Sense of Agency and Skills Learning in Virtual-Mediated Environment: A Systematic Review.

Agency is central to remote actions, and it may enhance skills learning due to a partial overlap between brain structures and networks, the promotion of confidence towards a telemanipulator, and the feeling of congruence of the motor choice to the motor plan. We systematically reviewed studies aiming to verify the role of agency in improving learning. Fifteen studies were selected from MEDLINE and Scopus®. When a mismatch is introduced between observed and performed actions, the decrease in agency and learning is proportional to the intensity of the mismatch, which is due to greater interference with the motor programming. Thanks to multisensory integration, agency and learning benefit both from sensory and performance feedback and from the timing of feedback based on control at the goal level or the perceptual-motor level. This work constitutes a bedrock for professional teleoperation settings (e.g., robotic surgery), with particular reference to the role of agency in performing complex tasks with remote control.

Sensory substitution increases robotic surgical performance and sets the ground for a mediating role of the sense of embodiment: a systematic review.

Sensory Substitution (SS) allows the elaboration of information via non preferential sensory modalities. This phenomenon occurs in robotic-assisted surgery (RAS), in which haptic feedback is lacking. It has been suggested that SS could sustain surgeons' proficiency by means of visual clues for inferring tactile information, that also promotes the feeling of haptic phantom sensations. A critical role in reaching a good performance in procedural tasks is also sustained by the Sense of Embodiment (SE), that is, the capacity to integrate objects into subjective bodily self-representation. As SE is enhanced by haptic sensations, we hypothesize a role of SS in promoting SE in RAS. Accordingly, the goal of this systematic review is to summarize the evidence pertaining the study of SS in RAS in order to highlight the impact on the performance, and to identify a mediating role of the SE in increasing dexterity in RAS. Eight studies selected from the MEDLINE and Scopus® databases met inclusion criteria for a qualitative synthesis. Results indicated that haptic to other modalities SS enhanced force consistency and accuracy, and decreased surgeon fatigue. Expert surgeons, as compared to novices, showed a better natural SS processing, testified by a proficient performance with and without SS aids. No studies investigated the mediating role of SE. These findings indicate that SS is subjected to learning and memory processes that help surgeons to rapidly derive haptic-correlates from visual clues, which are highly required for a good performance. Also, the higher ability of doing SS and the associated perception of haptic sensations might increase multisensory integration, which might sustain performance.

Functional correlates of subliminal stimulation in Posttraumatic Stress Disorder: Systematic review and meta-analysis.

Patients with Post-traumatic stress disorder (PTSD) exposed to traumatic reminders show hyperreactivity in brain areas (e.g., amygdala) belonging or related to the Innate Alarm System (IAS), allowing the rapid processing of salient stimuli. Evidence that IAS is activated by subliminal trauma-reminders could shed a new light on the factors precipitating and perpetuating PTSD symptomatology. Thus, we systematically reviewed studies investigating neuroimaging correlates of subliminal stimulation in PTSD. Twenty-three studies were selected from the MEDLINE and Scopus® databases for a qualitative synthesis, 5 of which allowed a further meta-analysis of fMRI data. The intensity of IAS responses to subliminal trauma-related reminders ranged from a minimum in healthy controls to a maximum in the PTSD patients with the most severe (e.g., dissociative) symptoms or the least responsiveness to treatment. Comparisons with other disorders (e.g., phobias) revealed contrasting results. Our findings demonstrate the hyperactivation of areas belonging or related to IAS in response to unconscious threats that should be integrated in diagnostic as well as in therapeutic protocols.

The effects of Transcranial Direct Current Stimulation on food craving and food intake in individuals affected by obesity and overweight: a mini review of the magnitude of the effects.

