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Tumor adaptation or selection is thought to underlie therapy resistance in glioma. To investigate longitudinal epigenetic evolution of gliomas in response to therapeutic pressure, we performed an epigenomic analysis of 132 matched initial and recurrent tumors from patients with IDH-wildtype (IDHwt) and IDH-mutant (IDHmut) glioma. IDHwt gliomas showed a stable epigenome over time with relatively low levels of global methylation. The epigenome of IDHmut gliomas showed initial high levels of genome-wide DNA methylation that was progressively reduced to levels similar to those of IDHwt tumors. Integration of epigenomics, gene expression, and functional genomics identified HOXD13 as a master regulator of IDHmut astrocytoma evolution. Furthermore, relapse of IDHmut tumors was accompanied by histologic progression that was associated with survival, as validated in an independent cohort. Finally, the initial cell composition of the tumor microenvironment varied between IDHwt and IDHmut tumors and changed differentially following treatment, suggesting increased neoangiogenesis and T-cell infiltration upon treatment of IDHmut gliomas. This study provides one of the largest cohorts of paired longitudinal glioma samples with epigenomic, transcriptomic, and genomic profiling and suggests that treatment of IDHmut glioma is associated with epigenomic evolution toward an IDHwt-like phenotype., Significance: Standard treatments are related to loss of DNA methylation in IDHmut glioma, resulting in epigenetic activation of genes associated with tumor progression and alterations in the microenvironment that resemble treatment-naïve IDHwt glioma., (©2023 The Authors; Published by the American Association for Cancer Research.)

Background: This study sought to investigate the diagnostic procedures, treatment modalities, and consequences of anastomotic leakage (AL) in low anterior resection rectal cancer patients. Methods: A retrospective analysis was conducted on 186 patients consecutively admitted and treated in the 1st Department of Surgery in Craiova, between January 2018 and June 2022, all of whom had undergone surgical interventions for adenocarcinoma of the rectum. Among this cohort, 106 patients who had undergone scheduled low and ultralow anterior rectal resections with total mesorectal excision were selected for further analysis. Twenty-four patients were diagnosed with postoperative AL and underwent diverse management strategies based on the severity of their condition. Results: The study revealed an incidence of 22.6% for postoperative AL, with all of them being classified as grade B and C, according to the 2010 International Study Group of Rectal Cancer Classification, which were associated with significant morbidity and mortality. Notably, patients exhibited various comorbidities, including obesity, arterial hypertension, type 2 diabetes mellitus, and kidney failure. The management approach depended on the severity of the clinical presentation and the availability of treatment options. Early diagnosis and conservative management constituted the initial therapeutic strategy for grade B AL, with surgical reintervention or transanal vacuum therapy being used in grade C AL. Conclusions: The incidence and mortality associated with AF following low anterior resections were notably elevated. Grade B AL were successfully managed through conservative treatment, whereas grade C AL required either surgical reintervention for drainage or diversion procedures, or transanal vacuum therapy., (Celsius.)

Background: While ventral hernia repair is a frequent surgical intervention, the possibility of complications remains present. The use of drains to mitigate complications is a topic of debate, with conflicting evidence. This study aimed to evaluate the association between drain usage and postoperative complications in ventral hernia repair. Materials and Methods: A single-center prospective study included patients undergoing ventral hernia repair from 2018 to 2022. Patient data and surgical techniques were recorded. Statistical analysis was performed to assess risk factors for drain insertion and complications. Results: Of the 216 patients included, 19.44% had diabetes, and 20% had cancer. Postoperative complications (Clavien Dindo grade IIIB) occurred in 9.3% of cases, resulting in a 3.7% mortality. Decision factors for drain insertion included older age, larger hernia size, bowel resection with anastomosis, emergency setting and the need for adhesiolysis. No differences were found between the two groups regarding seroma and hematoma formation and mesh infection. Patients with drains had a longer hospital stay and higher costs. Conclusion: The decision to use drains in ventral hernia repair was influenced by surgical complexity factors rather than patient characteristics. While drain usage did not correlate with postoperative morbidities, it was associated with longer hospitalization and higher costs. Individualized decision-making is crucial to balance complications and resource utilization in ventral hernia repair., (Celsius.)

