Future Therapies Research Articles

Synergistic antibacterial effect of Persicaria odorata synthesised silver nanoparticles with antibiotics on drug-resistant bacteria

For many years, combination therapies with two or more antimicrobial agents have gained interest to address the problem of emerging antibiotic resistance. Several studies have shown that the addition of AgNPs enhances the antimicrobial activity of antibiotics against resistant pathogens. However, the antibacterial activity of phytosynthesised silver nanoparticles (AgNPs) in combination with antibiotics has not been extensively studied. Therefore, this study aimed to investigate the antibacterial activity of the phytosynthesised AgNPs with different antibiotics against two common pathogenic bacteria, namely methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa. A green method was used to produce AgNPs, where Persicaria odorata extract acted as a reducing and stabilising agent in the aqueous synthesis process. The production of AgNPs was verified through various analytical techniques, including ultraviolet–visible spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, high-resolution transmission electron microscopy and energy-dispersive X-ray. Notably, strong absorption spectrum corresponding to the surface plasmon resonance peaks of AgNPs was found at 436 nm, with particles ranging from 10 to 35 nm. The disc diffusion technique was used to investigate the antibacterial activity of AgNPs, both alone and in combination with antibiotics. Initially, the MRSA and P. aeruginosa showed 100 % resistance to antibiotics, especially ampicillin. However, the disc diffusion result exhibit a significant increase in the inhibition zone when ampicillin-AgNPs, cefoxitin-AgNPs and chloramphenicol-AgNPs were used in combination. In short, the findings of this study showed that AgNPs exhibit a promising synergy when used in combination with antibiotics, opening up possibilities for future combination therapies against antibiotic-resistant pathogens.

EXTH-50. ACUTE AND LONG-TERM RESPONSES TO QUISINOSTAT TREATMENT IN PDX MODELS OF GLIOBLASTOMA

Abstract BACKGROUND Histone deacetylase inhibitors (HDACi) promote reactive oxygen species production, result in DNA damage, and trigger cell fate changes. Quisinostat is a Class I-II selective HDAC inhibitor with high specificity for HDAC1. Here, we evaluated acute and long-term quisinostat treated glioblastoma (GBM) PDX tissues and glioma stem cells for increase in reactive oxygen species and cell fate change. METHODS GBM PDX models (n = 6) and multiple PDX-derived glioma stem cells were treated with quisinostat, radiation, or a combination of quisinostat and radiation. Tissue and cells were processed via LC-MS/MS, RNA-seq, qRT-PCR, immunoblotting, and immunocytochemistry assays to examine increase in cellular reactive oxygen species, oxidative stress biomarkers, and markers of stemness and differentiation. RESULTS RNA sequencing of tumor tissue revealed that chronic co-treatment with quisinostat and radiation led to increased cellular expression of oxidative stress- and neuronal-related genes compared to control group. Immunoblotting confirmed upregulation of neuronal signaling proteins in the quisinostat treated tumor tissue in vivo. qRT-PCR and LC-MS/MS analysis demonstrated an increase in oxidative stress biomarkers following acute quisinostat treatment in vitro. CONCLUSIONS Treatment with quisinostat increased the expression of genes involved in promoting oxidative stress and neuronal signaling in vivo. LC-MS/MS, qRT-PCR, immunoblotting and immunocytochemistry assays confirmed increase in oxidative stress and neuronal markers upon acute treatment in vitro. Cancer cells often express higher levels of reactive oxygen species compared to normal tissue, and oxidative stress is linked to greater survival and proliferation of cancer cells. However, there is much interest in increasing oxidative stress in glioma cells to induce selective cell death in cancer cells. Ongoing studies are aimed at evaluating the significance of these cellular and molecular changes resulting from quisinostat treatment and whether the quisinostat-induced oxidative stress and cell fate changes can be exploited for future combination therapy for GBM.

Human antimicrobial peptide inactivation mechanism of enveloped viruses

HypothesisEnveloped viruses are pivotal in causing various illnesses, including influenza and COVID-19. The antimicrobial peptide LL-37, a critical part of the human innate immune system, exhibits potential as an antiviral agent capable of thwarting these viral threats. Its mode of action involves versatile and non-specific interactions that culminate in dismantling the viral envelope, ultimately rendering the viruses inert. However, the exact mechanism of action is not yet understood. ExperimentsHere, the mechanism of LL-37 triggered changes in the structure and function of an enveloped virus is investigated. The bacteriophage “Phi6” is used as a surrogate for pathogenic enveloped viruses. Small angle X-ray and neutron scattering combined with light scattering techniques demonstrate that LL-37 actively integrates into the virus’s lipid envelope. FindingsLL-37 addition to Phi6 leads to curvature modification in the lipid bilayer, ultimately separating the envelope from the nucleocapsid. Additional biological assays confirm the loss of virus infectivity in the presence of LL-37, which coincides with the structural transformations. The results provide a fundamental understanding of the structure-activity relationship related to enveloped viruses. The knowledge of peptide-virus interactions can guide the design of future peptide-based antiviral drugs and therapies.

