Search

Background: Acute pancreatitis (AP) presents a significant clinical challenge with limited therapeutic options. The complex etiology and pathophysiology of AP emphasize the need for innovative treatments. This study explores mRNA-based therapies delivering fibroblast growth factor 21 (FGF21) and apolipoprotein A1 (APOA1), alone and in combination, for treating experimental AP., Methods: Liver-targeted lipid nanoparticles (LNP)-mRNA formulations encoding FGF21, APOA1, and a chimeric APOA1-FGF21, were first tested for protein expression and bioavailability in vitro and in mice fed a high-fat diet. Efficacy studies were performed in the caerulein-induced AP (Cer-AP) model, and a new AP model combining ethanol feeding with ethanol binge plus palmitoleic acid administration, the EtOH/POA-AP model. A single dose of the APOA1, FGF21, and APOA1-FGF21 LNP-mRNAs formulations was administered in both models. Serum levels of pancreatic lipase (LIPC), amylase (AMYL), and aspartate aminotransferase (AST), along with pancreatic tissue analyses using two histopathological scores were performed to evaluate treatment effects., Results: In vitro studies demonstrated the translation and secretion of APOA1, FGF21, and APOA1-FGF21 proteins encoded by the LNP-mRNAs. In vivo, LNP-mRNA administration increased serum levels of the respective proteins in metabolically impaired (i.e. high fat diet-fed) mice. In the Cer-AP model, serum markers of pancreatic injury were similarly reduced when mice were treated with APOA1, FGF21, and APOA1-FGF21 LNP-mRNA, and this effect was also observed in the histopathological analyses. The EtOH/POA-AP model was more aggressive than the Cer-AP model. FGF21 and APOA1-FGF21 LNP-mRNAs were protective according to LIPC and AMYL serum levels, while APOA1 LNP-mRNA had little effect. On the other hand, histological improvements were more evident in mice receiving APOA1 LNP-mRNA. In the EtOH/POA-AP model, FGF21 and APOA1-FGF21 LNP-mRNAs reduced serum AST levels, indicating hepatoprotective activity., Discussion: This proof-of-concept study demonstrates the potential of mRNA-based therapies delivering FGF21 and APOA1 in experimental AP. While individual treatments effectively reduced pancreatic injury, the APOA1-FGF21 fusion molecule did not exhibit superior activity. Liver-targeted LNP-mRNA administration may offer a promising approach for treating AP, leveraging endogenous production pathways for therapeutic proteins. Further research is warranted to elucidate the mechanisms underlying their therapeutic efficacy and optimize treatment regimens for clinical translation., Competing Interests: Declarations. Ethics approval and consent to participate: The study received approval from the Ethics Committee for Animal Experimentation of the University of Navarra (protocol #065 − 22). Informed consent: Written informed consent was not applicable. Conflict of interest: Anne-Renee Graham holds stock in and is an employee of Moderna, Inc. The rest of the authors have no conflicts of interest to declare., (© 2025. The Author(s).)

