Specific Induction Research Articles

Phenomenological characteristics of auto-induced cognitive trance and Mahorikatan® trance.

Trance states include various practices characterized by a modulation of consciousness, but with their own specific characteristics and induction techniques. They have been very seldom scientifically studied, and their phenomenological similarities and differences are poorly documented. This paper will focus on two types of Western trances developed after the leaders were trained in traditional shamanic communities: the auto-induced cognitive trance (AICT) and the Mahorikatan® trance (MT). Twenty-five AICT and 26 MT participants who were able to self-induce the trance state completed questionnaires about their trance practice (e.g. context of the first trance episode, frequency of practice, and consequences on personal life) and the phenomenological characteristics (i.e. emotional, physical, and cognitive) of the trance episodes they experienced. These characteristics were compared to explore similarities and differences between the two trance states. AICT and MT are characterized by the expression of different emotions, modification of various perceptions, a feeling of unicity (i.e. being completely oneself), and an expansion of consciousness. AICT participants commonly reported body movements, vocalizations, as well as increased creativity, visions of entities and/or places, and feeling of interaction with the environment. MT participants commonly reported a feeling of body dissolution. Most participants in both groups reported positive effects of their trance practice on their personal life. These results helped characterize AICT and MT, as well as their similarities and differences. Further studies should continue to explore the characteristics of such trance states, as well as their potential clinical applications.

Single-cell landscape of the cellular microenvironment in three different colonic polyp subtypes in children.

The understanding of the heterogeneous cellular microenvironment of colonic polyps in paediatric patients with solitary juvenile polyps (SJPs), polyposis syndrome (PJS) and Peutz-Jeghers syndrome (PJS) remains limited. We conducted single-cell RNA sequencing and multiplexed immunohistochemistry (mIHC) analyses on both normal colonic tissue and different types of colonic polyps obtained from paediatric patients. We identified both shared and disease-specific cell subsets and expression patterns that played important roles in shaping the unique cellular microenvironments observed in each polyp subtype. As such, increased myeloid, endothelial and epithelial cells were the most prominent features of SJP, JPS and PJS polyps, respectively. Noticeably, memory B cells were increased, and a cluster of epithelial-mesenchymal transition (EMT)-like colonocytes existed across all polyp subtypes. Abundant neutrophil infiltration was observed in SJP polyps, while CX3CR1hi CD8+ T cells and regulatory T cells (Tregs) were predominant in SJP and JPS polyps, while GZMAhi natural killer T cells were predominant in PJS polyps. Compared with normal colonic tissues, myeloid cells exhibited specific induction of genes involved in chemotaxis and interferon-related pathways in SJP polyps, whereas fibroblasts in JPS polyps had upregulation of myofiber-associated genes and epithelial cells in PJS polyps exhibited induction of a series of nutrient absorption-related genes. In addition, the TNF-α response was uniformly upregulated in most cell subsets across all polyp subtypes, while endothelial cells and fibroblasts separately showed upregulated cell adhesion and EMT signalling in SJP and JPS polyps. Cell-cell interaction network analysis showed markedly enhanced intercellular communication, such as TNF, VEGF, CXCL and collagen signalling networks, among most cell subsets in polyps, especially SJP and JPS polyps. These findings strengthen our understanding of the heterogeneous cellular microenvironment of polyp subtypes and identify potential therapeutic approaches to reduce the recurrence of polyps in children.

In vitro differentiation of human CD45+ CD34+ induced hematopoietic progenitor cells from iPSCs using a monolayer system

Hematopoietic stem cells (HSCs) represent crucial target cells in the management of hematopoietic and immune system disorders. Unfortunately, the primary source of hematopoietic stem cells is limited. Hematopoietic stem cells derived from induced pluripotent stem cells (iPSCs) hold great promise for applications in cell therapy, disease modeling, and drug screening. To achieve a consistent induction method, one specific induction scheme capable of reliably generating CD34 and CD45 double-positive cells from iPSCs was optimized, employing a comparative analysis and screening of various induction methods. The comprehensive induction procedures are outlined in this document.

Intrinsically disordered regions of the Msn2 transcription factor encode multiple functions using interwoven sequence grammars.