Association of MRI Volume Parameters in Predicting Patient Outcome at Time of Initial Diagnosis of Glioblastoma.

Patients with glioblastoma (GBM) may demonstrate varying patterns of infiltration and relapse. Improving the ability to predict these patterns may influence the management strategies at the time of initial diagnosis. This study aims to examine the impact of the ratio (T2/T1) of the non-enhancing volume in T2-weighted images (T2) to the enhancing volume in MRI T1-weighted gadolinium-enhanced images (T1gad) on patient outcome. A retrospective audit was performed from established prospective databases in patients managed consecutively with radiation therapy (RT) for GBM between 2016 and 2019. Patient, tumour and treatment-related factors were assessed in relation to outcome. Volumetric data from the initial diagnostic MRI were obtained via the manual segmentation of the T1gd and T2 abnormalities. A T2/T1 ratio was calculated from these volumes. The initial relapse site was assessed on MRI in relation to the site of the original T1gad volume and surgical cavity. The major endpoints were median relapse-free survival (RFS) from the date of diagnosis and site of initial relapse (defined as either local at the initial surgical site or any distance more than 20 mm from initial T1gad abnormality). The analysis was performed for association between known prognostic factors as well as the radiological factors using log-rank tests for subgroup comparisons, with correction for multiple comparisons. One hundred and seventy-seven patients with GBM were managed consecutively with RT between 2016 and 2019 and were eligible for the analysis. The median age was 62 years. Seventy-four percent were managed under a 60Gy (Stupp) protocol, whilst 26% were on a 40Gy (Elderly) protocol. Major neuroanatomical subsites were Lateral Temporal (18%), Anterior Temporal (13%) and Medial Frontal (10%). Median volumes on T1gd and T2 were 20 cm3 (q1-3:8-43) and 37 cm3 (q1-3: 17-70), respectively. The median T2/T1 ratio was 2.1. For the whole cohort, the median OS was 16.0 months (95%CI:14.1-18.0). One hundred and forty-eight patients have relapsed with a median RFS of 11.4 months (95%CI:10.4-12.5). A component of distant relapse was evident in 43.9% of relapses, with 23.6% isolated relapse. Better ECOG performance Status (p = 0.007), greater extent of resection (p = 0.020), MGMT methylation (p < 0.001) and RT60Gy Dose (p = 0.050) were associated with improved RFS. Although the continuous variable of initial T1gd volume (p = 0.39) and T2 volume (p = 0.23) were not associated with RFS, the lowest T2/T1 quartile (reflecting a relatively lower T2 volume compared to T1gd volume) was significantly associated with improved RFS (p = 0.016) compared with the highest quartile. The lowest T2/T1 ratio quartile was also associated with a lower risk of distant relapse (p = 0.031). In patients diagnosed with GBM, the volumetric parameters of the diagnostic MRI with a ratio of T2 and T1gad abnormality may assist in the prediction of relapse-free survival and patterns of relapse. A further understanding of these relationships has the potential to impact the design of future radiation therapy target volume delineation protocols.

TMIC-01. PERICYTES ORCHESTRATE A TUMOR-SUPPRESSIVE MICROENVIRONMENT BY IMPINGING ON THE CROSSTALK BETWEEN MACROPHAGES AND TUMOR-INITIATING CELLS IN GLIOBLASTOMA MULTIFORME