Background: Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive cancer with limited treatment options and a poor prognosis. The critical role of epigenetic alterations such as changes in DNA methylation, histones modifications, and chromatin remodeling, in pancreatic tumors progression is becoming increasingly recognized. Moreover, in PDAC these aberrant epigenetic mechanisms can also limit therapy efficacy. This study aimed to investigate the expression and prognostic significance of a key epigenetic complex encompassing DNA methyltransferase-1 (DNMT1), the histone methyltransferase G9a, and the scaffold protein UHRF1 in PDAC. We also evaluated the therapeutic potential of an innovative inhibitor targeting these epigenetic effectors., Methods: Immunohistochemical analysis of DNMT1, G9a, and UHRF1 expression was conducted in human PDAC tissue samples. Staining was semi-quantitatively scored, and overexpression was defined as moderate to strong positivity. The prognostic impact was assessed by correlating protein expression with patient survival. The antitumoral effects of the dual DNMT1-G9a inhibitor CM272 were tested in PDAC cell lines, followed by transcriptomic analyses to identify underlying mechanisms. The in vivo antitumoral efficacy of CM272 was evaluated in PDAC xenograft and syngeneic mouse models, both alone and in combination with anti-PD1 immunotherapy., Results: DNMT1, G9a, and UHRF1 were significantly overexpressed in PDAC cells and stroma compared to normal pancreatic tissues. Simultaneous overexpression of the three proteins was associated with significantly reduced survival in resected PDAC patients. CM272 exhibited potent antiproliferative activity in PDAC cell lines, inducing apoptosis and altering key metabolic and cell cycle-related genes. CM272 also enhanced chemotherapy sensitivity and significantly inhibited tumor growth in vivo without detectable toxicity. Combination of CM272 with anti-PD1 therapy further improved antitumor responses and immune cell infiltration, particularly CD4 + and CD8 + T cells., Conclusions: The combined overexpression of DNMT1, G9a, and UHRF1 in PDAC is a strong predictor of poor prognosis. CM272, by targeting this epigenetic complex, shows promising therapeutic potential by inducing apoptosis, reprogramming metabolic pathways, and enhancing immune responses. The combination of CM272 with immunotherapy offers a novel, effective treatment strategy for PDAC., Competing Interests: Declarations. Ethics approval and consent to participate: The collection of PDAC patient samples was performed at two different research institutions while adhering to ethical guidelines and informed consent procedures. The study protocol was reviewed and approved by the local Ethical Committee of the University Hospital of Modena (Comitato Etico dell'Area Vasta Emilia Nord- Protocol Number: 0013043/19—Pratica 299/2019/OSS/AOUMO) and the Clinical Research Ethical Committee of Navarra (Pyto2015/120). All patients provided written informed consent. All animal procedures involving mice were performed in compliance with regulations concerning animals used in scientific research. The animal procedures in the VHIO were approved by the institution's Ethical Committee and the Catalan Regional Government., while the CIMA-University of Navarra approved the project under project approval #R-CP001-15GN. Consent for publication: All authors read this manuscript and approve for publication. Competing interests: The authors declare that they have no competing interest., (© 2025. The Author(s).)

Background: Hepatocellular carcinoma (HCC) genetic/transcriptomic signatures have been widely described. However, its proteomic characterization is incomplete. We performed non-targeted quantitative proteomics of HCC samples and explored its clinical, functional, and molecular consequences., Methods: Non-targeted quantitative proteomics were performed on cytosolic and nuclear fractions of liver samples [HCC vs. non-tumour adjacent tissue (NTAT), n = 42 patients]. Changes were confirmed in 7 in silico HCC cohorts. Functional and molecular implications were evaluated on HCC-derived cell lines after silencing/overexpressing VARS1 and/or MAGI1. VARS1-overexpressing Hep3B cells were used for in vivo studies [Extreme Limiting Dilution Assay (ELDA) and orthotopic tumour formation]. Quantitative proteomics were performed on VARS1-overexpressing HCC cell lines., Results: Quantitative proteomics revealed the dysregulation of the cytosolic and nuclear proteomes in HCC, and defined two proteomic HCC subgroups, the most aggressive associated to the dysregulation of the aminoacyl-tRNA synthetases (ARSs). ARSs dysregulation was corroborated in in silico HCC cohorts and associated to poor prognosis. Patients with ARSs upregulation had genomic/transcriptomic characteristics of the proliferative HCC. Valine tRNA-aminoacyl synthetase (VARS1) was the ARSs most consistently overexpressed and associated to aggressiveness. VARS1 modulation (silencing/overexpression) altered tumour establishment-associated parameters in vitro and/or in vivo. Quantitative proteomics on cells overexpressing VARS1 and rescue experiments identified the downregulation of MAGI1, a tumour suppressor in HCC, as a mediator of VARS1 function., Conclusions: Quantitative proteomics defines two prognosis-related proteomic HCC subgroups. ARSs machinery is dysregulated in the aggressive subgroup, bearing potential as prognostic biomarkers. VARS1 promotes aggressiveness through the modulation of MAGI1, representing a novel targetable vulnerability in HCC., Competing Interests: Declarations. Ethics approval and consent to participate: The Reina Sofia University Hospital Ethics Committee approved the protocol for this study, in agreement to institutional and Good Clinical Practice guidelines (Protocol numbers PI20/01301 and PI23/00652) and the declaration of Helsinki. Informed consent was obtained from all patients or their relatives. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2025. The Author(s).)