Obesity represents one of the wellness diseases concurring to increase the incidence of diabetes, cardiovascular diseases, and cancer. One of the main perpetuating factors of obesity is food craving, which is characterized by an urgent desire to eat a large and various amount of food, regardless of calories requirement or satiety signals, and it might be addressed to the alteration of the dorsolateral prefrontal cortex (DLPFC) activity. Despite most of the gold-standard therapies focus on symptom treatment only, non-invasive brain stimulation techniques such as transcranial direct current stimulation (tDCS) could help treat overeating by modulating specific neural pathways. The current systematic review was conducted to identify whether convergent evidence supporting the usefulness of tDCS to deal with food craving are present in the literature. The review was conducted by searching articles published up to January 1st 2022 on MEDLINE, Scopus and PsycInfo databases. We included studies investigating the effects of tDCS on food craving in subjects affected by overweight and obesity. According to eligibility criteria, 5 articles were included. Results showed that tDCS targeting left DLPFC with unipolar montage induced ameliorating effects on food craving. Controversial results were shown for the other studies, that might be ascribable to the use of bipolar montage, and the choice of other target areas. Further investigations including expectancy effect control, larger sample sizes and follow-up are needed to support more robust conclusions. To conclude, tDCS combined with the use of psychoeducative intervention, diet and physical activity, might represents a potential to manage food craving in individuals with overweight and obesity.

Intimate partner violence: psycho-physio-pathological sequelae for defining a holistic enriched treatment.

Intimate partner violence (IPV) is a health priority, which worldwide, mainly affects women. The consequences of IPV include several psychophysiological effects. These range from altered levels of hormones and neurotrophins to difficulties in emotion regulation and cognitive impairment. Mounting evidence from preclinical studies has shown that environmental enrichment, a form of sensory-motor, cognitive, and social stimulation, can induce a wide range of neuroplastic processes in the brain which consistently improve recovery from a wide variety of somatic and psychiatric diseases. To support IPV survivors, it is essential to ensure a safe housing environment, which can serve as a foundation for environmental enrichment-based interventions. However, some concerns have been raised when supportive housing interventions focus on the economic aspects of survivors' lives instead of the emotional ones. We thus propose a holistic intervention in which supportive housing is integrated with evidenced-based psychotherapies which could constitute an enriched therapeutic approach for IPV survivors.

Lavender aromatherapy: A systematic review from essential oil quality and administration methods to cognitive enhancing effects.

Modern society is reviving the practice of aromatherapy, and lavender is reported being the most worldwide purchased plant for essential oil (EO) extraction. Since recent studies reported cognitive enhancing effects of lavender besides the hypno-inducing effects, a literature review is needed. Considering EO quality and diffusion devices, we conducted a systematic review on the effects of lavender EO inhalation on arousal, attention and memory in healthy subjects. Starting from this new multidisciplinary perspective, cognitive effects were reviewed to link outcomes to effective and reproducible protocols. A systematic search on MEDLINE, ERIC, PsycInfo, Google Scholar, and Scopus databases using Cognitive Atlas and plant-related keywords was conducted. Among the 1,203 articles yielded, 11 met eligibility criteria. Subjects administered with lavender EO displayed arousal decrease and sustained attention increase. Controversial results emerged regarding memory. Lack of EO quality assessment and protocols heterogeneity did not allow assessing whether different EO composition differentially modulates cognition and whether placebo effect can be discerned from EO effect itself. However, GABAergic pathway modulation exerted by linalool, a major lavender EO constituent, might explain cognitive functions empowerment. We speculate aromatherapy could be a burgeoning cognition enhancing tool, although further investigation is required to reach robust conclusions.

Probiotics Modulate Mouse Gut Microbiota and Influence Intestinal Immune and Serotonergic Gene Expression in a Site-Specific Fashion.