Abstract Glioblastoma multiforme (GBM) is characterized by fast progression, infiltrative growth pattern and a high rate of relapse. A defining feature of GBM is the existence of spatially and functionally distinct cellular niches, i.e. a hypoxic niche, a leading-edge niche, and a peri-vascular niche, in which malignant cells engage in paracrine crosstalk with cell types comprising the tumor microenvironment, including immune cells, astrocytes, and vascular cells. Here, by analysis of single-cell transcriptomic data of human GBM and transgenic mouse models of GBM, we unexpectedly identified pericytes intimately associated with the endothelium as the most active paracrine signaling hub within the tumor parenchyma. Exclusive signaling axes emanating from pericytes were received by endothelial cells, malignant cells, astrocytes, and immune cells. Depletion of pericytes through genetic engineering in several different transgenic and orthotopic mouse models of GBM demonstrated accelerated tumor progression, a disrupted blood-brain-barrier, and premature death of pericyte-poor mice. Mechanistic studies revealed that pericyte deficiency altered the cellular composition of GBM, remodeled the endothelium, and impacted on the immune cell landscape. Specifically, endothelial cells deprived of pericyte association upregulated Colony Stimulating Factor (CSF)-1, which in turn attracted peri-vascular tumor-associated macrophages polarized towards an immune-suppressive phenotype. In the absence of pericytes, the recruited macrophages expressed Hepatocyte Growth Factor (HGF), which reinforced activation of its receptor tyrosine kinase c-Met on a subset of GBM cells harboring an extreme mesenchymal-like state. Indeed, orthotopic implantation of isolated c-Met-expressing GBM cells corroborated their superior tumor-initiating capability and invasive phenotype, compared to cells negative for c-Met. Taken together, we infer that the pericyte represents a critical modulator of GBM development by orchestrating a tumor-suppressive microenvironment; our findings thus highlight the importance of pericyte preservation in the face of current and future GBM therapies.

EXTH-80. A SYSTEMIC COMPARISON OF CAR T CELLS, CAR NK CELLS AND CAR MACROPHAGES FOR GLIOMA TREATMENT

Abstract Adoptive transfer of engineered immune cells expressing chimeric antigen receptors (CAR) is an emerging cancer immunotherapy that is also being investigated for glioblastoma. Many separate studies focus on either CAR T or CAR NK cells, and CAR macrophages are in early stages of development. However, a systemic cross-comparison of engineered immune effector cells against solid tumors, including glioblastoma, is lacking. Here, we generated second-generation NKG2D CAR mouse and human T cells, NK cells and macrophages and characterized them in vitro in co-culture assays with glioma cells and in vivo in orthotopic glioma mouse models. In vitro, mouse and human CAR T cells and CAR NK cells exhibited the highest tumor-killing activity. In contrast, in vivo, only CAR T cells exhibited anti-tumor activity in two immunocompetent orthotopic glioma-bearing mouse models, whereas CAR NK cells and CAR macrophages failed to prolong survival. More in-depth characterizations using flow cytometry, in vivo imaging and 3D microscopy, revealed that CAR T cells had the best tumor migration properties upon i.v. administration with distinct spatial distribution within the tumor compared to CAR NK cells and CAR macrophages. To profile the effects on bystander immune cells within the tumor microenvironment, we employed flow cytometry and single-cell RNA sequencing. This revealed effector cell-specific effects on the tumor microenvironment and provided insights into why the effective in vitro activity did not translate to in vivo. To unleash the anti-tumor potential of CAR immune effector cells, we engineered them to co-express the pro-inflammatory cytokines IL-12/IFNα2. This improved the in vivo anti-tumor activity of all effector cells and led to the highest fraction of long-term surviving mice with CAR NK cells. This study offers the first cross-comparison of different CAR immune effector cells against gliomas, providing valuable insights for the design of future adoptive cell therapies targeting malignant brain tumors.

B cell immune profiles in dysbiotic vermiform appendixes of pancreatic cancer patients.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the deadliest solid tumors and is resistant to immunotherapy. B cells play an essential role in PDAC progression and immune responses, both locally and systemically. Moreover, increasing evidence suggests that microbial compositions inside the tumor, as well as in the oral cavity and the gut, are important factors in shaping the PDAC immune landscape. However, the gut-associated lymphoid tissue (GALT) has not previously been explored in PDAC patients. In this study, we analyzed healthy vermiform appendix (VA) from 20 patients with PDAC and 32 patients with colon diseases by gene expression immune profiling, flow cytometry analysis, and microbiome sequencing. We show that the VA GALT of PDAC patients exhibits markers of increased inflammation and cytotoxic cell activity. In contrast, B cell function is decreased in PDAC VA GALT based on gene expression profiling; B cells express significantly fewer MHC class II surface receptors, whereas plasma cells express the immune checkpoint molecule HLA-G. Additionally, the vermiform appendix microbiome of PDAC patients is enriched with Klebsiella pneumoniae, Bifidobacterium animalis, and Adlercreutzia equolifaciens, while certain commensals are depleted. Our findings may suggest impaired B cell function within the GALT of PDAC patients, which could potentially be linked to microbial dysbiosis. Additional investigations are imperative to validate our observations and explore these potential targets of future therapies.