Background: To date, there are no studies in the literature that define the internal structure of the Tampa Scale for Kinesiophobia (TSK) in patients with chronic neck pain based on factorial analysis. As such, we aimed to verify and identify the best structure of the Brazilian version of the TSK in patients with chronic neck pain., Methods: We included Brazilian participants aged ≥18 years, both sexes, with self-reported neck pain for more than 3 months and pain intensity ≥3 on the Numerical Pain Rating Scale (NPRS). Dimensionality and number of TSK items were assessed using confirmatory factor analysis (CFA). We tested the following internal structures: structure 1 (1 domain and 17 items), structure 2 (1 domain and 11 items), structure 3 (2 domains and 11 items), and structure 4 (2 domains and 9 items). We used the Pain-Related Catastrophizing Thoughts Scale (PCTS) and the NPRS for construct validity. In addition, we assessed test-retest reliability for the seven-day interval using intraclass correlation coefficient (ICC 2,1 ), Cronbach's alpha to assess internal consistency, and ceiling and floor effects., Results: The study sample included of 335 patients. Most were women (77.6%), young adults (~ 34 years), single (48.4%), with complete primary education (57.3%), physically inactive (66.6%), with a mean pain duration of 46 months and a mean pain intensity of ~ 5 points on the NPRS. Redundancy was found in the following items: item 1 with item 2 (modification indices = 21.419) and item 13 with item 15 (modification indices = 13.641). Subsequently, based on these paired analyses, the items with the lowest factor loadings (items 2 and 15) were excluded. As such, TSK structure 4 was composed of two domains ("somatic focus" and "activity avoidance") and 9 items, which showed adequate fit indices and lower AIC and SABIC values. We observed significant values (p < 0.05) with a correlation magnitude greater than 0.142 to 0.657 between the two domains of the TSK-neck and the other instruments (PCTS and NPRS). We found excellent reliability (ICC 2,1  ≥ 0.96) and adequate internal consistency (Cronbach's alpha ≥0.98) of the TSK-neck. Finally, ceiling and floor effects were not observed., Conclusion: The TSK-neck structure with two domains (somatic focus and activity avoidance) and nine items is the most appropriate for patients with chronic neck pain., (© 2024. The Author(s).)

Background: Primary dysmenorrhea (PD) is an etiological cyclic pelvic pain related to the menstrual period; it can negatively impact women's quality of life and productivity. The aim of the present study was to estimate the prevalence of PD and analyze associated symptoms in Brazilian women., Methods: An online cross-sectional study was carried out in Brazil, with a structured questionnaire regarding dysmenorrhea and associated symptoms. PD intensity was measured with the Numerical Rating Scale for Pain and classified as mild (1-3), moderate (4-7) and severe (> 8). The association between qualitative variables was performed using Pearson's Chi-Square Test. The quantification of this association was measured using multinomial logistic regression models, with calculation of Odds Ratio and confidence interval. A significance level of 5% was considered., Results: A total of 10,070 women were included. Most participants classified PD intensity as moderate (40.4%, 41.9% and 49.7%) and severe (21.2%, 24.8% and 28.4%) in the previous month, 3 months and 5 years, respectively. The most common symptoms associated with PD were irritability, abdominal distension sensation, anxiety and feeling more emotional. The increased of the risk (OR > 1.0) for moderate and severe PD-related pain intensity is related to age, nulliparity and presence PD since adolescence., Conclusion: There is a high prevalence of PD among Brazilian women, and the most common symptoms reported were irritability, abdominal distension sensation, anxiety and feeling more emotional., (© 2024. The Author(s).)