Probiotic microorganisms may benefit the host by influencing diverse physiological processes, whose nature and underlying mechanisms are still largely unexplored. Animal models are a unique tool to understand the complexity of the interactions between probiotic microorganisms, the intestinal microbiota, and the host. In this regard, in this pilot study, we compared the effects of 5-day administration of three different probiotic bacterial strains (Bifidobacterium bifidum MIMBb23sg, Lactobacillus helveticus MIMLh5, and Lacticaseibacillus paracasei DG) on three distinct murine intestinal sites (ileum, cecum, and colon). All probiotics preferentially colonized the cecum and colon. In addition, probiotics reduced in the ileum and increased in the cecum and colon the relative abundance of numerous bacterial taxonomic units. MIMBb23sg and DG increased the inducible nitric oxide synthase (iNOS) in the ileum, which is involved in epithelial homeostasis. In addition, MIMBb23sg upregulated cytokine IL-10 in the ileum and downregulated the cyclooxygenase COX-2 in the colon, suggesting an anti-inflammatory/regulatory activity. MIMBb23sg significantly affected the expression of the main gene involved in serotonin synthesis (TPH1) and the gene coding for the serotonin reuptake protein (SERT) in the ileum and colon, suggesting a potential propulsive effect toward the distal part of the gut, whereas the impact of MIMLh5 and DG on serotonergic genes suggested an effect toward motility control. The three probiotics decreased the expression of the permeability marker zonulin in gut distal sites. This preliminary in vivo study demonstrated the safety of the tested probiotic strains and their common ability to modulate the intestinal microbiota. The probiotics affected host gene expression in a strain-specific manner. Notably, the observed effects in the gut were site dependent. This study provides a rationale for investigating the effects of probiotics on the serotonergic system, which is a topic still widely unexplored.

Unconscious processing of subliminal stimuli in panic disorder: A systematic review and meta-analysis.

Attentional biases to threat exist in panic disorder (PD), probably related to altered subliminal processing. We systematically reviewed studies investigating subliminal processing in PD. Studies were retrieved from MEDLINE and Scopus®. We meta-analytically compared PD (n = 167) and healthy controls (HC, n = 165) for processing of masked panic-related and neutral words. We also compared subliminal and supraliminal presentations of panic-related words relative to neutral words within PD subjects and HC. We found a significantly enhanced Stroop interference to masked panic-related words in PD vs HC (Hedges' g = 0.60, p = 0.03; Q = 14.83, I2 = 66.3 %, p = 0.01). While both PD subjects and HC tended to be slower to respond to supraliminal threat words than to neutral words, PD subjects only showed a marginally significant slower response to subliminal panic-related words vs neutral words. Findings remain inconclusive regarding comparison to other mental disorders, neural correlates, and the effect of psychotherapy. Even if possibly flawed by methodological weaknesses, our findings support the existence of a sensitivity to subliminal threat cues in PD, which could be targeted to improve treatment.

Enhancing Qualities of Consciousness during Online Learning via Multisensory Interactions.

Online-learning is a feasible alternative to in-person attendance during COVID-19 pandemic. In this period, information technologies have allowed sharing experiences, but have also highlighted some limitations compared to traditional learning. Learning is strongly supported by some qualities of consciousness such as flow (intended as the optimal state of absorption and engagement activity) and sense of presence (feeling of exerting control, interacting with and getting immersed into real/virtual environments), behavioral, emotional, and cognitive engagement, together with the need for social interaction. During online learning, feelings of disconnection, social isolation, distractions, boredom, and lack of control exert a detrimental effect on the ability to reach the state of flow, the feeling of presence, the feeling of social involvement. Since online environments could prevent the rising of these learning-supporting variables, this article aims at describing the role of flow, presence, engagement, and social interactions during online sessions and at characterizing multisensory stimulations as a driver to cope with these issues. We argue that the use of augmented, mixed, or virtual reality can support the above-mentioned domains, and thus counteract the detrimental effects of physical distance. Such support could be further increased by enhancing multisensory stimulation modalities within augmented and virtual environments.

Targeting temporal parietal junction for assessing and treating disembodiment phenomena: a systematic review of TMS effect on depersonalization and derealization disorders (DPD) and body illusions.

