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Abstract In Multiple Sclerosis (MS), inflammatory demyelinated lesions in the brain and spinal cord lead to neurodegeneration and progressive disability. Remyelination can restore fast saltatory conduction and neuroprotection but is inefficient in MS especially with increasing age, and is not yet treatable with therapies. Intrinsic and extrinsic inhibition of oligodendrocyte progenitor cell (OPC) function contributes to remyelination failure, and we hypothesised that the transplantation of ‘improved’ OPCs, genetically edited to overcome these obstacles, could improve remyelination. Here, we edit human(h) embryonic stem cell-derived OPCs to be unresponsive to a chemorepellent released from chronic MS lesions, and transplant them into rodent models of chronic lesions. Edited hOPCs display enhanced migration and remyelination compared to controls, regardless of the host age and length of time post-transplant. We show that genetic manipulation and transplantation of hOPCs overcomes the negative environment inhibiting remyelination, with translational implications for therapeutic strategies for people with progressive MS.

Abstract Neonatal sepsis is a major cause of childhood mortality. Limited diagnostic tools and mechanistic insights have hampered our abilities to develop prophylactic or therapeutic interventions. Biomarkers in human neonatal sepsis have been repeatedly identified as associated with dysregulation of angiopoietin signaling and altered arachidonic acid metabolism. We here provide the mechanistic evidence in support of the relevance for these observations. Angiopoetin-1 (Ang-1), which promotes vascular integrity, was decreased in blood plasma of human and murine septic newborns. In preclinical models, administration of Ang-1 provided prophylactic protection from septic death. Arachidonic acid metabolism appears to be functionally connected to Ang-1 via reactive oxygen species (ROS) with a direct role of nitric oxide (NO). Strengthening this intersection via oral administration of arachidonic acid and/or the NO donor L-arginine provided prophylactic as well as therapeutic protection from septic death while also increasing plasma Ang-1 levels among septic newborns. Our data highlight that targeting angiogenesis-associated pathways with interventions that increase Ang-1 activity directly or indirectly through ROS/eNOS provide promising avenues to prevent and/or treat severe neonatal sepsis.

Abstract Background Proximal humeral fractures (PHFs) are still controversial with regards to treatment and are difficult to classify. The study’s objective is to show that preoperative planning performed while handling a three-dimensional (3D) printed anatomical model of the fracture can ensure a better understanding of trauma for both surgeons and patients. Materials and methods Twenty patients (group A, cases) with complex PHF were evaluated preoperatively by reproducing life-size, full-touch 3D anatomical models. Intraoperative blood loss, radiographic controls, duration of surgery, and clinical outcomes of patients in group A were compared with 20 patients (group B, controls) who underwent standard preoperative evaluation. Additionally, senior surgeons and residents, as well as group A patients, answered a questionnaire to evaluate innovative preoperative planning and patient compliance. Cost analysis was evaluated. Results Intraoperative radiography controls and length of operation were significantly shorter in group A. There were no differences in clinical outcomes or blood loss. Patients claim a better understanding of the trauma suffered and the proposed treatment. Surgeons assert that the planning of the definitive operation with 3D models has had a good impact. The development of this tool has been well received by the residents. The surgery was reduced in length by 15%, resulting in savings of about EUR 400 for each intervention. Conclusions Fewer intraoperative radiography checks, shorter surgeries, and better patient compliance reduce radiation exposure for patients and healthcare staff, enhance surgical outcomes while reducing expenses, and lower the risk of medicolegal claims. Level of evidence Level I, prospective randomized case–control study.

