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Introduction: Adipose tissue represents an abundant source of mesenchymal stem-like cells. Adipose-derived stem cell progeny have been investigated and used in regenerative medicine for decades. In the last few years, they have been used to “enrich” lipoaspirates in fat grafting techniques, in an attempt to boost the regenerative potential of adipose tissue when used as autologous “filler”. Materials and Methods: 50 adipose tissue samples from lipoaspirates and subcutaneous breast tissue biopsies were used to generate adipose floating spheroid cell (ASphC) lines. ASphC were characterized for the expression of putative mesenchymal stem cell markers and used in vitro to test their multilineage potential. Furthermore, ASphCs were seeded on dermal regeneration template (Integra®) and implanted into the T8 vertebral laminectomy site of immunocompromised mice. Results: Here, we show that the majority of ASphCs are in a quiescent state and express the putative surface stem cell marker CD271. Unlike CD271- cells, CD271+ ASphCs grew indefinitely in vitro as undifferentiated spheres in serum-free medium, maintaining their multilineage differentiation potential ability. Importantly, p107, a functional adipose stem cell marker, is strictly expressed in ASphCs and barely present in their differentiated mesenchymal lineages. These sphere cells display an enhanced ability in vitro to differentiate into distinctive end-stage cell types, such as osteoblasts, chondrocytes and adipocytes. Gene expression profiling analysis indicated that ASphCs are endowed with stem cell potential that is gradually lost during specific differentiation. Finally, ASphCs facilitate bone inter body repair and regeneration after laminectomy. Conclusions: We conclude that ASphCs possess a pronounced in vivo activity to regenerate the bone injury. Overall, ASphCs represent a heterogeneous population of stem-like cells harbouring multilineage differential potential and representing a prospective promising tool in cell therapy and tissue engineering.

Metabolism sustains the stemness of Cancer Stem Cells (CSCs), affecting, in turn, tumor heterogeneity, metastatic potential, and therapy resistance. Therefore, it is appealing to target CSCs metabolism as a new therapeutic approach. Consequently, we paid considerable attention to the anti-apoptotic microRNA miR-483-3p, that we reported being regulated by glucose metabolism in liver cancer cells. We investigated the therapeutic potential of targeting miR-483-3p by using the anti-glucose metabolism 2-deoxyglucose (2-DG) molecule in tumor Xenograft mouse model originating from two different Colon-Cancer Stem Cell lines (CCSC lines). We show that 2-DG treatment does not affect CCSCs during tumor formation in immunocompromised mouse models despite its ability to increase the CCSCs apoptotic rate in vitro and decrease miR-483-3p expression in both in vitro and in vivo experimental conditions. The promising in vitro data contrast with in vivo results that show the inefficacy of the treatment. We think that, in our immuno-compromised mouse model, to inhibit the glucose metabolism could become not only ineffective but also counterproductive by creating a selective pressure on the most fitting tumoral clones.

ObjectiveCancer stem cells are responsible for tumour spreading and relapse. Human epidermal growth factor receptor 2 (HER2) expression is a negative prognostic factor in colorectal cancer (CRC) and a potential target in tumours carrying the gene amplification. Our aim was to define the expression of HER2 in colorectal cancer stem cells (CR-CSCs) and its possible role as therapeutic target in CRC resistant to anti- epidermal growth factor receptor (EGFR) therapy.DesignA collection of primary sphere cell cultures obtained from 60 CRC specimens was used to generate CR-CSC mouse avatars to preclinically validate therapeutic options. We also made use of the ChIP-seq analysis for transcriptional evaluation of HER2 activation and global RNA-seq to identify the mechanisms underlying therapy resistance.ResultsHere we show that in CD44v6-positive CR-CSCs, high HER2 expression levels are associated with an activation of the phosphatidylinositol 3-kinase (PI3K)/AKT pathway, which promotes the acetylation at the regulatory elements of the Erbb2 gene. HER2 targeting in combination with phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase kinase (MEK) inhibitors induces CR-CSC death and regression of tumour xenografts, including those carrying Kras and Pik3ca mutation. Requirement for the triple targeting is due to the presence of cancer-associated fibroblasts, which release cytokines able to confer CR-CSC resistance to PI3K/AKT inhibitors. In contrast, targeting of PI3K/AKT as monotherapy is sufficient to kill liver-disseminating CR-CSCs in a model of adjuvant therapy.ConclusionsWhile PI3K targeting kills liver-colonising CR-CSCs, the concomitant inhibition of PI3K, HER2 and MEK is required to induce regression of tumours resistant to anti-EGFR therapies. These data may provide a rationale for designing clinical trials in the adjuvant and metastatic setting.

