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Purpose: Meniscal injuries are common in knee surgery and often require preservation techniques to prevent secondary osteoarthritis. Despite advancements in repair techniques, some patients undergo partial meniscectomy, which can lead to postmeniscectomy syndrome. To address these challenges, meniscal substitution techniques like scaffolds have been developed. However, a comprehensive synthesis of the existing evidence through an umbrella review is lacking. Methods: A comprehensive search was conducted in the MEDLINE, Embase and Scopus databases to identify relevant systematic reviews and meta‐analyses. Studies were screened based on predefined inclusion and exclusion criteria. The quality of included studies was assessed using the AMSTAR‐2 tool. Results: A total of 17 studies met the inclusion criteria and were included in the review. Most studies focused on the use of collagen‐based scaffolds, with fewer studies evaluating synthetic scaffolds. The majority of studies (52.9%) were rated as having 'Critically Low' overall confidence, with only one study (5.9%) rated as 'High' confidence and most studies exhibiting methodological limitations, such as small sample sizes and lack of long‐term follow‐up. Despite these limitations, the majority of studies reported positive short‐term outcomes, including pain relief and functional improvement, following scaffold implantation. However, some studies noted a relatively high failure rate. Radiographically, outcomes also varied, with some studies reporting morphological deterioration of the implant seen on MRI, while others noted possible chondroprotective effects. Conclusions: Meniscal scaffold‐based approaches show promise in the management of meniscal deficiency; however, the current evidence is limited by methodological shortcomings. One notable gap in the literature is the lack of clear guidelines for patient selection and surgical technique. Future research should focus on conducting well‐designed randomized controlled trials with long‐term follow‐up to further elucidate the benefits and indications of these techniques in clinical practice. Additionally, efforts should be made to develop consensus guidelines to standardize the use of meniscal scaffolds and improve patient outcomes. Despite limited availability, synthesizing the literature on meniscal scaffold‐based approaches is crucial for understanding research, guiding clinical decisions and informing future directions. Level of Evidence: Level IV. [ABSTRACT FROM AUTHOR]

Background: Pediatric transplantation can be a stressful process for patients and caregivers. Some individuals may experience post‐traumatic stress symptoms (PTSS) and post‐traumatic growth (PTG) as a result. Although post‐traumatic stress disorder (PTSD) has been well‐studied in this population, the purpose of the present scoping review is to provide a first synthesis of the existing literature on PTG in pediatric transplant populations. Methods: We conducted a literature search of PsycINFO and Scopus in May 2023. Eligible articles must have included a sample of solid organ transplant (SOT) or stem cell transplant (SCT) recipients under age 18, siblings of recipients, or caregivers; and must have examined PTG. Results: Twenty‐three studies were identified, and nine studies met inclusion criteria and were included in the review (n = 5 cross sectional; n = 4 qualitative). Cross‐sectional studies examined demographic, mental health, and medical correlates of PTG in children and caregivers. PTG was correlated with PTSS among caregivers. Qualitative studies identified themes along each of the five factors of PTG. Conclusion: Findings overwhelmingly focused on caregiver PTG. Qualitative study findings align with the theoretical model of PTG. Additional research is needed to investigate PTG in siblings of children with a transplant and associations between PTG and medication adherence. This scoping review provides insight into positive change processes following a transplant among children and their caregivers. [ABSTRACT FROM AUTHOR]

The deformability of blood vessels in one‐dimensional blood flow models is typically described through a pressure‐area relation, known as the tube law. The most used tube laws take into account the elastic and viscous components of the tension of the vessel wall. Accurately parametrizing the tube laws is vital for replicating pressure and flow wave propagation phenomena. Here, we present a novel mathematical‐property‐preserving approach for the estimation of the parameters of the elastic and viscoelastic tube laws. Our goal was to estimate the parameters by using ovine and human in vitro data, while constraining them to meet prescribed mathematical properties. Results show that both elastic and viscoelastic tube laws accurately describe experimental pressure‐area data concerning both quantitative and qualitative aspects. Additionally, the viscoelastic tube law can provide a qualitative explanation for the observed hysteresis cycles. The two models were evaluated using two approaches: (i) allowing all parameters to freely vary within their respective ranges and (ii) fixing some of the parameters. The former approach was found to be the most suitable for reproducing pressure‐area curves. [ABSTRACT FROM AUTHOR]

Known for their high stability and surfactant properties, per‐ and polyfluoroalkyl substances (PFAS) have been widely used in a range of manufactured products. Despite being largely phased out due to concerns regarding their persistence, bioaccumulation, and toxicity, legacy PFAS such as perfluorooctanesulfonic acid (PFOS) and perfluorooctanoic acid continue to persist at high levels in the environment, posing risks to aquatic organisms. We used high‐resolution magic angle spinning nuclear magnetic resonance spectroscopy in intact zebrafish (Danio rerio) embryos to investigate the metabolic pathways altered by PFOS both before and after hatching (i.e., 24 and 72 h post fertilization [hpf], respectively). Assessment of embryotoxicity found embryo lethality in the parts‐per‐million range with no significant difference in mortality between the 24‐ and 72‐hpf exposure groups. Metabolic profiling revealed mostly consistent changes between the two exposure groups, with altered metabolites generally associated with oxidative stress, lipid metabolism, energy production, and mitochondrial function, as well as specific targeting of the liver and central nervous system as key systems. These metabolic changes were further supported by analyses of tissue‐specific production of reactive oxygen species, as well as nontargeted mass spectrometric lipid profiling. Our findings suggest that PFOS‐induced metabolic changes in zebrafish embryos may be mediated through previously described interactions with regulatory and transcription factors leading to disruption of mitochondrial function and energy metabolism. The present study proposes a systems‐level model of PFOS toxicity in early life stages of zebrafish, and also identifies potential biomarkers of effect and exposure for improved environmental biomonitoring. Environ Toxicol Chem 2024;43:896–914. © 2024 SETAC [ABSTRACT FROM AUTHOR]

