Single-cell guided prenatal derivation of primary fetal epithelial organoids from human amniotic and tracheal fluids.

Isolation of tissue-specific fetal stem cells and derivation of primary organoids is limited to samples obtained from termination of pregnancies, hampering prenatal investigation of fetal development and congenital diseases. Therefore, new patient-specific in vitro models are needed. To this aim, isolation and expansion of fetal stem cells during pregnancy, without the need for tissue samples or reprogramming, would be advantageous. Amniotic fluid (AF) is a source of cells from multiple developing organs. Using single-cell analysis, we characterized the cellular identities present in human AF. We identified and isolated viable epithelial stem/progenitor cells of fetal gastrointestinal, renal and pulmonary origin. Upon culture, these cells formed clonal epithelial organoids, manifesting small intestine, kidney tubule and lung identity. AF organoids exhibit transcriptomic, protein expression and functional features of their tissue of origin. With relevance for prenatal disease modeling, we derived lung organoids from AF and tracheal fluid cells of congenital diaphragmatic hernia fetuses, recapitulating some features of the disease. AF organoids are derived in a timeline compatible with prenatal intervention, potentially allowing investigation of therapeutic tools and regenerative medicine strategies personalized to the fetus at clinically relevant developmental stages.

The Benefit of Transvaginal Elastography in Detecting Deep Endometriosis: A Feasibility Study.

OBJECTIVES: This study aimed to evaluate elastography features of deep infiltrating endometriosis (DIE), and to define whether this technique may discriminate lesions from surrounding non-endometriotic tissue. METHODS: This was an exploratory observational study on women affected by DIE treated in a third-level academic hospital gynaecology outpatient facility between 2020 and 2021. Strain elastography (SE) was conducted via transvaginal probe. Tissue deformation of DIE and surrounding tissue was expressed as percentage tissue deformation or as subjective colour score (CS; from blue=stiff to red=soft, assigned numerical values from 0 to 3). Ratios of normal tissue/DIE were compared to ratio of normal tissue/stiffer normal tissue area. RESULTS: Evaluations were performed on 46 DIE nodules and surrounding tissue of the uterosacral ligaments (n=21), parametrium (n=7), rectum (n=14), and recto-vaginal septum (n =4). Irrespective of location, DIE strain ratio (3.09, IQR 2.38-4.14 vs. 1.25, IQR 1.11-1.48; p<0.001) and CS ratio (4.62, IQR 3.83-6.94 vs. 1.13, IQR 1.06-1.29; p<0.001) was significantly higher than that of normal tissue. ROC AUC of CS ratio was higher than ROC AUC of strain ratio (99.76%, CI.95 99.26-100% vs. 91.35%, CI.95 85.23-97.47%; p=0.007), and best ROC threshold for CS ratio was 1.82, with a sensitivity of 97.83% (CI.95 93.48-100%) and a specificity of 100% (CI.95 100-100%). CONCLUSIONS: Both strain and CS ratios accurately distinguish DIE nodules at various locations. Applications of elastography in improving the diagnosis DIE, in distinguishing different DIE lesions and in monitoring DIE evolution can be envisioned and are worthy of further evaluation.

Primary human organoids models: Current progress and key milestones.

During the past 10 years the world has experienced enormous progress in the organoids field. Human organoids have shown huge potential to study organ development, homeostasis and to model diseases in vitro. The organoid technology has been widely and increasingly applied to generate patient-specific in vitro 3D cultures, starting from both primary and reprogrammed stem/progenitor cells. This has consequently fostered the development of innovative disease models and new regenerative therapies. Human primary, or adult stem/progenitor cell-derived, organoids can be derived from both healthy and pathological primary tissue samples spanning from fetal to adult age. The resulting 3D culture can be maintained for several months and even years, while retaining and resembling its original tissue's properties. As the potential of this technology expands, new approaches are emerging to further improve organoid applications in biology and medicine. This review discusses the main organs and tissues which, as of today, have been modelled in vitro using primary organoid culture systems. Moreover, we also discuss the advantages, limitations, and future perspectives of primary human organoids in the fields of developmental biology, disease modelling, drug testing and regenerative medicine.

mRNA and Viral Vector COVID-19 Vaccines Do Not Affect Male Fertility: A Prospective Study.