Glioblastoma (GBM) is the most common and aggressive primary brain tumor. GBM contains a small subpopulation of glioma stem cells (GSCs) that are implicated in treatment resistance, tumor infiltration, and recurrence, and are thereby considered important therapeutic targets. Recent clinical studies have suggested that the choice of general anesthetic (GA), particularly propofol, during tumor resection, affects subsequent tumor response to treatments and patient prognosis. In this study, we investigated the molecular mechanisms underlying propofol's anti-tumor effects on GSCs and their interaction with microglia cells. Propofol exerted a dose-dependent inhibitory effect on the self-renewal, expression of mesenchymal markers, and migration of GSCs and sensitized them to both temozolomide (TMZ) and radiation. At higher concentrations, propofol induced a large degree of cell death, as demonstrated using microfluid chip technology. Propofol increased the expression of the lncRNA BDNF-AS, which acts as a tumor suppressor in GBM, and silencing of this lncRNA partially abrogated propofol's effects. Propofol also inhibited the pro-tumorigenic GSC-microglia crosstalk via extracellular vesicles (EVs) and delivery of BDNF-AS. In conclusion, propofol exerted anti-tumor effects on GSCs, sensitized these cells to radiation and TMZ, and inhibited their pro-tumorigenic interactions with microglia via transfer of BDNF-AS by EVs.

Diabetes mellitus (DM) and obesity are known to influence postoperative outcomes in surgical patients. This study aimed to analyze the divergence in outcomes between patients with and without DM who underwent ventral hernia repair, while also assessing the additional impact of obesity on postoperative complications, hospitalization, and costs. A prospective, non-interventional, observational study was conducted over five years (2018-2022) at the First Surgical Clinic of Craiova Emergency Clinical Hospital. The study included 216 patients, with 42 (19.44%) having DM, half of whom were also obese. There were no significant differences in age between patients with or without DM. However, diabetes and obesity were more prevalent in female patients. The mean duration of hospitalization was 10.7±8.6 days, with no significant differences observed between patients with or without DM or obesity. Statistical analysis revealed that patients with DM had a higher incidence of seroma formation, wound infection, and hematoma formation compared to patients without DM. However, there were no significant differences in mesh infection, hospitalization days, or costs between the two groups. Similarly, no significant differences were found between obese and non-obese patients in terms of complications, hospitalization days, or costs. In conclusion, this study highlights that DM is associated with an increased risk of specific complications in ventral hernia repair, including seroma formation, wound infection, and hematoma formation. However, the impact of obesity on these outcomes appears to be limited. Individualized preoperative optimization and targeted interventions are necessary to mitigate the risk of complications in patients with DM or obesity., Competing Interests: None to declare., (Copyright © 2023, Medical University Publishing House Craiova.)

We aimed to comparatively assess the prognostic preoperative value of the main peripheral blood components and their ratios-the systemic immune-inflammation index (SII), neutrophil-to-lymphocyte ratio (NLR), lymphocyte-to-monocyte ratio (LMR), and platelet-to-lymphocyte ratio (PLR)-to the use of artificial-neural-network analysis in determining undesired postoperative outcomes in colorectal cancer patients. Our retrospective study included 281 patients undergoing elective radical surgery for colorectal cancer in the last seven years. The preoperative values of SII, NLR, LMR, and PLR were analyzed in relation to postoperative complications, with a special emphasis on their ability to accurately predict the occurrence of anastomotic leak. A feed-forward fully connected multilayer perceptron network (MLP) was trained and tested alongside conventional statistical tools to assess the predictive value of the abovementioned blood markers in terms of sensitivity and specificity. Statistically significant differences and moderate correlation levels were observed for SII and NLR in predicting the anastomotic leak rate and degree of postoperative complications. No correlations were found between the LMR and PLR or the abovementioned outcomes. The MLP network analysis showed superior prediction value in terms of both sensitivity (0.78 ± 0.07; 0.74 ± 0.04; 0.71 ± 0.13) and specificity (0.81 ± 0.11; 0.69 ± 0.03; 0.9 ± 0.04) for all the given tasks. Preoperative SII and NLR appear to be modest prognostic factors for anastomotic leakage and overall morbidity. Using an artificial neural network offers superior prognostic results in the preoperative risk assessment for overall morbidity and anastomotic leak rate.

