Immunomodulatory effect of adipose-derived stem cells: the cutting edge of clinical application

Adipose-derived stem cells (ASCs) represent a promising tool for soft tissue engineering as well as for clinical treatment of inflammatory and autoimmune pathologies. The well-characterized multi-differentiation potential and self-renewal properties of ASCs are coupled with their immunomodulatory ability in providing therapeutic efficacy. Yet, their impact in immune or inflammatory disorders might rely both on cell contact-dependent mechanisms and paracrine effects, resulting in the release of various soluble factors that regulate immune cells functions. Despite the widespread use of ASCs in clinical trials addressing several pathologies, the pathophysiological mechanisms at the basis of their clinical use have been not yet fully investigated. In particular, a thorough analysis of ASC immunomodulatory potential is mandatory. Here we explore such molecular mechanisms involved in ASC immunomodulatory properties, emphasizing the relevance of the milieu composition. We review the potential clinical use of ASC secretome as a mediator for immunomodulation, with a focus on in vitro and in vivo environmental conditions affecting clinical outcome. We describe some potential strategies for optimization of ASCs immunomodulatory capacity in clinical settings, which act either on adult stem cells gene expression and local microenvironment. Finally, we discuss the limitations of both allogeneic and autologous ASC use, highlighting the issues to be fixed in order to significantly improve the efficacy of ASC-based cell therapy

Fibrinolysis protease receptors promote activation of astrocytes to express pro-inflammatory cytokines.

BACKGROUND:Astrocytes contribute to the crosstalk that generates chronic neuro-inflammation in neurological diseases; however, compared with microglia, astrocytes respond to a more limited continuum of innate immune system stimulants. Recent studies suggest that the fibrinolysis system may regulate inflammation. The goal of this study was to test whether fibrinolysis system components activate astrocytes and if so, elucidate the responsible biochemical pathway. METHODS:Primary cultures of astrocytes and microglia were prepared from neonatal mouse brains. The ability of purified fibrinolysis system proteins to elicit a pro-inflammatory response was determined by measuring expression of the mRNAs encoding tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and chemokine (C-C motif) ligand 2 (CCL2). IκBα phosphorylation also was measured. Plasminogen activation in association with cells was detected by chromogenic substrate hydrolysis. The activity of specific receptors was tested using neutralizing antibodies and reagents. RESULTS:Astrocytes expressed pro-inflammatory cytokines when treated with plasminogen but not when treated with agonists for Toll-like Receptor-4 (TLR4), TLR2, or TLR9. Microglia also expressed pro-inflammatory cytokines in response to plasminogen; however, in these cells, the response was observed only when tissue-type plasminogen activator (tPA) was added to activate plasminogen. In astrocytes, endogenously produced urokinase-type plasminogen activator (uPA) converted plasminogen into plasmin in the absence of tPA. Plasminogen activation was dependent on the plasminogen receptor, α-enolase, and the uPA receptor, uPAR. Although uPAR is capable of directly activating cell-signaling, the receptor responsible for cytokine expression and IκBα phosphorylation response to plasmin was Protease-activated Receptor-1 (PAR-1). The pathway, by which plasminogen induced astrocyte activation, was blocked by inhibiting any one of the three receptors implicated in this pathway with reagents such as εACA, α-enolase-specific antibody, uPAR-specific antibody, the uPA amino terminal fragment, or a pharmacologic PAR-1 inhibitor. CONCLUSIONS:Plasminogen may activate astrocytes for pro-inflammatory cytokine expression through the concerted action of at least three distinct fibrinolysis protease receptors. The pathway is dependent on uPA to activate plasminogen, which is expressed endogenously by astrocytes in culture but also may be provided by other cells in the astrocytic cell microenvironment in the CNS

Neuropilin 1mediates keratinocyte growth factor signaling in adipose-derived stem cells: potential involvement in adipogenesis