PURPOSE: To assess whether mRNA and viral vector coronavirus disease 2019 (COVID-19) vaccines detrimentally affected semen parameters. MATERIALS AND METHODS: In this prospective study, we enrolled 101 men vaccinated for COVID-19 (76% received mRNA vaccines, 20% viral vector vaccines, 2% a mixed formulation, and for 2 men no information about vaccine type was available) in 2021 and with a previous semen analysis. For each man we compared semen parameters before and after vaccination. RESULTS: Post-vaccine samples were obtained at a median of 2.3±1.5 months after the second dose. After vaccination, the median sample volume significantly decreased (from 3.0 to 2.6 mL, p=0.036), whereas the median sperm concentration, the progressive motility, and total motile sperm count increased (from 25.0 to 43.0 million/mL, p<0.0001; from 50% to 56%, p=0.022; from 34.8 to 54.6 million, p<0.0001, respectively). Thirty-four patients were oligospermic before the vaccine, and also in these patients we observed a significant increase of sperm parameters after vaccine. Finally, we confirmed the aforementioned results in men who received a mRNA or a viral vector vaccine. CONCLUSIONS: The semen parameters following COVID-19 vaccination did not reflect any causative detrimental effect from vaccination, and for the first time we demonstrated that this applies to both mRNA and viral-vector vaccines. The known individual variation in semen and the reduced abstinence time before the post-vaccine sample collection may explain the increases in sperm parameters.

Angle of Uterine Flexion and Adenomyosis.

The aim of this study was to assess the prevalence of adenomyosis in symptomatic women in relation to the angle of flexion of the uterus. A total of 120 patients referring to our Chronic Pelvic Pain Center were prospectively enrolled. Each woman scored menstrual pain, intermenstrual pain, and dyspareunia on a 10 cm visual analogue scale and underwent a clinical examination and transvaginal ultrasound. MUSA criteria were used for the diagnosis of adenomyosis. The angle of flexion of the uterus on the cervix was categorized as <150° (75% of cases), between 150° and 210° (6.7% of cases) and >210° (18.3% of cases). Adenomyosis was diagnosed in 76/120 women (63.3%). In women with adenomyosis, the VAS of intermenstrual pain was higher than in women without adenomyosis (4.04 ± 3.79 vs. 2.57 ± 3.34; p < 0.034). The angle of uterine flexion >210° was more prevalent in women with than without adenomyosis (25.0% vs. 6.8%; p < 0.015). The odds ratio of suffering from adenomyosis markedly increased in the presence of an angle of uterine flexion >210° (OR 5.8 95% CI 1.19, 28.3; p > 0.029). The data indicate that the ultrasound-estimated angle of uterine flexion >210° is related to a higher prevalence of adenomyosis.

Digital Twin for Civil Engineering Systems: An Exploratory Review for Distributed Sensing Updating.

We live in an environment of ever-growing demand for transport networks, which also have ageing infrastructure. However, it is not feasible to replace all the infrastructural assets that have surpassed their service lives. The commonly established alternative is increasing their durability by means of Structural Health Monitoring (SHM)-based maintenance and serviceability. Amongst the multitude of approaches to SHM, the Digital Twin model is gaining increasing attention. This model is a digital reconstruction (the Digital Twin) of a real-life asset (the Physical Twin) that, in contrast to other digital models, is frequently and automatically updated using data sampled by a sensor network deployed on the latter. This tool can provide infrastructure managers with functionalities to monitor and optimize their asset stock and to make informed and data-based decisions, in the context of day-to-day operative conditions and after extreme events. These data not only include sensor data, but also include regularly revalidated structural reliability indices formulated on the grounds of the frequently updated Digital Twin model. The technology can be even pushed as far as performing structural behavioral predictions and automatically compensating for them. The present exploratory review covers the key Digital Twin aspects-its usefulness, modus operandi, application, etc.-and proves the suitability of Distributed Sensing as its network sensor component.

Dataset of Sentinel-1 surface soil moisture time series at 1 km resolution over Southern Italy.