Intrinsically disordered regions (IDRs) are abundant in eukaryotic proteins, but their sequence-function relationship remains poorly understood. IDRs of transcription factors (TFs) can direct promoter selection and recruit coactivators, as shown for the budding yeast TF Msn2. To examine how IDRs encode both these functions, we compared genomic binding specificity, coactivator recruitment, and gene induction amongst a large set of designed Msn2-IDR mutants. We find that both functions depend on multiple regions across the>600AA IDR. Yet, transcription activity was readily disrupted by mutations that showed no effect on the Msn2 binding specificity. Our data attribute this differential sensitivity to the integration of a relaxed, composition-based code directing binding specificity with a more stringent, motif-based code controlling the recruitment of coactivators and transcription activity. Therefore, Msn2 utilizes interwoven sequence grammars for encoding multiple functions, suggesting a new IDR design paradigm of potentially general use.

Transient Autoreactive PF4 and Antiphospholipid Antibodies in COVID-19 Vaccine Recipients.

Vaccine-induced immune thrombotic thrombocytopenia (VITT) is a rare autoimmune condition associated with recombinant adenovirus (rAV)-based COVID-19 vaccines. It is thought to arise from autoantibodies targeting platelet factor 4 (aPF4), triggered by vaccine-induced inflammation and the formation of neo-antigenic complexes between PF4 and the rAV vector. To investigate the specific induction of aPF4 by rAV-based vaccines, we examined sera from rAV vaccine recipients (AZD1222, AD26.COV2.S) and messenger RNA (mRNA) based (mRNA-1273, BNT162b2) COVID-19 vaccine recipients. We compared the antibody fold change (FC) for aPF4 and for antiphospholipid antibodies (aPL) of rAV to mRNA vaccine recipients. We combined two biobanks of Dutch healthcare workers and matched rAV-vaccinated individuals to mRNA-vaccinated controls, based on age, sex and prior history of COVID-19 (AZD1222: 37, Ad26.COV2.S: 35, mRNA-1273: 47, BNT162b2: 26). We found no significant differences in aPF4 FCs after the first (0.99 vs. 1.08, mean difference (MD) = -0.11 (95% CI -0.23 to 0.057)) and second doses of AZD1222 (0.99 vs. 1.10, MD = -0.11 (95% CI -0.31 to 0.10)) and after a single dose of Ad26.COV2.S compared to mRNA-based vaccines (1.01 vs. 0.99, MD = 0.026 (95% CI -0.13 to 0.18)). The mean FCs for the aPL in rAV-based vaccine recipients were similar to those in mRNA-based vaccines. No correlation was observed between post-vaccination aPF4 levels and vaccine type (mean aPF difference -0.070 (95% CI -0.14 to 0.002) mRNA vs. rAV). In summary, our study indicates that rAV and mRNA-based COVID-19 vaccines do not substantially elevate aPF4 levels in healthy individuals.

Transformation of hydroxylated polychlorinated biphenyls by bacterial 2-hydroxybiphenyl 3-monooxygenase

Monohydroxylated PCBs (OH-PCBs) are an (eco)toxicologically significant group of compounds, as they arise from the oxidation of polychlorinated biphenyls (PCBs) and, at the same time, may exert even more severe toxic effects than their parent PCB molecules. Despite having been widely detected in environmental samples, plants, and animals, information on the fate of OH-PCBs in the environment is scarce, including on the enzymatic machinery behind their degradation. To date, only a few bacterial taxa capable of OH-PCB transformation have been reported. In this study, we aimed to obtain a deeper insight into the transformation of OH-PCBs in soil bacteria and isolated a Pseudomonas sp. strain P1B16 based on its ability to use o-phenylphenol (2-PP) which, when exposed to the Delor 103-derived OH-PCB mixture, depleted a wide spectrum of mono-, di, and trichlorinated OH-PCBs. In the P1B16 genome, a region designated as hbp was identified, which bears a set of putative genes involved in the transformation of OH-PCBs, namely hbpA encoding for a putative flavin-dependent 2-hydroxybiphenyl monooxygenase, hbpC (2,3-dihydroxybiphenyl-1,2-dioxygenase), hbpD (2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase), and the transcriptional activator-encoding gene hbpR. The hbpA coding sequence was heterologously expressed, purified, and its substrate specificity was investigated towards the Delor 103-derived OH-PCB mixture, individual OH-PCBs, and multiple (chlorinated) phenolics. Apart from 2-PP and 2-chlorophenol, HbpA was also demonstrated to transform a range of OH-PCBs, including a 3-hydroxy-2,2′,4′,5,5′-pentachlorobiphenyl. Importantly, this is the first direct evidence of HbpA homologs being involved in the degradation of OH-PCBs. Moreover, using a P1B16-based biosensor strain, the specific induction of hbp genes by 2-PP, 3-phenylphenol, 4-phenylphenol, and the OH-PCB mixture was demonstrated. This study provides direct evidence on the specific enzymatic machinery responsible for the transformation of OH-PCBs in bacteria, with many implications in ecotoxicology, environmental restoration, and microbial ecology in habitats burdened with PCB contamination.