Introduction: Erythroblastic island (EBI) macrophages play an essential role in the production and maturation of the vast numbers of red blood cells (RBCs) that are produced throughout life. Their location within the bone marrow makes it difficult to study the cellular and molecular interactions associated with their action so we have used an in vitro model of the EBI niche using macrophages derived from human induced pluripotent stem cells (hiPSCs). We previously demonstrated that the activation of the transcription factor KLF1 enhanced the activity of hiPSC-derived EBI macrophages.Methods: To elucidate the mechanisms associated with EBI-like activity we carried out a quantitative proteomic analysis and assessed the role of extracellular vesicles using Nanosight Tracking analyses and media filtration.Results and Discussion: Gene ontology analysis showed that many of the proteins upregulated by KLF1 were protein-binding factors, some of which were associated with the cell membrane or extracellular vesicles We demonstrated that filtration of macrophage-conditioned media resulted in a reduction in the supportive effects on erythroid cell viability and maturation implying a role for extracellular vesicles but this was not KLF1 dependent. Pathway analyses of the proteomic data revealed that proteins upregulated by KLF1 were associated with the citric acid cycle, pyruvate metabolism and ATP synthesis indicating that KLF1-activated macrophages had a metabolic profile comparable to a pro-reparative phenotype. This study has generated a proteomic dataset that could provide new insights into the role of macrophages within the EBI niche and has indicated a potential role for extracellular vesicles in the differentiation and maturation of RBCs in vitro. Further research will aid in the production of RBCs in vitro for use in disease modelling and cell therapy.

Abstract Uncovering the mechanisms regulating hematopoietic specification not only would overcome current limitations related to hematopoietic stem and progenitor cell (HSPC) transplantation, but also advance cellular immunotherapies. However, generating functional human induced pluripotent stem cell (hiPSC)-derived HSPCs and their derivatives has been elusive, necessitating a better understanding of the developmental mechanisms that trigger HSPC specification. Here, we reveal that early activation of the Nod1-Ripk2-NF-kB inflammatory pathway in endothelial cells (ECs) primes them to switch fate towards definitive hemogenic endothelium, a pre-requisite to specify HSPCs. Our genetic and chemical embryonic models show that HSPCs fail to specify in the absence of Nod1 and its downstream kinase Ripk2 due to a failure on hemogenic endothelial (HE) programming, and that small Rho GTPases coordinate the activation of this pathway. Manipulation of NOD1 in a human system of definitive hematopoietic differentiation indicates functional conservation. This work establishes the RAC1-NOD1-RIPK2-NF-kB axis as a critical intrinsic inductor that primes ECs prior to HE fate switch and HSPC specification. Manipulation of this pathway could help derive a competent HE amenable to specify functional patient specific HSPCs and their derivatives for the treatment of blood disorders.

• Purpose: Partial wrist denervation can be performed by isolated posterior interosseous nerve (PIN) or combined PIN plus (+) anterior interosseous nerve (AIN) neurectomy procedures. • The purpose of the current systematic review is to investigate any differences in clinical outcomes and failures in patients undergoing AIN + PIN vs isolated PIN neurectomy. • Methods: A review of the English Literature was performed on Medline, WOS and Scopus according to PRISMA protocol combining ‘wrist denervation’, ‘PIN neurectomy’, ‘AIN neurectomy’, anterior interosseous nerve neurectomy’ and ‘posterior interosseous nerve neurectomy’. Studies were assessed with a modified Coleman Methodology Score (CMS). The primary outcome for meta-analysis was ‘Failures’, including all patients who have required a second surgery or those who are left with pain (defined as ‘bad’). • Results: Overall, 10 studies totalling 347 wrists were included in this systematic review, with a ‘moderate’ CMS. The isolated PIN neurectomy technique showed a 15.1% pooled failure rate at a median follow-up of 22 months, while the combined AIN+PIN denervation had a pooled failure rate of 23.6% at a follow-up with a median of 29 months. The combined analysis of both procedures did not show significantly better results in favour of either technique, with a general failure rate of 21.6% (P = 0.0501). • Conclusion: Partial denervation for chronic wrist pain is a salvage procedure that leads to an overall success of 78.4% for pain relief, with no substantial complications. Apparently, performing the neurectomy also of the AIN does not offer greater advantages compared to the isolated PIN neurectomy.