Introduction: Inguinal hernia is one of the most common conditions in surgical departments and diabetes is known to have a significant impact on both patients' health and healthcare system. The current study aims to evaluate extensively the differences in costs for different subgroups of diabetic patients undergoing inguinal hernia repair in a tertiary care medical center in South-West Romania., Material and Methods: A total number of 195 patients underwent hernia repair for primary or recurrent inguinal hernia between 2015 and 2020 and were included in the analysis. The group was divided into diabetic/non-diabetic patients and each of these subgroups was studied separately according to the presence or absence of comorbidities, sex and place of origin., Results: Statistically significant differences were found in the division of the age group, highlighting the fact that in diabetic patients the average age was around 74.76 years, unlike those without diabetes in which the average age was approximately 61.31 years. Another statistically significant difference was observed in the classification by hospitalization days of the group of patients with incarcerated inguinal hernia, in a sense that diabetic patients required a prolonged hospitalization by 1.5 days as opposed to non-diabetics., Conclusions: The costs of hospitalization for diabetic patients undergoing hernia repair surgery were marginally higher than in non-diabetic patients, but no statistical difference could be observed between any of the costs subsets in the two patients groups., Competing Interests: None to declare., (Copyright © 2014, Medical University Publishing House Craiova.)

Background: There is a compelling evidence from animal models that early exposure to clinically relevant general anesthetics (GAs) interferes with brain development, resulting in long-lasting cognitive impairments. Human studies have been inconclusive and are challenging due to numerous confounding factors. Here, we employed primary human neural cells to analyze ketamine neurotoxic effects focusing on the role of glial cells and their activation state. We also explored the roles of astrocyte-derived extracellular vesicles (EVs) and different components of the brain-derived neurotrophic factor (BDNF) pathway. Methods: Ketamine effects on cell death were analyzed using live/dead assay, caspase 3 activity and PARP-1 cleavage. Astrocytic and microglial cell differentiation was determined using RT-PCR, ELISA and phagocytosis assay. The impact of the neuron-glial cell interactions in the neurotoxic effects of ketamine was analyzed using transwell cultures. In addition, the role of isolated and secreted EVs in this cross-talk were studied. The expression and function of different components of the BDNF pathway were analyzed using ELISA, RT-PCR and gene silencing. Results: Ketamine induced neuronal and oligodendrocytic cell apoptosis and promoted pro-inflammatory astrocyte (A1) and microglia (M1) phenotypes. Astrocytes and microglia enhanced the neurotoxic effects of ketamine on neuronal cells, whereas neurons increased oligodendrocyte cell death. Ketamine modulated different components in the BDNF pathway: decreasing BDNF secretion in neurons and astrocytes while increasing the expression of p75 in neurons and that of BDNF-AS and pro-BDNF secretion in both neurons and astrocytes. We demonstrated an important role of EVs secreted by ketamine-treated astrocytes in neuronal cell death and a role for EV-associated BDNF-AS in this effect. Conclusions: Ketamine exerted a neurotoxic effect on neural cells by impacting both neuronal and non-neuronal cells. The BDNF pathway and astrocyte-derived EVs represent important mediators of ketamine effects. These results contribute to a better understanding of ketamine neurotoxic effects in humans and to the development of potential approaches to decrease its neurodevelopmental impact., Competing Interests: AB was employed by Precise Cell Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Penning, Cazacu, Brodie, Jevtovic-Todorovic, Kalkanis, Lewis and Brodie.)