Venetoclax Combined with Homoharringtonine,Cytarabine and Aclacinomycin (HAAV) As Induction Therapy in Newly Diagnosed Young Adult Acute Myeloid Leukemia

Background: The remission rate of acute myeloid leukemia (AML) patients using traditional induction chemotherapy is approximately 60-70%. Venetoclax (Ven), a b-cell lymphoma 2 (BCL-2) inhibitor, in combination with chemotherapy or hypomethylating agents, has shown a significant increase in remission rates and improvement in patient survival for elderly or unfit for intensive chemotherapy newly diagnosed AML patients. The HAA regimen (consisting of Homoharringtonine, Cytarabine and Aclacinomycin), a specific traditional induction therapy for acute myeloid leukemia in China. The HAA regimen achieved complete remission rates about 70% as an induction regimen for newly diagnosed AML. Whether the combination of Ven with HAA regimen as an induction treatment can further enhance the complete remission rate in newly diagnosed adult AML patients requires investigation through well-designed clinical trials. Objective: To evaluate the efficacy and safety of HAA combined with Ven as induction regimen (HAAV) in young adult patients with newly diagnosed AML. Design, setting and participants: Single-arm, prospective clinical trial conducted at 8 hospitals in China. Eligible patients (16-60 years old) with newly diagnosed AML (exclude acute promyelocytic leukemia) were enrolled. The trial was registered on ClinicalTrials.gov (NCT05893472) and continues to accrue patients. Induction regimen: The induction regimen consisted of Homoharringtonine (2.5mg/㎡/d，d3-7 ), cytarabine (100mg/㎡/d by continuous intravenous infusion daily on d3-7) and Aclacinomycin (20mg/d, d3-7) combined with venetoclax (100mg d1, 200mg d2, 400mg d3-8). Main outcomes and measures: The primary endpoint was the overall response rate (ORR) after once cycle of HAAV regimen, which included complete remission (CR), complete remission with incomplete count recovery (CRi), morphologic leukemia-free state (MLFS) and partial remission (PR). Secondary endpoints included minimal residual disease (MRD), overall survival (OS), event-free survival (EFS) and adverse events. Results: Untill June 30, 2023, 40 patients were enrolled. The median age is 43 years (range, 18-60), with 60% (24/40) was male. According to the European Leukemia Network prognostic group (2022), 14 (35.0%), 14 (35.0%), and 12 (30.0%) patients were considered to belong to the favorable, intermediate, and adverse groups, respectively. Further details of the patients' characteristics are provided in Table 1. The ORR (CR+PR) was 97.5% (39/40 patients) after one cycle of the induction regimen with the CR rate was 95.0% (38/40). According to the ELN prognostic group (2022), the CR was 100% in favorable-risk patients (14/14), 100% in intermediate-risk patients (14/14), and 83.3% in adverse-risk patients (10/12). For patients who achieved CR, MRD negative rate was 88.9% (32/36) by flow cytometry. During the induction treatment, all patients experienced ≥Grade 3 hematological toxicity, the remaining common adverse reactions were infections and fever. None of patients died during the induction therapy. In patients who achieved response, the median time to recovery of the white blood count (WBC) to ≥1.0 × 10 9/L and the platelet count to ≥20 × 10 9/L after induction therapy was 13 (range: 10-52) and 13 days (range: 2-33), respectively. ( Table 1) Conclusions: Ven combined with HAA (HAAV) is a highly effective and safe induction therapy for young adult patients with de novo AML. To the best of our knowledge, this is the first report about Ven combined with HAA.