Background: miRNA‐based strategies have recently emerged as a promising therapeutic approach in several neurodegenerative diseases. Unregulated cation influx is implicated in several cellular mechanisms underlying neural cell death during ischemia. The brain constitutively active isoform of transient receptor potential melastatin 7 (TRPM7) represents a glutamate excitotoxicity‐independent pathway that significantly contributes to the pathological Ca2+ overload during ischemia. Aims: In the light of these premises, inhibition of TRPM7 may be a reasonable strategy to reduce ischemic injury. Since TRPM7 is a putative target of miRNA135a, the aim of the present paper was to evaluate the role played by miRNA135a in cerebral ischemia. Therefore, the specific objectives of the present paper were: (1) to evaluate miR135a expression in temporoparietal cortex of ischemic rats; (2) to investigate the effect of the intracerebroventricular (icv) infusion of miR135a on ischemic damage and neurological functions; and (3) to verify whether miR135a effects may be mediated by an alteration of TRPM7 expression. Methods: miR135a expression was evaluated by RT‐ PCR and FISH assay in temporoparietal cortex of ischemic rats. Ischemic volume and neurological functions were determined in rats subjected to transient middle cerebral artery occlusion (tMCAo) after miR135a intracerebroventricular perfusion. Target analysis was performed by Western blot. Results: Our results demonstrated that, in brain cortex, 72 h after ischemia, miR135a expression increased, while TRPM7 expression was parallelly downregulated. Interestingly, miR135a icv perfusion strongly ameliorated the ischemic damage and improved neurological functions, and downregulated TRPM7 protein levels. Conclusions: The early prevention of TRPM7 activation is protective during brain ischemia. [ABSTRACT FROM AUTHOR]

Glioblastoma multiforme (GBM) is a lethal disease characterized by an overall survival of about 1 year, making it one of the most aggressive tumours, with very limited therapeutic possibilities. Specific biomarkers for early diagnosis as well as innovative therapeutic strategies are urgently needed to improve the management of this deadly disease. In this work, we demonstrated that vesicular galectin‐3‐binding protein (LGALS3BP), a glycosylated protein overexpressed in a variety of human malignancies, is a potential GBM disease marker and can be efficiently targeted by a specific antibody–drug conjugate (ADC). Immunohistochemical analysis on patient tissues showed that LGALS3BP is highly expressed in GBM and, compared with healthy donors, the amount of vesicular but not total circulating protein is increased. Moreover, analysis of plasma‐derived extracellular vesicles from mice harbouring human GBM revealed that LGALS3BP can be used for liquid biopsy as a marker of disease. Finally, an ADC targeting LGALS3BP, named 1959‐sss/DM4, specifically accumulates in tumour tissue, producing a potent and dose‐dependent antitumor activity. In conclusion, our work provides evidence that vesicular LGALS3BP is a potential novel GBM diagnostic biomarker and therapeutic target deserving further preclinical and clinical validation. [ABSTRACT FROM AUTHOR]

Juvenile idiopathic arthritis (JIA) is the most common chronic rheumatic condition in childhood. The disease etiology remains largely unknown; however, a key role in JIA pathogenesis is surely mediated by T cells. T‐lymphocytes activity is controlled via signals, known as immune checkpoints. Delivering an inhibitory signal or blocking a stimulatory signal to achieve immune suppression is critical in autoimmune diseases. However, the role of immune checkpoints in chronic inflammation and autoimmunity must still be deciphered. In this study, we investigated at the single‐cell level the feature of T cells in JIA chronic inflammation, both at the transcriptome level via single‐cell RNA sequencing and at the protein level by flow cytometry. We found that despite the heterogeneity in the composition of synovial CD4+ and CD8+ T cells, those characterized by PD‐1 expression were clonally expanded tissue‐resident memory (Trm)‐like cells and displayed the highest proinflammatory capacity, suggesting their active contribution in sustaining chronic inflammation in situ. Our data support the concept that novel therapeutic strategies targeting PD‐1 may be effective in the treatment of JIA. With this approach, it may become possible to target overactive T cells regardless of their cytokine production profile. [ABSTRACT FROM AUTHOR]

Background: Understanding the role of substance use is important in the equitable allocation of solid organs and may present an opportunity for improving outcomes among substance users who receive transplants. This scoping review presents findings related to substance use among pediatric and young adult transplant populations and suggests future directions. Methods: A scoping review was conducted seeking studies related to substance use in pediatric and young adult transplant populations under the age of 39 years. Studies were deemed eligible if they collected data or addressed policy concerns and participants' mean age was below 39 years. Results: Twenty‐nine studies were identified as eligible for this review. Overall, policies around substance use are largely inconsistent throughout both pediatric and adult transplant centers. Findings indicated that substance use among pediatric and young adult transplant recipients is similar to or lower than healthy peers. Few studies addressed marijuana use and opioid misuse, among other substances. Conclusions: There is a general dearth of research on substance use in this population. The current findings suggest that substance use, although relatively less common, affects eligibility for transplant, may lead to poor outcomes, and affects medication adherence. Inconsistent substance use policies in transplant centers have the potential to result in bias. However, more research is needed on the effects of substance use among pediatric and young adult transplant candidates and recipients as well as on equitable policies for organ allocation for individuals who use substances. [ABSTRACT FROM AUTHOR]

Glioblastoma (GBM) cancer stem cells (GSCs) contribute to GBM's origin, recurrence, and resistance to treatment. However, the understanding of how mRNA expression patterns of GBM subtypes are reflected at global proteome level in GSCs is limited. To characterize protein expression in GSCs, we performed in‐depth proteogenomic analysis of patient‐derived GSCs by RNA‐sequencing and mass‐spectrometry. We quantified > 10 000 proteins in two independent GSC panels and propose a GSC‐associated proteomic signature characterizing two distinct phenotypic conditions; one defined by proteins upregulated in proneural and classical GSCs (GPC‐like), and another by proteins upregulated in mesenchymal GSCs (GM‐like). The GM‐like protein set in GBM tissue was associated with necrosis, recurrence, and worse overall survival. Through proteogenomics, we discovered 252 non‐canonical peptides in the GSCs, i.e., protein sequences that are variant or derive from genome regions previously considered non‐protein‐coding, including variants of the heterogeneous ribonucleoproteins implicated in RNA splicing. In summary, GSCs express two protein sets that have an inverse association with clinical outcomes in GBM. The discovery of non‐canonical protein sequences questions existing gene models and pinpoints new protein targets for research in GBM. [ABSTRACT FROM AUTHOR]

