Miracle-dz/UniFAR-FTUs huggingface dataset:
Population Ecology and Genetic Diversity of the Invasive Alien Species Procambarus clarkii in Lake Trasimeno (Italy)

Simple Summary The population of Procambarus clarkii was surveyed twenty years after its introduction into Lake Trasimeno (Central Italy), investigating both ecological and genetic aspects. Our results confirmed that P. clarkii is well acclimatized with a stable population structure, mainly characterized by a unique mitochondrial lineage, suggesting that a single introduction event may have occurred in the lake, complemented by secondary events. Abstract The deliberate or accidental introduction of invasive alien species (IAS) causes negative ecological and economic impacts altering ecosystem processes, imperiling native species and causing damage to human endeavors. A monthly monitoring program was performed in Lake Trasimeno (Central Italy) from July 2018 to July 2019 in order to provide an upgrade of the population ecology of Procambarus clarkii and to assess the genetic diversity by analyzing the relationships among mitochondrial DNA diversity. Our results confirmed that P. clarkii is well acclimatized in the lake, revealing a stable population structure favored by the resources and conditions typical of this ecosystem, which seem to be optimal for the maintenance of the species. Four distinct mitochondrial haplotypes were detected, but one of them was clearly overrepresented (76%), suggesting that a single predominant introduction event may have occurred in this area, likely followed by secondary events. The identification of the typical genetic variants provides a better understanding of the evolutionary scenarios of P. clarkii in this biotope and it can be helpful in management plans concerning the expanding populations of this invasive alien species.
Aberrant Mechanical Efficiency during Exercise Relates to Metabolic Health and Exercise Intolerance in Adolescents with Obesity

Background. Mechanical efficiency (ME) might be an important parameter evaluating cardiometabolic health and the effectiveness of physical activity interventions in individuals with obesity. However, whether these cardiometabolic risk factors may relate to ME in adolescents with obesity is not known yet. Therefore, this study aims to compare the mechanical efficiency during maximal exercise testing between adolescents with obesity and lean adolescents, and to examine associations with exercise tolerance and metabolic health. Methods. Twenty-nine adolescents with obesity (BMI SDS: 2.11 ± 0.32, age: 13.4 ± 1.1 years, male/female: 15/14) and 29 lean (BMI SDS: −0.16 ± 0.84, age: 14.0 ± 1.5 years, male/female: 16/13) adolescents performed a maximal cardiopulmonary exercise test from which the net mechanical efficiency (MEnet) and substrate oxidation (carbohydrates and lipids) were calculated. Indicators for peak performance were collected. Biochemistry (lipid profile, glycaemic control, inflammation, leptin) was studied in fasted blood samples. Regression analyses were applied to examine relations between MEnet and exercise tolerance or blood variables in the total group. Results. Peak work rate (WRpeak), oxygen uptake (V˙O2peak)/WRpeak, ME, and MEnet were significantly lower (p < 0.05) in adolescents with obesity compared to their lean counterparts (p < 0.05). Furthermore, a reduced MEnet was independently related to a lower WRpeak (SC β = 2.447; p < 0.001) and elevated carbohydrate oxidation during exercise (SC β = −0.497; p < 0.001), as well as to elevated blood low-density lipoprotein cholesterol (SC β = −0.275; p = 0.034) and fasting glucose (SC β = −0.256; p = 0.049) concentration. Conclusion. In adolescents with obesity, the mechanical efficiency is lowered during exercise and this relates to exercise intolerance and a worse metabolic health.
The SZT2 Interactome Unravels New Functions of the KICSTOR Complex