IKS03, a Novel CD19-Targeted Antibody Drug Conjugate, Induces Target Dependent In Vivo Cell Killing of B-Cell Lymphoma Xenografts By DNA Crosslinking

CD19 is widely expressed in B-cell non-Hodgkin lymphoma including diffuse large B-cell lymphoma (DLBCL) and mantle cell lymphoma (MCL). IKS03 is a CD19-targeting antibody drug conjugate (ADC) with a novel linker payload design for tumor-selective payload release. Site-specific conjugation using farnesyltransferase-catalysed ConjuALL technology to a DNA crosslinking agent yields a homogenous ADC with drug-to-antibody ratio (DAR) of 2. To minimize systemic release of the highly potent payload in human plasma, IKS03 incorporates a linker specifically cleavable by the lysosomal enzyme beta-glucuronidase. In addition, the payload is a pro-drug with a protecting moiety cleavable by the same enzyme. Therefore, following CD19-dependent binding and uptake, IKS03 requires intracellular lysosomal processing in the target cell to both release and fully activate the free pyrrolobenzodiazepine (PBD) dimer payload which induces DNA crosslinking, blocks DNA replication and ultimately leads to tumor selective cell death. In vivo efficacy was evaluated in CD19-expressing preclinical human xenograft models representing different subtypes of B-NHL. IKS03 was highly active against Farage and OCI-LY10 DLBCL xenografts as well as Granta-519 MCL xenografts in SCID mice with durable tumor regressions observed following treatment with a single dose of 0.1 to 0.3 mg/kg. Likewise, in a Ramos lymphoma model a single IKS03 dose of 0.3 mg/kg significantly inhibited tumor growth. IKS03 demonstrated greater in vivo efficacy than the clinical benchmark ADCs polatuzumab vedotin and loncastuximab tesirine approved for the treatment of patients with B-cell non-Hodgkin lymphomas. PBD cytotoxic agents have been described to induce DNA crosslinking leading to DNA damage. To demonstrate that IKS03 in vivo activity is the result of targeted payload release, Ramos xenografts were established in immunodeficient mice and treated with up to 1 mg/kg of IKS03 in comparison with a non-targeted ADC utilizing the same linker payload format. Tumors were excised at 1 day and 3 days post-treatment and assessed for the presence of phospho-H2AX positive foci, a marker of DNA damage, by immunohistochemistry. IKS03 treatment increased both frequency and staining intensity of nuclear phospho-H2AX foci in Ramos xenografts in a dose- and time-dependent manner. In contrast, the non-targeted control ADC did not result in any changes as compared to a vehicle control. This confirms the specific induction of DNA damage in CD19 positive lymphomas as a pharmacodynamic response to IKS03 treatment in vivo. In conclusion, IKS03 is highly effective in an array of preclinical MCL and DLBCL xenograft models at doses that are well tolerated. In vivo, IKS03 treatment increased the level of phospho-H2AX indicative of irreversible DNA damage in a target-dependent manner. This is consistent with the proposed mechanism of action of CD19-mediated delivery of a PBD payload specifically to B-cell tumors. Clinical investigation in patients with advanced B-Cell Non-Hodgkin Lymphomas is being initiated (NCT05365659).

HPB P14 Setting the Foundation: Empowering Junior Doctors in Complex Specialties through Induction Excellence

Abstract Background The newly qualified junior doctors in the UK face challenges due to limited experience and unfamiliarity with their rotations. We aim to share the experience of establishing a Hepatobiliary and Pancreatic (HPB) surgery specific induction program at University Hospitals of Leicester NHS Trust and assess its impact on doctors' knowledge and experience. Methods A booklet was distributed to new junior doctors and a two-hour structured teaching session was also conducted, with pre- and post-session assessments using multiple-choice questions and a feedback survey. The survey measured understanding of HPB anatomy, interventions, and satisfaction with the teaching methodology. Results Prior to the teaching session, levels 4 and 5 understanding of HPB anatomy were reported by 18.1% of participants. After the teaching session, there was a significant improvement in understanding, with levels 4 and 5 reported by 95% of participants, and only 5% reporting level 3. Similar improvements were observed in other tested categories. Factual knowledge also improved, with a 38% increase in correct responses on basic HPB and surgical knowledge questions. Participants found the session helpful and well-structured, with 60% strongly agreeing and an additional 30% agreeing. The delivery method was deemed appropriate by the majority. Conclusions Specialty-specific induction programs are crucial for providing support and ensuring the development of competent doctors. Efforts should be made to create supportive working environments for junior doctors to alleviate stress and improve their well-being.