This paper describes the specifications of the surface soil volumetric water content ( Θ ) [m3/m3] product derived from Sentinel-1 (S-1) data and assessed in the study "Sentinel-1 soil moisture at 1 km resolution: a validation study" [1]. The S-1 Θ product consists of Θ mean and standard deviation values at 1 km spatial resolution and is expected to support applications in agriculture and hydrology as well as the Numerical Weather Prediction at regional scale [2]. The retrieval algorithm is a time series based short term change detection that is implemented in the "Soil MOisture retrieval from multi-temporal SAR data" (SMOSAR) code (v2.0). The provided dataset represents an example of the developed S-1 Θ product and consists of a time series of 183 S-1 Θ images over Southern Italy from January 2015 to December 2018. The maps were produced for each ascending S-1 acquisition date on the Relative Orbit Number (RON) 146 and the temporal gap between consecutive maps is 6 days (when both S-1A and S-1B data are available) or 12 days.

A Review of Recent Distributed Optical Fiber Sensors Applications for Civil Engineering Structural Health Monitoring.

The present work is a comprehensive collection of recently published research articles on Structural Health Monitoring (SHM) campaigns performed by means of Distributed Optical Fiber Sensors (DOFS). The latter are cutting-edge strain, temperature and vibration monitoring tools with a large potential pool, namely their minimal intrusiveness, accuracy, ease of deployment and more. Its most state-of-the-art feature, though, is the ability to perform measurements with very small spatial resolutions (as small as 0.63 mm). This review article intends to introduce, inform and advise the readers on various DOFS deployment methodologies for the assessment of the residual ability of a structure to continue serving its intended purpose. By collecting in a single place these recent efforts, advancements and findings, the authors intend to contribute to the goal of collective growth towards an efficient SHM. The current work is structured in a manner that allows for the single consultation of any specific DOFS application field, i.e., laboratory experimentation, the built environment (bridges, buildings, roads, etc.), geotechnical constructions, tunnels, pipelines and wind turbines. Beforehand, a brief section was constructed around the recent progress on the study of the strain transfer mechanisms occurring in the multi-layered sensing system inherent to any DOFS deployment (different kinds of fiber claddings, coatings and bonding adhesives). Finally, a section is also dedicated to ideas and concepts for those novel DOFS applications which may very well represent the future of SHM.

Menstrual Pain and Elasticity of Uterine Cervix.

Menstrual pain is consequent to intense uterine contraction aimed to expel menstrual flow through downstream uterine cervix. Herein it was evaluated whether characteristics of uterine cervix are associated with intensity of menstrual pain. Ultrasound elastography was used to analyze cervix elasticity of 75 consecutive outpatient women. Elasticity was related to intensity of menstrual pain defined by a Visual Analogue Scale (VAS). Four regions of interest (ROI) were considered: internal uterine orifice (IUO), anterior (ACC) and posterior cervical (PCC) compartment and middle cervical canal (MCC). Tissue elasticity, evaluated by color score (from 0.5 = blue/violet (low elasticity) to 3.0 = red (high elasticity), and percent tissue deformation was analyzed. Elasticity of IUO was lower (p = 0.0001) than that of MCC or ACC, and it was negatively related (R2 = 0.428; p = 0.0001) to menstrual VAS (CR -2.17; 95%CI -3.80, -0.54; p = 0.01). Presence of adenomyosis (CR 3.24; 95% CI 1.94, 4.54; p = 0.0001) and cervix tenderness at clinical examination (CR 2.74; 95% CI 1.29, 4.20; p = 0.0004), were also independently related to menstrual VAS. At post hoc analysis, women with vs. without menstrual pain had lower IUO elasticity, expressed as color score (0.72 ± 0.40 vs. 0.92 ± 0.42; p = 0.059), lower percent tissue deformation at IUO (0.09 ± 0.05 vs. 0.13 ± 0.08; p = 0.025), a higher prevalence of cervical tenderness at bimanual examination (36.2% vs. 9.5%; p = 0.022) and a higher prevalence of adenomyosis (46.5% vs. 19.9%; p = 0.04). These preliminary data indicate that IUO elasticity is associated with the presence and the intensity of menstrual pain. Mechanisms determining IUO elasticity are useful to be explored.

Long-Term Concrete Shrinkage Influence on the Performance of Reinforced Concrete Structures.

