 1); (d) the child with typical development showed an increase in the frequency of correct and spontaneous responses, and (c) the intervention was positively evaluated by the caregivers. Adaptations of the protocol, as well as its limitations, are discussed.

Sustained overexpression of spliced X-box-binding protein-1 in neurons leads to spontaneous seizures and sudden death in mice

The underlying etiologies of seizures are highly heterogeneous and remain incompletely understood. While studying the unfolded protein response (UPR) pathways in the brain, we unexpectedly discovered that transgenic mice (XBP1s-TG) expressing spliced X-box–binding protein-1 (Xbp1s), a key effector of UPR signaling, in forebrain excitatory neurons, rapidly develop neurologic deficits, most notably recurrent spontaneous seizures. This seizure phenotype begins around 8 days after Xbp1s transgene expression is induced in XBP1s-TG mice, and by approximately 14 days post induction, the seizures evolve into status epilepticus with nearly continuous seizure activity followed by sudden death. Animal death is likely due to severe seizures because the anticonvulsant valproic acid could significantly prolong the lives of XBP1s-TG mice. Mechanistically, our gene profiling analysis indicates that compared to control mice, XBP1s-TG mice exhibit 591 differentially regulated genes (mostly upregulated) in the brain, including several GABAA receptor genes that are notably downregulated. Finally, whole-cell patch clamp analysis reveals a significant reduction in both spontaneous and tonic GABAergic inhibitory responses in Xbp1s-expressing neurons. Taken together, our findings unravel a link between XBP1s signaling and seizure occurrence.

Pre-existing TGF-β-specific T-cell immunity in patients with pancreatic cancer predicts survival after checkpoint inhibitors combined with radiotherapy

BackgroundCirculating transforming growth factor-β (TGF-β)-specific T cells that recognize TGF-β-expressing immune regulatory cells have been described in patients with cancer. TGF-β-derived peptide vaccination modulates the tumor microenvironment and has shown...

An arginase1- and PD-L1-derived peptide-based vaccine for myeloproliferative neoplasms: A first-in-man clinical trial.

Arginase-1 (ARG1) and Programed death ligand-1 (PD-L1) play a vital role in immunosuppression in myeloproliferative neoplasms (MPNs) and directly inhibit T-cell activation and proliferation. We previously identified spontaneous T-cell responses towards PD-L1 and ARG1 derived peptide epitopes in patients with MPNs. In the present First-in-Man study we tested dual vaccinations of ARG1- derived and PD-L1-derived peptides, combined with Montanide ISA-51 as adjuvant, in patients with Janus Kinase 2 (JAK2) V617F-mutated MPN. Safety and efficacy of vaccination with ARG1- derived and PD-L1-derived peptides with montanide as an adjuvant was tested in 9 patients with MPN The primary end point was safety and toxicity evaluation. The secondary end point was assessment of the immune response to the vaccination epitope (www.clinicaltrials.gov identifier NCT04051307). The study included 9 patients with JAK2-mutant MPN of which 8 received all 24 planned vaccines within a 9-month treatment period. Patients reported only grade 1 and 2 vaccine related adverse events. No alterations in peripheral blood counts were identified, and serial measurements of the JAK2V617F allelic burden showed that none of the patients achieved a molecular response during the treatment period. The vaccines induced strong immune responses against both ARG1 and PD-L1- derived epitopes in the peripheral blood of all patients, and vaccine-specific skin-infiltrating lymphocytes from 5/6 patients could be expanded in vitro after a delayed-type hypersensitivity test. In two patients we also detected both ARG1- and PD-L1-specific T cells in bone marrow samples at the end of trial. Intracellular cytokine staining revealed IFNγ and TNFγ producing CD4+- and CD8+- T cells specific against both vaccine epitopes. Throughout the study, the peripheral CD8/CD4 ratio increased significantly, and the CD8+ TEMRA subpopulation was enlarged. We also identified a significant decrease in PD-L1 mRNA expression in CD14+ myeloid cells in the peripheral blood in all treated patients and a decrease in ARG1 mRNA expression in bone marrow of 6 out of 7 evaluated patients. Overall, the ARG1- and PD-L1-derived vaccines were safe and tolerable and induced strong T-cell responses in all patients. These results warrant further studies of the vaccine in other settings or in combination with additional immune-activating treatments.