Background: There are wide variations in antibiotic use in neonatal intensive care units (NICUs). Limited data are available on antimicrobial stewardship (AS) programs and long-term maintenance of AS interventions in preterm very-low-birth-weight (VLBW) infants. Methods: We extended a single-centre observational study carried out in an Italian NICU. Three periods were compared: I. “baseline” (2011–2012), II. “intervention” (2016–2017), and III. “maintenance” (2020–2021). Intensive training of medical and nursing staff on AS occurred between periods I and II. AS protocols and algorithms were maintained and implemented between periods II and III. Results: There were 111, 119, and 100 VLBW infants in periods I, II, and III, respectively. In the “intervention period”, there was a reduction in antibiotic use, reported as days of antibiotic therapy per 1000 patient days (215 vs. 302, p < 0.01). In the “maintenance period”, the number of culture-proven sepsis increased. Nevertheless, antibiotic exposure of uninfected VLBW infants was lower, while no sepsis-related deaths occurred. Our restriction was mostly directed at shortening antibiotic regimens with a policy of 48 h rule-out sepsis (median days of early empiric antibiotics: 6 vs. 3 vs. 2 in periods I, II, and III, respectively, p < 0.001). Moreover, antibiotics administered for so-called culture-negative sepsis were reduced (22% vs. 11% vs. 6%, p = 0.002), especially in infants with a birth weight between 1000 and 1499 g. Conclusions: AS is feasible in preterm VLBW infants, and antibiotic use can be safely reduced. AS interventions, namely, the shortening of antibiotic courses in uninfected infants, can be sustained over time with periodic clinical audits and daily discussion of antimicrobial therapies among staff members.

Methods to evaluate soil water repellency (SWR) require extensive studies on field soils and are subject to the heterogeneity of SWR throughout the soil profile as well as plant/soil interactions. The objectives of this study were to develop a synthetic method to create hydrophobic sand, and to determine if that hydrophobic sand would affect the establishment of bermudagrass (Cynodon dactylon L. Pers. × C. transvaalensis Burtt-Davy, cv. Tifeagle) sprigs. Two techniques were developed to render sand hydrophobic: soap:sand method (hydrophobic sand; HSS) and sand:peat method (hydrophobic sand and read sedge peat; HSP). Both HSS and HSP remained severely hydrophobic at 0 cm depth for only 7 d, and at the 1- to 6-cm depth for 77 continuous days, as determined by water drop penetration time. Bermudagrass establishment, root growth, or shoot growth in two greenhouse experiments with four root zone substrates–HSS, HSP, WSAND (wettable sand), and WSP (wettable sand and reed sedge peat)—were not consistent. In conclusion, both HSS and HSP were shown to be safe and effective methods to synthetically produce hydrophobic sand for potential use in laboratory research, but further evaluation is needed to determine the feasibility of using HSS and HSP for turfgrass growth evaluations.

BackgroundThe aim of this randomized clinical trial (RCT) was to compare immunological changes in virally suppressed people living with HIV (PLWH) switching from a three-drug regimen (3DR) to a two-drug regimen (2DR).MethodsAn open-label, prospective RCT enrolling PLWH receiving a 3DR who switched to bictegravir/emtricitabine/tenofovir alafenamide (B/F/TAF) or dolutegravir/lamivudine (DTG/3TC) was performed. Blood was taken at baseline and months 6 and 12. The primary outcome was the change in CD4+ or CD8+ T-cell counts and CD4/CD8 ratio over time points. The secondary outcomes were the changes in immunological and inflammatory parameters. Parametric mixed-linear models with random intercepts and slopes were fitted separately for each marker after controlling for potential confounders.ResultsBetween the two arms (33 PLWH each), there was no difference in CD4+ or CD8+ T cells, CD4/CD8 ratio, and IL-6 trajectories. PLWH switching to DTG/3TC had increased levels of both transitional memory and terminally differentiated CD4+ T cells (arm–time interaction p-value = 0.02) and to a lesser extent for the corresponding CD8+ T-cell subsets (p = 0.09). Significantly lower levels of non-classical monocytes were detected in the B/F/TAF arm at T6 (diff = −6.7 cells/mm3; 95% CI; −16, +2.6; p-value for interaction between arm and time = 0.03). All differences were attenuated at T12.ConclusionNo evidence for a difference in absolute CD4+ and CD8+ T-cell counts, CD4/CD8 ratio, and IL-6 trajectories by study arm over 12 months was found. PLWH on DTG/3TC showed higher levels of terminally differentiated and exhausted CD4+ and CD8+ T lymphocytes and non-classical monocytes at T6. Further studies are warranted to better understand the clinical impact of our results.Clinical Trial Registrationhttps://clinicaltrials.gov, identifier NCT04054089.