Seizure threshold 2 (SZT2) is a component of the KICSTOR complex which, under catabolic conditions, functions as a negative regulator in the amino acid-sensing branch of mTORC1. Mutations in this gene cause a severe neurodevelopmental and epileptic encephalopathy whose main symptoms include epilepsy, intellectual disability, and macrocephaly. As SZT2 remains one of the least characterized regulators of mTORC1, in this work we performed a systematic interactome analysis under catabolic and anabolic conditions. Besides numerous mTORC1 and AMPK signaling components, we identified clusters of proteins related to autophagy, ciliogenesis regulation, neurogenesis, and neurodegenerative processes. Moreover, analysis of SZT2 ablated cells revealed increased mTORC1 signaling activation that could be reversed by Rapamycin or Torin treatments. Strikingly, SZT2 KO cells also exhibited higher levels of autophagic components, independent of the physiological conditions tested. These results are consistent with our interactome data, in which we detected an enriched pool of selective autophagy receptors/regulators. Moreover, preliminary analyses indicated that SZT2 alters ciliogenesis. Overall, the data presented form the basis to comprehensively investigate the physiological functions of SZT2 that could explain major molecular events in the pathophysiology of developmental and epileptic encephalopathy in patients with SZT2 mutations.
Atypical Teratoid Rhabdoid Tumours Are Susceptible to Panobinostat-Mediated Differentiation Therapy

Simple Summary Atypical teratoid rhabdoid tumour (ATRT) is an aggressive undifferentiated malignancy of the central nervous system in children. A defining feature of ATRT is the loss of the SMARCB1 gene that is essential for regulating gene expression required for normal developmental processes. We show that treatment of human ATRT cell models with the histone deacetylate inhibitor, panobinostat, inhibits tumour growth, reactivates the expression of developmental genes, and drives neuronal differentiation. These results demonstrate the therapeutic potential of panobinostat for the treatment of ATRT. Abstract Atypical teratoid rhabdoid tumour (ATRT) is a rare but highly aggressive undifferentiated solid tumour arising in the central nervous system and predominantly affecting infants and young children. ATRT is exclusively characterized by the inactivation of SMARCB1, a member of the SWI/SNF chromatin remodelling complex that is essential for the regulation of large sets of genes required for normal development and differentiation. Histone deacetylase inhibitors (HDACi) are a promising anticancer therapy and are able to mimic the normal acetylation functions of SMARCB1 in SMARCB1-deficient cells and drive multilineage differentiation in extracranial rhabdoid tumours. However, the potential efficacy of HDACi in ATRT is unknown. Here, we show that human ATRT cells are highly responsive to the HDACi panobinostat and that sustained treatment leads to growth arrest, increased cell senescence, decreased clonogenicity and induction of a neurogenesis gene-expression profile. Furthermore, in an orthotopic ATRT xenograft model, continuous panobinostat treatment inhibits tumour growth, increases survival and drives neuronal differentiation as shown by the expression of the neuronal marker, TUJ1. Collectively, this preclinical study supports the therapeutic potential of panobinostat-mediated differentiation therapy for ATRT.
Expanding the Phenotypic Spectrum of ECEL1-Associated Distal Arthrogryposis

Distal arthrogryposis type 5D (DA5D), a rare autosomal recessive disorder, is caused by mutations in ECEL1. We describe two consanguineous families (three patients) with novel ECEL1 gene mutations detected by next-generation sequencing (NGS). A 12-year-old boy (patient 1) presented with birth asphyxia, motor developmental delay, multiple joint contractures, pes planus, kyphoscoliosis, undescended testis, hypophonic speech with a nasal twang, asymmetric ptosis, facial weakness, absent abductor pollicis brevis, bifacial, and distal lower limb weakness. Muscle MRI revealed asymmetric fatty infiltration of tensor fascia lata, hamstring, lateral compartment of the leg, and gastrocnemius. In addition, 17-year-old monozygotic twins (patients 2 and 3) presented with motor development delay, white hairlock, hypertelorism, tented upper lip, bulbous nose, tongue furrowing, small low set ears, multiple contractures, pes cavus, prominent hyperextensibility at the knee, hypotonia of lower limbs, wasting and weakness of all limbs (distal > proximal), areflexia, and high steppage gait. One had perinatal insult, seizures, mild intellectual disability, unconjugated eye movements, and primary optic atrophy. In the twins, MRI revealed extensive fatty infiltration of the gluteus maximus, quadriceps, hamstrings, and anterior and posterior compartment of the leg. Electrophysiology showed prominent motor axonopathy. NGS revealed rare homozygous missense variants c.602T > C (p.Met201Thr) in patient 1 and c.83C > T (p.Ala28Val) in patients 2 and 3, both localized in exon 2 of ECEL1 gene. Our three cases expand the clinical, imaging, and molecular spectrum of the ECEL1-mutation-related DA5D.
Development of a Spinal Cord Injury Model Permissive to Study the Cardiovascular Effects of Rehabilitation Approaches Designed to Induce Neuroplasticity