Background: Fibromyalgia guidelines indicate that exercise is critical in the management of fibromyalgia, and there is evidence that patients with fibromyalgia can perform resistance training at moderate and high intensities. However, despite the biological plausibility that progression of intensity provides greater benefit to individuals, no studies have compared different intensities (progressive versus constant intensities) of the same exercise in this population., Objective: To compare the effect of 24 sessions of resistance training (progressive vs. constant intensity) on impact of fibromyalgia, sleep quality, anxiety, depression, pain, walking ability, and musculoskeletal capacity., Methods: A protocol for a blinded randomized controlled trial. The sample will be randomized into three groups: group 1 (progressive intensity, experimental), group 2 (constant intensity, control A), and group 3 (walking, control B). Group 1 will perform resistance training at moderate intensity (50% of maximum dynamic strength), previously determined by the 1 repetition maximum (1-RM) test in the proposed exercises. The strength of each individual will be reassessed every 4 weeks (by 1-RM) and the intensity of each exercise will be positively adjusted by 20% of the value observed in kg (i.e., first month 50%; second month 70%; third month 90% of the maximum dynamic strength). Group 2 will perform the same procedure, but the intensity will be maintained at 50% of the maximum dynamic strength throughout the treatment (i.e., constant intensity from the first to the third month). Group 3 will perform a 40-minute treadmill walk at low intensity, defined by a walking speed corresponding to 60-70% of the maximum heart rate, which we will control with a heart rate monitor. All groups will receive a 45-minute pain education session prior to the exercise program, covering the pathophysiologic mechanisms of chronic pain, strategies for coping with pain, avoiding hypervigilance, and deconstructing beliefs and myths about chronic pain., Discussion: The results of the present study may help health care professionals adjust the intensity of resistance training and thus plan the most effective intervention (progressive or constant intensity) to reduce the impact of fibromyalgia on patients' lives., Trial Registration: Brazilian Registry of Clinical Trials (ReBEC) ID: RBR-9pbq9fg, date of registration: October 06, 2022., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Background: Chronic non-specific neck pain (CNNP) is a prevalent musculoskeletal disorder known for its significant disability and economic burden, ranking second only to low back pain in musculoskeletal conditions. Physical therapy offers effective interventions for CNNP, including low-level laser therapy (LLLT). High-intensity laser therapy (HILT) is a recent treatment for musculoskeletal pain, but studies that support its use in CNNP are limited. The objective of this study is to assess the effect of high-intensity laser therapy on pain intensity in patients with CNNP, given the existing evidence on LLLT for this condition., Methods: This is a 2-arm, randomized, placebo-controlled trial with blinded evaluators. The research will be carried out in the laboratory of physical agents at the Andrés Bello University, Campus Casona de las Condes. Eligible participants include the entire internal and external community associated with Andrés Bello University suffering from chronic non-specific NP. Participants will be stratified by sex (4 subgroups) and randomized into 2 study groups: group 1 (HILT and stretching exercises) and group 2 (sham HILT and stretching exercises). Treatments will be performed twice a week for 4 weeks with 3 assessments: before treatment (T0), at the end of treatment (T1), and 12 weeks after treatment (follow-up) (T2). The main outcomes will be pain intensity at rest, pain intensity at movement (active cervical movements: flexion, extension, right and left side bending, and right and left rotation), and pain pressure threshold (average obtained for six evaluation points). Secondary outcome measures will include neck range of motion in the sagittal, coronal, and transverse planes and neck disability., Discussion: In this study, HILT's effects on patients with non-specific NP will be compared to those of a sham laser intervention. This RCT will offer new evidence regarding the potential benefits of HILT in terms of pain intensity, range of movement, and disability in people suffering with non-specific NP., Trial Registration: ClinicalTrials.gov NCT05689788. January 19, 2023., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Background: As with fibromyalgia, several musculoskeletal disorders are characterized by chronic pain, raising a clinical question - do the instruments used to assess fibromyalgia symptoms according to ACR criteria (ACR criteria) generate similar scores in other chronic musculoskeletal pain?, Objective: To compare the symptoms among fibromyalgia and other chronic musculoskeletal pain. Additionally, we also compared the most researched outcomes in fibromyalgia (i.e., present pain at rest and after movement; fatigue; pain severity and impact; function, global impact, and fibromyalgia symptom)., Methods: A cross-sectional study. Participants over 18 years old were included if they presented report of chronic musculoskeletal pain (≥ 3 months) and after that, they were divided into two groups (fibromyalgia and chronic pain). They answered the Fibromyalgia Impact Questionnaire-Revised (FIQ-R), Brief Pain Inventory (BPI), Numerical Pain Rating Scale (NPRS) for pain and fatigue, WPI, and SSS., Results: A total of 166 participants were included in this study into two independent groups (chronic pain, n = 83; fibromyalgia, n = 83). We observed significant differences (p < 0.05) and large effect sizes (Cohen's d, ≥ 0.7) in clinical outcomes comparisons between groups (i.e., widespread pain; symptom severity; present pain at rest and after movement; fatigue; pain severity and impact; function, global impact, and fibromyalgia symptoms)., Conclusion: Fibromyalgia patients (2016 ACR criteria) compared to other chronic musculoskeletal pain patients have higher levels of pain (at rest or after movement) and fatigue, greater impairment in both functionality and global impact, and worse symptoms. Therefore, the WPI and SSS instruments should be used exclusively to assess fibromyalgia symptoms., (© 2023. The Author(s).)