Adipocyte Glucocorticoid Receptor Activation With High Glucocorticoid Doses Impairs Healthy Adipose Tissue Expansion by Repressing Angiogenesis.

In Human, glucocorticoids (GC) are commonly prescribed because of their anti-inflammatory and immunosuppressive properties. However, high doses of GC often lead to adverse side effects including diabetes and lipodystrophy. We recently reported that adipocyte glucocorticoid receptor (GR)-deficient (AdipoGR-KO) mice under corticosterone (CORT) treatment exhibited a massive adipose tissue (AT) expansion associated with a paradoxical improvement of metabolic health compared to control mice. However, whether GR may control adipose development remains unclear. Here, we show a specific induction of the hypoxiainducible factor HIF-1α and the pro-angiogenic Vascular Endothelial Growth Factor-A expression in GR-deficient adipocytes of AdipoGR-KO mice as compared to control mice, together with an increased adipose vascular network, as assessed by 3D-analysis imaging. GR activation reduced HIF-1α recruitment on Vegfa promoter resulting from Hif-1α downregulation at the transcriptional and post-translational levels. Importantly, in CORT-treated AdipoGR-KO mice, the blockade of VEGFA by a soluble decoy receptor prevented AT expansion and the healthy metabolic phenotype. Finally, in subcutaneous AT from Cushing patients, higher VEGFA expression was associated with a better metabolic profile. Collectively, these results highlight that adipocyte GR negatively controls AT expansion and metabolic health through the down-regulation of the major angiogenic effector VEGFA and inhibiting vascular network development. (LIPOCUSH, NCT01688349).

Stromal cell models of arrhythmogenic cardiomyopathy

Abstract Background Arrhythmogenic cardiomyopathy (ACM) is an inherited heart disease mainly associated with mutations in genes encoding cardiac desmosomes, most frequently Plakophilin-2 (PKP2). Primary cardiac mesenchymal stromal cells (CMSCs) were shown to contribute to the aberrant ACM cardiac remodeling based on their ability to differentiate into adipocytes and myofibroblasts, thus representing a useful in vitro model for the disease. The limitations associated with access to biopsies and finite passages in culture have been overcome by the advancement of stem cell technology, especially human induced pluripotent stem cells (hiPSCs) and their ability to differentiate towards cardiac stromal population. Specifically, CMSCs from hiPSCs provide a virtually unlimited cell sources that are genetically identical to patient cells of origin. Purpose The study compares hiPSC-derived stromal cells (hiPSC-D) and primary stromal cells (PRs), to explore the feasibility to use iPSC-D as an alternative in vitro model to study ACM. Methods PRs were isolated from ACM patients and healthy donors (HC). hiPSCs from a PKP2 mutated ACM patient (c.2013delC) and a sex/aged matched HC were used to produce iPSC-D. Flow cytometry analysis and specific differentiation induction (osteogenic, chondrogenic, adipogenic and pro-fibrotic) were performed to confirm mesenchymal lineage and multipotency. qRT-PCR and western blot analysis were performed to evaluate the expression of desmosomes, both gene and protein levels. Illumina transcriptomic profiling of both PRs and iPSC-D were compared and enriched pathways were identified by GSEA. Results Both PR and iPSC-D cells expressed mesenchymal surface markers and were able to differentiate towards osteogenic, chondrogenic, adipogenic and myofibroblast lineages. Desmosomal genes and proteins were expressed by both populations, showing a general trend toward lower expression of desmosomal markers in ACM compared to HC cells. Overall, higher expression of desmosomes was observed in iPSC-D stromal cells compared to their primary counterparts, independently on the genotype. As occurs for PRs, ACM iPSC-D exhibited an increased propensity to accumulate lipid droplets and collagen compared to HC cells. Moreover, both cell populations shared a highly similar transcriptomic profile, showing a significant linear correlation (R=0.85). Interestingly, PRs were enriched for genes associated with the immune response and differentiation processes, hinting to the hypothesis that an inflammatory stimulus received from the cardiac tissue might activate the innate immunity and drive the cell differentiation fates in the PRs. Conclusion(s) Both PR and iPSC-D cardiac stromal cells obtained from ACM patients can be used to model the disease, where the choice for the most suitable cell populations can be determined by the experimental strategy and/or the availability of human primary material.