The temporal-parietal junction (TPJ) is a key structure for the embodiment, term referred to as the sense of being localized within one's physical body and is a fundamental aspect of the self. On the contrary, the sense of disembodiment, an alteration of one's sense of self or the sense of being localized out of one's physical body, is a prominent feature in specific dissociative disorders, namely depersonalization/derealization disorders (DPD). The aims of the study were to provide: 1) a qualitative synthesis of the effect of Transcranial Magnetic Stimulation (TMS), taking into account its use for therapeutic and experimental purposes; 2) a better understanding on whether the use of TMS could support the treatment of DPD and other clinical conditions in which depersonalization and derealization are displayed. To identify suitable publications, an online search of the PubMed, Cochrane Library, Web of science and Scopus databases was performed using relevant search terms. In addition, an in-depth search was performed by screening review articles and the references section of each included articles. Our search yielded a total of 108 records through multiple databases searching and one additional record was identified through other sources. After duplicates removal, title and abstract reading, we retained 16 records for the assessment of eligibility. According to our inclusion criteria, we retained 8 studies. The selected studies showed that TMS targeting the TPJ is a promising technique for treating disembodiment phenomena DPD and for inducing reversible disembodiment states in healthy subjects. These data represent the first step towards a greater understanding of possible treatments to be used in disembodiment disorders. The use of TMS over the TPJ appears to be promising for treating disembodiment phenomena.

Attention-Dependent Physiological Correlates in Sleep-Deprived Young Healthy Humans.

Cognitive functions could be specifically altered but masked from the unspecific effect of workload, a common factor affecting cognitive functions that modulate peripheral outputs. To identify workload-related and specific, task-dependent components, physiological correlates of cognitive functioning were derived by studying 15 healthy volunteers performing attentional tasks in baseline and post-sleep-deprivation conditions (one week interval). Sleep deprivation was introduced to increase workload. We performed recordings of heart pulse, facial temperature, and head movements during tasks assessing attentional network efficiency (ANT, Attentional Network Task; CCT, Continuous Compensatory Tracker) workload assessments after execution of tasks. Changes in cognitive and physiological indices were studied in both conditions; physiological correlates of cognitive performance were identified by correlating changes from baseline to post-sleep-deprivation condition of task indices with those of physiological measures after correction for between-conditions workload changes. We found that mental and physical demands of workload increased after sleep deprivation. We identified no changes in cognitive and physiological indices across conditions; specific physiological correlates of attentional systems, as indicated by the negative correlation between changes in ANT-alerting and changes in amplitude of head movements and the positive correlation between changes in CCT-speed indexing alertness and changes in facial temperature.

The clinical application of transcranial direct current stimulation in patients with cerebellar ataxia: a systematic review.

AIM: The aim of this review was to investigate the effects of transcranial direct current stimulation (tDCS) on motor function in patients with cerebellar ataxia. MATERIALS AND METHODS: Our systematic review has been performed by searching full-text articles on Pubmed and Scopus. Only studies investigating the motor effects of tDCS in patients with cerebellar ataxias were considered. A qualitative analysis of data was performed, as the methodology of the selected studies was highly heterogeneous. RESULTS: Our search yielded a total of twenty-seven hits. Based on the inclusion criteria, 19 of these were excluded and 89 were retained (number of patients = 81).The results reviewed so far suggest that tDCS over cerebellum combined or not with extra-cerebellar areas might be promising approach to improve motor outcomes, with a greater success in patients less impaired. In particular, it is been shown an improvement in both clinical measures assessing cerebellar deficits (i.e. gait, stance and oculomotor disorders) and performance measures (finger dexterity, upper limb coordination and gait speed). Some of the assessed investigations highlighted a restore effect of cerebellar brain inhibition pathway and resting motor threshold after tDCS. CONCLUSIONS: tDCS could be considered an effective approach to promote plasticity in patient with cerebellar ataxia with significant motor effects. Future studies, with larger sample sizes are needed in order to evaluate the effective tDCS benefits on motor functionality. Due to the limited number of studies available so far, conclusions on the effectiveness of the reported approaches are premature.

Task-independent Electrophysiological Correlates of Motor Imagery Ability from Kinaesthetic and Visual Perspectives.