Simple Summary People living with high-level spinal cord injury experience worse cardiovascular health than the general population. In most spinal cord injuries, there are some remaining functioning pathways leading from the brain through the spinal cord to the organs and muscles, but not enough to sustain normal levels of function. Recently, therapies that aim to increase the strength of connections in these remaining pathways have shown great potential in restoring walking, hand, and breathing function in the spinal cord injured population. In order to test these therapies for their effects on cardiovascular function, we developed a new type of spinal cord injury rat model that spares enough pathways for these therapies to act upon but still produces measurable reductions in heart and blood vessel function that can be targeted with interventions/treatments. Abstract As primary medical care for spinal cord injury (SCI) has improved over the last decades there are more individuals living with neurologically incomplete (vs. complete) cervical injuries. For these individuals, a number of promising therapies are being actively researched in pre-clinical settings that seek to strengthen the remaining spinal pathways with a view to improve motor function. To date, few, if any, of these interventions have been tested for their effectiveness to improve autonomic and cardiovascular (CV) function. As a first step to testing such therapies, we aimed to develop a model that has sufficient sparing of descending sympathetic pathways for these interventions to target yet induces robust CV impairment. Twenty-six Wistar rats were assigned to SCI (n = 13) or naïve (n = 13) groups. Animals were injured at the T3 spinal segment with 300 kdyn of force. Fourteen days post-SCI, left ventricular (LV) and arterial catheterization was performed to assess in vivo cardiac and hemodynamic function. Spinal cord lesion characteristics along with sparing in catecholaminergic and serotonergic projections were determined via immunohistochemistry. SCI produced a decrease in mean arterial pressure of 17 ± 3 mmHg (p < 0.001) and left ventricular contractility (end-systolic elastance) of 0.7 ± 0.1 mmHg/µL (p < 0.001). Our novel SCI model produced significant decreases in cardiac and hemodynamic function while preserving 33 ± 9% of white matter at the injury epicenter, which we believe makes it a useful pre-clinical model of SCI to study rehabilitation approaches designed to induce neuroplasticity.
Gravity-Based Flow Efficient Perfusion Culture System for Spheroids Mimicking Liver Inflammation

The spheroid culture system provides an efficient method to emulate organ-specific pathophysiology, overcoming the traditional two-dimensional (2D) cell culture limitations. The intervention of microfluidics in the spheroid culture platform has the potential to enhance the capacity of in vitro microphysiological tissues for disease modeling. Conventionally, spheroid culture is carried out in static conditions, making the media nutrient-deficient around the spheroid periphery. The current approach tries to enhance the capacity of the spheroid culture platform by integrating the perfusion channel for dynamic culture conditions. A pro-inflammatory hepatic model was emulated using a coculture of HepG2 cell line, fibroblasts, and endothelial cells for validating the spheroid culture plate with a perfusable channel across the spheroid well. Enhanced proliferation and metabolic capacity of the microphysiological model were observed and further validated by metabolic assays. A comparative analysis of static and dynamic conditions validated the advantage of spheroid culture with dynamic media flow. Hepatic spheroids were found to have improved proliferation in dynamic flow conditions as compared to the static culture platform. The perfusable culture system for spheroids is more physiologically relevant as compared to the static spheroid culture system for disease and drug analysis.
Using Transfer Learning Method to Develop an Artificial Intelligence Assisted Triaging for Endotracheal Tube Position on Chest X-ray