Prevalence and analysis of acquired and transmitted integrase strand transfer inhibitor-associated HIV-1 drug resistance in Chongqing, China

ABSTRACT In this study, we examined the occurrence of acquired and transmitted drug resistance to integrase strand transfer inhibitor (INSTI) in HIV-1 strains in Chongqing (China) for guiding for the routine testing of INSTI-associated HIV-1 genotype resistance. Plasma samples were obtained from HIV-1 patients at Chongqing Public Health Medical Center from July 2019 to August 2022. Besides, amplification, sequence, and analysis of the portion of the HIV-1 pol gene that encodes the integrase protein were implemented to identify INSTI resistance. Integrase sequence data was harvested for a comprehensive cohort of 1032 patients infected with HIV-1. This cohort consisted of 564 ART-naive patients, 465 ART-treated patients, and 3 patients with an unknown treatment history. Within the study group, we identified INSTI resistance in 21 patients (2.03%, 21/1032), including 17 ART-treated patients (3.66%, 17/465). Among the ART-treated patients, 12 were INSTI-treated (11.76%, 12/102), 5 were INSTI-naive (1.38%, 5/363), and 4 were ART-ineffective patients (0.71%, 4/564). The prevalent major resistance mutation was Q148R (0.48%, 5/1032), while the most prevalent accessory resistance mutation was E157Q (1.65%, 17/1032). In light of the above, it is recommended that the incidence of accessory genotype analysis should be considered before starting any future INSTI-based therapy, especially in patients with drug resistance to NRTIs and NNRTIs and the reduction of INSTI sensitivity should be carefully monitored and investigated. Regular monitoring for resistance should be implemented after the use of INSTIs, and, importantly, ongoing monitoring of the decreasing susceptibility to INSTIs is crucial following the initiation of treatment with INSTIs.

Coronary flow reserve predicts response to anti-ischaemic therapy in patients with angina with non-obstructive coronary arteries

Abstract Background Half of all patients with angina have non-obstructive coronary arteries (ANOCA) and therapy is often empirical. The CorMicA trial showed improved symptom scores with a physiology-guided approach but the mechanistic relationship between physiological classification and outcome remains unclear. Purpose To assess if invasively assessed coronary flow reserve (CFR) can predict change in exercise time (ET) and angina-specific quality of life (QOL) in response to anti-ischaemic therapy in patients with ANOCA. Methods Prospective, randomised, crossover trial of consecutively evaluated patients with ANOCA. Inclusion criteria: (i) typical and limiting angina; (ii) non-obstructive coronary arteries (fractional flow reserve &amp;gt;0.80); (iii) normal left ventricular systolic function. Coronary physiological assessment was performed using a pressure and Doppler dual sensor-tipped wire in the left anterior descending artery and microvascular dysfunction defined a priori as CFR&amp;lt;2.5 in response to intravenous Adenosine. Patients, clinicians, and researchers were blinded to coronary physiology data. All patients were then sequentially treated with Amlodipine and Ranolazine (in a randomised order), in 4 week blocks with 1 week washout (Figure 1). The outcome measures are 1) Primary (objective): change in ET on exercise stress testing and 2) Secondary (subjective): change in angina-specific QOL on Seattle Angina Questionnaire (SAQ). Data are presented as mean±SD or median (IQR). Results Sixty-six patients with ANOCA were enrolled into the study [62±8 years, 64% females, CCS 3 (2, 3)]; eight patients were excluded. Forty patients had an impaired CFR (2.0±0.3), whilst 18 patients had normal CFR (3.1±0.6), with similar baseline ET (324±133 vs 321±136 seconds, respectively; p=0.94). Patients with an impaired CFR improved their exercise capacity with Ranolazine (ΔET 70±108 seconds; p&amp;lt;0.01) and Amlodipine (ΔET 70±82 seconds; p&amp;lt;0.01). However, there was no such improvement in patients with normal CFR, with either Ranolazine (ΔET 16±88 seconds; p=0.49) or Amlodipine (ΔET 21±59 seconds; p=0.18) (Figure 2). There was no evidence of a training effect (ΔET between visit 1 and visit 4 was 15±112 seconds; p=0.57). Angina-specific QOL improved in patients with impaired CFR [Ranolazine ΔSAQ 12±18 (p&amp;lt;0.01) and Amlodipine ΔSAQ 8±13 (p&amp;lt;0.01)]; the change in SAQ was less marked in patients with normal CFR [Ranolazine ΔSAQ 7±14 (p=0.07) and Amlodipine ΔSAQ 6±10 (p=0.03)]. Conclusion Patients with impaired CFR demonstrated an improved exercise capacity, whereas patients with normal CFR did not. CFR-guided management of ANOCA is likely to be superior to an empirical strategy. This type of physiology-blinded study design could act as a template for future therapy trials in patients with stable angina.Study design for therapeutic trialResponse to anti-ischaemic therapy