De nombreuses classes visitent chaque année avec leur enseignant·e le site d’Augusta Raurica. Le présent article traite dans sa deuxième partie d’une activité en particulier réalisée lors de la visite d’un groupe composé de plusieurs classes : les élèves plus âgés ont aidé les plus jeunes à déchiffrer l’épitaphe du fondateur de la colonie romaine. Le scénario à la base de cette activité est présenté dans la première partie, tout comme les notions et concepts didactiques sur lesquels il se fonde.

Resumen: Este trabajo busca visibilizar la diversidad religiosa en Buenos Aires, ciudad que al igual que el resto de la Argentina estuvo históricamente marcada por el catolicismo. Esta visibilización se indaga a través de tres perspectivas. En primer lugar, se detectan diferencias y especificidades, comparando indicadores estadísticos de creencia, autoafiliación y práctica religiosa de territorios al interior de la ciudad, entre Buenos Aires y con otros aglomerados del país. Intenta luego profundizar la detección de la diversidad religiosa local desde una óptica socio-histórica. Por último, se problematizan los modos de cuantificar la relevancia de los diversos grupos religiosos, a través de un abordaje territorial por medio de una contabilidad de los lugares de cultos presentes actualmente en la ciudad y luego desde quienes asisten de modo frecuente al culto.

A systematic monograph of the Trembleya s.s. clade is presented, a Brazilian endemic lineage of Melastomataceae comprising 11 species and currently recognised as part of Microlicia s.l. (Melastomataceae). First, we investigate phylogenetic relationships within Lavoisiereae using two nuclear markers and two sampling datasets (102 and 134 terminals). Then, we provide a systematic revision and new circumscription of the Trembleya s.s. clade, including line drawings, photos of living specimens, leaves and floral parts, distribution maps, a key to the 11 accepted species, comments on morphology, reproductive biology, richness, endemism, biogeography and recommended conservation assessments. A nomenclatural update of all taxa previously treated in Trembleya is also provided, including the designation of 45 lectotypes and the proposal of 38 new synonyms.

Biofilms are multicellular aggregates of bacteria immersed in an extracellular matrix that forms on various surfaces, including biological tissues and artificial surfaces. However, more and more reports point out the fact that even biological fluids and semifluid, such as synovial liquid, blood, urine, or mucus and feces, harbor “non-attached” biofilm aggregates of bacteria, which represent a significant phenomenon with critical clinical implications that remain to be fully investigated. In particular, biofilm aggregates in biological fluid samples have been shown to play a relevant role in bacterial count and in the overall accuracy of microbiological diagnosis. In line with these observations, the introduction in the clinical setting of fluid sample pretreatment with an antibiofilm chemical compound called dithiothreitol (DTT), which is able to dislodge microorganisms from their intercellular matrix without killing them, would effectively improve the microbiological yield and increase the sensitivity of cultural examination, compared to the current microbiological techniques. While other ongoing research continues to unveil the complexity of biofilm formation in biological fluids and its impact on infection pathogenesis and diagnosis, we here hypothesize that the routine use of a chemical antibiofilm pretreatment of fluid and semi-solid samples may lead to a paradigm shift in the microbiological approach to the diagnosis of biofilm-related infections and should be further investigated and eventually implemented in the clinical setting.

Patients over the age of 70 show inflammaging and a weaker anti-viral response to SARS-CoV-2, pointing at the immunological changes associated with COVID-19 severity and outcome for aged patients.