Motor imagery (MI) ability is highly subjective, as indicated by the individual scores of the MIQ-3 questionnaire, and poor imagers compensate for the difficulty in performing MI with larger cerebral activations, as demonstrated by MI studies involving hands/limbs. In order to identify general, task-independent MI ability correlates, 16 volunteers were stratified with MIQ-3. The scores in the kinaesthetic (K) and 1st-person visual (V) perspectives were associated with EEG patterns obtained during K-MI and V-MI of the same complex MIQ-3 movements during these MI tasks (Spearman's correlation, significance at <0.05, SnPM corrected). EEG measures were relative to rest (relaxation, closed eyes), and based on six electrode clusters both for band spectral content and connectivity (Granger causality). Lower K-MI ability was associated with greater theta decreases during tasks in fronto-central clusters and greater inward information flow to prefrontal clusters for theta, high alpha and beta bands. On the other hand, power band relative decreases were associated with V-MI ability in fronto-central clusters for low alpha and left fronto-central and both centro-parietal clusters for beta bands. The results thus suggest different computational mechanisms for MI-V and MI-K. The association between low alpha/beta desynchronization and V-MIQ scores and between theta changes and K-MIQ scores suggest a cognitive effort with greater cerebral activation in participants with lower V-MI ability. The association between information flow to prefrontal hub and K-MI ability suggest the need for a continuous update of information to support MI-related executive functions in subjects with poor K-MI ability.

Amniotic MSCs reduce pulmonary fibrosis by hampering lung B-cell recruitment, retention, and maturation.

Growing evidence suggests a mechanistic link between inflammation and the development and progression of fibrotic processes. Mesenchymal stromal cells derived from the human amniotic membrane (hAMSCs), which display marked immunomodulatory properties, have been shown to reduce bleomycin-induced lung fibrosis in mice, possibly by creating a microenvironment able to limit the evolution of chronic inflammation to fibrosis. However, the ability of hAMSCs to modulate immune cells involved in bleomycin-induced pulmonary inflammation has yet to be elucidated. Herein, we conducted a longitudinal study of the effects of hAMSCs on alveolar and lung immune cell populations upon bleomycin challenge. Immune cells collected through bronchoalveolar lavage were examined by flow cytometry, and lung tissues were used to study gene expression of markers associated with different immune cell types. We observed that hAMSCs increased lung expression of T regulatory cell marker Foxp3, increased macrophage polarization toward an anti-inflammatory phenotype (M2), and reduced the antigen-presentation potential of macrophages and dendritic cells. For the first time, we demonstrate that hAMSCs markedly reduce pulmonary B-cell recruitment, retention, and maturation, and counteract the formation and expansion of intrapulmonary lymphoid aggregates. Thus, hAMSCs may hamper the self-maintaining inflammatory condition promoted by B cells that continuously act as antigen presenting cells for proximal T lymphocytes in injured lungs. By modulating B-cell response, hAMSCs may contribute to blunting of the chronicization of lung inflammatory processes with a consequent reduction of the progression of the fibrotic lesion.

Combination Therapy of Inhaled Indacaterol/Glycopyrronium for Chronic Obstructive Pulmonary Disease in the Very Elderly: Is It Safe? An Electrocardiographic Evaluation.

Cardiovascular (CV) comorbidities in patients with chronic obstructive pulmonary disease (COPD) are associated with increased morbidity and mortality, especially in old and very old subjects. The question if long-acting beta-agonist and long-acting muscarinic antagonist could be associated with the increased prevalence of CV-related adverse effects has puzzled, particularly in the past, specialists involved in the management of respiratory diseases. The safety of these compounds has scarcely been tested in patients aged ≥ 65 years with CV comorbidities, since randomized controlled trials rarely include this subpopulation. However, the fixed combination indacaterol/glycopyrronium has shown a favorable CV safety profile in both healthy volunteers and COPD patients. Thus, we aimed to assess the CV safety pro-- file of the fixed combination indacaterol/glycopyrronium 110/50 µg in a series of COPD patients aged ≥ 80 years with several comorbidities. Our results indicate that this combination is safe in the comorbid elderly, since no significant electrocardiographic abnormalities were recorded after the administration of the inhaled therapy. Only rare and nonclinically significant changes in heart rate and corrected QT interval duration were evident, mainly in females and in patients with concomitant impaired kidney function.

Taurine Administration Recovers Motor and Learning Deficits in an Angelman Syndrome Mouse Model.