LmNaTx15, a novel scorpion toxin, enhances the activity of Nav channels and induces pain in mice

Polypeptide toxins are major bioactive components found in venomous animals. Many polypeptide toxins can specifically act on targets, such as ion channels and voltage-gated sodium (Nav) channels, in the nervous, muscle, and cardiovascular systems of the recipient to increase defense and predation efficiency. In this study, a novel polypeptide toxin, LmNaTx15, was isolated from the venom of the scorpion Lychas mucronatus, and its activity was analyzed. LmNaTx15 slowed the fast inactivation of Nav1.2, Nav1.3, Nav1.4, Nav1.5, and Nav1.7 and inhibited the peak current of Nav1.5, but it did not affect Nav1.8. In addition, LmNaTx15 altered the voltage-dependent activation and inactivation of these Nav channel subtypes. Furthermore, like site 3 neurotoxins, LmNaTx15 induced pain in mice. These results show a novel scorpion toxin with a modulatory effect on specific Nav channel subtypes and pain induction in mice. Therefore, LmNaTx15 may be a key bioactive component for scorpion defense and predation. Besides, this study provides a basis for analyzing structure-function relationships of the scorpion toxins affecting Nav channel activity.

Using an episodic specificity induction to improve children's future thinking.

Episodic future thinking (EFT) is the ability to subjectively pre-experience a specific future event. Future-oriented cognition in young children positively predicts physical health and financial status later in life. Can EFT be improved in children, even temporarily? Developmental research emphasizes the importance of thinking about one's own near future to enhance EFT, whereas research in adults suggests benefits reside in constructing a richly detailed event. We bridged the two perspectives to examine whether a procedure, the "episodic specificity induction" (ESI), could be adapted to encourage an episodic mode of thinking in children, benefitting performance on a variety of subsequent EFT tasks. The present study implemented a child-friendly ESI in which children mentally simulated a future event and were probed for specific details about it. We randomly assigned 66 children aged 6 and 7 years to one of two conditions: (1) ESI, in which children imagined "having breakfast tomorrow" in detail, describing surroundings, people, and actions, or (2) a Control condition (i.e., no construction), in which children simply viewed and described a picture of another child having breakfast. Children then completed a series of future thinking tasks assessing prospective memory, recollection/imagination of events, delay of gratification, and planning. Our ESI was successful in promoting the construction of a detailed event, and subsequently increasing the number of details of recollected and imagined events on an outcome task as compared to a control condition. Nonetheless, the effect of ESI was smaller than expected - a finding that fits with recent work suggesting that such interventions may be too cognitively taxing for young children and/or that benefits may hinge on further development in episodic processes. We discuss possible modifications to the induction and implications for EFT amelioration in young children.

Insulin-degrading enzyme (IDE) as a modulator of microglial phenotypes in the context of Alzheimer’s disease and brain aging

The insulin-degrading enzyme (IDE) is an evolutionarily conserved zinc-dependent metallopeptidase highly expressed in the brain, where its specific functions remain poorly understood. Besides insulin, IDE is able to cleave many substrates in vitro, including amyloid beta peptides, making this enzyme a candidate pathophysiological link between Alzheimer's disease (AD) and type 2 diabetes (T2D). These antecedents led us to address the impact of IDE absence in hippocampus and olfactory bulb. A specific induction of microgliosis was found in the hippocampus of IDE knockout (IDE-KO) mice, without any effects in neither hippocampal volume nor astrogliosis. Performance on hippocampal-dependent memory tests is influenced by IDE gene dose in 12-month-old mice. Furthermore, a comprehensive characterization of the impact of IDE haploinsufficiency and total deletion in metabolic, behavioral, and molecular parameters in the olfactory bulb, a site of high insulin receptor levels, reveals an unambiguous barcode for IDE-KO mice at that age. Using wildtype and IDE-KO primary microglial cultures, we performed a functional analysis at the cellular level. IDE absence alters microglial responses to environmental signals, resulting in impaired modulation of phenotypic states, with only transitory effects on amyloid-β management. Collectively, our results reveal previously unknown physiological functions for IDE in microglia that, due to cell-compartment topological reasons, cannot be explained by its enzymatic activity, but instead modulate their multidimensional response to various damaging conditions relevant to aging and AD conditions.

