In-vivo Exposure Research Articles

Diagnosis and Management of Knee Arthrofibrosis and Associated Pain-Related Fear Using Multidimensional Clinical Reasoning and Exposure In Vivo Concept: A Case Report

BACKGROUND: A plethora of treatment approaches are used to manage persistent pain, disability, and fear associated with restricted knee range of motion following knee surgery. We used the pain and disability driver model and exposure in vivo concepts to manage pain-related fear and disability. CASE PRESENTATION: A 68-year-old male who underwent arthroscopic partial meniscectomy presented to physical therapy with a medical diagnosis of Complex Regional Pain Syndrome Type I and knee range-of-motion restriction. Despite extensive medical care and physical therapy for 3 years (202 visits), he continued to experience debilitating knee pain and demonstrated a high level of fear. To ambulate, he wore a knee brace and used a trekking pole. OUTCOME AND FOLLOW UP: Interventions to address the knee stiffness included combined tibiofemoral joint mobilization and high-velocity low-amplitude thrust manipulation targeting tibial internal rotation and low-load long-duration sustained capsular stretching using total end range time principle for extension. His fear was managed using techniques inspired by in vivo exposure concepts. Total knee range of motion improved 110 degrees and the patient was able to ambulate for 30 minutes (0.5 mile) without an assistive device with <2/10 pain intensity. DISCUSSION: Outside-the-box clinical reasoning suggested that poor outcomes with previous interventions were likely due to hypervigilance and pain-related fear. This case delineates the value of nonreductionist clinical reasoning in diagnosis and management of musculoskeletal conditions. It also outlines how the exposure in vivo approach helped the patient overcome his long-established avoidance behaviors. JOSPT Cases 2024;4(3):121-132. Epub 2 July 2024. doi:10.2519/josptcases.2024.0034

In Vivo Exposure Pathways of Ambient Magnetite Nanoparticles Revealed by Machine Learning-Aided Single-Particle Mass Spectrometry.

Nanosized ultrafine particles (UFPs) from natural and anthropogenic sources are widespread and pose serious health risks when inhaled by humans. However, tracing the inhaled UFPs in vivo is extremely difficult, and the distribution, translocation, and metabolism of UFPs remain unclear. Here, we report a label-free, machine learning-aided single-particle inductively coupled plasma mass spectrometry (spICP-MS) approach for tracing the exposure pathways of airborne magnetite nanoparticles (MNPs), including external emission sources, and distribution and translocation in vivo using a mouse model. Our results provide quantitative analysis of different metabolic pathways in mice exposed to MNPs, revealing that the spleen serves as the primary site for MNP metabolism (84.4%), followed by the liver (11.4%). The translocation of inhaled UFPs across different organs alters their particle size. This work provides novel insights into the in vivo fate of UFPs as well as a versatile and powerful platform for nanotoxicology and risk assessment.

NAMPT-targeting PROTAC and nicotinic acid co-administration elicit safe and robust anti-tumor efficacy in NAPRT-deficient pan-cancers

Nicotinamide phosphoribosyltransferase (NAMPT) catalyzes the biosynthesis of nicotinamide adenine dinucleotide (NAD+), making it a potential target for cancer therapy. Two challenges hinder its translation in the clinic: targeting the extracellular form of NAMPT (eNAMPT) remains insufficient, and side effects are observed in normal tissues. We previously utilized proteolysis-targeting chimera (PROTAC) to develop two compounds capable of simultaneously degrading iNAMPT and eNAMPT. Unfortunately, the pharmacokinetic properties were inadequate, and toxicities similar to those associated with traditional inhibitors arose. We have developed a next-generation PROTAC molecule 632005 to address these challenges, demonstrating exceptional target selectivity and bioavailability, improved invivo exposure, extended half-life, and reduced clearance rate. When combined with nicotinic acid, 632005 exhibits safety and robust efficacy in treating NAPRT-deficient pan-cancers, including xenograft models with hematologic malignancy and prostate cancer and patient-derived xenograft (PDX) models with liver cancer. Our findings provide clinical references for patient selection and treatment strategies involving NAMPT-targeting PROTACs.