Understanding the impact of international music therapy student placements on music therapy practice and professional identity

ABSTRACT Introduction In this article, authors present and contextualise findings from a research project examining the experience and impact of participation in an international music therapy student placement for nine Australian music therapists. Research participants reflect on the impact both at the time of the student placement, and subsequently on their professional lives as music therapists. Method Researchers used qualitative thematic analysis to analyse nine open-ended interviews with the research participants. Themes were articulated over three iterations of analysis and were developed into a series of narratives that reflected various perspectives on key emerging themes. Results Researchers interpreted five broad areas of learning from the data: Placement structure; challenges and supporting factors on placement; learning opportunities identified within the placement; future impact on individuals; and broader insights for the international development music therapy literature. Discussion Learning from across these five areas is considered in relation to the context of international development as an emerging practice area in music therapy. Researchers suggest how key data points may inform future approaches to music therapy practice. A critical lens is used to consider some ethical considerations related to this area of work, and to propose ways that research learning may inform future music therapy practice in international development.

Human Skeletal myopathy myosin mutations disrupt myosin head sequestration.

Myosin heavy chains encoded by MYH7 and MYH2 are abundant in human skeletal muscle, and important for muscle contraction. However, it is unclear how mutations in these genes disrupt myosin structure and function leading to skeletal muscle myopathies termed myosinopathies. Here, we used multiple approaches to analyse the effects of common MYH7 and MYH2 mutations in the light meromyosin region of myosin (LMM). Analyses of expressed and purified MYH7 and MYH2 LMM mutant proteins combined with in-silico modelling showed that myosin coiled-coil structure and packing of filaments in vitro are commonly disrupted. Using muscle biopsies from patients, and Mant-ATP chase protocols to estimate the proportion of myosin heads that were super-relaxed, together with X-ray diffraction measurements to estimate myosin head order we found that basal myosin ATP consumption was increased and the myosin super-relaxed state was decreased in vivo. In addition, myofibre mechanics experiments to investigate contractile function showed myofibre contractility was not affected. These findings indicate that the structural remodelling associated with LMM mutations induces a pathogenic state in which formation of shutdown heads is impaired, thus increasing myosin head ATP demand in the filaments, rather than affecting contractility. These key findings will help design future therapies for myosinopathies.

Abstract 14056: The Role of Immune Activation in Trametinib-Induced Cardiomyopathy

Introduction: Trametinib (Trm) is a highly selective inhibitor of MEK1/2, downstream targets of the RAS signaling pathway that has been used widely for the treatment of BRAF V600E/K-mutant melanoma. Multiple clinical trials are also underway investigating its use in colorectal, prostate cancer, leukemia, and triple negative breast cancer. Trm is generally well tolerated but can be associated with potentially serious cardiotoxicity in 5-20% of treated patients. The specific mechanisms driving Trm-induced cardiomyopathy (TIC) are largely unexplored. The goal of this project is to investigate the role that resident and infiltrating inflammatory cells play in TIC. Hypothesis: Our data suggest that Trm provokes cardiomyocyte immune responses including increased production of inflammatory cytokines, NF-KB signaling, and macrophage infiltration. We hypothesize that this activation occurs through DAMP-driven activation of the TLR and cGAS-STING pathway. Methods: C57BL/6 mice were gavaged with Trm 3mg/kg/day for 3 days prior to sacrifice. Heart tissue was analyzed via RNAseq, confocal microscopy, and flow cytometry. In vitro assays were completed using neonatal rat ventricular myocytes (NRVM). Results: RNAseq data of whole heart tissue from Trm treated mice surprisingly demonstrated increased expression of transcripts in inflammatory pathways. In vitro stimulation of NRVMs with Trm demonstrated increased production of inflammatory cytokines including IFNy and increased NF-KB signaling. Confocal microscopy of Trm treated heart tissue demonstrated increased CD68+ macrophage recruitment compared to control. None of these findings would have been predicted by our previous knowledge of cardiomyocyte MEK biology. Conclusions: Trm treatment is associated with cardiomyopathy in up to 20% of patients. Understanding the mechanisms driving this disease pathogenesis may help reduce likelihood of adverse cardiac events associated with future MEK inhibitor therapy used alone or in combination with other agents. Our novel findings suggest that Trm causes immune dysregulation and inflammation in the heart. Future studies will focus on further characterization of immune cell subsets involved and mechanisms of innate immune activation in TIC.