The SPECTRA Study: Validating a New Memory Training Program based on the Episodic Specificity Induction to Promote Transfer in Older Adults.

Some complex cognitive activities impacted by aging (future thinking, problem-solving, creative thinking) have been shown to rely on episodic retrieval, suggesting that cognitive interventions aiming to improve retrieval have the potential to induce transfer effects to these activities. Prior studies have shown that a brief one-session technique called Episodic Specificity Induction (ESI) can transiently improve episodic retrieval and induce transfer effects to complex tasks that rely on episodic retrieval in older adults. In the present proof-of-concept study, we assessed whether a training program consisting of repeated practice of the ESI technique can improve episodic retrieval and transfer to complex tasks. Fifteen healthy older adults completed a six-session intervention where they received repeated ESI practice. Before and after the intervention, nearest transfer effects were assessed using free recall, near transfer effects using recognition and associative recognition, and far-transfer effects using mean-ends problem-solving and divergent creative thinking. Before the intervention, typical ESI effects were observed (better performance after an ESI than after a control task), indicating that the ESI operated as expected in our sample. When examining the intervention effects, performance was increased after the intervention on free recall and recognition (nearest- and near-transfer) as well as problem-solving and divergent creative thinking (far transfer). These results indicate that an intervention relying on the ESI technique can produce both near and far transfer. These findings support the use of the ESI in the design of interventions that could improve retrieval and have a broad impact on a range of complex tasks.

SIRT1 induction in the skeletal muscle of male mice partially attenuates changes to whole-body metabolism in response to androgen deprivation

In males, androgens regulate whole body metabolism. The components in androgen target organs contributing to whole-body metabolic function remain ill defined. Sirtuin1 (SIRT1) protein levels are lower in the limb muscle of male mice subjected to androgen deprivation. Because SIRT1 can influence whole-body metabolism, the purpose was to assess whether muscle specific SIRT1 induction attenuated changes to whole-body metabolism in response to androgen deprivation. Physically mature male mice containing an inducible muscle specific SIRT1 transgene (SIRT1) were subjected to a sham or castration surgery and compared to sham and castrated male mice where the SIRT1 transgene was not induced (WT). The respiratory exchange ratio (RER), energy expenditure, and carbohydrate and fat oxidation rates were determined using metabolic cages. Castration lowered RER in WT mice and the lower RER coincided with lower energy expenditure, lower carbohydrate oxidation rates, and higher fat oxidation rates. SIRT1 induction attenuated the castration-induced changes to RER and fat oxidation rates. Changes to energy expenditure and glucose oxidation rates were not affected by SIRT1. Decreases in muscle SIRT1 protein in males may partially contribute to the dysregulation of whole-body metabolism in response to androgen deprivation.

Multi-scale cellular imaging of DNA double strand break repair

Live-cell and high-resolution fluorescence microscopy are powerful tools to study the organization and dynamics of DNA double-strand break repair foci and specific repair proteins in single cells. This requires specific induction of DNA double-strand breaks and fluorescent markers to follow the DNA lesions in living cells. In this review, where we focused on mammalian cell studies, we discuss different methods to induce DNA double-strand breaks, how to visualize and quantify repair foci in living cells., We describe different (live-cell) imaging modalities that can reveal details of the DNA double-strand break repair process across multiple time and spatial scales. In addition, recent developments are discussed in super-resolution imaging and single-molecule tracking, and how these technologies can be applied to elucidate details on structural compositions or dynamics of DNA double-strand break repair.

Identification of Potential Protein Targets in Extracellular Vesicles Isolated from Chemotherapy-Treated Ovarian Cancer Cells.

