Targeting AMP-activated kinase impacts hepatocellular cancer stem cells induced by long-term treatment with sorafenib

Hepatocellular carcinoma (HCC) is the third leading cause of cancer death worldwide. HCC treatment is hindered by the frequent emergence of chemoresistance to the multikinase inhibitor sorafenib, which has been related to the presence of cancer stem cells (CSCs) that self‐renew and often escape therapy. The key metabolic sensor AMP‐activated kinase (AMPK) has recently been recognized as a tumour growth regulator. In this study, we aimed to elucidate the role of AMPK in the development of a stem cell phenotype in HCC cells. To this end, we enriched the CSC population in HCC cell lines that showed increased expression of drug resistance (ALDH1A1, ABCB1A) and stem cell (CD133, Nanog, Oct4, alpha fetoprotein) markers and demonstrated their stemness phenotype. These cells were refractory to sorafenib‐induced cell death. We report that sorafenib‐resistant cells had lower levels of total and phosphorylated AMPK as well as its downstream substrate, ACC, compared with the parental cells. Interestingly, AMPK knockdown with siRNA or inhibition with dorsomorphin increased the expression of stem cell markers in parental cells and blocked sorafenib‐induced cell death. Conversely, the upregulation of AMPK, either by transfection or by pharmacological activation with A‐769662, decreased the expression of ALDH1A1, ABCB1A, CD133, Nanog, Oct4, and alpha fetoprotein, and restored sensitivity to sorafenib. Analysis of the underlying mechanism points to hypoxia‐inducible factor HIF‐1α as a regulator of stemness. In vivo studies in a xenograft mouse model demonstrated that stem‐like cells have greater tumourigenic capacity. AMPK activation reduced xenograft tumour growth and decreased the expression of stem cell markers. Taken together, these results indicate that AMPK may serve as a novel target to overcome chemoresistance in HCC

Effects of JWH015 in cytokine secretion in primary human keratinocytes and fibroblasts and its suitability for topical/transdermal delivery

BACKGROUND: JWH015 is a cannabinoid (CB) receptor type 2 agonist that produces immunomodulatory effects. Since skin cells play a key role in inflammatory conditions and tissue repair, we investigated the ability of JWH015 to promote an anti-inflammatory and pro-wound healing phenotype in human primary skin cells. METHODS: Human primary keratinocytes and fibroblasts were stimulated with lipopolysaccharide. The mRNA expression of cannabinoid receptors was determined using RT-PCR. The effects of JWH015 (0.05, 0.1, 0.5, and 1 µM) in pro- and anti-inflammatory factors were tested in lipopolysaccharide-stimulated cells. A scratch assay, using a co-culture of keratinocytes and fibroblasts, was used to test the effects of JWH015 in wound healing. In addition, the topical and transdermal penetration of JWH015 was studied in Franz diffusion cells using porcine skin and LC-MS. RESULTS: The expression of CB1 and CB2 receptors (mRNA) and the production of pro- and anti-inflammatory factors enhanced in keratinocytes and fibroblasts following lipopolysaccharide stimulation. JWH015 reduced the concentration of major pro-inflammatory factors (IL-6 and MCP-1) and increased the concentration of a major anti-inflammatory factor (TGF-β) in lipopolysaccharide-stimulated cells. JWH015 induced a faster scratch gap closure. These JWH015’seffects were mainly modulated through both CB1 and CB2 receptors. Topically administered JWH015 was mostly retained in the skin and displayed a sustained and low level of transdermal permeation. CONCLUSIONS: Our findings suggest that targeting keratinocytes and fibroblasts with cannabinoid drugs could represent a therapeutic strategy to resolve peripheral inflammation and promote tissue repair

The effect of photoaging on the accumulation of somatic mutations in cancer-free human skin

This is a pre-print of an article published in Annals of Oncology. The final authenticated version is available online at: https://doi.org/10.1016/j.annonc.2020.11.023Background: The accumulation of somatic mutations contributes to ageing and cancer. Sunlight is the principal aetiological factor associated with skin cancer development. However, genetic and phenotypic factors also contribute to skin cancer risk. This study aimed at exploring the role of photoaging, as well as other well-known epidemiological risk factors, in the accumulation of somatic mutations in cancer-free human epidermis. Material and methods: We deeply sequenced 46 genes in normal skin biopsies from 123 healthy donors, from which phenotypic data (including age, pigmentation-related genotype and phenotype) and sun exposure habits were collected. We determined the somatic mutational burden, mutational signatures, clonal selection and frequency of driver mutations in all samples. Results: Our results reveal an exponential accumulation of UV-related somatic mutations with age, matching skin cancer incidence. The increase of mutational burden is in turn modified by an individual's skin phototype. Somatic mutations preferentially accumulated in cutaneous squamous cell carcinoma cancer genes and clonally expanded with age, with distinct mutational processes underpinning different age groups. Our results suggest a loss of fidelity in transcription-coupled repair later in life. Conclusion: Our findings reveal that ageing is not only associated with an exponential increase in the number of somatic mutations accumulated in normal epidermis, but also with selection and expansion of cancer-associated mutations. Aged, sun-exposed normal skin is thus an extended mosaic of multiple clones with driver mutations, poised for the acquisition of transforming events