Neuroligin-2 controls the establishment of fast GABAergic transmission in adult-born granule cells.

GABAergic inhibition is critical for the precision of neuronal spiking and the homeostatic regulation of network activity in the brain. Adult neurogenesis challenges network homeostasis because new granule cells (GCs) integrate continuously in the functional dentate gyrus. While developing, adult-born GCs undergo a transient state of enhanced excitability due to the delayed maturation of perisomatic GABAergic inhibition by parvalbumin interneurons (PV-INs). The mechanisms underlying this delayed synaptic maturation remain unknown. We examined the morphology and function of synapses formed by PV-INs onto new GCs over a 2-month interval in young adult mice, and investigated the influence of the synaptic adhesion molecule neuroligin-2 (NL2). Perisomatic appositions of PV-IN terminals onto new GCs were conspicuous at 2 weeks and continued to grow in size to reach a plateau over the fourth week. Postsynaptic knockdown of NL2 by expression of a short-hairpin RNA (shNL2) in new GCs resulted in smaller size of synaptic contacts, reduced area of perisomatic appositions of the vesicular GABA transporter VGAT, and the number of presynaptic active sites. GCs expressing shNL2 displayed spontaneous GABAergic responses with decreased frequency and amplitude, as well as slower kinetics compared to control GCs. In addition, postsynaptic responses evoked by optogenetic stimulation of PV-INs exhibited slow kinetics, increased paired-pulse ratio and coefficient of variation in GCs with NL2 knockdown, suggesting a reduction in the number of active synapses as well as in the probability of neurotransmitter release (Pr ). Our results demonstrate that synapses formed by PV-INs on adult-born GCs continue to develop beyond the point of anatomical growth, and require NL2 for the structural and functional maturation that accompanies the conversion into fast GABAergic transmission.

Improving Social Interactions for Young Children on the Autism Spectrum Through Parent-Mediated LEGO Play Activities

Given the impact of the global COVID-19 pandemic on children’s schooling and learning, evidence for interventions implemented by parents is urgently needed. This study aimed to evaluate the effects of a parent-mediated LEGO intervention on the social interactions of four children on the autism spectrum in China. The intervention involved four sets of two parents engaging with their child in collaborative LEGO construction, with each person assigned a role as Engineer, Supplier, or Builder. Using a multiple probe design across the four family triads, the intervention effectively increased spontaneous social initiations and responses in all four children, and these behaviors were maintained for 6 weeks following the intervention. The effect on inappropriate interactions was not detected; parental prompts increased in initial intervention sessions but decreased over time. Parents implemented the intervention at home with high fidelity. Efficacy of the intervention and potential benefits of virtual parent training are discussed.

The cognitive neural mechanism of response inhibition and error processing to fearful expressions in adolescents with high reactive aggression.

Reactive aggression in adolescents is characterized by high levels of impulsivity. This is associated with deficits in response inhibition and error processing and spontaneous emotion-driven responses to a perceived threat. However, the characteristics and cognitive neural mechanisms of response inhibition and error processing to indirect threat in adolescents with high levels of reactive aggression are unclear. This study explored the characteristics and cognitive neural mechanisms of response inhibition and error processing to fearful expressions in adolescents with high levels of reactive aggression using an emotional Go/No-Go paradigm combined with ERP recordings. Adolescents with high levels of reactive aggression (n = 31) and a control group (n = 30) took part in this study. Results showed that when presented with fearful expressions, adolescents with high levels of reactive aggression showed a smaller No-Go P3 effect and smaller ERN amplitudes following commission errors on the No-Go task than the control group. Results suggested that when presented with fearful expressions, adolescents with high levels of reactive aggression have impaired response inhibition in the later stage of actual inhibitory control of the motor system and impaired error processing in the early stage of fast and automatic initial error detection.