The contribution of concrete to the tensile stiffness (tension stiffening) of a reinforced concrete (RC) member is a key governing factor for structural serviceability analyses. However, among the current tension stiffening models, few consider the effect brought forth by concrete shrinkage, and none studies take account of the effect for very long-term shrinkage. The present work intends to tackle this exact issue by testing multiple RC tensile elements (with different bar diameters and reinforcement ratios) after a five-year shrinking time period. The experimental deformative and tension stiffening responses were subjected to a mathematical process of shrinkage removal aimed at assessing its effect on the former. The results showed shrinkage distinctly lowered the cracking load of the RC members and caused an apparent tension stiffening reduction. Furthermore, both of these effects were exacerbated in the members with higher reinforcement ratios. The experimental and shrinkage-free behaviors of the RC elements were finally compared to the values predicted by the CEB-fib Model Code 2010 and the Euro Code 2. Interestingly, as a consequence of the long-term shrinkage, the codes expressed a smaller relative error when compared to the shrinkage-free curves versus the experimental ones.

Analysis of quality palliative care in a specialized medicine setting: an observational study.

BACKGROUND: Recently health economic expenditure has increased considerably determining an increased awareness of excessive use of "low-value care" treatments, especially for chronic-degenerative diseases. AIM: Describing Quality of life (QoL) of patients and palliative care's skills of nurses in internal medicine unit. METHODS: Observational study conducted in an internal medicine unit that admit patients with chronic critical illness. Patients' QoL was measured with the integrated Palliative Care Outcome Scale (IPOS), and nursing staff skills regarding palliative care with the Questionnaire of Professional Skills Nurses in Palliative Care (QVPICP). IPOS was administered at admission and one week later. RESULTS: All the patients (n=13) accepted to participate to the study (mean age 80 years; SD 15). Eight (61%) were female. All the nurses invited (n=17) completed the QCPICP. Most of them was female (70%). The mean age was 40 years SD 14. Nurses perceived they have skills, knowledge, and abilities to approach patient's palliative care path (mean QVPICP: 7). IPOS means scores were worse in the following variables: pain (before: 3; after: 4); lack of mobility (before: 3; after: 4); sleepiness (before: 3; after: 4); depression (before: 3; after: 4). CONCLUSION: Patient's QoL showed a worsening in the after assessment. Although nurses perceived themselves skilled in taking care of patients with palliative care needs, organizations may not allow nurses to perform their role to the best of their competencies. NURSING IMPLICATION: Team briefing and debriefings in internal medicine unit may promote a holistic approach aimed at improving patients' QoL.

Distributed Optical Fiber Sensing Bonding Techniques Performance for Embedment inside Reinforced Concrete Structures.

Distributed optical fiber sensors (DOFS) are modern-day cutting-edge monitoring tools that are quickly acquiring relevance in structural health monitoring engineering. Their most ambitious use is embedded inside plain or reinforced concrete (RC) structures with the scope of comprehending their inner-workings and the functioning of the concrete-reinforcement interaction. Yet, multiple studies have shown that the bonding technique with which the DOFS are bonded to the reinforcement bars has a significant role on the quality of the extracted strain data. Whilst this influence has been studied for externally bonded DOFS, it has not been done for embedded ones. The present article is set on performing such study by monitoring the strain measurement quality as sampled by DOFS bonded to multiple rebars with different techniques and adhesives. These instrumented rebars are used to produce differently sized RC ties later tested in tension. The discussion of the test outputs highlights the quasi-optimal performance of a DOFS/rebar bonding technique consisting of incising a groove in the rebar, positioning the DOFS inside it, bonding it with cyanoacrylate and later adding a protective layer of silicone. The resulting data is mostly noise-free and anomalies-free, yet still presents a newly diagnosed hitch that needs addressing in future research.

Decellularized skeletal muscles display neurotrophic effects in three-dimensional organotypic cultures.