Inactivating mutations in the Methyl-CpG Binding Protein 2 (MECP2) gene are the main cause of Rett syndrome (RTT). Despite extensive research into MECP2 function, no treatments for RTT are currently available. Here, we used an evolutionary genomics approach to construct an unbiased MECP2 gene network, using 1028 eukaryotic genomes to prioritize proteins with strong co-evolutionary signatures with MECP2. Focusing on proteins targeted by FDA-approved drugs led to three promising targets, two of which were previously linked to MECP2 function (IRAK, KEAP1) and one that was not (EPOR). The drugs targeting these three proteins (Pacritinib, DMF, and EPO) were able to rescue different phenotypes of MECP2 inactivation in cultured human neural cell types, and appeared to converge on Nuclear Factor Kappa B (NF-κB) signaling in inflammation. This study highlights the potential of comparative genomics to accelerate drug discovery, and yields potential new avenues for the treatment of RTT., Competing Interests: IU, IB, SC, GK, BB, BB, CB, YT No competing interests declared, (© 2021, Unterman et al.)

Objectives: To determine if patients with coronavirus disease 2019 had a greater number of unplanned extubations resulting in reintubations than in patients without coronavirus disease 2019., Design: Retrospective cohort study comparing the frequency of unplanned extubations resulting in reintubations in a group of coronavirus disease 2019 patients to a historical (noncoronavirus disease 2019) control group., Setting: This study was conducted at Henry Ford Hospital, an academic medical center in Detroit, MI. The historical noncoronavirus disease 2019 patients were treated in the 68 bed medical ICU. The coronavirus disease 2019 patients were treated in the coronavirus disease ICU, which included the 68 medical ICU beds, 18 neuro-ICU beds, 32 surgical ICU beds, and 40 cardiovascular ICU beds, as the medical ICU was expanded to these units at the peak of the pandemic in Detroit, MI., Patients: The coronavirus disease 2019 cohort included patients diagnosed with coronavirus disease 2019 who were intubated for respiratory failure from March 12, 2020, to April 13, 2020. The historic control (noncoronavirus disease 2019) group consisted of patients who were admitted to the medical ICU in the year spanning from November 1, 2018 to October 31, 2019, with a need for mechanical ventilation that was not related to surgery or a neurologic reason., Interventions: None., Measurements and Main Results: To identify how many patients in each cohort had unplanned extubations, an electronic medical records query for patients with two intubations within 30 days was performed, in addition to a review of our institutional quality and safety database of reported self-extubations. Medical charts were manually reviewed by board-certified anesthesiologists to confirm each event was an unplanned extubation followed by a reintubation within 24 hours. There was a significantly greater incidence of unplanned extubations resulting in reintubation events in the coronavirus disease 2019 cohort than in the noncoronavirus disease 2019 cohort (coronavirus disease 2019 cohort: 167 total admissions with 22 events-13.2%; noncoronavirus disease 2019 cohort: 326 total admissions with 14 events-4.3%; p < 0.001). When the rate of unplanned extubations was expressed per 100 intubated days, there was not a significant difference between the groups (0.88 and 0.57, respectively; p = 0.269)., Conclusions: Coronavirus disease 2019 patients have a higher incidence of unplanned extubation that requires reintubation than noncoronavirus disease 2019 patients. Further study is necessary to evaluate the variables that contribute to this higher incidence and clinical strategies that can reduce it., Competing Interests: The authors have disclosed that they do not have any potential conflicts of interest., (Copyright © 2020 The Authors. Published by Wolters Kluwer Health, Inc. on behalf of the Society of Critical Care Medicine.)