Self-amplifying mRNA vaccines offer the benefit of driving potent immune responses at low doses, as the mRNA replicates intracellularly. Here, the authors report the preclinical evaluation of a self-amplifying mRNA SARS-CoV-2 vaccine in non-human primates.

IntroductionA growing number of evidences suggest that the combination of hyperinflammation, dysregulated T and B cell response and cytokine storm play a major role in the immunopathogenesis of severe COVID-19. IL-6 is one of the main pro-inflammatory cytokines and its levels are increased during SARS-CoV-2 infection. Several observational and randomized studies demonstrated that tocilizumab, an IL-6R blocker, improves survival in critically ill patients both in infectious disease and intensive care units. However, despite transforming the treatment options for COVID-19, IL-6R inhibition is still ineffective in a fraction of patients.MethodsIn the present study, we investigated the impact of two doses of tocilizumab in patients with severe COVID-19 who responded or not to the treatment by analyzing a panel of cytokines, chemokines and other soluble factors, along with the composition of peripheral immune cells, paying a particular attention to T and B lymphocytes.ResultsWe observed that, in comparison with non-responders, those who responded to tocilizumab had different levels of several cytokines and different T and B cells proportions before starting therapy. Moreover, in these patients, tocilizumab was further able to modify the landscape of the aforementioned soluble molecules and cellular markers.ConclusionsWe found that tocilizumab has pleiotropic effects and that clinical response to this drug remain heterogenous. Our data suggest that it is possible to identify patients who will respond to treatment and that the administration of tocilizumab is able to restore the immune balance through the re-establishment of different cell populations affected by SARS-COV-2 infection, highlighting the importance of temporal examination of the pathological features from the diagnosis.

Introduction: Congenital dyserythropoietic anaemia (CDA) type IV has been associated with an amino acid substitution, Glu325Lys (E325K), in the transcription factor KLF1. These patients present with a range of symptoms, including the persistence of nucleated red blood cells (RBCs) in the peripheral blood which reflects the known role for KLF1 within the erythroid cell lineage. The final stages of RBCs maturation and enucleation take place within the erythroblastic island (EBI) niche in close association with EBI macrophages. It is not known whether the detrimental effects of the E325K mutation in KLF1 are restricted to the erythroid lineage or whether deficiencies in macrophages associated with their niche also contribute to the disease pathology.Methods: To address this question, we generated an in vitro model of the human EBI niche using induced pluripotent stem cells (iPSCs) derived from one CDA type IV patient as well as two iPSC lines genetically modified to express an KLF1-E325K-ERT2 protein that could be activated with 4OH-tamoxifen. The one patient iPSC line was compared to control lines from two healthy donors and the KLF1-E325K-ERT2 iPSC line to one inducible KLF1-ERT2 line generated from the same parental iPSCS.Results: The CDA patient-derived iPSCs and iPSCs expressing the activated KLF1-E325K-ERT2 protein showed significant deficiencies in the production of erythroid cells with associated disruption of some known KLF1 target genes. Macrophages could be generated from all iPSC lines but when the E325K-ERT2 fusion protein was activated, we noted the generation of a slightly less mature macrophage population marked by CD93. A subtle trend in their reduced ability to support RBC enucleation was also associated with macrophages carrying the E325K-ERT2 transgene.Discussion: Taken together these data support the notion that the clinically significant effects of the KLF1-E325K mutation are primarily associated with deficiencies in the erythroid lineage but it is possible that deficiencies in the niche might have the potential to exacerbate the condition. The strategy we describe provides a powerful approach to assess the effects of other mutations in KLF1 as well as other factors associated with the EBI niche.