Endotracheal tubes (ETTs) provide a vital connection between the ventilator and patient; however, improper placement can hinder ventilation efficiency or injure the patient. Chest X-ray (CXR) is the most common approach to confirming ETT placement; however, technicians require considerable expertise in the interpretation of CXRs, and formal reports are often delayed. In this study, we developed an artificial intelligence-based triage system to enable the automated assessment of ETT placement in CXRs. Three intensivists performed a review of 4293 CXRs obtained from 2568 ICU patients. The CXRs were labeled “CORRECT” or “INCORRECT” in accordance with ETT placement. A region of interest (ROI) was also cropped out, including the bilateral head of the clavicle, the carina, and the tip of the ETT. Transfer learning was used to train four pre-trained models (VGG16, INCEPTION_V3, RESNET, and DENSENET169) and two models developed in the current study (VGG16_Tensor Projection Layer and CNN_Tensor Projection Layer) with the aim of differentiating the placement of ETTs. Only VGG16 based on ROI images presented acceptable performance (AUROC = 92%, F1 score = 0.87). The results obtained in this study demonstrate the feasibility of using the transfer learning method in the development of AI models by which to assess the placement of ETTs in CXRs.
Empirical Investigation of the Verbal Cues Involved in Delivering Experiential Metaphors

Delivering metaphors experientially has been emphasized in several psychotherapies, such as acceptance and commitment therapy. However, few research has analyzed the variables involved in the efficacy of metaphors. This experimental analog study aims to advance in this topic by analyzing the effect of two components involved in the experiential delivery of metaphors in psychotherapy. The first component is presenting the metaphor by asking the individual to imagine herself as the protagonist of the story versus presenting the metaphor in the third person (Self vs. Other). The second component is the inclusion of verbal cues prompting the relational elaboration of the rules derived from the metaphor content versus not including these prompts (Elaboration vs. No Elaboration). The effect of these components was tested in a double-blind, randomized, 2 × 2 factorial experiment that used the cold pressor task (CPT). Eighty-four participants were exposed to the CPT at the pretest. Afterward, participants were randomly assigned to four experimental protocols. The protocols were audiotaped and consisted of the same metaphor presented in four slightly different ways. Specifically, the protocol of Condition A involved a metaphor with Self and Elaboration, Condition B involved Self and No Elaboration, Condition C involved Other and Elaboration, and Condition D involved Other and No Elaboration. Then, participants were re-exposed to the CPT in the posttest. We hypothesized that Condition A (Self and Elaboration) would show a higher mean increase in pain tolerance than the remaining conditions, which would show similar results. The results were consistent with this hypothesis because Condition A showed a higher percentual increase in pain tolerance (Condition A: M = 268.21, SD = 167.47; Condition B: M = 180.86, SD = 73.01; Condition C: M = 204.81, SD = 100.19; Condition D: M = 175.41, SD = 76.00). A Bayesian informative hypothesis evaluation showed that this hypothesis obtained the highest posterior model probability. Thus, the results indicate that introducing metaphors by asking the individual to imagine herself as the protagonist of the story and providing prompts for relational elaboration might increase the therapeutic effect of the metaphor.
Antecedents of Workplace Bullying among Employees in Germany: Five-Year Lagged Effects of Job Demands and Job Resources

Objectives: The aim of the present study was to examine the long-term association of job demands and job resources with self-reported exposure to workplace bullying in a representative sample of employees in Germany. Methods: We analysed a nation-wide representative cohort of employees working in the same workplace with a 5-year follow-up (S-MGA; N = 1637). The study contained self-reported measures of psychosocial working conditions, including work pace, amount of work, influence at work, role clarity and quality of leadership, and workplace bullying, and of organisational factors, including organisational restructuring and layoffs. Results: After controlling for bullying and occupational level at baseline, higher baseline levels of organisational restructuring (OR 1.73; 95% CI 1.10–2.70), work pace (1.30; 95% CI 1.01–1.66), and amount of work (1.55; 95% CI 1.21–1.99), and lower baseline levels of influence at work (0.70; 95% CI 0.55–0.90) and quality of leadership (0.64; 95% CI 0.50–0.82), were associated with an elevated risk of workplace bullying at follow-up. In all, 90% of cases of self-reported workplace bullying could be attributed to these factors. Conclusions: The study suggests that employees reporting higher demands and lower resources, as well as organisational factors such as restructuring, are at a higher long-term risk of being targets of workplace bullying. Interventions aimed at preventing workplace bullying could benefit from a focus on psychosocial working conditions and organisational factors.
Synthesis and Characterization of Novel Succinyl Chitosan-Dexamethasone Conjugates for Potential Intravitreal Dexamethasone Delivery