We present a flux vector splitting method for the one and two‐dimensional shallow water equations following the approach first proposed by Toro and Vázquez for the compressible Euler equations. The resulting first‐order schemes turn out to be exceedingly simple, with accuracy and robustness comparable to that of the sophisticated Godunov upwind method used in conjunction with complete non‐linear Riemann solvers. The technique splits the full system into two subsystems, namely an advection system and a pressure system. The sought numerical flux results from fluxes for each of the subsystems. As to the source terms, there is potential for treating general source terms by incorporating them into either subsystem. In this article we show preliminary results for the case of a discontinuous bottom, incorporated into the pressure system. Results show that the resulting method is well balanced. The basic methodology, extended on 2D unstructured meshes, constitutes the building block for the construction of numerical schemes of very high order of accuracy following the ADER approach. The presented numerical schemes are systematically assessed on a carefully selected suite of test problems with reference solutions, in one and two space dimensions. The applicability of the schemes is illustrated through simulations of tsunami wave propagation in the Pacific Ocean. [ABSTRACT FROM AUTHOR]

Background: There is evidence that, after severe acute respiratory syndrome coronavirus 2 infection, male reproductive function and semen quality may be damaged Objectives: To evaluate a panel of inflammatory mediators in semen in patients recovered from coronavirus disease 2019. Material and methods: Sexually active men with previous severe acute respiratory syndrome coronavirus 2 infection and proven recovery from coronavirus disease 2019 were enrolled in a prospective cohort study. Clinical, uro‐andrological data and semen specimens were prospectively collected. For previously hospitalized coronavirus disease 2019 patients, data on serum inflammatory markers were retrospectively collected. Results: A total of 43 men were enrolled in the study. Of these, 32 men were normozoospermic, three were oligozoospermic, and eight were crypto‐azoospermic. Serum inflammatory markers (procalcitonin and C‐reactive protein) were analyzed in previously hospitalized patients both at admission and at peak of infection. Levels at admission were statistically significantly higher in patients resulting in crypto‐azoospermic with respect to those resulting in normozoospermic (p = 0.05; p = 0.03 and p = 0.02, respectively) after healing. Seminal cytokine levels were similar among all groups. Interleukin‐1β and tumor necrosis factor‐α levels were significantly negatively related to sperm total number and concentration, whereas interleukin‐4 was correlated with sperm motility. Discussion and conclusion: Negative correlations between interleukin‐1β and tumor necrosis factor‐α and sperm number and the overall high levels of semen cytokines indicate a potential detrimental role of severe acute respiratory syndrome coronavirus 2 driven inflammation on spermatogenesis. Overall, our results indicate that male patients recovering from coronavirus disease 2019 deserve accurate follow‐up for their fertility status. [ABSTRACT FROM AUTHOR]

Background: Since the start of the COVID‐19 pandemic and consequent lockdowns, the use of telehealth interventions has rapidly increased both in the general population and among transplant recipients. Among pediatric transplant recipients, this most frequently takes the form of interventions on mobile devices, or mHealth, such as remote visits via video chat or phone, phone‐based monitoring, and mobile apps. Telehealth interventions may offer the opportunity to provide care that minimizes many of the barriers of in‐person care. Methods: The present review followed the PRISMA guidelines. Sources up until October 2020 were initially identified through searches of PsycInfo® and PubMed®. Results: We identified ten papers that reported findings from adult interventions and five studies based in pediatrics. Eight of the adult publications stemmed from the same two trials; within the pediatric subset, this was the case for two papers. Studies that have looked at mHealth interventions have found high acceptability rates over the short run, but there is a general lack of data on long‐term use. Conclusions: The literature surrounding pediatric trials specifically is sparse with all findings referencing interventions that are in early stages of development, ranging from field tests to small feasibility trials. The lack of research highlights the need for a multi‐center RCT that utilizes robust measures of medication adherence and other outcome variables, with longer‐term follow‐up before telehealth interventions should be fully embraced. [ABSTRACT FROM AUTHOR]

Keywords: COVID-19; HIV; CD4; immunocompromised; monoclonal; convalescent plasma EN COVID-19 HIV CD4 immunocompromised monoclonal convalescent plasma 352 355 4 02/07/22 20220201 NES 220201 A late presenter AIDS patient with severe T cell depletion presented non-severe COVID-19 symptoms, with prolonged viral shedding. AIDS patient with severe T cell depletion achieved control but not clearance of SARS-CoV-2 infection PBMNC from the patient and one COVID-19 recovered subject were cultured for 6 h in vitro in medium alone or in presence of peptide pools covering Nucleoprotein, Membrane, and Spike proteins (SARS-CoV-2 antigens). Several cases of fully recovered SARS-CoV-2 infection in patients with genetic or pharmacological B cell depletion for the unrelated condition have been reported, suggesting that a protective immune response against SARS-CoV-2 can be produced even without humoral immunity [16, 17]. [Extracted from the article]

The third edition of Flow Cytometry Guidelines provides the key aspects to consider when performing flow cytometry experiments and includes comprehensive sections describing phenotypes and functional assays of all major human and murine immune cell subsets. Notably, the Guidelines contain helpful tables highlighting phenotypes and key differences between human and murine cells. Another useful feature of this edition is the flow cytometry analysis of clinical samples with examples of flow cytometry applications in the context of autoimmune diseases, cancers as well as acute and chronic infectious diseases. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid. All sections are written and peer‐reviewed by leading flow cytometry experts and immunologists, making this edition an essential and state‐of‐the‐art handbook for basic and clinical researchers. [ABSTRACT FROM AUTHOR]