Objective The purpose of this review is to evaluate the use and effectiveness of the local administration of tramadol in reducing post-operative pain during surgical interventions. Methods The PubMed, EMBASE and Cochrane Central Register of Controlled Trials (CENTRAL) databases will be searched for this review. This systematic review will include studies evaluating the clinical efficacy of the local infiltration of tramadol, with no study design restrictions. Only studies that present clear descriptions of local tramadol administration are published in peer-reviewed journals in the English, Italian, Spanish, French, Portuguese or German language and are published in full will be taken into consideration. A meta-analysis will be performed when there is sufficient clinical homogeneity among the retrieved studies, and only randomized controlled studies and quasi-randomized controlled studies will be included. The Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach will be used to assess the certainty in the evidence. If a quantitative analysis cannot be conducted, a qualitative description of the results of the retrieved studies will be provided. Results A high-quality synthesis of the current evidence on the local administration of tramadol for managing post-surgical pain will be illustrated using subjective reports and objective measures of performance. The primary outcomes will include the magnitude of post-operative pain intensity improvement, with improvement being as defined by a reduction by at least 2 points in the visual analogue scale (VAS) score or numerical rating scale (NRS) score. The secondary outcomes will be the magnitude of reduction in tramadol rescue doses and in other analgesic drug doses. Conclusion This protocol will present evidence on the efficacy of tramadol in relieving post-surgical pain. Systemic review registration PROSPERO CRD42018087381

Carbapenem antibiotics were first introduced in the 1980s and have long been considered the most active agents for the treatment of multidrug-resistant gram-negative bacteria. Over the last decade, carbapenem-resistant Enterobacteriaceae (CRE) have emerged as organisms causing spontaneous bacterial peritonitis. Infections caused by CRE have shown a higher mortality rate than those caused by bacteria sensitive to carbapenem antibiotics. Current antibiotic guidelines for the treatment of spontaneous bacterial peritonitis are insufficient, and rapid de-escalation of empiric antibiotic treatment is not widely recognized. This review summarizes the molecular characteristics, epidemiology and possible treatment of spontaneous bacterial peritonitis caused by CRE.

Ceftazidime-avibactam (CZA) is a novel beta-lactam beta-lactamase inhibitor combination approved for the treatment of complicated urinary tract infections, complicated intra-abdominal infections, and for hospital-acquired/ventilator-associated pneumonia. The aim of this systematic review (PROSPERO registration number: CRD42019128927) was to evaluate the effectiveness of CZA combination therapy versus CZA monotherapy in the treatment of severe infections. The databases included in the search, until February 12th, 2020, were MEDLINE by PubMed, EMBASE, and The Cochrane Central Register of Controlled Trials. We included both randomized controlled trials (RCTs) and non-randomized studies published in peer-reviewed journals and in the English language. The primary outcome was all-cause mortality (longest follow-up) evaluated in patients with the diagnosis of infection with at least one pathogen, secondary outcomes were clinical and microbiological improvement/cure. Thirteen studies were included in the qualitative synthesis: 7 RCTs and 6 retrospective studies All the six retrospective studies identified carbapenamase-producing Enterobacteriaceae (CRE) as the cause of infection and for this reason were included in the network meta-analysis (NMA), the quality of the studies, assessed using the New Castle-Ottawa Scale, was moderate-high. In all the six retrospective studies included in the NMA, CZA was used in large part for off-label indications (mostly blood stream infections: 80&ndash, 100% of patients included). No difference in mortality rate was observed in patients undergoing CZA combination therapy compared to CZA monotherapy [n = 503 patients, direct evidence OR: 0.96, 95% CI: 0.65&ndash, 1.41].

Colorectal cancer is the fourth most common cause of cancer-related death. Although many advances in the treatment of this disease have been made, a large number of patients develop metastasis and resistance to current therapies. The current evidence indicates that cancer stem cells (CSCs) and epithelial-to-mesenchymal transition (EMT) have crucial roles in colorectal carcinogenesis and metastasis. It is also very important to understand the mechanisms that allow the survival of CSCs, such as metabolic reprogramming, which permits them to obtain specific properties or the activation of alternative signaling pathways in response to first-line therapies. In this review, we discuss the failure of conventional therapies for colorectal cancer and provide a brief analysis of new therapeutic strategies for targeting non-responsive CSC. We highlight the use of combination therapies, such as horizontal or vertical targeting, as the most efficient strategy for eradicating these subpopulations of cancer cells.