Diclofenac Prolongs Repolarization in Ventricular Muscle with Impaired Repolarization Reserve

Background: The aim of the present work was to characterize the electrophysiological effects of the non-steroidal anti- inflammatory drug diclofenac and to study the possible proarrhythmic potency of the drug in ventricular muscle. Methods: Ion currents were recorded using voltage clamp technique in canine single ventricular cells and action potentials were obtained from canine ventricular preparations using microelectrodes. The proarrhythmic potency of the drug was investigated in an anaesthetized rabbit proarrhythmia model. Results: Action potentials were slightly lengthened in ventricular muscle but were shortened in Purkinje fibers by diclofenac (20 mM). The maximum upstroke velocity was decreased in both preparations. Larger repolarization prolongation was observed when repolarization reserve was impaired by previous BaCl 2 application. Diclofenac (3 mg/kg) did not prolong while dofetilide (25 mg/kg) significantly lengthened the QT c interval in anaesthetized rabbits. The addition of diclofenac following reduction of repolarization reserve by dofetilide further prolonged QT c . Diclofenac alone did not induce Torsades de Pointes ventricular tachycardia (TdP) while TdP incidence following dofetilide was 20%. However, the combination of diclofenac and dofetilide significantly increased TdP incidence (62%). In single ventricular cells diclofenac (30 mM) decreased the amplitude of rapid (I Kr ) and slow (I Ks ) delayed rectifier currents thereby attenuating repolarization reserve. L-type calcium current (I Ca ) was slightly diminished, but the transient outward (I to ) and inward rectifier (I K1 ) potassium currents were not influenced. Conclusions: Diclofenac at therapeutic concentrations and even at high dose does not prolong repolarization markedly and does not increase the risk of arrhythmia in normal heart. However, high dose diclofenac treatment may lengthen repolarization and enhance proarrhythmic risk in hearts with reduced repolarization reserve

A socio-ecological approach to reduce the physical activity drop-out ratio in primary care-based patients with type 2 diabetes: the SENWI study protocol for a randomized control trial

BACKGROUND: Physical activity (PA) is a key behaviour for patients with type 2 diabetes (T2DM). However, healthcare professionals' (HCP) recommendations (walking advice), which are short-term and individually focused, did not reduce the PA drop-out ratio in the long run. Using a socio-ecological model approach may contribute to reducing patient dropout and improving adherence to PA. The aim of this study is threefold: first, to evaluate the effectiveness of a theory-driven Nordic walking intervention using a socio-ecological approach with T2DM patients in Spain; second, to explore the feasibility on the PA adherence process in T2DM patients while participating in the SENWI programme; and third, to understand the HCPs' opinion regarding its applicability within the Spanish healthcare system. METHODS: A three-arm randomized control trial (n = 48 each group) will assess the efficacy of two primary care-based PA interventions (Nordic walking vs. Nordic walking plus socio-ecological approach; two sessions per week for twelve weeks) compared to a control group (usual HCPs' walking advice on PA). Inclusion criteria will include physically inactive patients with T2DM, older than 40 years and without health contraindications to do PA. PA levels and drop-out ratio, quality of life and metabolic and health outcomes will be assessed at baseline, post-intervention and at 9- and 21-month follow-ups. The effect of the different interventions will be assessed by a two-factor analysis of variance: treatment group vs time. Also, a two-factor ANOVA test will be performed with linear mixed models for repeated measures. A qualitative analysis using focus groups will explore the reasons for the (in)effectiveness of the new PA interventions. Qualitative outcomes will be assessed at post-intervention using thematic analysis. DISCUSSION: Compared with the general PA walking advice and Nordic walking prescriptions, integrating a socio-ecological approach into Spanish primary care visits could be an effective way to reduce the PA drop-out ratio and increase PA levels in patients with T2DM. Such interventions are necessary to understand the role that multiple socio-complex process in day-to-day PA behaviour has in patients with T2DM in the Spanish context. TRIAL REGISTRATION: ClinicalTrials.gov NCT05159089. Physical Activity Drop-out Ratio in Patients' Living with Type 2 Diabetes. Prospectively registered on 15 December 2021