Meniscus injuries are common and a major cause of long‐term joint degeneration and disability. Current treatment options are limited, so novel regenerative therapies or tissue engineering strategies are urgently needed. The development of new therapies is hindered by a lack of knowledge regarding the cellular biology of the meniscus and a lack of well‐established methods for studying meniscus cells in vitro. The goals of this study were to (1) establish baseline expression profiles and dedifferentiation patterns of inner and outer zone primary meniscus cells, and (2) evaluate the utility of poly(ethylene glycol) diacrylate (PEGDA) and gelatin methacrylate (GelMA) polymer hydrogels to reverse dedifferentiation trends for long‐term meniscus cell culture. Using reverse transcription‐quantitative polymerase chain reaction, we measured expression levels of putative meniscus phenotype marker genes in freshly isolated meniscus tissue, tissue explant culture, and monolayer culture of inner and outer zone meniscus cells from porcine knees to establish baseline dedifferentiation characteristics, and then compared these expression levels to PEGDA/GelMA embedded passaged meniscus cells. COL1A1 showed robust upregulation, while CHAD, CILP, and COMP showed downregulation with monolayer culture. Expression levels of COL2A1, ACAN, and SOX9 were surprisingly similar between inner and outer zone tissue and were found to be less sensitive as markers of dedifferentiation. When embedded in PEGDA/GelMA hydrogels, expression levels of meniscus cell phenotype genes were significantly modulated by varying the ratio of polymer components, allowing these materials to be tuned for phenotype restoration, meniscus cell culture, and tissue engineering applications. [ABSTRACT FROM AUTHOR]

(A) ILC2 percentages in LTP-allergic patients (LTP-AP, N = 10) and tolerant controls (TC, N = 10); (B) Correlation of the IL-4+ and IL-13+ILC2 percentages vs clinical parameters. Lipid transfer protein allergy induces complex clinical manifestations, often severe or life-threatening in food allergy (FA).1 It is characterised by an immunologic tolerance breakdown to ingested food associated with the immunological response type 2. We report for the first time ILC2 implication in LTP allergy and SLIT-Prup3 ability to modulate effector Th2 cells and ILC2 in LTP-allergic patients (LTP-AP). [Extracted from the article]

The spread of two‐dimensional numerical hydrodynamic tools for ecohydraulic applications allowed for the development of automatic habitat detection methods, adopted as predicting tools for river habitat analysis. These automatic approaches differ for the employed identification rules, such as preference curves, fuzzy rules and clustering methods. Previous research has shown promising results in the automatic identification of mesoscale habitat patches by using clustering algorithms together with numerical hydrodynamic model results. These algorithms attempt to implement and simulate some of the expert‐based requirements adopted in the field to delineate habitat patches. Spatial contiguity is one of such expert‐based requirements that has not been enforced and exploited in automatic mesohabitat identification so far. In this work, we propose a novel tool (BASEmeso) based on an agglomerative hierarchical clustering algorithm where we enforced a spatial contiguity criteria. We compare our approach with a more established method without spatial constraints, considering a synthetic river reach where the composition of mesohabitat patches is known a priori, and on three experimental river reaches, to investigate the effects of different river morphologies. Our results show that when employing a contiguity constraint, a patch's extent is better captured, different patches can be distinguished better and the distribution of patch characteristics is smoother. This holds for all investigated morphologies. Together, it suggests that including a spatial contiguity constraint can improve the automatic delineation of river mesohabitat patches. The proposed methodology could positively contribute in the development of automatic, objective and predictive meso‐scale habitat assessment workflows. [ABSTRACT FROM AUTHOR]

Objective: Although the adaptive immune response to SARS‐CoV‐2 has been characterised in the acute and early convalescent phase of the disease, few studies explore whether natural infection elicits long‐lasting immunological memory in recovered individuals. In this work, we aimed to assess the maintenance of immunological memory to SARS‐CoV‐2. Methods: We evaluated the long‐term virus‐specific cellular and humoral immune response in the members of an Italian Serie A football team, who experienced a cluster of COVID‐19 in March 2020, which was strictly evaluated in the following months. Results: Our results highlight a heterogeneous magnitude of immunological memory at 5 months after infection. Indeed, 20% of the subjects displayed a weak cellular and humoral memory to SARS‐CoV‐2, suggesting that they may be at higher risk of reinfection. In addition, a history of symptomatic COVID‐19 was associated with higher levels of SARS‐CoV‐2‐reactive CD4+ T cells and specific antibody levels than in asymptomatic individuals. Conclusion: Collectively, these data demonstrate that immunity to SARS‐CoV‐2 is maintained five months postinfection even if the magnitude of response is heterogeneous among individuals. This finding suggests that some COVID‐19‐recovered subjects may benefit from vaccination. [ABSTRACT FROM AUTHOR]

T lymphocytes accumulate in inflamed tissues of patients with chronic inflammatory diseases (CIDs) and express pro‐inflammatory cytokines upon re‐stimulation in vitro. Further, a significant genetic linkage to MHC genes suggests that T lymphocytes play an important role in the pathogenesis of CIDs including juvenile idiopathic arthritis (JIA). However, the functions of T lymphocytes in established disease remain elusive. Here we dissect the transcriptional and the clonal heterogeneity of synovial T lymphocytes in JIA patients by single‐cell RNA sequencing combined with T cell receptor profiling on the same cells. We identify clonally expanded subpopulations of T lymphocytes expressing genes reflecting recent activation by antigen in situ. A PD‐1+TOX+EOMES+ population of CD4+ T lymphocytes expressed immune regulatory genes and chemoattractant genes for myeloid cells. A PD‐1+TOX+BHLHE40+ population of CD4+, and a mirror population of CD8+ T lymphocytes expressed genes driving inflammation, and genes supporting B lymphocyte activation in situ. This analysis points out that multiple types of T lymphocytes have to be targeted for therapeutic regeneration of tolerance in arthritis. [ABSTRACT FROM AUTHOR]