Molybdenum disulphide (MoS2) is currently regarded as a promising material for the next generation of electronic and optoelectronic devices. However, several issues need to be addressed to fully exploit its potential for field effect transistor (FET) applications. In this context, the contact resistance, RC, associated with the Schottky barrier between source/drain metals and MoS2 currently represents one of the main limiting factors for suitable device performance. Furthermore, to gain a deeper understanding of MoS2 FETs under practical operating conditions, it is necessary to investigate the temperature dependence of the main electrical parameters, such as the field effect mobility (μ) and the threshold voltage (Vth). This paper reports a detailed electrical characterization of back-gated multilayer MoS2 transistors with Ni source/drain contacts at temperatures from T = 298 to 373 K, i.e., the expected range for transistor operation in circuits/systems, considering heating effects due to inefficient power dissipation. From the analysis of the transfer characteristics (ID−VG) in the subthreshold regime, the Schottky barrier height (ΦB ≈ 0.18 eV) associated with the Ni/MoS2 contact was evaluated. The resulting contact resistance in the on-state (electron accumulation in the channel) was also determined and it was found to increase with T as RC proportional to T3.1. The contribution of RC to the extraction of μ and Vth was evaluated, showing a more than 10% underestimation of μ when the effect of RC is neglected, whereas the effect on Vth is less significant. The temperature dependence of μ and Vth was also investigated. A decrease of μ proportional to 1/Tα with α = 1.4 ± 0.3 was found, indicating scattering by optical phonons as the main limiting mechanism for mobility above room temperature. The value of Vth showed a large negative shift (about 6 V) increasing the temperature from 298 to 373 K, which was explained in terms of electron trapping at MoS2/SiO2 interface states.

Introduction: Spontaneous bacterial peritonitis (SBP) is a main infectious complication in end-stage liver disease (ESLD) patients. The increasing trend of bacterial resistance in ESLD patients with SBP has been associated with low treatment efficacy of traditional therapy. Cephalosporin use has been restricted to community-acquired infections and in areas/health care settings with low rates of multidrug-resistant (MDR) bacteria. To date, several changes are necessary with regard to empiric therapy recommendations in areas/health care settings with high rates of MDR bacteria. Areas covered: An overview of the epidemiology and antimicrobial treatments of SBP caused by Gram-negative bacteria. Expert opinion: Broad-spectrum antibiotics have been recommended as empiric therapy for suspected SBP in areas/health care settings with high rates of MDR bacteria and secondary treatment, with newer antibiotics, for SBP caused by MDR-Gram-negative bacteria (i.e. new beta-lactam/beta-lactamase inhibitor combinations, cefiderocol, plazomicin, and eravacycline) either alone or in combination.

// Bregje van Oorschot 1 , Giovanna Granata 1 , Simone Di Franco 2 , Rosemarie ten Cate 1 , Hans M. Rodermond 1 , Matilde Todaro 3 , Jan Paul Medema 1 , Nicolaas A.P. Franken 1 1 Laboratory for Experimental Oncology and Radiobiology (LEXOR), Center for Experimental Molecular Medicine, Department of Radiation Oncology, Academic Medical Center, Cancer Genomics Center, Amsterdam, The Netherlands 2 Department of Surgical, Oncological and Stomatological Sciences (DICHIRONS), Cellular and Molecular Pathophysiology Laboratory, University of Palermo, Palermo, Italy 3 Biomedical Department of Internal and Specialistic Medicine (DIBIMIS), University of Palermo, Palermo, Italy Correspondence to: Nicolaas A.P. Franken, email: n.a.franken@amc.uva.nl Keywords: radiation oncology, DNA repair, hyperthermia, double-strand breaks Received: May 10, 2016 Accepted: August 24, 2016 Published: September 01, 2016 ABSTRACT Radiotherapy is based on the induction of lethal DNA damage, primarily DNA double-strand breaks (DSB). Efficient DSB repair via Non-Homologous End Joining or Homologous Recombination can therefore undermine the efficacy of radiotherapy. By suppressing DNA-DSB repair with hyperthermia (HT) and DNA-PKcs inhibitor NU7441 (DNA-PKcs i ), we aim to enhance the effect of radiation. The sensitizing effect of HT for 1 hour at 42°C and DNA-PKcs i [1 μM] to radiation treatment was investigated in cervical and breast cancer cells, primary breast cancer sphere cells (BCSCs) enriched for cancer stem cells, and in an in vivo human tumor model. A significant radio-enhancement effect was observed for all cell types when DNA-PKcs i and HT were applied separately, and when both were combined, HT and DNA-PKcs i enhanced radio-sensitivity to an even greater extent. Strikingly, combined treatment resulted in significantly lower survival rates, 2 to 2.5 fold increase in apoptosis, more residual DNA-DSB 6 h post treatment and a G2-phase arrest. In addition, tumor growth analysis in vivo showed significant reduction in tumor growth and elevated caspase-3 activity when radiation was combined with HT and DNA-PKcs i compared to radiation alone. Importantly, no toxic side effects of HT or DNA-PKcs i were found. In conclusion, inhibiting DNA-DSB repair using HT and DNA-PKcs i before radiotherapy leads to enhanced cytotoxicity in cancer cells. This effect was even noticed in the more radio-resistant BCSCs, which are clearly sensitized by combined treatment. Therefore, the addition of HT and DNA-PKcs i to conventional radiotherapy is promising and might contribute to more efficient tumor control and patient outcome.