A Systematic Review on The Efficacy of VRET For Agoraphobia

This systematic review paper aims to evaluate the efficacy of Virtual Reality Exposure Therapy (VRET) for Agoraphobia, a severe psychological disorder characterized by avoidance behavior and sometimes panic attacks. The paper presents a qualitative assessment of 15 studies published after 2000, using Scopus databases. The experiments were evaluated for eligibility based on their abstracts and full content, and only English-language published articles were included. The review shows that VRET is a promising therapy for Agoraphobia, with similar efficacy to In Vivo Exposure (IVE), but with certain advantages such as better control over exposure content and higher patient satisfaction. However, implementing VRET may be difficult for some patients due to cybersickness and higher costs. The paper concludes with a diagram of the 15 experiments evaluated and emphasizes the importance of further research on the efficacy of VR therapy for anxiety disorders, including Agoraphobia.

SPHK1/S1P/S1PR pathway promotes the progression of peritoneal fibrosis by mesothelial-mesenchymal transition.

Long-term exposure to non-physiologically compatible dialysate inevitably leads to peritoneal fibrosis (PF) in patients undergoing peritoneal dialysis (PD), and there is no effective prevention or treatment for PF. Sphingosine-1-phosphate (S1P) is a bioactive sphingolipid produced after catalysis by sphingosine kinase (SPHK) 1/2 and activates signals through the S1P receptor (S1PR) via autocrine or paracrine. However, the role of SPHK1/S1P/S1PR signaling has never been elucidated in PF. In our research, we investigated S1P levels in peritoneal effluents and demonstrated the role of SPHK1/S1P/S1PR pathway in peritoneal fibrosis. It was found that S1P levels in peritoneal effluents were positively correlated with D/P Cr (r = 0.724, p < .001) and negatively correlated with 4 h ultrafiltration volume (r = -0.457, p < .001). S1PR1 and S1PR3 on peritoneal cells were increased after high glucose exposure invivo and invitro. Fingolimod was applied to suppress S1P/S1PR pathway. Fingolimod restored mouse peritoneal function by reducing interstitial hyperplasia, maintaining ultrafiltration volume, reducing peritoneal transport solute rate, and mitigating the protein expression changes of fibronectin, vimentin, α-SMA, and E-cadherin induced by PD and S1P. Fingolimod preserved the morphology of the human peritoneal mesothelial cells, MeT-5A, and moderated the mesothelial-mesenchymal transition (MMT) process. We further delineated that SPHK1 was elevated in peritoneal cells after high glucose exposure and suppression of SPHK1 in MeT-5A cells reduced S1P release. Overexpression of SPHK1 in MeT-5A cells increased S1P levels in the supernatant and fostered the MMT process. PF-543 treatment, targeting SPHK1, alleviated deterioration of mouse peritoneal function. In conclusion, S1P levels in peritoneal effluent were correlated with the deterioration of peritoneal function. SPHK1/S1P/S1PR pathway played an important role in PF.

The Efficacy and Therapeutic Alliance of Augmented Reality Exposure Therapy in Treating Adults With Phobic Disorders: Systematic Review.

Phobic disorders are characterized by excessive fear of a stimulus that can affect the quality of a patient's life. The lifetime prevalence in adults is 7.7% to 12.5%. The current literature provides evidence-based inferences about the effectiveness of in-vivo exposure therapy (IVET) in treating phobia. However, this method can put the therapist and the client in danger, with high drop out and refusal rates. A newer approach for exposure therapy using augmented reality technology is under assessment. This systematic review investigated the novel technology's efficacy, cost-efficacy, and therapeutic alliance in treating adults with phobia. An extensive search was conducted using 4 major databases (MEDLINE, PsycINFO, Embase, and Scopus) using a comprehensive list of synonyms for augmented reality exposure therapy (ARET) and phobic disorders. The search targeted any randomized control trial testing ARET in adults with phobic disorders up to August 8, 2022. A total of 6 studies were included, with 208 participants providing results. Studies investigating the efficacy of ARET compared to no intervention showed significant results (P<.05) in the ARET group improvement. Head-to-head comparative studies comparing ARET to IVET showed no significant difference (P>.05) in the effectiveness and therapeutic alliance between both therapies. Further, the results demonstrated that the ARET group had a better long-term effect than IVET, with the ability to put the patients in more situations to face the feared object. The current data suggest clinically significant efficacy and a promising therapeutic alliance of ARET. However, no data are available investigating the cost-effectiveness of ARET. Further research is warranted to ascertain ARET's cost-effectiveness and examine its efficacy in other populations and anxiety conditions.