Background: There is an association of dysmenorrhea with human functioning and disability. However, no patient-reported outcome measure has been developed to assess this construct in women with dysmenorrhea. WHODAS 2.0 has been recognized as an important generic patient-reported outcome information of physical function and disability. Thus, the aim of this study was to assess the measurement properties of the WHODAS 2.0 in women with dysmenorrhea., Methods: This is an online and cross-sectional study conducted with Brazilian women aged 14 to 42 years with self-report of dysmenorrhea in the last three months. According to COSMIN, structural validity was evaluated by exploratory and confirmatory factor analysis; internal consistency by Cronbach's Alpha; measurement invariance by multigroup confirmatory factor analysis between geographic regions of Brazil; and construct validity by correlating WHODAS 2.0 to the Numerical Rating Scale for pain severity., Results: One thousand three hundred and eighty-seven women (24.7 ± 6.5 years) with dysmenorrhea participated in the study. WHODAS 2.0 presented a single factor by exploratory factor analysis and adequate model by confirmatory factor analysis (CFI = 0.924, TLI = 0.900, RMSEA = 0.038), excellent internal consistence (α = 0.892) for all items and an invariancy across geographic regions (ΔCFI ≤ 0.01 and ΔRMSEA < 0.015). Correlation between WHODAS 2.0 and numerical rating scale was positive and moderate (r = 0.337)., Conclusion: WHODAS 2.0 has a valid structure to assess functioning and disability related to dysmenorrhea in women., (© 2023. The Author(s).)

Background: The Roland-Morris Disability Questionnaire (RMDQ) is one of the most used instruments to measure self-reported disability in patients with low back pain, however, the uncertainty on which version to use may lead to inadequate disability measurement and consequently, improper management of patients with chronic low back pain., Objective: To propose a short version of the RMDQ, compare it with the other short versions presented by the specialized literature, and identify the best internal structure of the RMDQ for the Brazilian population., Methods: This is a cross-sectional study in which we used confirmatory factor analysis to identify the best structure of the RMDQ. We assessed 545 participants, most of which were women, aged ≥ 30 years old, single, with mean low back pain intensity ~ 5 points, and mean pain chronicity ~ 72 months. We used lavaan and semPlot packages, with implementation of a tetrachoric matrix and the robust diagonally weighted least squares extraction method. We also used fit indices chi-square/degree of freedom, comparative fit index, Tucker-Lewis index, root mean square error of approximation, and standardized root mean squared residual. For the comparison between models, we considered the structure with the lowest values of the Akaike information criterion and Bayesian information criterion. In addition, we assessed criterion validity via Spearman's correlation coefficient to correlate the long and short versions. In this study, the 15-item structure was created through the use of modification indices to identify redundant items (9 items were excluded)., Results: RMDQ structure with one domain and 15 items and the structure with two domains and 16 items showed all fit indices with adequate values, but the one-dimensional version showed the lowest Akaike information criterion and Bayesian information criterion values. Regarding criterion validity, correlation between the RMDQ with 24 items and 15 items is adequate (rho = 0.954, p < 0.001)., Conclusion: The RMDQ-15 is a short version of the RMDQ instrument with the most adequate internal structure and satisfactorily correlated with the long version of the instrument., (© 2022. The Author(s).)