Adipogenesis is regulated by a complex network of molecules, including fibroblast growth factors. Keratinocyte growth factor (KGF) has been previously reported to promote proliferation on rat preadipocytes, although the expression of its specific receptor, FGFR2-IIIb/KGFR, is not actually detected in mesenchymal cells. Here, we demonstrate that human adipose-derived stem cells (ASCs) show increased expression of KGF during adipogenic differentiation, especially in the early steps. Moreover, KGF is able to induce transient activation of ERK and p38 MAPK pathways in these cells. Furthermore, KGF promotes ASC differentiation and supports the activation of differentiation pathways, such as those of PI3K/Akt and the retinoblastoma protein (Rb). Notably, we observed only a low amount of FGFR2-IIIb in ASCs, which seems not to be responsible for KGF activity. Here, we demonstrate for the first time that Neuropilin 1 (NRP1), a transmembrane glycoprotein expressed in ASCs acting as a coreceptor for some growth factors, is responsible for KGF-dependent pathway activation in these cells. Our study contributes to clarify the molecular bases of human adipogenesis, demonstrating a role of KGF in the early steps of this process, and points out a role of NRP1 as a previously unknown mediator of KGF action in ASCs

Radiation risks knowledge in resident and fellow in paediatrics: A questionnaire survey

Background: Analyse through a multi-choice anonymous questionnaire the knowledge's level in paediatric residents and fellows in two different main Italian hospital, looking mainly to the information to patients and relatives related to risks of ionizing radiation used in common radiological investigations in children. Methods: 65 multi choice questionnaires were distributed to paediatric residents and fellows of two different hospitals, an University Hospital (A.O.U.P. "P. Giaccone"- University of Palermo) and a national reference centre for paediatrics (Ospedale Pediatrico Bambino Gesù - Rome). The questionnaire included twelve multiple-choice questions with the aim of analyzing the knowledge about ionizing radiation related risks in infants and children who undergo common diagnostic radiology investigations. The data obtained were processed using software Stata/MP version 11.2. In order to measure the level of expertise of each interviewee a binary indicator was built. The value 1 was assigned if the percentage of correct answers exceeds the median of the distribution and 0 for values not exceeding the median. The association between the level of competence and demographic characteristics (gender, age) and training experience was measured by means of α2 test. Results: 51/65 questionnaires were completed, returned and analysed (87.7%). Only 18 surveyed (35%), (95% IC=[22%-48%]) can be defined as competent in radiation risk knowledge for common radiological investigations, considering the percentage of correct answers at least of 50% (sufficient knowledge was given with a minimum score of 8 correct answers out of 12). Conclusions: The study demonstrates an urgent need to implement the radiation protection knowledge in the training programme of paediatricians, that improve if just a short targeted training is performed

Multiscale environmental heterogeneity in a large river-floodplain system

River-floodplain systems host very high biodiversity and provide a great variety of ecosystem services due largely to their high environmental heterogeneity, which is strongly dependent on spatial and temporal scales of observation. We aimed to describe the main patterns of environmental variation of the subtropical system of the Middle Paraná River at different scales of analysis using univariate and multivariate statistics. Physical, chemical and biological variables were analyzed during different hydro-sedimentological phases along four consecutive years in 31 aquatic environments. We considered four aquatic habitat types with different degrees of hydrological connectivity and water flow characteristics (major rivers, secondary channels, floodplain lakes permanently connected to the major rivers, and floodplain lakes temporarily connected to the major rivers) from two geomorphologic units with different drainage patterns: anastomosing and meandering. The environmental heterogeneity was mainly associated with the habitat types and relative contributions of lateral tributaries with different water qualities to each unit, in combination with the influence of the hydro-sedimentological regime. The heterogeneity of the system was significantly higher during low water than during high water phases. However, although the extreme flood reduced dissimilarities between habitat types within each geomorphologic unit, it increased differences between major rivers and between units. In conclusion, at different spatial scales, floods may simultaneously have a homogenization effect due to increased hydrological connectivity and a differentiation effect due to enhanced inputs of water from regions with variable geological characteristics and land uses. Finally, geomorphologic units can play a key role in maintaining the environmental heterogeneity during floods of high magnitude, even when land barriers among aquatic environments disappear.Fil: Mayora, Gisela Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; ArgentinaFil: Scarabotti, Pablo Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; ArgentinaFil: Schneider, Berenice. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; ArgentinaFil: Alvarenga Mariani, Patricio Francisco. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; ArgentinaFil: Marchese Garello, Mercedes Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto Nacional de Limnología. Universidad Nacional del Litoral. Instituto Nacional de Limnología; Argentin

Evaluation of the activity of natural phenolic antioxidants, extracted from industrial coffee residues, on the stability of poly(1,4‐butylene succinate) formulations