Background The use of stem cells, including mesenchymal stem cells (MSCs), for regenerative medicine is gaining interest for the clinical benefits so far obtained in patients. This study investigates the use of adipose autologous tissue in combination with platelet-rich plasma (PRP) to improve the clinical outcome of patients affected by systemic sclerosis (SSc). Methods Adipose-derived mesenchymal stem cells (AD-MSCs) and PRPs were purified from healthy donors and SSc patients. The multilineage differentiation potential of AD-MSCs and their genotypic–phenotypic features were investigated. A cytokine production profile was evaluated on AD-MSCs and PRPs from both healthy subjects and SSc patients. The adipose tissue-derived cell fraction, the so-called stromal vascular fraction (SVF), was coinjected with PRP in the perioral area of SSc patients. Results Histopathological and phenotypical analysis of adipose tissue from SSc patients revealed a disorganization of its distinct architecture coupled with an altered cell composition. Although AD-MSCs derived from SSc patients showed high multipotency, they failed to sustain a terminally differentiated progeny. Furthermore, SVFs derived from SSc patients differed from healthy donors in their MSC-like traits coupled with an aberrant cytokine production profile. Finally, the administration of PRP in combination with autologous SVF improved buccal’s rhyme, skin elasticity and vascularization for all of the SSc patients enrolled in this study. Conclusions This innovative regenerative therapy could be exploited for the treatment of chronic connective tissue diseases, including SSc. Electronic supplementary material The online version of this article (doi:10.1186/s13287-017-0690-3) contains supplementary material, which is available to authorized users.

Contacts with MoS2 are currently the object of many investigations, since current injection through metal/MoS2 interfaces represents one of the limiting factors to the performance of MoS2 thin film transistors. In this paper, we employed conductive atomic force microscopy (CAFM) to investigate the current injection mechanisms from a nanometric contact (a Pt coated tip) to the surface of MoS2 thin films exfoliated on SiO2. The analysis of local current-voltage (I-V) characteristics on a large array of tip positions provided high spatial resolution information on the lateral homogeneity of the tip/MoS2 Schottky barrier Phi(B) and of the ideality factor n. From the histograms of the measured Phi(B) and n values, an average Schottky barrier height of 297 meV with standard deviation of 22 meV and an average ideality factor of 1.65 with a standard deviation is 0.15 have been estimated. The implications of these lateral variations of Phi(B) and n in MoS2 nano-Schottky diodes on the electrical properties of macroscopic contacts to MoS2 have been discussed also in relation with recent literature results. (C) 2015 Elsevier Ltd. All rights reserved.

P63 is a transcription factor belonging to the family of p53, essential for the development and differentiation of epithelia. In recent years, it has become clear that altered expression of the different isoforms of this gene can play an important role in carcinogenesis. The p63 gene encodes for two main isoforms known as TA and ΔN p63 with different functions. The role of these different isoforms in sustaining tumor progression and metastatic spreading however has not entirely been clarified. Here we show that breast cancer initiating cells express ΔNp63 isoform that supports a more mesenchymal phenotype associated with a higher tumorigenic and metastatic potential. On the contrary, the majority of cells within the tumor appears to express predominantly TAp63 isoform. While ΔNp63 exerts its effects by regulating a PI3K/CD44v6 pathway, TAp63 modulates this pathway in an opposite fashion. As a result, tumorigenicity and invasive capacity of breast cancer cells is a balance of the two isoforms. Finally, we found that tumor microenvironmental cytokines significantly contribute to the establishment of breast cancer cell phenotype by positively regulating ΔNp63 and CD44v6 expression.

It is widely accepted by the scientific community that cancer, including colon cancer, is a “stem cell disease”. Until a few years ago, common opinion was that all neoplastic cells within a tumor contained tumorigenic growth capacity, but recent evidences hint to the possibility that such a feature is confined to a small subset of cancer-initiating cells, also called cancer stem cells (CSCs). Thus, malignant tumors are organized in a hierarchical fashion in which CSCs give rise to more differentiated tumor cells. CSCs possess high levels of ATP-binding cassette (ABC) transporters and anti-apoptotic molecules, active DNA-repair, slow replication capacities and they produce growth factors that confer refractoriness to antineoplastic treatments. The inefficacy of conventional therapies towards the stem cell population might explain cancer chemoresistance and the high frequency of relapse shown by the majority of tumors. Nowadays, in fact all the therapies available are not sufficient to cure patients with advanced forms of colon cancer since they target differentiated cancer cells which constitute most of the tumor mass and spare CSCs. Since CSCs are the entities responsible for the development of the tumor and represent the only cell population able to sustain tumor growth and progression, these cells represent the elective target for innovative therapies.