Glioblastoma (GBM) is a highly aggressive tumor with poor prognosis. A small subpopulation of glioma stem cells (GSCs) has been implicated in radiation resistance and tumor recurrence. In this study we analyzed the expression of miRNAs associated with the functions of GSCs using miRNA microarray analysis of these cells compared with human neural stem cells. These analyses identified gene clusters associated with glioma cell invasiveness, axonal guidance, and TGF-β signaling. miR-504 was significantly downregulated in GSCs compared with NSCs, its expression was lower in GBM compared with normal brain specimens and further decreased in the mesenchymal glioma subtype. Overexpression of miR-504 in GSCs inhibited their self-renewal, migration and the expression of mesenchymal markers. The inhibitory effect of miR-504 was mediated by targeting Grb10 expression which acts as an oncogene in GSCs and GBM. Overexpression of exogenous miR-504 resulted also in its delivery to cocultured microglia by GSC-secreted extracellular vesicles (EVs) and in the abrogation of the GSC-induced polarization of microglia to M2 subtype. Finally, miR-504 overexpression prolonged the survival of mice harboring GSC-derived xenografts and decreased tumor growth. In summary, we identified miRNAs and potential target networks that play a role in the stemness and mesenchymal transition of GSCs and the miR-504/Grb10 pathway as an important regulator of this process. Overexpression of miR-504 exerted antitumor effects in GSCs as well as bystander effects on the polarization of microglia via delivery by EVs.

Duchenne muscular dystrophy (DMD) is a progressive, lethal, X-linked disease of skeletal and cardiac muscles caused by mutations in the dystrophin gene. Loss of dystrophin leads to muscle fiber damage and impairment of satellite cell asymmetric division, which are essential for muscle regeneration. These processes ultimately result in muscle wasting and the replacement of the degenerating muscles by fibrogenic cells, a process that leads to the generation of fibrotic tissues. Preimplantation factor (PIF) is an evolutionary conserved 15-amino acid peptide secreted by viable mammalian embryos. Synthetic PIF (sPIF) reproduces the protective/regenerative effects of the endogenous peptide in immune disorders and transplantation models. In this study, we demonstrated that sPIF treatment promoted mouse and human myoblast differentiation and inhibited the expression of collagen 1A1, collagen 1A2, and TGF-β in DMD patient-derived myoblasts. Additionally, sPIF increased the expression of utrophin, a homolog of dystrophin protein. sPIF effects were mediated via the upregulation of lncRNA H19 and miR-675 and downregulation of let-7. sPIF also inhibited the expression of miR-21, a major fibrosis regulator. The administration of sPIF in mdx mice significantly decreased serum creatine kinase and collagen I and collagen IV expression in the diaphragm, whereas it increased utrophin expression in the diaphragm, heart and quadriceps muscles. In conclusion, sPIF promoted the differentiation of DMD myoblasts, increased utrophin expression via the H19/miRNA-675/let-7 pathway, and reduced muscle fibrosis possibly via the upregulation of miR-675 and inhibition of miR-21 expression. These findings strongly support pursuing sPIF as a potential therapeutic agent for DMD. Moreover, the completion of an sPIF phase I safety trial will further promote the use of sPIF for the treatment of muscular dystrophies.

Duchenne muscular dystrophy (DMD) is a degenerative lethal, X-linked disease of skeletal and cardiac muscles caused by mutations in the dystrophin gene. Cell therapy using different cell types, including mesenchymal stromal cells (MSCs), has been considered as a potential approach for the treatment of DMD. MSCs can be obtained from autologous sources such as bone marrow and adipose tissues or from allogeneic placenta and umbilical cord. The safety and therapeutic impact of these cells has been demonstrated in pre-clinical and clinical studies and their functions are attributed to paracrine effects that are mediated by secreted cytokines and extracellular vesicles. Here, we studied the therapeutic effects of placenta-derived MSCs (PL-MSCs) and their secreted exosomes using mouse and human myoblasts from healthy controls, Duchenne patients and mdx mice. Treatment of myoblasts with conditioned medium or exosomes secreted by PL-MSCs increased the differentiation of these cells and decreased the expression of fibrogenic genes in DMD patient myoblasts. In addition, these treatments also increased the expression of utrophin in these cells. Using a quantitative miR-29c reporter, we demonstrated that the PL-MSC effects were partly mediated by the transfer of exosomal miR-29c. Intramuscular transplantation of PL-MSCs in mdx mice resulted in decreased creatine kinase levels. PL-MSCs significantly decreased the expression of TGF-β and the level of fibrosis in the diaphragm and cardiac muscles, inhibited inflammation and increased utrophin expression. In vivo imaging analyses using MSCs labeled with gold nanoparticles or fluorescent dyes demonstrated localization of the cells in the muscle tissues up to 3 weeks post treatment. Altogether, these results demonstrate that PL-MSCs and their secreted exosomes have important clinical applications in cell therapy of DMD partly via the targeted delivery of exosomal miR-29c., (Copyright © 2018. Published by Elsevier Ltd.)