ПАТОГЕНЕТИЧЕСКИЕ ОСОБЕННОСТИ ФОРМИРОВАНИЯ СЕРДЕЧНО-СОСУДИСТОГО КОНТИНУУМА ПРИ ХРОНИЧЕСКОЙ ОБСТРУКТИВНОЙ БОЛЕЗНИ ЛЕГКИХ

Chronic obstructive pulmonary disease (COPD) is currently considered as a systemic disease with multiple extrapulmonary manifestations which can negatively affect the prognosis. Cardiovascular abnormalities are main systemic manifestations of COPD. Cardiovascular diseases and COPD have common pathogenic features that include the systemic inflammatory response, oxidative stress and endothelial dysfunction. Main pathogenic mechanisms of this comorbidity were discussed in this review.В настоящее время хроническая обструктивная болезнь легких рассматривается как системное заболевание с множественными внелегочными проявлениями, которые в ряде случаев и определяют прогноз. При этом сердечно-сосудистые осложнения относятся к основным системным проявлениям заболевания. Появление кардиореспираторной коморбидности, сопровождающейся синдромом взаимного отягощения, формирует определенные особенности клинической картины ввиду общности некоторых звеньев патогенеза, которые включают развитие системного воспалительного ответа, оксидативного стресса и эндотелиальной дисфункции. В обзоре представлены основные патогенетические механизмы, характерные для данной коморбидной патологии

Pancreatic adenocarcinoma in type 2 progressive familial intrahepatic cholestasis

<p>Abstract</p> <p>Background</p> <p>BSEP disease results from mutations in ABCB11, which encodes the bile salt export pump (BSEP). BSEP disease is associated with an increased risk of hepatobiliary cancer.</p> <p>Case Presentation</p> <p>A 36 year old woman with BSEP disease developed pancreatic adenocarcinoma at age 36. She had been treated with a biliary diversion at age 18. A 1.7 × 1.3 cm mass was detected in the pancreas on abdominal CT scan. A 2 cm mass lesion was found at the neck and proximal body of the pancreas. Pathology demonstrated a grade 2-3 adenocarcinoma with invasion into the peripancreatic fat.</p> <p>Conclusions</p> <p>Clinicians should be aware of the possibility of pancreatic adenocarcinoma in patients with BSEP disease.</p

Impaired Spleen Formation Perturbs Morphogenesis of the Gastric Lobe of the Pancreas

Despite the extensive use of the mouse as a model for studies of pancreas development and disease, the development of the gastric pancreatic lobe has been largely overlooked. In this study we use optical projection tomography to provide a detailed three-dimensional and quantitative description of pancreatic growth dynamics in the mouse. Hereby, we describe the epithelial and mesenchymal events leading to the formation of the gastric lobe of the pancreas. We show that this structure forms by perpendicular growth from the dorsal pancreatic epithelium into a distinct lateral domain of the dorsal pancreatic mesenchyme. Our data support a role for spleen organogenesis in the establishment of this mesenchymal domain and in mice displaying perturbed spleen development, including Dh +/−, Bapx1−/− and Sox11−/−, gastric lobe development is disturbed. We further show that the expression profile of markers for multipotent progenitors is delayed in the gastric lobe as compared to the splenic and duodenal pancreatic lobes. Altogether, this study provides new information regarding the developmental dynamics underlying the formation of the gastric lobe of the pancreas and recognizes lobular heterogeneities regarding the time course of pancreatic cellular differentiation. Collectively, these data are likely to constitute important elements in future interpretations of the developing and/or diseased pancreas

Electrocortical evidence for long-term incidental spatial learning through modified navigation instructions

© Springer Nature Switzerland AG 2018. The use of Navigation Assistance Systems for spatial orienting has become increasingly popular. Such automated navigation support, however, comes with a reduced processing of the surrounding environment and often with a decline of spatial orienting ability. To prevent such deskilling and to support spatial learning, the present study investigated incidental spatial learning by comparing standard navigation instructions with two modified navigation instruction conditions. The first modified instruction condition highlighted landmarks and provided additional redundant information regarding the landmark (contrast condition), while the second highlighted landmarks and included information of personal interest to the participant (personal-reference condition). Participants’ spatial knowledge of the previously unknown virtual city was tested three weeks later. Behavioral and electroencephalographic (EEG) data demonstrated enhanced spatial memory performance for participants in the modified navigation instruction conditions without further differentiating between modified instructions. Recognition performance of landmarks was better and the late positive complex of the event-related potential (ERP) revealed amplitude differences reflecting an increased amount of recollected information for modified navigation instructions. The results indicate a significant long-term spatial learning effect when landmarks are highlighted during navigation instructions