Abstract 14779: Whole Exome Sequencing Study in Pediatric Postural Orthostatic Tachycardia Syndrome Unveils a Path Forward From Genes to Drugs

Introduction: Postural orthostatic tachycardia syndrome (POTS) is more common in Caucasian individuals with a positive family history, while the risk in females is ~4-times of that in males. To date, the etiology of POTS is poorly understood. Hypothesis: Integrative sequencing and translational approaches to identify its genetic underpinning may better understand its pathogenesis and molecular mechanisms for the development of novel precision-based therapies. Methods: We adopted exome sequencing to evaluate the etiology and molecular mechanisms of POTS. We examined a unique collection of 87 unrelated pediatric POTS cases (61 females and 26 males, 6~21 years old), using exome sequencing. Non-relationship between individuals were validated by genome wide genotyping and identity by descent (IBD) analysis. Results: All the individuals were classified as European by principal component analysis (Figure 1). We identified 184 pathogenic (P) or likely pathogenic (LP) variants (in 162 genes). Sixty-eight (78%) cases have at least one P/LP variant. Each P/LP mutation is supported by a minimum of two databases, including ClinVar, InterVar or HGMD_Pro_2023.1. Over-representation analysis (ORA) by the DisGeNET approach showed the enriched gene sets of muscle hypotonia and epilepsy/seizures with statistical significance (FDR&lt;E-10). Clinically, among the 87 WES patients, 41 (47%) suffered from chronic fatigue, and 13 (15%) patients have the symptoms of epilepsy, seizure or convulsion. A number of drug targets in these gene sets are highlighted, including vitamin D receptor, carbamoyl-phosphate synthase 1, methylmalonyl-CoA mutase, androgen receptor, and phenylalanine hydroxylase. Conclusions: Our results suggests that chronic fatigue and epilepsy/seizure symptoms are comorbid symptoms in POTS patients, resulting from common genetic defects. Future precision-based therapies targeting these specific genetic defects may lead to improved outcome in POTS.

Abstract 14969: Association Between Apolipoprotein A-I and Ischemia Risk at Different Diabetes Statuses in Patients Undergoing Percutaneous Coronary Intervention: A Large-Scale Real-World Study

Introduction: Recently, growing attention has shifted to apolipoprotein A-I (ApoA-I), revealing compelling relationship between low ApoA-I and ischemia risk. Diabetes mellitus (DM) frequently coexists with abnormal ApoA-I levels. However, the association between ApoA-I and ischemia risk under different diabetes statuses in patients undergoing percutaneous coronary intervention (PCI) remains unclear. Hypothesis: We hypothesized that diabetes status could modify the relationship between ApoA-I levels and ischemia risk in PCI patients. Methods: This prospective study included consecutive 10724 patients treated with PCI throughout 2013. Patients were categorized into 3 subgroups: DM, pre-DM, and normal group. The primary endpoint was major adverse cardiovascular and cerebrovascular events (MACCE, including all-cause death, myocardial infarction, stroke, and revascularization). Results: Among 10232 PCI patients finally included, 2139 (20.9%) MACCEs occurred at 5-year follow-up. There was a significant interaction between ApoA-I and diabetes status (P for interaction = 0.044). In the DM group, an L-shaped curve was observed (Figure 1A), and multivariate Cox regression analysis revealed that only the lowest quintile level of ApoA-I exhibited an increased risk of MACCE (hazard ratio [HR], 1.336; 95% CI, 1.101-1.620; P = 0.003) (Figure 1B). However, neither the normal group nor pre-DM groups could observe the association between ApoA-I and MACCE (both P &gt;0.05). Further sensitivity analysis, stratifying ApoA-I into two groups based on median level, also indicated the highest MACCE in those combined with lower ApoA-I levels and DM (Figure 1C). Conclusions: In a large-scale 5-years follow-up study, we were the first to demonstrate that diabetes status modified the ischemia risk of low ApoA-I levels, highlighting the clinical importance of low ApoA-I levels for risk stratification and focusing on diabetes status in future therapy targeting ApoA-I.