The early diagnosis of esophageal cancer is necessary for improving the surviving of patients with this disease. To ensure an accurate staging, there are necessary imaging tests to establish the local and regional extension, as well as excluding the metastases. Computed tomography (CT), endoscopic ultrasonography (EUS), and positron emission computed tomography (PET-CT) constitute standard methods for esophageal cancer staging. These techniques are complementary; using only one of these tests is not suitable for correct staging. The role of EUS has improved the doctors' ability to evaluate and select the patients to undergo surgery, radiotherapy, or chemotherapy.

Recent studies have proposed that abnormal glutamatergic neurotransmission and glial pathology play an important role in the etiology and manifestation of depression. It was postulated that restoration of normal glutamatergic transmission, by enhancing glutamate uptake, may have a beneficial effect on depression. We examined this hypothesis using unique human glial-like mesenchymal stem cells (MSCs), which in addition to inherent properties of migration to regions of injury and secretion of neurotrophic factors, were differentiated to express high levels of functional glutamate transporters (excitatory amino acid transporters; EAAT). Additionally, gold nanoparticles (GNPs), which serve as contrast agents for CT imaging, were loaded into the cells for non-invasive, real-time imaging and tracking of MSC migration and final location within the brain. MSC-EAAT (2×10 5 ; 10 μl) were administered (i.c.v.) to Flinder Sensitive Line rats (FSLs), a genetic model for depression, and longitudinal behavioral and molecular changes were monitored. FSL rats treated with MSC-EAAT showed attenuated depressive-like behaviors (measured by the forced swim test, novelty exploration test and sucrose self-administration paradigm), as compared to controls. CT imaging, Flame Atomic Absorption Spectroscopy analysis and immunohistochemistry showed that the majority of MSCs homed specifically to the dentate gyrus of the hippocampus, a region showing structural brain changes in depression, including loss of glial cells. mRNA and protein levels of EAAT1 and BDNF were significantly elevated in the hippocampus of MSC-EAAT-treated FSLs. Our findings indicate that MSC-EAATs effectively improve depressive-like manifestations, possibly in part by increasing both glutamate uptake and neurotropic factor secretion in the hippocampus., Competing Interests: Competing Interests: The authors have declared that no competing interest exists.

Despite advances in novel therapeutic approaches for the treatment of glioblastoma (GBM), the median survival of 12-14 months has not changed significantly. Therefore, there is an imperative need to identify molecular mechanisms that play a role in patient survival. Here, we analyzed the expression and functions of a novel lncRNA, TALNEC2 that was identified using RNA seq of E2F1-regulated lncRNAs. TALNEC2 was localized to the cytosol and its expression was E2F1-regulated and cell-cycle dependent. TALNEC2 was highly expressed in GBM with poor prognosis, in GBM specimens derived from short-term survivors and in glioma cells and glioma stem cells (GSCs). Silencing of TALNEC2 inhibited cell proliferation and arrested the cells in the G1\S phase of the cell cycle in various cancer cell lines. In addition, silencing of TALNEC2 decreased the self-renewal and mesenchymal transformation of GSCs, increased sensitivity of these cells to radiation and prolonged survival of mice bearing GSC-derived xenografts. Using miRNA array analysis, we identified specific miRNAs that were altered in the silenced cells that were associated with cell-cycle progression, proliferation and mesenchymal transformation. Two of the downregulated miRNAs, miR-21 and miR-191, mediated some of TALNEC2 effects on the stemness and mesenchymal transformation of GSCs. In conclusion, we identified a novel E2F1-regulated lncRNA that is highly expressed in GBM and in tumors from patients of short-term survival. The expression of TALNEC2 is associated with the increased tumorigenic potential of GSCs and their resistance to radiation. We conclude that TALNEC2 is an attractive therapeutic target for the treatment of GBM.