Angelman syndrome (AS, MIM 105830) is a rare neurodevelopmental disorder affecting 1:10-20,000 children. Patients show moderate to severe intellectual disability, ataxia and absence of speech. Studies on both post-mortem AS human brains and mouse models revealed dysfunctions in the extra synaptic gamma-aminobutyric acid (GABA) receptors implicated in the pathogenesis. Taurine is a free intracellular sulfur-containing amino acid, abundant in brain, considered an inhibiting neurotransmitter with neuroprotective properties. As taurine acts as an agonist of GABA-A receptors, we aimed at investigating whether it might ameliorate AS symptoms. Since mice weaning, we orally administered 1 g/kg/day taurine in water to Ube3a-deficient mice. To test the improvement of motor and cognitive skills, Rotarod, Novel Object Recognition and Open Field tests were assayed at 7, 14, 21 and 30 weeks, while biochemical tests and amino acid dosages were carried out, respectively, by Western-blot and high-performance liquid chromatography (HPLC) on frozen whole brains. Treatment of Ube3am-/p+ mice with taurine significantly improved motor and learning skills and restored the levels of the post-synaptic PSD-95 and pERK1/2-ERK1/2 ratio to wild type values. No side effects of taurine were observed. Our study indicates taurine administration as a potential therapy to ameliorate motor deficits and learning difficulties in AS.

Effect of human amniotic epithelial cells on pro-fibrogenic resident hepatic cells in a rat model of liver fibrosis.

Myofibroblasts are key fibrogenic cells responsible for excessive extracellular matrix synthesis characterizing the fibrotic lesion. In liver fibrosis, myofibroblasts derive either from activation of hepatic stellate cells (HSC) and portal fibroblasts (PF), or from the activation of fibroblasts that originate from ductular epithelial cells undergoing epithelial-mesenchymal transition. Ductular cells can also indirectly promote myofibroblast generation by activating TGF-ß, the main fibrogenic growth factor, through αvß6 integrin. In addition, after liver injury, liver sinusoidal cells can lose their ability to maintain HSC quiescence, thus favouring HSC differentiation towards myofibroblasts. The amniotic membrane and epithelial cells (hAEC) derived thereof have been shown to decrease hepatic myofibroblast levels in rodents with liver fibrosis. In this study, in a rat model of liver fibrosis, we investigated the effects of hAEC on resident hepatic cells contributing to myofibroblast generation. Our data show that hAEC reduce myofibroblast numbers with a consequent reduction in fibronectin and collagen deposition. Interestingly, we show that hAEC strongly act on specific myofibroblast precursors. Specifically, hAEC reduce the activation of PF rather than HSC. In addition, hAEC target reactive ductular cells by inhibiting their proliferation and αvß6 integrin expression, with a consequent decrease in TGF-ß activation. Moreover, hAEC counteract the transition of ductular cells towards fibroblasts, while it does not affect injury-induced and fibrosis-promoting sinusoidal alterations. In conclusion, among the emerging therapeutic applications of hAEC in liver diseases, their specific action on PF and ductular cells strongly suggests their application in liver injuries involving the expansion and activation of the portal compartment.

Mouse aldehyde-oxidase-4 controls diurnal rhythms, fat deposition and locomotor activity.

Aldehyde-oxidase-4 (AOX4) is one of the mouse aldehyde oxidase isoenzymes and its physiological function is unknown. The major source of AOX4 is the Harderian-gland, where the enzyme is characterized by daily rhythmic fluctuations. Deletion of the Aox4 gene causes perturbations in the expression of the circadian-rhythms gene pathway, as indicated by transcriptomic analysis. AOX4 inactivation alters the diurnal oscillations in the expression of master clock-genes. Similar effects are observed in other organs devoid of AOX4, such as white adipose tissue, liver and hypothalamus indicating a systemic action. While perturbations of clock-genes is sex-independent in the Harderian-gland and hypothalamus, sex influences this trait in liver and white-adipose-tissue which are characterized by the presence of AOX isoforms other than AOX4. In knock-out animals, perturbations in clock-gene expression are accompanied by reduced locomotor activity, resistance to diet induced obesity and to hepatic steatosis. All these effects are observed in female and male animals. Resistance to obesity is due to diminished fat accumulation resulting from increased energy dissipation, as white-adipocytes undergo trans-differentiation towards brown-adipocytes. Metabolomics and enzymatic data indicate that 5-hydroxyindolacetic acid and tryptophan are novel endogenous AOX4 substrates, potentially involved in AOX4 systemic actions.