Alloantigen Infusion Activates the Transcriptome of Type 2 Conventional Dendritic Cells.

Recent studies have revealed novel molecular mechanisms by which innate monocytic cells acutely recognize and respond to alloantigen with significance to allograft rejection and tolerance. What remains unclear is the single-cell heterogeneity of the innate alloresponse, particularly the contribution of dendritic cell (DC) subsets. To investigate the response of these cells to exposure of alloantigen, C57BL/6J mice were administered live allogenic BALB/cJ splenic murine cells versus isogenic cells. In parallel, we infused apoptotic allogenic and isogenic cells, which have been reported to modulate immunity. Forty-eight hours after injection, recipient spleens were harvested, enriched for DCs, and subjected to single-cell mRNA sequencing. Injection of live cells induced a greater transcriptional change across DC subsets compared with apoptotic cells. In the setting of live cell infusion, type 2 conventional DCs (cDC2s) were most transcriptionally responsive with a Ccr2+ cDC2 subcluster uniquely responding to the presence of alloantigen compared with the isogenic control. In vitro experimentation confirmed unique activation of CCR2+ cDC2s following alloantigen exposure. Candidate receptors of allorecognition in other innate populations were interrogated and A type paired Ig-like receptors were found to be increased in the cDC2 population following alloexposure. These results illuminate previously unclear distinctions between therapeutic infusions of live versus apoptotic allogenic cells and suggest a role for cDC2s in innate allorecognition. More critically, these studies allow for future interrogation of the transcriptional response of immune cells in the setting of alloantigen exposure invivo, encouraging assessment of novel pathways and previously unexamined receptors in this setting.

Overcoming Social Anxiety: How Virtual Reality and Game-Based Elements are Revolutionizing Patient Therapy.

In this paper, we reflect upon the design process and the game-elements for developing a tool for patients. These reflections are based upon a case in where we developed six different Virtual Reality (VR) scenarios as a combined tool for treatment of social anxiety disorder (SAD) for usage within the Danish healthcare system. The core theory for this development is based upon behavioral therapy as a standardized treatment method and the exposure that occur within this. This research is a part of a larger funded project titled VR8, which also contains integration of biofeedback and the application of the practice in where the tool should be applied. The main goal of this paper is to explain the development of the six scenarios, to provide a deeper understanding for the patients on how their anxiety triggers unfolds when exposed to a variety of different scenarios. And to provide a tool for both patients and practitioners to use as an addition to existing treatment methods. The reason for this development is due to (among others) the cost-reduction, time-reduction, heighten of effectivity and user experience in using VR as a tool for treatment compared to regular in-vivo exposure therapy in where the practitioner guides the patient in a physical environment. In this environment a lot of factors can’t be controlled and often results in the patient giving up and abandoning the treatment. With VR it is a controlled and closed environment making it more suitable for the patient to interact and level out their anxiety. The combination of iterative design thinking, filmmaking methods, game-based elements and agile software development are used as the core methods to develop the six scenarios. The six scenarios are developed with the aid of specific game-based elements, such as: mapping, level-design, audio control and remote action triggers.

Clinician perspectives on technology-enhanced in vivo exposures during prolonged exposure therapy for PTSD.

To investigate clinicians'perceptions regarding the use of mobile technology tools during prolonged exposure (PE) therapy to allow for monitoring and enhancing in-vivo exposures (IVEs). Clinicians with training in PE therapy (N = 32; average of 9 years of practice) completed surveys asking about their perspectives on the utility of virtually attending IVEs with patients while simultaneously having access to real-time subjective and physiological data (i.e., heart rate, galvanic skin conductance) to guide exposure exercises and assure optimal stimulus engagement. Findings showed clinicians to have a favorable view of applying technology devices and systems to enhance IVEs of PE therapy. Most clinicians (93.8%) believed that real-time monitoring of IVEs-particularly monitoring patients'subjective distress and completion of and duration of time in the IVE-would be useful and significantly enhance PE therapy. The positive perceptions toward integrating technology into IVEs in this study have important implications for the development and implementation of technology-enhanced PE therapy. A mobile technology system that incorporates real-time indicators of engagement (i.e., both subjective and physiological) during IVEs and allows clinicians to review recordings of, or virtually accompany, patients during IVEs has the potential to innovate and transform PE and other exposure-based treatments. Clinicians also believed that technology-enhanced IVEs may help reduce early termination from PE.