In this work, the evaluation of the antioxidant activity of natural phenolic compounds is performed and compared to that of a conventional antioxidative agent. Phenolic molecules, extracted from industrial processing coffee residues, are added to a matrix of poly(1,4-butylene succinate) (PBS). The apparent activation energy (Ea) of the thermo-oxidative degradation is calculated by employing different methods like Kissinger-Akahira-Sunose, Flynn-Wall-Ozawa and Friedman. The results are compared with the antioxidant activity evaluation obtained through the ABTS radical scavenging assay. From the average activation energies, it is observed that the addition of the natural antioxidants led to an increase in the activation energy of the degradation process as a function of the phenolic compound content. This trend is confirmed by the results of the ABTS assay. Hence, this study proves that the active molecules extracted from agri-food waste could be employed to improve the antioxidant capacity of the biopolymer, even if the composition of the extract must be evaluated in order to mitigate the effects of other components

Prevalence, clinical correlates, and use of glucose-lowering drugs among older patients with type 2 diabetes living in long-term care facilities

Prevalence, clinical correlates, and use of glucose-lowering drugs were comprehensively evaluated among 863 nursing home older patients with diabetes (mean age 82.9 ± 2.1 years): functional dependence and cognitive impairment were present in 84.1% and 68% of patients, respectively, and 66.3% of patients had 2–4 comorbidities. HbA1c values < 7.0% were documented in 54.9% of diabetic; significantly lower HbA1c levels were observed in demented patients than in nondemented subjects. Documented hypoglycemic episodes were reported for 57 patients (6.6%), without significant association with age, functional dependence, cognitive impairment, or HbA1c levels. About one-fifth of older long-term facilities residents have diabetes, with concomitant poor health conditions and high prevalence of cognitive impairment and functional dependence. Roughly three-fourths of these older and frail diabetic patients have HbA1c values lower than optimal, suggesting a potential for hypoglycemic harm especially among patients with severe cognitive impairment

From winery waste to bioactive compounds and new polymeric biocomposites: a contribution to the circular economy concept

The paper aims at optimising and validating possible routes toward the full valorisation of grape agrowaste to produce bioactive molecules and new materials. Starting from Merlot red pomace, phenol complex mixtures were successfully extracted by using two different approaches. Extracts obtained by solvent-based (SE) technique contained up to 46.9 gGAeq/kgDW of total phenols. Depending on the used solvent, the prevalence of compounds belonging to different phenol families was achieved. Pressurized liquid extraction (PLE) gave higher total phenol yields (up to 79 gGAeq/kgDW) but a lower range of extracted compounds. All liquid extracts exerted strong antioxidant properties. Moreover, both SE and PLE extraction solid residues were directly exploited (between 5 and 20% w/w) to prepare biocomposite materials by direct mixing via an eco-friendly approach with PHBV polymer. The final composites showed mechanical characteristics similar to PHVB matrix. The use of pomace residues in biocomposites could therefore bring both to the reduction of the cost of the final material, as a lower amount of costly PHBV is used. The present research demonstrated the full valorisation of grape pomace, an agrowaste produced every year in large amounts and having a significant environmental impact

Successful treatment of long-standing post-stroke dysphagia with botulinum toxin and rehabilitation

Cricopharyngeal myotomy is the most common treatment used to restore normal swallowing in patients with persistent ( /6 months) cricopharyngeal muscle dysfunction post-stroke. We describe 2 patients whose dysphagia was due to cricopharyngeal muscle over-activity and who significantly improved after a percutaneous botulinum toxin injection in the cricopharyngeal muscle in combination with a rehabilitation treatment (dietary modifications, postural techniques, airflow protection manoeuvres). Swallowing was assessed clinically and by fibreoptic endoscopic evaluation of swallowing and videofluoroscopy; the degree of dysphagia was scored using the penetration-aspiration scale. Two months after the botulinum toxin injection the patients, who were previously fed via percutaneous endoscopic gastrostomy, returned to independent oral feeding and at 6, 12 and 24 month follow-up, both were still able to maintain an adequate oral intake with no signs of aspiration (by videofluoroscopy) or clinical complications. No further botulinum toxin injections or rehabilitation treatments were required. Our findings strongly suggest that even longstanding dysphagia can improve dramatically in selected patients. To the best of our knowledge, there are no other reports with such a long follow-up