The development of intravitreal glucocorticoid delivery systems is a current global challenge for the treatment of inflammatory diseases of the posterior segment of the eye. The main advantages of these systems are that they can overcome anatomical and physiological ophthalmic barriers and increase local bioavailability while prolonging and controlling drug release over several months to improve the safety and effectiveness of glucocorticoid therapy. One approach to the development of optimal delivery systems for intravitreal injections is the conjugation of low-molecular-weight drugs with natural polymers to prevent their rapid elimination and provide targeted and controlled release. This study focuses on the development of a procedure for a two-step synthesis of dexamethasone (DEX) conjugates based on the natural polysaccharide chitosan (CS). We first used carbodiimide chemistry to conjugate DEX to CS via a succinyl linker, and we then modified the obtained systems with succinic anhydride to impart a negative ζ-potential to the polymer particle surface. The resulting polysaccharide carriers had a degree of substitution with DEX moieties of 2–4%, a DEX content of 50–85 μg/mg, and a degree of succinylation of 64–68%. The size of the obtained particles was 400–1100 nm, and the ζ-potential was −30 to −33 mV. In vitro release studies at pH 7.4 showed slow hydrolysis of the amide and ester bonds in the synthesized systems, with a total release of 8–10% for both DEX and succinyl dexamethasone (SucDEX) after 1 month. The developed conjugates showed a significant anti-inflammatory effect in TNFα-induced and LPS-induced inflammation models, suppressing CD54 expression in THP-1 cells by 2- and 4-fold, respectively. Thus, these novel succinyl chitosan-dexamethasone (SucCS-DEX) conjugates are promising ophthalmic carriers for intravitreal delivery.
Competitive Coherence Generates Qualia in Bacteria and Other Living Systems

Simple Summary Subjective experiences, sensations, or feelings, alias qualia, include the experiences of seeing the colour blue, hearing the sound of middle C, smelling a rose, touching an ice cube and feeling a pain in one’s knee. The origin of these qualia and their relationship with the material world have long been subject to intense debate. Here, I propose that qualia arise from—and can even determine—the operation of competitive coherence. The idea behind competitive coherence is that (1) the behaviour of a system depends on the activity of only a subset of the elements of the system and (2) the selection of elements belonging to this subset depends on both the history and the present state of the system (which includes its present environment). In arguing that competitive coherence is characteristic of living systems at every level, I contend that it is to bacteria that we should turn if we are to understand qualia. This is because bacteria were here first, constitute much of the biomass, are very complex, made the world as we know it, and help control its denizens. Finally, I propose that qualia are important in the functioning of high-level systems such as ecosystems. Abstract The relevance of bacteria to subjective experiences or qualia is underappreciated. Here, I make four proposals. Firstly, living systems traverse sequences of active states that determine their behaviour; these states result from competitive coherence, which depends on connectivity-based competition between a Next process and a Now process, whereby elements in the active state at time n+1 are chosen between the elements in the active state at time n and those elements in the developing n+1 state. Secondly, bacteria should help us link the mental to the physical world given that bacteria were here first, are highly complex, influence animal behaviour and dominate the Earth. Thirdly, the operation of competitive coherence to generate active states in bacteria, brains and other living systems is inseparable from qualia. Fourthly, these qualia become particularly important to the generation of active states in the highest levels of living systems, namely, the ecosystem and planetary levels.