Despite the ongoing clinical trials and the introduction of novel treatments over the past few decades, ovarian cancer remains one of the most fatal malignancies in women worldwide. Platinum- and paclitaxel-based chemotherapy is effective in treating the majority of patients with ovarian cancer. However, more than 70% of patients experience recurrence and eventually develop chemoresistance. To improve clinical outcomes in patients with ovarian cancer, novel technologies must be developed for identifying molecular alterations following drug-based treatment of ovarian cancer. Recently, extracellular vesicles (EVs) have gained prominence as the mediators of tumor progression. In this study, we used mass spectrometry to identify the changes in EV protein signatures due to different chemotherapeutic agents used for treating ovarian cancer. By examining these alterations, we identified the specific protein induction patterns of cisplatin alone, paclitaxel alone, and a combination of cisplatin and paclitaxel. Specifically, we found that drug sensitivity was correlated with the expression levels of ANXA5, CD81, and RAB5C in patients receiving cisplatin with paclitaxel. Our findings suggest that chemotherapy-induced changes in EV protein signatures are crucial for the progression of ovarian cancer.

Investigating the impact of tea mosquito bug on the phytochemical profile and quality of Indian tea cultivars using HPLC and LC-MS-based metabolic profiling

The impact of biotic and abiotic stress on tea not only reduces yield but also influences flavour, aroma, and phytochemical composition. The tea mosquito bug (TMB) is the major biotic stress that affects the growth, yield, and quality of tea. In this study, we have studied the impact of biotic stress induced by TMB on three Indian tea cultivars: UPASI-2, CR-6017, and ATK-1. The content of antioxidant enzymes and phytochemical composition were found to be increased upon TMB infestation in all tea clones compared to the control. The UPASI-2 tea cultivar showed an effectively increased level of antioxidant enzymes and phytochemicals. HPLC and LC-MS analysis showed a differential production of various secondary metabolites like catechin ("flavan-3-ols") and phenolics in control and infected tea clones. Altogether, the specific induction of various antioxidant enzymes, catechins, and phenolics contribute to the defense response of tea plants toward TMB infection. In addition, the analysis of transcriptional variation of key flavonoid genes (PAL, DFR, CHI, F3H, and 4CL) showed induction in infected plants compared to the control.

Abstract P3087: Macrophage Fatty Acid Metabolism Promotes Splenic Hematopoiesis Through Mitochondrial Signaling During Heart Failure With Preserved Ejection Fraction

Rationale: Diastolic dysfunction, including during heart failure with preserved ejection fraction (HFpEF), is a widespread morbidity with limited therapeutic options. Though heterogeneous in etiology, major risk factors include hyperlipidemia, hypertension, and inflammation. Recent studies have also shown significant changes in systemic metabolite availability during HFpEF, further suggesting that metabolic changes may be an important indicator of HFpEF pathophysiology. Macrophages, which have previously been implicated in the promotion of diastolic dysfunction, utilize specific metabolic pathways to drive inflammation or tissue repair. However, little is understood about myeloid metabolism during HFpEF. Objective: We set out to define the functional and metabolic heterogeneity of the cardiac immune response during diastolic dysfunction. Methods/Results: Diastolic dysfunction was induced by HFD and eNOS inhibition in mice. scRNAseq showed a specific induction of pro-fibrotic and lipid handling pathways in cardiac macrophages, including the fatty acid receptor Cd36. In live animals, myeloid deficiency of Cd36 prevented diastolic dysfunction and was associated with dampened cardiac inflammation. Investigating immune cell sourcing during HFpEF revealed that a splenic hematopoietic niche formed in wild-type mice. Further, mitochondrial metabolism in splenic macrophages was associated with increased expression of VCAM-1, the latter of which has been shown to play a role in retention of HSCs in the spleen. Indeed, blocking VCAM-1 or genetic deletion of VCAM-1 in myeloid cells was sufficient to disrupt retention of HSCs within the spleen during HFpEF. Culturing splenic macrophages with saturated fatty acids (SFAs) in vitro was sufficient to induce expression of Vcam1. Increased cellular citrate levels were correlated with increased histone acetylation at the Vcam1 promoter in splenic macrophages cultured with SFAs. Inhibition of mitochondrial uptake of fatty acids through CPT1a siRNA or inhibition of ACLY were sufficient to prevent this upregulation of Vcam1 expression. Conclusions: Our data suggest a critical role for myeloid metabolism in the sourcing of detrimental-monocyte-derived macrophages from the spleen to the heart.