Infection is a rare occurrence after revision anterior cruciate ligament reconstruction (rACLR). Because of the low rates of infection, it has been difficult to identify risk factors for infection in this patient population. The purpose of this study was to report the rate of infection following rACLR and assess whether infection is associated with patient‐ and surgeon‐dependent risk factors. We reviewed two large prospective cohorts to identify patients with postoperative infections following rACLR. Age, sex, body mass index (BMI), smoking status, history of diabetes, and graft choice were recorded for each patient. The association of these factors with postoperative infection following rACLR was assessed. There were 1423 rACLR cases in the combined cohort, with 9 (0.6%) reporting postoperative infections. Allografts had a higher risk of infection than autografts (odds ratio, 6.8; 95% CI, 0.9–54.5; p =.045). Diabetes (odds ratio, 28.6; 95% CI, 5.5–149.9; p =.004) was a risk factor for infection. Patient age, sex, BMI, and smoking status were not associated with risk of infection after rACLR. [ABSTRACT FROM AUTHOR]

How T‐helper (Th) lymphocyte subpopulations identified in synovial fluid from patients with juvenile idiopathic arthritis (JIA) (Th17, classic Th1, or nonclassic Th1) drive joint damage is of great interest for the possible use of biological drugs that inhibit the specific cytokines. Our objective was to clarify the role of such Th subpopulations in the pathogenesis of articular cartilage destruction by synovial fibroblasts (SFbs), and the effect of Th17 blockage in an animal model. SFbs were isolated from healthy subjects and patients with JIA, and peripheral blood Th lymphocytes subsets were obtained from healthy subjects. Fragments of human cartilage from healthy subjects in a collagen matrix containing JIA or normal SFbs grafted underskin in SCID mice were used to measure cartilage degradation under the effects of Th supernatants. JIA SFbs overexpress MMP9 and MMP2 and Th17 induce both MMPs in normal SFbs, while nonclassic Th1 upregulate urokinase plasminogen activator (uPA) activity. In vitro invasive phenotype of normal SFbs is stimulated with conditioned medium of Th17 and nonclassic‐Th1. In the in vivo "inverse wrap" model, normal SFbs stimulated with supernatants of Th17‐lymphocytes and nonclassic Th1 produced a cartilage invasion and degradation similar to JIA SFbs. Secukinumab inhibits the cartilage damage triggered by factors produced by Th17. [ABSTRACT FROM AUTHOR]

Summary: SARS‐CoV‐2 is responsible for a new infectious disease (COVID‐19) in which individuals can either remain asymptomatic or progress from mild to severe clinical conditions including acute respiratory distress syndrome and multiple organ failure. The immune mechanisms that potentially orchestrate the pathology in SARS‐CoV‐2 infection are complex and only partially understood. There is still paucity of data on the features of myeloid cells involved in this viral infection. For this reason, we investigated the different activation status profiles and the subset distribution of myeloid cells and their correlation with disease progression in 40 COVID‐19 patients at different stages of disease. COVID‐19 patients showed a decrease in the absolute number of plasmacytoid and myeloid dendritic cells, different subset distribution of monocytes and different activation patterns of both monocytes and neutrophils, coupled to a significant reduction of HLA‐DR monocyte levels. We found that some of these alterations are typical of all COVID‐19 patients, while some others vary at different stages of the disease and correlate with biochemical parameters of inflammation. Collectively, these data suggest that not only the lymphoid, but also the myeloid compartment, is severely affected by SARS‐CoV‐2 infection. [ABSTRACT FROM AUTHOR]

The characterization of cell‐mediated and humoral adaptive immune responses to SARS‐CoV‐2 is fundamental to understand COVID‐19 progression and the development of immunological memory to the virus. In this study, we detected T‐cells reactive to SARS‐CoV‐2 proteins M, S, and N, as well as serum virus‐specific IgM, IgA, IgG, in nearly all SARS‐CoV‐2 infected individuals, but not in healthy donors. Virus‐reactive T cells exhibited signs of in vivo activation, as suggested by the surface expression of immune‐checkpoint molecules PD1 and TIGIT. Of note, we detected antigen‐specific adaptive immune response both in asymptomatic and symptomatic SARS‐CoV‐2 infected subjects. More importantly, symptomatic patients displayed a significantly higher magnitude of both cell‐mediated and humoral adaptive immune response to the virus, as compared to asymptomatic individuals. These findings suggest that an uncontrolled adaptive immune response contribute to the development of the life‐threatening inflammatory phase of the disease. Finally, this study might open the way to develop effective vaccination strategies. [ABSTRACT FROM AUTHOR]

Tissue‐resident memory (Trm) cells are specialized components of both CD4+ and CD8+ T cell subsets that persist in peripheral nonlymphoid tissues following infections and provide fast response in case of a secondary invasion by the same pathogen. Trm cells express the surface markers CD69, CD103, and the immune checkpoint molecule PD‐1. Trm cells develop not only in the context of infections but also in tumors, where they can provide a line of defense as suggested by the positive correlation between the frequency of tumor‐infiltrating Trm cells and patients' survival. Trm cells persistence in peripheral tissues depends on their adaptation to the local microenvironment and the presence of survival factors, mainly IL‐7, IL‐15, and Notch ligands. However, the cell sources of these factors are largely unknown, especially in the context of tumors. Here, we show that head–neck squamous cell carcinoma (HNSCC) is enriched in CD4+ and CD8+ T cells with a Trm phenotype. Moreover, we show that mesenchymal stromal cells that accumulate in HNSCC are a source of survival factors and allow proper expression of Trm‐typical markers in a VCAM1‐dependent manner. [ABSTRACT FROM AUTHOR]