Brown patch (Rhizoctonia spp.) is a major disease of turf‐type tall fescue (TF) [Schedonorus arundinaceus (Schreb.) Dumort., nom. cons.]. Many cool‐season turfgrass lawns consist of species mixtures or cultivar blends, but the exact proportion of resistant cultivars in blends and mixtures to effectively reduce disease has not been well documented. A field study was conducted in West Lafayette, IN, and Blacksburg, VA, during 2022 and 2023 to determine the brown patch severity of various blend ratios (0%, 25%, 50%, 75%, and 100% by weight) using a brown patch susceptible and resistant TF cultivar. Additionally, mixtures (90% and 10% by weight, respectively) of TF and Kentucky bluegrass (Poa pratensis L.) with a susceptible and resistant TF cultivar were evaluated. Seasonal appearance/turf quality and brown patch severity were visually determined, and area under the disease progress curve (AUDPC) was calculated. Turf quality and brown patch severity were similar at both locations. Additionally, blends and mixtures containing ≥75% of the resistant cultivar maintained higher average visual quality across both locations compared to the susceptible cultivar alone. Between the two mixtures, the inclusion of a resistant TF cultivar maintained higher canopy density and increased the proportion of TF at both locations. Blends and mixtures containing ≥75% of a resistant cultivar reduced brown patch AUDPC by 71% and 83% in 2022 and 2023, respectively, when compared to the 100% susceptible cultivar. This field study reinforces the importance of selecting resistant TF cultivars to reduce seasonal brown patch symptoms in cool‐season turfgrass lawns. Core Ideas: Increasing the proportion of a resistant tall fescue cultivar in a blend significantly reduced brown patch severity.Turfgrass quality increased as the proportion of a resistant tall fescue cultivar increased in a blend.A brown patch‐resistant cultivar in tall fescue: Kentucky bluegrass mix increased density and fescue composition.The proportion of Kentucky bluegrass significantly increased over time in tall fescue: Kentucky bluegrass mixtures. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate clinical and instrumental outcomes of the autologous matrix-induced chondrogenesis (AMIC) technique for the treatment of isolated traumatic condyle and femoropatellar cartilage lesions. A total of 25 patients (12 males, 13 females, mean age 47.3 years) treated between 2018 and 2021 were retrospectively reviewed and subdivided into two groups based on age (Group A, age < 45 years; Group B, age > 45 years). A clinical evaluation was performed using the International Knee Documentation Committee (IKDC), Lysholm score and Visual Analogue Score (VAS). Cartilage regeneration was evaluated via magnetic resonance (1.5 Tesla) and classified according to a Magnetic resonance Observation of CArtilage Repair Tissue (MOCART) scoring system. At a minimum follow-up of 2 years, Group A patients obtained greater instrumental results in comparison to group B: in fact, the MOCART score was statistically significantly correlated with IKDC (r = 0.223) (p < 0.001) exclusively in group A. Nevertheless, a significant improvement in clinical functionality was shown in Group B (p < 0.001), demonstrating that this technique is safe, reproducible and capable of offering satisfactory clinical results regardless of age.

Abstract Background High tibial osteotomy (HTO) provides reliable and good long-term results, if performed with correct indications, but different techniques and types of fixation have been described. The purpose of this study is to present a novel modified biplanar medial opening-wedge (MOW) HTO technique where the osteotomies are performed in a Z-shaped fashion, and to present the medium-term clinical and radiographic results. Materials and methods We present a case series of 75 patients (80 knees) with mean age of 45.8 years, affected by isolated medial knee osteoarthritis and symptomatic varus knee malalignment, who underwent novel biplanar Z-shaped MOWHTO. Clinical and radiological outcomes were collected, retrospectively before surgery and at median follow-up of 7.2 years (95% CI 5.6–9.2 months) after surgery. Clinical results and satisfaction were assessed by visual analog scale (VAS), Western Ontario and McMaster University Osteoarthritis Index (WOMAC), and Likert scale. Radiological assessment involved the evaluation of the medial proximal tibial angle (MPTA), tibial slope (TS), Caton–Deschamps index, and knee osteoarthritis grade according to Ahlbäck classification. Pre- and postoperative results were compared using the two-tailed t-test or Wilcoxon’s test of independent samples for paired data or nonparametric analog. P