Prognostic Value of Histopathological Thrombus Age in Large Vessel Occlusion-Related Stroke

Introduction: Acute mechanical thrombectomy (MT) is the preferred treatment for large vessel occlusion-related stroke. Histopathological research on the obtained occlusive embolic thrombus may provide information regarding the aetiology and pathology of the lesion to predict prognosis and propose possible future acute ischaemic stroke therapy. Methods: A total of 75 consecutive patients who presented to the Amphia Hospital with acute large vessel occlusion-related stroke and underwent MT were included in the study. The obtained thrombus materials were subjected to standard histopathological examination. Based on histological criteria, they were considered fresh (<1 day old) or old (>1 day old). Patients were followed for 2 years for documentation of all-cause mortality. Results: Thrombi were classified as fresh in 40 patients (53%) and as older in 35 patients (47%). Univariate Cox regression analysis showed that thrombus age, National Institutes of Health Stroke Scale at hospital admission, and patient age were associated with long-term mortality (p < 0.1). Multivariable Cox hazards and Kaplan-Meier analysis demonstrated that after extensive adjustment for clinical and procedural variables, thrombus age persisted in being independently associated with higher long-term mortality (hazard ratio: 3.34; p = 0.038, log-rank p = 0.013). Conclusion: In this study, older thromboemboli are responsible for almost half of acute large ischaemic strokes. Moreover, the presence of an old thrombus is an independent predictor of mortality in acute large vessel occlusion-related stroke. More research is warranted regarding future therapies based on thrombus composition.

Multimodal Ophthalmic Imaging in Spinocerebellar Ataxia Type 7.

The aim of this case series and narrative literature review is to highlight the importance of multimodal imaging in the ophthalmological examination of patients with spinocerebellar ataxia type 7 and provide a summary of the most relevant imaging techniques. Three patients with SCA7 were included in this case series. A literature review revealed twenty-one publications regarding ocular manifestations of SCA7, and the most relevant aspects are summarized. The role of different imaging techniques in the follow-up of SCA7 patients is analyzed, including color vision testing, corneal endothelial topography, color fundus photography (CFP) and autofluorescence, near infrared reflectance imaging, spectral domain optical coherence tomography (SDOCT), visual field examination, and electrophysiological tests. SDOCT provides a rapid and non-invasive imaging evaluation of disease progression over time. Additional examination including NIR imaging can provide further information on photoreceptor alteration and subtle disruption of the RPE, which are not evident with CFP at an early stage. Electrophysiological tests provide essential results on the state of cone and rod dystrophy, which could be paramount in guiding future genetic therapies. Multimodal imaging is a valuable addition to comprehensive ophthalmological examination in the diagnosis and management of patients with SCA7.

Mesenchymal stem cell‐derived extracellular vesicles for metabolic syndrome therapy: Assessing efficacy with nuclear magnetic resonance spectroscopy

AbstractMetabolic syndrome (MetS) represents a cluster of metabolic abnormalities. The prevalence of MetS has surged, transforming it into a pressing public health concern that could potentially affect around 20%–25% of the global population. As MetS continues its ascent, diverse interventions, pharmacological, nonpharmacological and combined have been deployed. Yet, a comprehensive remedy that fully eradicates MetS symptoms remains elusive, compounded by the risks of polypharmacy's emergence. Acknowledging the imperative to grasp MetS's intricate pathologies, deeper insights for future research and therapy optimisation become paramount. Conventional treatments often target specific syndrome elements. However, a novel approach emerges in mesenchymal stem cell‐derived extracellular vesicles (MSC‐EVs) therapy, promising a holistic shift. MSC‐EVs, tiny membranous vesicles secreted by mesenchymal stem cells, have garnered immense attention for their multifaceted bioactivity and regenerative potential. Their ability to modulate inflammation, enhance tissue repair and regulate metabolic pathways has prompted researchers to explore their therapeutic application in MetS. This review primarily aims to provide an overview of how MSC‐EVs therapy can improve metabolic parameters in subjects with MetS disease and also introduce the usefulness of NMR spectroscopy in assessing the efficacy of MSC‐EVs therapy for treating MetS.