In-vivo exposure of a plant model organism for the assessment of the ability of PM samples to induce oxidative stress

This study aims to propose an innovative, simple, rapid, and cost-effective method to study oxidative stress induced by PM through in-vivo exposure of the plant model organism Arabidopsis thaliana. A. thaliana seedlings were exposed to urban dust certified for its elemental content and to PM2.5 samples collected in an urban-industrial area of Northern Italy. An innovative technique for the detachment and suspension in water of the whole intact dust from membrane filters was applied to expose the model organism to both the soluble and insoluble fractions of PM2.5, which were analyzed for 34 elements by ICP-MS. Oxidative stress induced by PM on A. thaliana was assessed by light microscopic localization and UV–Vis spectrophotometric determination of superoxide anion (O2−) content on the exposed seedlings by using the nitro blue tetrazole (NBT) assay. The results showed a good efficiency and sensitivity of the method for PM mass concentrations >20 μg m−3 and an increase in O2− content in all exposed seedlings, which mainly depends on the concentration, chemical composition, and sources of the PM administered to the model organism. Particles released by biomass burning appeared to contribute more to the overall toxicity of PM. This method was found to be cost-effective and easy to apply to PM collected on membrane filters in intensive monitoring campaigns in order to obtain valuable information on the ability of PM to generate oxidative stress in living organisms.

Increased In Vivo Exposure of N-(4-Hydroxyphenyl) Retinamide (4-HPR) to Achieve Plasma Concentrations Effective against Dengue Virus.

N-(4-hydroxyphenyl) retinamide (4-HPR, or fenretinide) has promising in vitro and in vivo antiviral activity against a range of flaviviruses and an established safety record, but there are challenges to its clinical use. This study evaluated the in vivo exposure profile of a 4-HPR dosage regime previously shown to be effective in a mouse model of severe dengue virus (DENV) infection, comparing it to an existing formulation for human clinical use for other indications and developed/characterised self-emulsifying lipid-based formulations of 4-HPR to enhance 4-HPR in vivo exposure. Pharmacokinetic (PK) analysis comprising single-dose oral and IV plasma concentration-time profiles was performed in mice; equilibrium solubility testing of 4-HPR in a range of lipids, surfactants and cosolvents was used to inform formulation approaches, with lead formulation candidates digested in vitro to analyse solubilisation/precipitation prior to in vivo testing. PK analysis suggested that effective plasma concentrations could be achieved with the clinical formulation, while novel lipid-based formulations achieved > 3-fold improvement. Additionally, 4-HPR exposure was found to be limited by both solubility and first-pass intestinal elimination but could be improved through inhibition of cytochrome P450 (CYP) metabolism. Simulated exposure profiles suggest that a b.i.d dosage regime is likely to maintain 4-HPR above the minimum effective plasma concentration for anti-DENV activity using the clinical formulation, with new formulations/CYP inhibition viable options to increase exposure in the future.

Scoping review of the hardware and software features of virtual reality exposure therapy for social anxiety disorder, agoraphobia, and specific phobia

Introduction: The use of virtual reality technology to deliver exposure therapy in the treatment of phobic anxiety (i.e., social anxiety disorder, agoraphobia, and specific phobia) has been proposed to be advantageous compared with in-vivo exposure therapy. These supposed advantages depend on the features of the virtual reality technology and how it is used therapeutically. Therefore, the aim of this study was to provide a comprehensive overview of the features of the hardware and software used in studies examining virtual reality exposure therapy studies for phobic anxiety disorders.Methods: 70 studies using virtual reality exposure therapy to treat social anxiety disorder, agoraphobia and/or specific phobia, were systematically reviewed for 46 data points relating to these features.Results: We found that studies generally did not utilize contemporary virtual reality technology and that hardware and software features were inconsistently delineated.Discussion: The implications of these findings are that the use of modern virtual reality technology represents a relevant frontier in anxiety treatment and that a framework for reporting technical features of virtual reality exposure interventions would benefit the field.