Skeletal muscle decellularization allows the generation of natural scaffolds that retain the extracellular matrix (ECM) mechanical integrity, biological activity, and three-dimensional (3D) architecture of the native tissue. Recent reports showed that in vivo implantation of decellularized muscles supports muscle regeneration in volumetric muscle loss models, including nervous system and neuromuscular junctional homing. Since the nervous system plays pivotal roles during skeletal muscle regeneration and in tissue homeostasis, support of reinnervation is a crucial aspect to be considered. However, the effect of decellularized muscles on reinnervation and on neuronal axon growth has been poorly investigated. Here, we characterized residual protein composition of decellularized muscles by mass spectrometry and we show that scaffolds preserve structural proteins of the ECM of both skeletal muscle and peripheral nervous system. To investigate whether decellularized scaffolds could per se attract neural axons, organotypic sections of spinal cord were cultured three dimensionally in vitro, in presence or in absence of decellularized muscles. We found that neural axons extended from the spinal cord are attracted by the decellularized muscles and penetrate inside the scaffolds upon 3D coculture. These results demonstrate that decellularized scaffolds possess intrinsic neurotrophic properties, supporting their potential use for the treatment of clinical cases where extensive functional regeneration of the muscle is required.

Combined Notch and PDGF Signaling Enhances Migration and Expression of Stem Cell Markers while Inducing Perivascular Cell Features in Muscle Satellite Cells.

Satellite cells are responsible for skeletal muscle regeneration. Upon activation, they proliferate as transient amplifying myoblasts, most of which fuse into regenerating myofibers. Despite their remarkable differentiation potential, these cells have limited migration capacity, which curtails clinical use for widespread forms of muscular dystrophy. Conversely, skeletal muscle perivascular cells have less myogenic potential but better migration capacity than satellite cells. Here we show that modulation of Notch and PDGF pathways, involved in developmental specification of pericytes, induces perivascular cell features in adult mouse and human satellite cell-derived myoblasts. DLL4 and PDGF-BB-treated cells express markers of perivascular cells and associate with endothelial networks while also upregulating markers of satellite cell self-renewal. Moreover, treated cells acquire trans-endothelial migration ability while remaining capable of engrafting skeletal muscle upon intramuscular transplantation. These results extend our understanding of muscle stem cell fate plasticity and provide a druggable pathway with clinical relevance for muscle cell therapy.

Two subtypes of symptomatic joint hypermobility: a descriptive study using latent class analysis.

OBJECTIVE: To investigate a cohort of children with symptomatic joint hypermobility. METHODS: Case notes for 318 children with joint hypermobility attending a rheumatology clinic were reviewed for clinical presentation, medical history, psychosocial factors and physical examination findings. Seven key variables were extracted and used as indicator variables in a latent class analysis to estimate the presence and number of subgroups of children with symptomatic joint hypermobility. RESULTS: Two subgroups with differing clinical presentations were identified accounting for age and gender: an 'athletic-persistent' class (62%) characterised by higher probabilities for recurrent and chronic musculoskeletal pain, and less severe hypermobility; and a 'systemic-profound' class (38%) characterised by generalised hypermobility, recurrent musculoskeletal pain, gastro-oesophageal reflux and motor delay. CONCLUSION: Findings suggest the presence of two distinct presentations of children with hypermobility. This finding may be important for clinical decision-making and management of this group of children.

Performance of Rayleigh-Based Distributed Optical Fiber Sensors Bonded to Reinforcing Bars in Bending.

Distributed Optical Fiber Sensors (DOFSs), thanks to their multiple sensing points, are ideal tools for the detection of deformations and cracking in reinforced concrete (RC) structures, crucial as a means to ensure the safety of infrastructures. Yet, beyond a certain point of most DOFS-monitored experimental tests, researchers have come across unrealistic readings of strain which prevent the extraction of further reliable data. The present paper outlines the results obtained through an experimental test aimed at inducing such anomalies to isolate and identify the physical cause of their origin. The understanding of such a phenomenon would enable DOFS to become a truly performant strain sensing technique. The test consists of gradually bending seven steel reinforcement bars with a bonded DOFS under different conditions such as different load types, bonding adhesives, bar sections and more. The results show the bonding adhesives having an influence on the DOFS performance but not on the rise of anomalies while the reasons triggering the latter are narrowed down from six to two, reaching a strain threshold and a change in structure's deformative behavior. Further planned research will allow identification of the cause behind the rise of strain-reading anomalies.

Bed Rest for Sciatica: A Closer Look at the Evidence.