Glioblastoma (GBM) is the most aggressive primary brain tumor with poor prognosis. Here, we studied the effects of phenformin, a mitochondrial complex I inhibitor and more potent chemical analog of the diabetes drug metformin on the inhibition of cell growth and induction of apoptosis of glioma stem cells (GSCs) using both in vitro and in vivo models. Phenformin inhibited the self-renewal of GSCs, decreased the expression of stemness and mesenchymal markers and increased the expression of miR-124, 137 and let-7. Silencing of let-7 abrogated phenformin effects on the self-renewal of GSCs via a pathway associated with inhibition of H19 and HMGA2 expression. Moreover, we demonstrate that phenformin inhibited tumor growth and prolonged the overall survival of mice orthotopically transplanted with GSCs. Combined treatments of phenformin and temozolomide exerted an increased antitumor effect on GSCs in vitro and in vivo. In addition, dichloroacetate, an inhibitor of the glycolysis enzyme pyruvate dehydrogenase kinase, that decreases lactic acidosis induced by biguanides, enhanced phenformin effects on the induction of cell death in GSCs and prolonged the survival of xenograft-bearing mice. Our results demonstrate for the first time that phenformin targets GSCs and can be efficiently combined with current therapies for GBM treatment and GSC eradication., Competing Interests: Conflicts of Interest: None declared.

Aims: It is well known that endoscopic ultrasound guided fine needle aspiration (EUS-FNA) has a high sensitivity (over 85%) and specificity (100%) for diagnosis of pancreatic cancer. The aim of the study was to establish a EUS based clinical diagnostic algorithm in patients with pancreatic masses and negative cytopathology after EUS-FNA, based on previously published results and cut-offs of real-time elastographic (RTE) EUS and contrast-enhanced harmonic (CEH) EUS., Material and Methods: We included in the study a subgroup of 50 consecutive patients with focal pancreatic masses which underwent EUS examinations with negative EUS-FNA. RTE-EUS and CEH-EUS were performed sequentially in all patients. The sensitivity, specificity and accuracy of these methods were calculated separately. A clinical decision algorithm based on elastography followed by CEH was established., Results: For the diagnosis of possible malignancy, the sensitivity, specificity and accuracy of RTE-EUS were: 97.7%, 77.4%, and 84% respectively. CEH-EUS had similar results: 89.5%, 80.7%, and 84%, respectively. In 25 patients with soft/mixed appearance during elastography,sequential assessment using contrast-enhanced EUSwas performed. The specificity of CEH-EUS for detection of chronic pancreatitis in this sub-set of patients was excellent (100%). In other 25 patients with hard appearance in elastography (low strain) CEH-EUS had an excellent specificity (100%) and accuracy (93%) in the detection of pancreatic cancer., Conclusions: The proposed algorithm with sequential use of elastography followed by CEH could be a good clinical tool in the set of patients with negative EUS-FNA results for the differentiation between benign and malignant focal pancreatic masses.