In vitro, in-vivo, and in-silico investigation of physicochemical interactions between pioglitazone and rifampicin

There is a possibility of in-situ physicochemical interactions between concomitantly administered drugs. This study aimed to investigate such physicochemical interactions between pioglitazone and rifampicin. Pioglitazone exhibited significantly higher dissolution in the presence of rifampicin, while the dissolution of rifampicin remained unaffected. The solid-state characterization of precipitates recovered after pH-shift dissolution experiments revealed the conversion of pioglitazone into an amorphous form in the presence of rifampicin. The Density Function Theory (DFT) calculations showed the intermolecular hydrogen bonding between rifampicin and pioglitazone. In-situ conversion of pioglitazone in amorphous form and subsequent supersaturation of GIT milieu translated into significantly higher in-vivo exposure of pioglitazone and its metabolites (M-III and M-IV) in Wistar rats. Therefore, it is advisable to consider the possibility of physicochemical interactions between concomitantly administered drugs. Our findings may be beneficial in tailoring the dose of concomitantly administered drugs, particularly for chronic conditions that entail polypharmacy.

Four Days Exposure And Reprocessing Therapy For PTSD

IntroductionPost-traumatic stress disorder (PTSD) is a debilitating disorder affecting approximately 6% of the population. Current treatments have been shown to efficaciously reduce symptom burden between 30%-50%. However, due to the high intensity of treatment over a long period of time. drop-out rates are as high as 50%.ObjectivesAssess the effect of one-time ketamine infusion in subanasthetic dosage on PTSD psychotherapyAssess feasibility and effect of massed, four days, esposure focused psychotherapy for PTSdMethodsHere, we tested the efficacy of a four-day exposure and processing-focused psychotherapy at reducing PTSD severity. Twenty-seven participants with chronic PTSD were randomized to two groups, one receiving a one-time infusion of ketamine in a subanesthetic dose (0.5mg/kg for 40 minutes), the other receiving midazolam. Both groups underwent four 90-120 minutes of daily psychotherapy sessions a day after infusion, followed by in-vivo exposure practice. The severity of PTSD was assessed with the PCL-5 before and at the end of treatment, and at 30 and 90 days follow-up. Brain reactivation to the trauma reminders was measured using fMRIResultsPTSD severity in both treatment groups decreased by 13, 16, and 15 points on the PCL-5 at the end of treatment, 30 days follow-up, and 90 days respectively, surpassing the minimum clinical difference of 7.9 points. There was no significant difference in symptom reductions between the treatment groups. However, brain reactivation to trauma stories differed between the groups, with the ketamine group showing a decline in the amygdala and hippocampus reactivation compared to the midazolam group, at the end of treatment.Image:ConclusionsOur results imply comparable efficacy of this short-term intervention to standard trauma-focused psychotherapies, emphasizing its clinical usefulness as a short and effective intervention.Disclosure of InterestNone Declared

Client and staff perspectives regarding effective work injury rehabilitation.

Objective Early, targeted treatment is critical to recovery and overall health following a work-related illness or injury. Limited research has explored the important dimensions of work-specific injury rehabilitation from both client and staff perspectives. Methods A total of 17 participants (13 clients with work-related injuries, 3 physiotherapists, 1 project manager) involved in a unique program providing allied health treatment in combination with return-to-work services, were interviewed. Data were analysed using reflexive thematic analysis. Results Four themes were generated: (1) a biopsychosocial approach to rehabilitation; (2) a self-paced environment where client outcomes are optimised through transparent and collaborative team processes; (3) comprehensive care aids client recovery and return to work; and (4) a desire for service expansion is hampered by systemic barriers. Conclusions Injured workers and staff provided very positive feedback about the biopsychosocial supports needed for successful return to work, particularly the use of in-house work-specific simulation tasks as gradual in-vivo exposure and collaboration with scheme stakeholders. How to best provide this holistic care within current legislative requirements remains a challenge.