The underlying theme throughout this series of studies authored by the Multicenter anterior cruciate ligament (ACL) Revision Study consortium has been to determine the modifiable predictors or risk factors of long‐term outcomes of revision ACL reconstruction. The observational studies described and summarized in the manuscript are both clinically relevant and of great interest in finding out the long‐term consequences of the intervention and its relationship to the original injury. The successful completion of these studies has important implications for both therapy and future clinical trials. The identification of modifiable risk factors will play an important role in secondary prevention, while the identification of nonmodifiable risk factors will aid us in counseling our patients and making surgical decisions. Thus, we expect a profound clinical impact on patients' care. More importantly, this project represents an important step forward in bringing evidence to bear in clinical decision making in orthopedic surgery. [ABSTRACT FROM AUTHOR]

Genetically engineered mouse models (GEMMs) of cancer have proven to be of great value for basic and translational research. Although CRISPR‐based gene disruption offers a fast‐track approach for perturbing gene function and circumvents certain limitations of standard GEMM development, it does not provide a flexible platform for recapitulating clinically relevant missense mutations in vivo. To this end, we generated knock‐in mice with Cre‐conditional expression of a cytidine base editor and tested their utility for precise somatic engineering of missense mutations in key cancer drivers. Upon intraductal delivery of sgRNA‐encoding vectors, we could install point mutations with high efficiency in one or multiple endogenous genes in situ and assess the effect of defined allelic variants on mammary tumorigenesis. While the system also produces bystander insertions and deletions that can stochastically be selected for when targeting a tumor suppressor gene, we could effectively recapitulate oncogenic nonsense mutations. We successfully applied this system in a model of triple‐negative breast cancer, providing the proof of concept for extending this flexible somatic base editing platform to other tissues and tumor types. Synopsis: Most human cancers, including breast cancer, are predominantly characterized by missense mutations in driver genes. This study describes a genetically engineered mouse model, in which conditional expression of the BE3 cytidine base editor in the adult mammary gland permits installation of missense or nonsense mutations at one or multiple genomic loci. Introduction of point mutations in endogeneous genes by CRISPR‐Cas9‐mediated base editing allows for assessment of their contribution to mammary tumor formation.Rapid somatic engineering of allelic series of oncogenic missense mutations determines the relative effect size of each genetic perturbation.Biallelic inactivation of endogenous tumor suppressor genes by base editing, although bystander indels are also observed.Simultaneous base editing at multiple genomic loci using arrayed sgRNA vectors. [ABSTRACT FROM AUTHOR]

Mature T helper (Th) effector cells originate following antigen recognition by naive T precursors. The maturation process is accompanied by the acquisition of specific effector functions that distinguish at least three different T helper subsets: Th1, Th2 and Th17. In general, maturation of somatic cells is accompanied by terminal differentiation. However, accumulating evidence shows that effector T cells retain a certain degree of plasticity. This is especially true for Th17 cells, which have been shown to converge towards other phenotypes in response to specific microenvironmental pressure. In this review we will discuss the experimental evidence that supports the hypothesis of Th17 plasticity, with particular emphasis on the generation of Th17‐derived 'non‐classic' Th1 cells, and the molecular networks that control it. Moreover, we will consider why Th17 plasticity is important for host protection, but also why it can have pathogenic functions during chronic inflammation. Regarding the last point, we will discuss a possible role for biological drugs in the control of Th17 plasticity and disease course. [ABSTRACT FROM AUTHOR]

Celiac disease (CD) is characterized by a spectrum of intestinal inflammatory lesions. Most patients have villous atrophy (overt‐CD), while others have a morphologically normal mucosa, despite the presence of CD‐specific autoantibodies (potential‐CD). As the mechanism responsible for villous atrophy is not completely elucidated, we investigated biomarkers specific for the different celiac lesions. Phenotype and cytokine production of intestinal mucosa cells were analyzed by flow cytometry in gut biopsies of children with overt‐ or potential‐CD and in healthy controls. Density of TCRγδ+ T cells was found markedly enhanced in intestinal mucosa of children with overt‐CD compared to potential‐CD or controls. By contrast, very few IL4+ T cells infiltrated the mucosa with villous atrophy compared to morphologically normal mucosa. IL4+ T cells were classical CD4+ T‐helper cells (CD161−), producing or not IFN‐γ, and negative for IL17A. Our study demonstrated that the transition to villous atrophy in CD patients is characterized by increased density of TCRγδ+ T cells, and concomitant disappearance of IL4+ cells. These findings suggest that immunomodulatory mechanisms are active in potential‐CD to counteract the inflammatory cascade responsible of villous atrophy. Further studies are required to validate the use of IL4+ and TCRγδ+ T cells as biomarkers of the different CD forms. [ABSTRACT FROM AUTHOR]

These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer‐reviewed by leading experts in the field, making this an essential research companion. [ABSTRACT FROM AUTHOR]

During the last decades, progression of research has led to great achievements for treatment and therapy of several disabling disorders, particularly in the field of chronic inflammatory diseases. The increased knowledge of the molecular mechanisms operating in such diseases has represented the first step in this process, and the discovery of molecules able to interfere with the natural history of the diseases, has been the second. This review is focused on the effects of biologics on type 2 diseases such as asthma, chronic urticaria and atopic dermatitis, both biologics just approved for clinical application and also those that are currently undergoing clinical trials. We will also discuss aspects and emphasize clinical trials and recently published studies, as well as research that is currently in the progress, which will be highly relevant for basic immunologists. Likewise, we will cover aspects that are pertinent for clinical immunologists and highlight translational studies that are evaluating novel biologicals in animal models. [ABSTRACT FROM AUTHOR]