Bioinformatics analyses of potentially common pathogenic networks for primary Sjögren’s syndrome complicated with acute myocardial infarction

Both primary Sjögren’s syndrome (pSS) and acute myocardial infarction (AMI) are intricately linked. However, their common mechanism is not fully understood. Herein, we examined the underlying network of molecular action associated with developing this complication. Datasets were downloaded from the GEO database. We performed enrichment and protein–protein interaction analyses and screened key genes. We used external datasets to confirm the diagnostic performance for these hub genes. Transcription factor and microRNA regulatory networks were constructed for the validated hub genes. Finally, drug prediction and molecular docking validation were performed. We identified 62 common DEGs, many of which were enriched regarding inflammation and immune response. 5 DEGs were found as key hub genes (IGSF6, MMP9, S100A8, MNDA, and NCF2). They had high diagnostic performance in external datasets. Functional enrichment of these five hub genes showed that they were associated with the adaptive immune response. The Type 1T helper cell showed the most association among all cell types related to AMI and pSS. We identified 36 common TFs and 49 identical TF-miRNAs. The drugs, including Benzo, dexamethasone, and NADP, were predicted as potential therapeutic agents. Herein, we revealed common networks involving pSS and AMI etiologies. Knowledge of these networks and hub genes can enhance research into their associated mechanism and the development of future robust therapy.

Five families of diverse DNA viruses comprehensively restructure the nucleus.

Many viruses have evolved ways to restructure their host cell's nucleus profoundly and unexpectedly upon infection. In particular, DNA viruses that need to commandeer their host's cellular synthetic functions to produce their progeny can induce the condensation and margination of host chromatin during productive infection, a phenomenon known as virus-induced reorganization of cellular chromatin (ROCC). These ROCC-inducing DNA viruses belong to 5 families (herpesviruses, baculoviruses, adenoviruses, parvoviruses, and geminiviruses) that infect a wide range of hosts and are important for human and ecosystem health, as well as for biotechnology. Although the study of virus-induced ROCC is in its infancy, investigations are already raising important questions, such as why only some DNA viruses that replicate their genomes in the nucleus elicit ROCC. Studying the shared and distinct properties of ROCC-inducing viruses will provide valuable insights into viral reorganization of host chromatin that could have implications for future therapies that target the viral life cycle.

Bone marrow from periacetabular osteotomies as a novel source for human mesenchymal stromal cells

BackgroundBone marrow-derived mesenchymal stromal cells (BM-MSCs) are used in regenerative medicine and related research involving immunomodulatory, anti-inflammatory, anti-fibrotic and regenerative functions. Isolation of BM-MSCs from samples obtained during total hip arthroplasty (THA) is routinely possible. Advanced age and comorbidities of the majority of patients undergoing THA limit their applicability. Our study aimed to evaluate the potential of bone marrow obtained during periacetabular osteotomy (PAO) as a novel source of BM-MSCs from young donors by analyzing cell yield and cell characteristics.MethodsBone samples were obtained from the anterior Os ilium or superior Os pubis during PAO and from the femoral cavity during primary THA. Isolation of bone marrow-derived mononuclear cells (BM-MNCs) was performed by density gradient centrifugation. The samples from PAO and THA patients were compared in terms of BM-MSC yield, colony formation and the proportion of BM-MSCs within the BM-MNC population using flow cytometry analysis. The cells were characterized based on the expression of BM-MSC-specific surface markers. The functionality of the cells was compared by quantifying post-thaw viability, metabolic activity, proliferation capacity, senescence-associated beta galactosidase (SA-β-gal) expression, trilineage differentiation potential and major secretome proteins.ResultsIsolation of BM-MNCs was possible in a reliable and reproducible manner when using bone from PAO containing more than 0.24 g bone marrow. PAO patients were younger than patients of the THA group. Bone obtained during PAO contained less bone marrow and led to a lower BM-MSC number after the first cell culture passage compared to BM-MSCs obtained during THA. BM-MSCs from PAO samples are characterized by a higher proliferation capacity. This results in a higher yield in cell culture passage two, when normalized to the sample weight. BM-MSCs from PAO patients showed increased secretion of TGF-β1, TIMP2, and VEGF upon osteogenic differentiation. BM-MSCs from PAO and THA patients revealed similar results regarding the onset of SA-β-gal expression and trilineage differentiation capacity.ConclusionsWe suggest that bone obtained during PAO is a promising novel source for BM-MSCs from young donors. Limited absolute cell yield due to low sample weight must be considered in early cell culture passages and might be critical for the range of clinical applications possible for BM-MSCs from this source. The higher proliferation capacity and increased growth factor secretion of BM-MSCs from young donors may be beneficial for future regenerative cell therapies, in vitro models, and tissue engineering.