Changes in avoidance and distress related to trauma reminders in PTSD psychotherapy

Background and objectivesResearch has examined reductions in patient distress recounting trauma narratives in Prolonged Exposure (PE) for posttraumatic stress disorder (PTSD). It remains unclear whether changes in distress and avoidance related to environmental trauma reminders matter in PE and other PTSD treatments, including non-exposure Interpersonal Psychotherapy (IPT). MethodsData came from adults with chronic PTSD (N = 92) who completed a treatment trial comparing PE, IPT, and Relaxation Therapy (RT). We employed the Self-Initiated In-Vivo Exposure Scale (SIIVES), which measures patient distress from and avoidance of situational trauma reminders, to calculate reliable change in distress and avoidance. PTSD symptoms, depression, quality of life, and functioning assessments were collected before and after 14 weeks of treatment. ResultsOverall, 48.1% of patients experienced reliable change in avoidance, while 51.9% showed reliable change in distress. Rates of reliable change did not differ by treatment group, although PE appeared to achieve reliable change earlier. Only one baseline characteristic predicted reliable change: patients with comorbid depression were less likely to reliably change in avoidance. At post-treatment, patients achieving reliable change had lower PTSD severity and depression and increased quality of life and social functioning. Statistical modeling revealed that changes in distress and avoidance related to subsequent reduction in PTSD symptoms in all three treatment groups, though this relationship appeared strongest in PE. LimitationsThe sample was relatively small. ConclusionsChange in avoidance and distress associated with situational trauma reminders was associated with a range of clinical outcomes and may represent important factors in multiple PTSD psychotherapies.

Technology-enhanced in vivo exposures in Prolonged Exposure for PTSD: A pilot randomized controlled trial

In vivo exposures (IVEs) are a key component of exposure-based treatments, during which patients approach fear-provoking, yet safe, situations in “real life.” This pilot study assessed the use of a wearable technology (Bio Ware) during IVEs to enhance Prolonged Exposure (PE) therapy for PTSD. Bio Ware provides a clinician dashboard with real-time physiological and subjective data for clinicians to use for virtually guided IVEs. Participants (N = 40) were randomized to a Guided group that received standard PE and virtual, clinician-guided IVEs with the Bio Ware device, or a Non-Guided group that received standard PE and used the Bio Ware device on their own for IVEs. Multilevel linear models with bootstrapping were completed on the intent-to-treat (ITT; N = 39) and per-protocol samples (PP; n = 23), defined as completing at least eight sessions of PE and using the Bio Ware system during ≥ 1 IVEs. In the PP sample, there were significant effects of treatment condition (b = −14.55, SE = 1.47, 95% CI [-17.58, −11.78], p < .001) and time (b = −1.98, SE = 0.25, 95% CI [-2.47, −1.48], p < .001). While both groups showed reductions in PTSD symptoms, the Guided group evidenced significantly greater reductions than the Non-Guided group. These findings demonstrate the feasibility and safety of leveraging Bio Ware for virtual, clinician-guided IVEs during PE therapy for PTSD and suggest that virtual, clinician-guided exposures may enhance treatment outcomes. Clinical trial registrationNCT04471207.

PMON12 Effects of Endocrine Disruptors on Autophagy Markers Gene Expression in Immortalized GnRH Neurons and Hypothalami of Adult Male Mice