High-sensitivity BRAF mutation analysis: BRAF V600E is acquired early during tumor development but is heterogeneously distributed in a subset of papillary thyroid carcinomas.

CONTEXT: The homogeneous distribution of BRAF V600E in papillary thyroid carcinoma (PTC) has been called into question by recent reports. These studies claim that BRAF V600E is heterogeneous and is limited to tumor cell subsets in the majority of PTCs. OBJECTIVE: The objective of the study was to understand the allele distribution of BRAF V600E by evaluating the percentage of mutated neoplastic cells in a group of PTCs using two different highly sensitive analytical approaches: allele-specific locked nucleic acid PCR and 454 next-generation sequencing targeted to BRAF exon 15. STUDY DESIGN: BRAF V600E was investigated using allele-specific locked nucleic acid PCR on 155 consecutive samples of PTC. Mutated cases were reanalyzed by 454 next-generation sequencing and immunohistochemistry. Because the evaluation of genetic heterogeneity in tumor samples can be profoundly biased by contamination with normal cells, all mutation frequency data were normalized to the real amount of neoplastic cells within each tumor. RESULTS: Eighty-five of 155 PTCs (54.8%) were BRAF V600E mutated. The distribution of mutated neoplastic cells within the tumor was as follows: greater than 80% in 37 of 85 (43.5%), 30-80% in 39 of 85 (45.9%), and less than 30% in 9 of 85 (10.6%). In most of the PTCs with less than 80% BRAF V600E-positive neoplastic cells, the mutation was present in large neoplastic cell subpopulations. Tumors with less than 30% mutated neoplastic cells were smaller than tumors with a percentage of mutated cells greater than 80% or between 30% and 80% (P < .05). CONCLUSIONS: BRAF V600E is heterogeneously distributed in some PTCs. The large BRAF V600E neoplastic cell subpopulations found in mutated cases is consistent with the view that the BRAF V600E is acquired early during PTC development.

Conditioned medium from amniotic membrane-derived cells prevents lung fibrosis and preserves blood gas exchanges in bleomycin-injured mice-specificity of the effects and insights into possible mechanisms.

BACKGROUND AND AIMS: We recently demonstrated that injection of conditioned medium (CM) generated from cells of the mesenchymal region of human amniotic membrane (AMTCs) reduces bleomycin-induced lung fibrosis in mice, suggesting a crucial role of paracrine factor(s) secreted by AMTCs in these beneficial effects. We further investigated this hypothesis, the mechanisms involved, the effects on some lung functional parameters and whether AMTC-secreted effector(s) are specific to these cells and not produced by other cell types, extending the time of analysis up to 28 days after treatment. METHODS: Bleomycin-challenged mice were either treated with AMTC-CM or CM generated from human skin fibroblasts, human peripheral blood mononuclear cells or Jurkat cells, or were left untreated. Mouse lungs were analyzed for content of pro-inflammatory and pro-fibrotic molecules, presence of lymphocytes and macrophages and for fibrosis level (through histological semi-quantitative evaluation and quantitative measurement of collagen content). Arterial blood gas analysis was also performed. RESULTS: Up to 28 days after delivery, AMTC-CM-treated mice developed reduced lung fibrosis with respect to mice treated with other CM types. AMTC-CM-treated mice had comparatively better preservation of blood gas parameters and showed lower lung content of interleukin-6, tumor necrosis factor-α, macrophage inflammatory protein-1α, monocyte chemoattractant protein-1 and transforming growth factor-ß associated with reduced lung macrophage levels. CONCLUSIONS: AMTC-CM prevents lung fibrosis in bleomycin-challenged mice, improving survival and preserving lung functional parameters such as blood gas exchanges. The specificity of AMTC-CM action was indicated by the absence of fibrosis reduction when other CM types were used. Finally, we provide some insights into the possible mechanisms underlying AMTC-CM-mediated control of fibrosis.