Objective: The Fibromyalgia Rapid Screening Tool (FiRST) was developed to screen people with chronic pain for Fibromyalgia (FM), especially in primary health care settings. This study aimed to translate the FiRST into Brazilian Portuguese and evaluate its measurement properties for an online application., Methods: After the process of translation and backtranslation, the FiRST was applied online in 483 adults with chronic pain (FM group n = 395; Chronic pain group n = 88), along with the Numerical Rating Scale for pain and fatigue, the Brief Pain Inventory, and the Fibromyalgia Impact Questionnaire-Revised. A Receiver Operating Characteristics (ROC) curve was computed and the area under the curve (AUC) was used to determine the sensibility, specificity, and cut-off score for the FiRST. The Mann-Whitney test was used for quantitative variables and the Chi-square and the Fisher's exact test, for the categorical variables with level of significance of 5%. Fleiss' Kappa, Gwet's AC1 and percentage of agreement were also calculated between test and retest., Results: For all the questionnaires, the FM group presented higher scores, which mean a worst condition. The FiRST presented a sensitivity of 92.3%, and a specificity of 61.6% with 5 as the cut-off score. AUC, Fleiss' Kappa, Gwet's AC1 and percentage of agreement were, respectively, 0.82, 0.38, 0.63 and 71.8%., Conclusion: The FiRST was translated into Brazilian Portuguese and the online version presented a good content validity and adequate measurement errors that allow FM patients to be screened among people with chronic pain., (© 2022. The Author(s).)

Background: To verify the use of pain drawing to assess multisite pain in with primary dysmenorrhea (PD) and to assess its divergent validity, test-retest reliability, intra- and inter-rater reliability and measurement errors., Methods: Cross-sectional study. Adult women with self-reported PD three months prior to the study. Women answered the Numerical Rating Scale (NRS) and the pain drawing during two consecutive menstruations. The pain drawings were digitalized and assessed for the calculation of total pain area (%). Intra- and inter-rater reliability and the test-retest reliability between the first and the second menstruations were assessed with the intraclass correlation coefficient (ICC). Measurement errors were calculated with the standard error of measurement (SEM), smallest detectable change (SDC) and the Bland-Altman plot. Spearman correlation (rho) was used to check the correlation between the total pain area and pain intensity of the two menstruations., Results: Fifty-six women (24.1 ± 3.1 years old) participated of the study. Their average pain was 6.2 points and they presented pain in the abdomen (100%), low back (78.6%), head (55.4%) and lower limbs (50%). All reliability measures were considered excellent (ICC > 0.75) for the total pain area; test-retest SEM and SDC were 5.7% and 15.7%, respectively. Inter-rater SEM and SDC were 8% and 22.1%, respectively. Correlation between total pain area and pain intensity was moderate in the first (rho = 0.30; p = 0.021) and in the second menstruations (rho = 0.40; p = 0.002)., Conclusion: Women with PD presented multisite pain, which could be assessed with the pain drawing, considered a reliable measurement., (© 2022. The Author(s).)

Background: Febrile Neutropenia (FN) is a common and serious condition related to cancer chemotherapy. Human recombinant Granulocyte-Colony Stimulating Factor (G-CSF) prevents and attenuates the severity and duration of FN. We evaluated the use and predictors of G-CSF adherence among women with breast cancer with a high risk of FN in Puerto Rico., Methods: This retrospective cohort study used the Puerto Rico Central Cancer Registry-Health Insurance Linkage Database. Women with invasive breast cancer diagnosed during 2009-2015 who received selected chemotherapy regimens (n = 816) were included. The risk of FN was categorized as high and low risk based on the chemotherapy regimens according to the National Comprehensive Cancer Network guidelines and literature. Adherence was defined as the use or no use of G-CSF at the start of the first chemotherapy cycle among women with breast cancer based on the risk of developing FN. We used a multivariate logistic model to identify factors associated with G-CSF use in women classified at high risk for FN., Results: Adherence to G-CSF clinical practice guidelines was low (38.2%) among women with a high risk of FN. Women at high risk of FN with Medicaid (aOR: 0.14; CI 95%: 0.08, 0.24) and Medicare/Medicaid (aOR: 0.33; CI 95%: 0.15, 0.73) were less likely to receive G-CSF than women with private health insurance. Women with regional stage (aOR: 1.82; CI 95%: 1.15, 2.88) were more likely to receive G-CSF than women with localized cancers., Conclusions: Adherence to clinical practice guidelines was poor among women with a high risk of FN. Furthermore, disparities in the adherence to G-CSF use in terms of health insurance, health region, and cancer stage granted the opportunity to implement strategies to follow the recommended guidelines for using G-CSF as part of cancer treatment., (© 2022. The Author(s).)