It is well accepted that Th17 cells are a highly plastic cell subset that can be easily directed toward the Th1 phenotype in vitro and also in vivo during inflammation. However, there is an ongoing debate regarding the reverse plasticity (conversion from Th1 to Th17). We show here that ectopic ROR‐γt expression can restore or initiate IL‐17 expression by non‐classic or classic Th1 cells, respectively, while common pro‐Th17 cytokine cocktails are ineffective. This stability of the Th1 phenotype is at least partially due to the presence of a molecular machinery governed by the transcription factor Eomes, which promotes IFN‐γ secretion while inhibiting the expression of ROR‐γt and IL‐17. By using a mouse model of T cell‐dependent colitis we demonstrate that Eomes controls non‐classic Th1 cell development also in vivo and promotes their pathogenic potential. Eomes expression associates to a highly inflammatory phenotype also in patients with juvenile idiopathic arthritis. Indeed, it favors the acquisition of a cytotoxic signature, and promotes the development of IFN‐γ+GM‐CSF+ cells that have been described to be pathogenic in chronic inflammatory disorders. Chronic inflammation leads to conversion of Th17 cells towards a non‐classic Th1 phenotype. We show in vitro and in vivo in a mouse model of colitis and in juvenile idiopathic arthritis patients that this process is Eomes‐dependent. Eomes acts inhibiting RORC2 and IL17 expression, while contemporary promoting IFNG and CSF2. [ABSTRACT FROM AUTHOR]

Even if omalizumab is broadly used in the treatment of severe, allergic asthma, the immunological effects in long‐term treated patients have not been fully elucidated. To this aim, a cohort of 15 allergic asthmatic patients treated with omalizumab for at least three years was compared with 12 allergic asthma patients treated with standard therapy. Omalizumab treated asthmatic patients showed lower frequencies of circulating plasmacytoid DCs, and lower CD154 expression on CD4 T‐helper cells than the control group. Moreover, basophils and DCs from omalizumab‐treated patients had lower surface expression of IgE compared to the control group. In a longitudinal evaluation of two patients that started omalizumab treatment, we show that FcεRI free of IgE were evident on basophils just after four weeks of drug administration. Finally, in vitro experiments with basophils obtained from healthy donors confirm that omalizumab is able to detach IgE from high affinity IgE receptors. Collectively these data indicate that long‐term omalizumab treatment dampens type 2 inflammation acting on different cell types that play a pivotal role in the pathogenesis of allergic asthma. Moreover, we have identified a further mechanism of action of omalizumab, such as the ability to detach IgE from its receptor. In order to define the immunological effects of the anti‐IgE mAb omalizumab, a group severe allergic asthma patients treated with omalizumab (OTP), was compared with a group of allergic asthma patients treated with standard therapy (STP). We found that omalizumab dampens type 2 inflammation in OTP. Moreover, beyond the well‐known ability of Omalizumab to bind to circulating IgE and prevent their interactions with FcεRI, we showed that omalizumab dissociate prebound IgE from its receptor on basophils. Both these mechanisms participate to the reduction of FcεRI bound IgE and FcεRI downregulation. [ABSTRACT FROM AUTHOR]

Abstract: Objectives: We employed the correspondence analysis (CA) biplot to estimate correlations between gender–age levels of cardiovascular disease patients and their psychiatric and physical symptoms. Utilization of this correlation estimation can inform clinical practice by elucidating associations between certain psychiatric or physical symptoms and specific gender–age levels. Method: The CA biplot utilized here was designed to visually inspect row–column category associations in a 2‐dimensional plane and then to numerically estimate the category associations with correlations. To do so, we (a) estimated dimensions from row and column categories with CA; (b) verified statistical significance of dimensions with a permutation test; (c) projected row and column categories in a plan constructed with the first 2 dimensions that were statistically significant; (d) visually inspected category associations in the plane; and (e) numerically estimated category associations with correlations. Results: Consistent with the previous results, female cardiovascular disease patients were more likely to experience psychiatric symptoms than the male patients. However, when examining the results by gender and age, both female and male patients in their 50s and 60s tended to experience elevated rates of the psychiatric symptoms. Conclusions: The CA biplot can be useful for isolating key clinical concerns among any medical populations. [ABSTRACT FROM AUTHOR]

Abstract: We investigated differential treatment effects on specific eating disorder (ED) indicators to enhance conclusions about treatment efficacy. Profile Analysis via Multidimensional Scaling, which identifies core profiles in a population and interprets person profiles with core profile information, was utilized to identify core profiles from a sample of 5,177 patients who were repeatedly measured with the ED inventory‐2 at admission and at discharge. To assess differential treatment effects for individual ED indicators, we compared the core profiles at admission with those at discharge. Three core profiles were identified and labeled as High Body Dissatisfaction with Low Bulimia (Core Profile 1), High Interoceptive Awareness with Low Body Dissatisfaction (Core Profile 2), and High Ineffectiveness with Low Bulimia (Core Profile 3). Treatment had the greatest effects on Core Profile 2. The patients whose profile patterns were similar to that of Core Profiles 1 and 2 were positively related with weight gain. However, treatment was least on Core Profile 3, and the patients whose profile patterns were like that of Core Profile 3 were negatively related with weight gain. In conclusion, those patients who fit Core Profile 3 may benefit from different treatment modalities than those that are standard in inpatient settings. [ABSTRACT FROM AUTHOR]

Abstract: Non‐adherence to immunosuppressant medications is an important risk factor for graft dysfunction. To evaluate the effectiveness of adherence‐enhancing interventions, we reviewed adherence intervention studies in solid organ transplant recipients (all ages). Using the following databases: PsycINFO, PubMed, Scopus, and ScienceDirect, we identified 41 eligible studies. Only three non‐randomized trials showed a possible positive effect on objective indicators of transplant outcomes (such as rejection, liver enzyme levels, kidney function). None of the 21 RCTs showed an improvement in transplant outcomes. Three studies showed a higher rate of adverse events in the intervention group as compared with controls, although this may be related to ascertainment bias. Improvement in adherence as measured indirectly (eg, with electronic monitoring devices) was not aligned with effects on transplant outcomes. We conclude that adherence interventions, to date, have largely been ineffective in improving transplant outcomes. To improve this track record, intervention efforts may wish to concentrate on non‐adherent patients (rather than use convenience sampling, which excludes many of the patients who need the intervention), use direct measures of adherence to guide the interventions, and employ strategies that are intensive and yet engaging enough to ensure that non‐adherent patients are able to participate. [ABSTRACT FROM AUTHOR]