Abstract Benzophenones (BP) and Bisphenol A (BPA) are endocrine disrupting chemicals (EDC). Previously, we showed that the in-vitro exposure to EDC inhibited Kiss-induced gene expression in GT1-7 and GN11 cells (donated by Pamela Mellon, UCSD), and altered expression of inflammatory markers in GT1-7 cells and hypothalami of adult male mice.In the present study, we evaluated the effects of the in-vivo exposure to BP2 and BP3 (250 µg/kg/day, orally, for 5 days) on the expression of autophagy markers in hypothalamic nuclei in C57Bl/6 adult male mice, and the in-vitro exposure to BPA, BP2 or BP3 in immortalized GnRH neurons. Animals were sacrificed, brains rapidly dissected and conserved -80°C until processed. Micropunches containing Anteroventral Periventricular (AVPV) or Arcuate (ARC) nucleus were obtained. Effects of the in-vitro exposure for 12 or 24 hours to BPA, BP2 or BP3 (1×10-9M, Sigma), or DMSO as control, were evaluated in GT1-7 and GN11 cell lines, mature and immature GnRH neurons. ARN was obtained using Tri-Reagent (Molecular Research Center, Inc), reverse-transcribed and gene expression analyzed by qPCR, using Ppib as control gene. Results were presented as Media±SE and analyzed by ANOVA (Statistica, StatSoft Inc, USA).In-vivo, five-day exposure to BP2 or BP3 did not alter Lamp2 or p62 gene expression in the AVPV or ARC in adult male mice (ANOVA: ns). In GT1-7 cells, Ulk1 was not significantly altered by any of the stimuli (ANOVA: ns; n=5), whereas in GN11 cells, BP3 significantly increased Ulk1 gene expression after 12 and 24 h exposure [DMSO-24h: 0.93±0.21, DMSO-12h: 0.73± 0.19, BPA-24h: 1.78±0.31, BPA-12h: 1.47±0.33, BP2-24h: 1.51±0.49, BP2-12h: 1.06±0.23, BP3-24h: 2.41±0.89, BP3-12h: 2.45±0.82, Repeated Measures ANOVA: Main effect "TREATMENT" p&amp;lt;0.05, BP3 different from DMSO, p&amp;lt;0.05, n=4]. Our results reinforce the notion that the exposure to EDC have different effects depending on the experimental model and time of exposure, with developing cells being more susceptible to disruption that mature cells. We need more experiments to fully understand if exposure to EDC alters other markers of the autophagy process in mouse hypothalamus and GnRH neurons.Funding: CONICET, ANPCYT, International Society for Neurochemistry, Fund. Williams, Fund. R. Barón. Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

From hemolymph to in-vivo: The potential of a 1 mm microlitre flow probe with separate lock chamber for NMR metabolomics in mass limited environmental samples

Metabolomics is one of the leading approaches for understanding the toxic-mode-of-action of environmental contaminants. Nuclear Magnetic Resonance (NMR) spectroscopy has been commonly used in metabolomic studies; however, its main drawback is its relatively low sensitivity, making it challenging to study mass limited but environmentally crucial samples. In this work a 1 mm microlitre probe modified with a separate lock chamber to address this challenge, provided substantial improvements in mass sensitivity relative to conventional 5 mm NMR probes. The 1 mm probe is used to analyze various components of the model organism Daphnia magna, including hemolymph, parthenogenetic eggs, dormant eggs, and neonates. A μL volume flow system is designed for the 1 mm probe to perform an in-vivo exposure of neonates to high salt concentrations. The metabolic investigation of these samples was only achieved due to the minimum sample requirements and high salt tolerance of the probe, demonstrating that the 1 mm microlitre probe modified with a separate lock chamber holds significant potential for future metabolomic studies of mass limited samples.

Intensive Cognitive-Behavioral Therapy for Anxiety Disorders in Adolescents: A Case Study

Anxiety disorders are one of the most common psychiatric conditions in youth and can contribute to impairment in social, academic, and family functioning. Cognitive-behavioral therapy (CBT) has been shown to be efficacious in treating youth anxiety disorders; however, for a multitude of reasons, fewer than 20% of adolescents with anxiety disorders receive services for anxiety-related problems. Intensive treatments, which rely on the same traditional components of CBT but are delivered over a shorter period of time or in a fewer number of sessions, may be particularly helpful for anxiety disorders and can offer a number of advantages over standard CBT. Despite emerging evidence supporting the advantages of the intensive approach, there are few established intensive treatment programs for youth with anxiety disorders. Further, no treatment to date has comprehensively targeted the entire spectrum of comorbid adolescent anxiety disorders in a combined intensive and transdiagnostic format, even though non-intensive (i.e., weekly delivered) CBT has been tested using a transdiagnostic approach. We developed an intensive, six-session intervention based on Angelosante and colleagues’ 2009 The Adolescent Panic Control Treatment with In-Vivo Exposures (Angelosante et al., 2009) and other empirically-supported treatments for youth to target all anxiety disorders in adolescents. We present a case study on an adolescent with multiple comorbid anxiety and related disorders who received intensive CBT treatment as a way to illustrate the clinical benefit and utility of an intensive, transdiagnostic approach. Findings support the acceptability and feasibility of transdiagnostic treatment of youth anxiety.