Background: Cholangiocarcinoma (CCA) is still a deadly tumour. Histological and molecular aspects of thioacetamide (TAA)-induced intrahepatic CCA (iCCA) in rats mimic those of human iCCA. Carcinogenic changes and therapeutic vulnerabilities in CCA may be captured by molecular investigations in bile, where we performed bile proteomic and metabolomic analyses that help discovery yet unknown pathways relevant to human iCCA., Methods: Cholangiocarcinogenesis was induced in rats (TAA) and mice (Jnk Δhepa  + CCl 4  + DEN model). We performed proteomic and metabolomic analyses in bile from control and CCA-bearing rats. Differential expression was validated in rat and human CCAs. Mechanisms were addressed in human CCA cells, including Huh28-KRAS G12D cells. Cell signaling, growth, gene regulation and [U- 13 C]-D-glucose-serine fluxomics analyses were performed. In vivo studies were performed in the clinically-relevant iCCA mouse model., Results: Pathways related to inflammation, oxidative stress and glucose metabolism were identified by proteomic analysis. Oxidative stress and high amounts of the oncogenesis-supporting amino acids serine and glycine were discovered by metabolomic studies. Most relevant hits were confirmed in rat and human CCAs (TCGA). Activation of interleukin-6 (IL6) and epidermal growth factor receptor (EGFR) pathways, and key genes in cancer-related glucose metabolic reprogramming, were validated in TAA-CCAs. In TAA-CCAs, G9a, an epigenetic pro-tumorigenic writer, was also increased. We show that EGFR signaling and mutant KRAS G12D can both activate IL6 production in CCA cells. Furthermore, phosphoglycerate dehydrogenase (PHGDH), the rate-limiting enzyme in serine-glycine pathway, was upregulated in human iCCA correlating with G9a expression. In a G9a activity-dependent manner, KRAS G12D promoted PHGDH expression, glucose flow towards serine synthesis, and increased CCA cell viability. KRAS G12D CAA cells were more sensitive to PHGDH and G9a inhibition than controls. In mouse iCCA, G9a pharmacological targeting reduced PHGDH expression., Conclusions: In CCA, we identified new pro-tumorigenic mechanisms: Activation of EGFR signaling or KRAS mutation drives IL6 expression in tumour cells; Glucose metabolism reprogramming in iCCA includes activation of the serine-glycine pathway; Mutant KRAS drives PHGDH expression in a G9a-dependent manner; PHGDH and G9a emerge as therapeutic targets in iCCA., (© 2022. The Author(s).)

Hepatocellular carcinoma (HCC) is the most frequent primary liver cancer, being the sixth most commonly diagnosed cancer and the fourth leading cause of cancer-related death. As other heterogeneous solid tumours, HCC results from a unique synergistic combination of genetic alterations mixed with epigenetic modifications.In HCC the patterns and frequencies of somatic variations change depending on the nearby chromatin. On the other hand, epigenetic alterations often induce genomic instability prone to mutations. Epigenetics refers to heritable states of gene expression without alteration to the DNA sequence itself and, unlike genetic changes, the epigenetic modifications are reversible and affect gene expression more extensively than genetic changes. Thus, studies of epigenetic regulation and the involved molecular machinery are greatly contributing to the understanding of the mechanisms that underline HCC onset and heterogeneity. Moreover, this knowledge may help to identify biomarkers for HCC diagnosis and prognosis, as well as future new targets for more efficacious therapeutic approaches.In this comprehensive review we will discuss the state-of-the-art knowledge about the epigenetic landscape in hepatocarcinogenesis, including evidence on the diagnostic and prognostic role of non-coding RNAs, modifications occurring at the chromatin level, and their role in the era of precision medicine.Apart from other better-known risk factors that predispose to the development of HCC, characterization of the epigenetic remodelling that occurs during hepatocarcinogenesis could open the way to the identification of personalized biomarkers. It may also enable a more accurate diagnosis and stratification of patients, and the discovery of new targets for more efficient therapeutic approaches., (© 2022. The Author(s).)