Gliomas are the most common primary brain tumor and one of the most lethal solid tumors. Mechanistic studies into identification of novel biomarkers are needed to develop new therapeutic strategies for this deadly disease. The objective for this study was to explore the potential direct impact of IL-17-IL-17R interaction in gliomas. Immunohistochemistry and flow cytometry analysis of 12 tumor samples obtained from patients with high grade gliomas revealed that a considerable population (2-19%) of cells in all malignant gliomas expressed IL-17RA, with remarkable co-expression of the glioma stem cell (GSC) markers CD133, Nestin, and Sox2. IL-17 enhanced the self-renewal of GSCs as determined by proliferation and Matrigel® colony assays. IL-17 also induced cytokine/chemokine (IL-6, IL-8, interferon-γ-inducible protein [IP-10], and monocyte chemoattractant protein-1 [MCP-1]) secretion in GSCs, which were differentially blocked by antibodies against IL-17R and IL-6R. Western blot analysis showed that IL-17 modulated the activity of signal transducer and activator of transcription 3 (STAT3), nuclear factor κ-light-chain-enhancer of activated B cells (NF-κB), glycogen synthase kinase-3β (GSK-3β) and β-catenin in GSCs. While IL-17R-mediated secretion of IL-6 and IL-8 were significantly blocked by inhibitors of NF-κB and STAT3; NF-κB inhibitor was more potent than STAT3 inhibitor in blocking IL-17-induced MCP-1 secretion. Overall, our results suggest that IL-17-IL-17R interaction in GSCs induces an autocrine/paracrine cytokine feedback loop, which may provide an important signaling component for maintenance/self-renewal of GSCs via constitutive activation of both NF-κB and STAT3. The results also strongly implicate IL-17R as an important functional biomarker for therapeutic targeting of GSCs.

Background: Crohn's disease and ulcerative colitis are inflammatory bowel diseases (IBD) associated with colorectal cancer risk in long-standing diseases. In order to assess the colonic mucosa and to discover dysplastic or neoplastic lesions, advanced endoscopic techniques are needed. Such techniques are detailed in this review: chromoendoscopy, autofluorescence imaging (AFI), narrow band imaging (NBI), i-SCAN, Fujinon Intelligent Color Enhancement (FICE) and confocal laser endomicroscopy (CLE)., Aim: The aim of the review is to describe and establish the clinical impact of advanced endoscopic techniques, that could be used in IBD patients'examination in order to assess mucosal healing, microscopic inflammation, dysplasia or neoplasia., Materials and Methods: A literature research about new endoscopic approaches of patients with IBD was made., Results: A lot of studies have been performed to reveal which imaging technique might be used for IBD surveillance. Regarding dysplasia or neoplasia detection and mucosal healing or inflammation assessment, CE proved to be superior to white light endoscopy (WLE), while NBI and AFI did not show an encouraging result. I-SCAN did not improve the colonoscopy quality while FICE has been used in a few studies. CLE could be used to characterize a lesion, providing the same results as conventional histology., Conclusion: At the moment, CE is the only technique which has been included in guidelines for IBD surveillance. CLE can be used to assess any lesion detected with WLE during surveillance, while the other imaging techniques require.more studies to determine their efficacy or inefficacy.

Fibrosis assessment is a necessary component of liver disease evaluation not only for prognosis but also for future therapeutic management. Our study objective was to evaluate the accuracy of quantitative assessment of liver fibrosis in patients with chronic viral hepatitis B or C, relying on RTSE results, a method approved and acknowledged in Europe., Material and Methods: Sixty-three consecutive patients diagnosed with chronic viral hepatitis B or C between January 2014 and December 2014 at the Emergency County Hospital of Craiova were enrolled in the study. Patients underwent both TE and RTSE., Results: The reference method used for staging liver fibrosis was TE, based on its recognition and validation by the European guidelines. Fibrosis was classified as follows: 17.47% of patients were staged as F0, 11.11% of patients staged as F1, 14.28% as F2, 17.47% as F3, and 39.68% as F4. Correlation coefficients between measurements for each parameter was done with ANOVA test, in order to identify any differences, according to the fibrosis stage. Valuable information was obtained suggesting that MEAN, SD, %AREA, COMP, Skewness, IDM and Contrast had highly significant differences when related to the Fibrosis Stage (FS) (p<0,001) and ASM had significant differences (p<0,05). As for Kurtosis, ENT and Correlation parameters no significant differences with the FS was found., Conclusions: Imaging methods of assessing liver fibrosis are of special interest in chronic liver fibrosis assessment. RTSE comes as a potential new technology based on elastogram evaluation which may prove to be more efficient along with larger prospective studies.