ABSTRACT Interfacial polymerization is used to prepare biobased furan polyamides from the carbohydrate-derived monomer, 2,5-furan dicarboxylic acid, aromatic diamines, and varying chain length aliphatic diamines. The molecular weights of the furan polyamides variations range 10,000-70,000 g/mol. These biobased polyamides have improved solubility relative to petroleum-derived polyamides affording enhanced processability options. The glass transition temperatures ( Tg) of the biobased furan polyamides are higher than that of aliphatic analogs, but lower than phenyl-aromatic analogs. The Tg for these furan polyamides are as high as 280 °C. Also, the furan polyamide glass transition temperatures increase with decreasing aliphatic diamine chain length similar to results exemplified in petroleum-based nylons. Group contribution parameters are determined for furans to enable simple prediction of the glass transition temperature and decomposition temperature of furan polyamides. The molar glass transition function for the furan is calculated to be 27.6 ± 1.5 K kg/mol. Thermal analysis measurements of the biobased furan polyamides have maximum thermal degradation temperatures at 350 °C or higher, similar to that of aliphatic polyamides when scaled with the number average molecular weight. The molar decomposition temperature functions are determined to be 37 K kg/mol for furans bonded to aliphatic units and 42 K kg/mol for furans bonded to phenyl units. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017, 134, 45514. [ABSTRACT FROM AUTHOR]

Medication adherence is an important determinant of transplant outcomes. Attempts to investigate adherence are frequently undermined by selection bias: It is very hard to recruit and retain non-adherent patients in research efforts. This manuscript presents recruitment strategies and results from the MALT (Medication Adherence in children who had a Liver Transplant) multisite prospective cohort study. MALT sites recruited 400 pediatric liver transplant patients who agreed to be followed for 2 years. The primary purpose was to determine whether a marker of adherence, the Medication Level Variability Index (MLVI), predicts rejection outcomes. The present manuscript describes methods used in MALT to ensure that a representative sample was recruited, and presents detailed recruitment results. MALT sites were able to recruit a nationally representative sample, as determined by a comparison between the MALT cohort and a national sample of transplant recipients. Strategies that helped ensure that the sample was representative included monitoring of the outcome measure in comparison with a national sample, drastically limiting patient burden, and specific recruitment methods. We discuss the importance of a representative sample in adherence research and recommend that future efforts to study adherence pay special attention to sample characteristics. [ABSTRACT FROM AUTHOR]

We recently demonstrated that human T-helper (Th) 17 cells, unlike Th1 cells, do not proliferate in response to T-cell receptor stimulation, mainly because of their reduced capacity to produce and respond to IL-2. In this study, we show that their lower responsiveness to IL-2 is due to the selective expression of Musculin (MSC), a member of the basic helix-loop-helix transcription factors. We show that MSC expression in human Th17 cells is retinoic acid orphan receptor (ROR)γt-dependent, and allows the upregulation of PPP2R2B, a regulatory member of the protein phosphatase 2A (PP2A) enzyme. High PPP2R2B levels in human Th17 cells were responsible for the reduced STAT5B Ser-193 phosphorylation upon IL-2 signalling and, therefore, impaired STAT5B DNA binding and transcriptional activity on IL-2 target genes. PP2A, observed in Th17 cells, controls also STAT3, dephosphorylating Ser727, thus increasing its activity that plays a crucial role in Th17 development and/or maintenance. Thus, our findings identify an additional mechanism responsible for the limited expansion of human Th17 cells, and could provide a further explanation for the rarity of these cells in inflamed tissues. [ABSTRACT FROM AUTHOR]

A growing number of older adults are undergoing liver transplantation ( LT) in the United States. In some settings, it is thought that adherence declines with age. This retrospective study examined adherence and clinical outcomes in older vs younger adult LT recipients. Medical records of adult LT recipients from 2009 to 2012 from a single urban center were reviewed. The medication level variability index ( MLVI) was the predefined primary outcome, with nonadherence defined as MLVI >2.5. The secondary outcome was incidence of rejection. Outcomes were evaluated starting 1 year post- LT until 2015. A total of 42 of 248 patients were ≥65 at transplant. Older adults had significantly better adherence than younger ones (65%≥65 were adherent vs 42% younger adults; chi-square two-tailed P=.02). Survival analyses of rejection between age groups censored by time since transplant showed no difference among the four age groups (χ2=0.84, P=.84). Older age was not found to be a risk factor for reduced adherence or graft rejection in patients surviving at least 1 year post- LT. [ABSTRACT FROM AUTHOR]

Lysosomal storage disorders ( LSDs) are inherited diseases characterized by lysosomal dysfunction and often showing a neurodegenerative course. There is no cure to treat the central nervous system in LSDs. Moreover, the mechanisms driving neuronal degeneration in these pathological conditions remain largely unknown. By studying mouse models of LSDs, we found that neurodegeneration develops progressively with profound alterations in presynaptic structure and function. In these models, impaired lysosomal activity causes massive perikaryal accumulation of insoluble α-synuclein and increased proteasomal degradation of cysteine string protein α ( CSPα). As a result, the availability of both α-synuclein and CSPα at nerve terminals strongly decreases, thus inhibiting soluble NSF attachment receptor ( SNARE) complex assembly and synaptic vesicle recycling. Aberrant presynaptic SNARE phenotype is recapitulated in mice with genetic ablation of one allele of both CSPα and α-synuclein. The overexpression of CSPα in the brain of a mouse model of mucopolysaccharidosis type IIIA, a severe form of LSD, efficiently re-established SNARE complex assembly, thereby ameliorating presynaptic function, attenuating neurodegenerative signs, and prolonging survival. Our data show that neurodegenerative processes associated with lysosomal dysfunction may be presynaptically initiated by a concomitant reduction in α-synuclein and CSPα levels at nerve terminals. They also demonstrate that neurodegeneration in LSDs can be slowed down by re-establishing presynaptic functions, thus identifying synapse maintenance as a novel potentially druggable target for brain treatment in LSDs. [ABSTRACT FROM AUTHOR]