Background: Urgency is a complaint of sudden, compelling desire to pass urine, which is difficult to defer, caused by involuntary contraction of the detrusor muscle during the bladder-filling stage. To enable detrusor inhibition, electrotherapy resources such as transcutaneous tibial nerve stimulation (TTNS) and parasacral transcutaneous electrical stimulation (PTES) have been used. The objective this study is to publish the study protocol that aims to investigate whether urgency decreases after treatment with both of the techniques., Methods: This randomized controlled clinical trial will include 99 women, aged more than 18 years old, with urgency (score ≥ 8 in the Overactive Bladder-Validated 8-Question Awareness Tool [OAB-V8]). Women will be randomly allocated into three groups: TTNS, PTES, and placebo. The following questionnaires will be applied: the Anamnesis Record, the Incontinence Questionnaire Overactive Bladder, the King's Health Questionnaire, the 24-Hour Voiding Diary, and the OAB-V8, at four different time points: at baseline prior to the first session, at the 6th session, the 12th session and at follow-up. The current used for the transcutaneous electrical stimulation will be a symmetrical balanced biphasic pulsed current, for 12 sessions, twice a week, for 20 minutes. Qualitative variables will be displayed as frequency and percentage, quantitative variables as mean and standard deviation. Comparison of urgency severity among groups will be performed with a repeated measures ANOVA, considering the effect of the three groups and the four evaluations, and interactions among them., Discussion: The present study aims to contribute evidence for a more in-depth discussion on electrode positioning for electrostimulation used in urgency treatment. It should be emphasized that, based on the possibility of confirming the hypothesis that urgency will decrease in a similar way after both treatments (TTNS and PTES), the PTES will be used as an option for positioning the electrodes alternatively to the tibial nerve region in special populations, such as amputees or people with severe lower limb sensory impairment., Trial Registration: Brazilian Registry of Clinical Trials (ReBEC) ID: RBR-9rf33n, date of registration: 17 May 2018.

Background: Melanoma is the most deadly form of skin cancer without effective treatment. Methylthioadenosine (MTA) is a naturally occurring nucleoside with differential effects on normal and transformed cells. MTA has been widely demonstrated to promote anti-proliferative and pro-apoptotic responses in different cell types. In this study we have assessed the therapeutic potential of MTA in melanoma treatment., Methods: To investigate the therapeutic potential of MTA we performed in vitro proliferation and viability assays using six different mouse and human melanoma cell lines wild type for RAS and BRAF or harboring different mutations in RAS pathway. We also have tested its therapeutic capabilities in vivo in a xenograft mouse melanoma model and using variety of molecular techniques and tissue culture we investigated its anti-proliferative and pro-apoptotic properties., Results: In vitro experiments showed that MTA treatment inhibited melanoma cell proliferation and viability in a dose dependent manner, where BRAF mutant melanoma cell lines appear to be more sensitive. Importantly, MTA was effective inhibiting in vivo tumor growth. The molecular analysis of tumor samples and in vitro experiments indicated that MTA induces cytostatic rather than pro-apoptotic effects inhibiting the phosphorylation of Akt and S6 ribosomal protein and inducing the down-regulation of cyclin D1., Conclusions: MTA inhibits melanoma cell proliferation and in vivo tumor growth particularly in BRAF mutant melanoma cells. These data reveal a naturally occurring drug potentially useful for melanoma treatment.