When managing acute nonspecific low back pain (LBP), bed rest is commonly discouraged as a form of therapy. However, when the picture includes radiating leg pain arising from the lumbar spine, the evidence is less robust. In this Viewpoint, the authors explore evidence pertaining to sciatica, defined as "low back pain with verified neurological deficits," and bed rest as part of the management of radiating leg pain. J Orthop Sports Phys Ther 2018;48(6):436-438. doi:10.2519/jospt.2018.0609.

Perfusion decellularization of a human limb: A novel platform for composite tissue engineering and reconstructive surgery.

Muscle and fasciocutaneous flaps taken from autologous donor sites are currently the most utilized approach for trauma repair, accounting annually for 4.5 million procedures in the US alone. However, the donor tissue size is limited and the complications related to these surgical techniques lead to morbidities, often involving the donor sites. Alternatively, recent reports indicated that extracellular matrix (ECM) scaffolds boost the regenerative potential of the injured site, as shown in a small cohort of volumetric muscle loss patients. Perfusion decellularization is a bioengineering technology that allows the generation of clinical-scale ECM scaffolds with preserved complex architecture and with an intact vascular template, from a variety of donor organs and tissues. We recently reported that this technology is amenable to generate full composite tissue scaffolds from rat and non-human primate limbs. Translating this platform to human extremities could substantially benefit soft tissue and volumetric muscle loss patients providing tissue- and species-specific grafts. In this proof-of-concept study, we show the successful generation a large-scale, acellular composite tissue scaffold from a full cadaveric human upper extremity. This construct retained its morphological architecture and perfusable vascular conduits. Histological and biochemical validation confirmed the successful removal of nuclear and cellular components, and highlighted the preservation of the native extracellular matrix components. Our results indicate that perfusion decellularization can be applied to produce human composite tissue acellular scaffolds. With its preserved structure and vascular template, these biocompatible constructs, could have significant advantages over the currently implanted matrices by means of nutrient distribution, size-scalability and immunological response.

Inflammation converts human mesoangioblasts into targets of alloreactive immune responses: implications for allogeneic cell therapy of DMD.

Stem cell therapy is a promising approach to regenerate healthy tissues starting from a limited amount of self-renewing cells. Immunological rejection of cell therapy products might represent a major limitation. In this study, we investigated the immunological functional profile of mesoangioblasts, vessel-associated myogenic stem cells, currently tested in a phase 1-2a trial, active in our Institute, for the treatment of Duchenne muscular dystrophy. We report that in resting conditions, human mesoangioblasts are poorly immunogenic, inefficient in promoting the expansion of alloreactive T cells and intrinsically resistant to T-cell killing. However, upon exposure to interferon-γ or differentiation into myotubes, mesoangioblasts acquire the ability to promote the expansion of alloreactive T cells and acquire sensitivity to T-cell killing. Resistance of mesoangioblasts to T-cell killing is largely due to the expression of the intracellular serine protease inhibitor-9 and represents a relevant mechanism of stem cell immune evasion.

Influence of Kinesio Taping applied over biceps brachii on isokinetic elbow peak torque. A placebo controlled study in a population of young healthy subjects.

OBJECTIVES: To investigate the effect of Kinesio Taping (KT) applied over the biceps brachii on maximal isokinetic elbow torque. DESIGN: This study followed a single-blinded, placebo controlled, repeated measures design. METHODS: Isokinetic eccentric and concentric elbow peak torques were evaluated without taping (NT), with KT or placebo taping (PT) in 20 healthy participants. Furthermore, all the participants were required to perform a proprioceptive task of elbow joint position sense (JPS) in the same experimental conditions. RESULTS: A significant effect of taping condition was found for concentric elbow peak torque (p=0.01). Post hoc analysis revealed a statistically significant concentric elbow peak torque improvement between NT and KT (p<0.05) but not between NT and PT. As regards eccentric elbow peak torque, we found a significant effect of taping condition (p<0.0001). Significant eccentric elbow peak torque differences were observed between NT and PT (p<0.01) and between KT and PT (p<0.001), while the increase observed from NT to KT conditions failed to reach significance at a post hoc analysis. CONCLUSIONS: When applied over the biceps brachii, KT increases concentric elbow peak torque in a population of healthy participants, if compared with a PT.